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    Application of the CRISPR/Cas system in gene editing and nucleic acid detection of parasitic diseases: a review
    YAN Shuning, YANG Shuo, YANG Hanyin, XIN Yi, XU Bin, HU Wei, LU Yan, ZHENG Bin
    Chinese Journal of Schistosomiasis Control    2024, 36 (3): 314-320.  
    Abstract601)      PDF(pc) (900KB)(2597)       Save
    CRISPR/Cas system, an adaptive immune system with clustered regularly interspaced short palindromic repeats, may interfere with exogenous nucleic acids and protect prokaryotes from external damages, is an effective gene editing and nucleic acid detection tools. The CRISPR/Cas system has been widely applied in virology and bacteriology; however, there is relatively less knowledge about the application of the CRISPR/Cas system in parasitic diseases. The review summarizes the mechanisms of action of the CRISPR/Cas system and provides a comprehensive overview of their application in gene editing and nucleic acid detection of parasitic diseases, so as to provide insights into future studies on parasitic diseases.
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    Progress of schistosomiasis control in People’s Republic of China in 2023
    ZHANG Lijuan, HE Junyi, YANG Fan, DANG Hui, LI Yinlong, GUO Suying, LI Shizhen, CAO Chunli, XU Jing, LI Shizhu
    Chinese Journal of Schistosomiasis Control    2024, 36 (3): 221-227.  
    Abstract2411)      PDF(pc) (942KB)(2411)       Save
    To understand the progress of national schistosomiasis elimination program of China in 2023 and summarize the lessons and experiences, data on the endemic status of schistosomiasis and national schistosomiasis surveillance results in the People’s Republic of China were collected and analyzed at a national level. By the end of 2023, Shanghai Municipality, Zhejiang Province, Fujian Province, Guangdong Province and Guangxi Zhuang Autonomous Region continued to consolidate the achievements of schistosomiasis elimination, and Sichuan and Jiangsu provinces maintained the criteria of transmission interruption, while Yunnan and Hubei provinces were identified to achieve the criteria of transmission interruption in 2020, and Anhui, Jiangxi and Hunan provinces achieved the criteria of transmission interruption in 2023. A total of 451 counties (cites, districts) were found to be endemic for schistosomiasis in China in 2023, including 26 250 endemic villages covering 73 034 500 residents at risk of infections. Among the 451 endemic counties (cities, districts), 78.49% (354/451) achieved the criteria of schistosomiasis elimination and 21.51% (97/451) achieved the criteria of transmission interruption, respectively. In 2023, a total of 4 216 643 individuals received immunological tests, with 47 794 sero⁃positives identified, and a total of 184 216 individuals received parasitological examinations, with 4 egg⁃positives detected. A total of 27 768 cases with advanced schistosomiasis were documented in China by the end of 2023. In 2023, 539 548 bovines were raised in schistosomiasis⁃endemic areas of China, and 125 440 bovines received immunological tests, with 124 sero⁃positives detected, while no egg⁃positives were identified among the 133 508 bovines receiving parasitological examinations. In 2023, snail survey was performed at an area of 641 339.53 hm2 and 184 819.77 hm2 snail habitats were identified, including 51.53 hm2 emerging snail habitats and 642.25 hm2 reemerging snail habitats. In 2023, there were 20 198 schistosomiasis patients receiving praziquantel chemotherapy, and 598 183 person⁃time individuals and 283 954 herd⁃time bovines were given expanded chemotherapy. In 2023, snail control with chemical treatment was performed in 116 347.95 hm2 snail habitats, and the actual area of chemical treatment was 65 690.89 hm2, while environmental improvements were performed in snail habitats covering an area of 1 334.62 hm2. The national schistosomiasis surveillance results showed that the mean prevalence of Schistosoma japonicum infections were both zero among humans and bovines in 2023, and no S. japonicum infection was detected in snails. These data demonstrated that transmission interruption of schistosomiasis had been achieved across all endemic provinces in China in 2023, and the endemic status of schistosomiasis tended to be stable, while advanced cases were predominant among all schistosomiasis cases. However, the areas of snail habitats remained high and cattle re⁃raising was very common in some regions. Intensified schistosomiasis surveillance and forecast and snail control in high⁃risk areas are needed.
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    Risk factors of visceral leishmaniasis in the world: a review
    CHEN Xi, SHI Yue, ZHOU Sheng, GENG Mengjie, TU Hong, SONG Jiandong, ZHENG Canjun, SUN Junling
    Chinese Journal of Schistosomiasis Control    2024, 36 (4): 412-421,427.  
    Abstract443)      PDF(pc) (1023KB)(2399)       Save
    Visceral leishmaniasis is a zoonotic parasitic disease caused by viscerotropic Leishmania species and transmitted by bites of infected phlebotomine sandflies, which is predominantly prevalent in the Indian subcontinent, eastern Africa and South America. Currently, visceral leishmaniasis is the second most fatal parasitic disease in the world. Because of climate changes, urban development and individual conditions, there are changes in the density of visceral leishmaniasis vector sandflies and the likelihood of contact with humans, resulting in a visceral leishmaniasis transmission risk. The review summarizes natural, social and biological factors affecting the transmission of visceral leishmaniasis, so as to provide insights into formulation of targeted control measures for visceral leishmaniasis.
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    Current status and challenges of zoonosis prevention and control: a One Health perspective
    LI Cihuai, CHEN Feng, LÜ Shan
    Chinese Journal of Schistosomiasis Control    2025, 37 (1): 98-103.  
    Abstract1217)      PDF(pc) (1059KB)(2334)       Save
    Zoonosis prevention and control is a complex public health concern, which requires the collaboration of multiple regions, disciplines, and departments to enhance the effectiveness. The One Health concept aims to achieve the joint health security of humans, animals and environments through cross⁃disciplinary, cross⁃sector and cross⁃field collaborations. This review summarizes the development of One Health and the successful practices in the prevention and control of echinococcosis, rabies, COVID⁃19 and schistosomiasis, as well as explores the challenges faced in applying this concept to the prevention and control of zoonoses, so as to provide insights into formulation of the integrated zoonoses control strategy and implementation of zoonoses control interventions at the human⁃animal⁃environment interface.
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    Progress of schistosomiasis control in the People's Republic of China in 2024
    HE Junyi, ZHANG Lijuan, YANG Fan, DANG Hui, LI Yinlong, GUO Suying, LI Shizhen, CAO Chunli, XU Jing, LI Shizhu
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 223-231.  
    Abstract1196)      PDF(pc) (1146KB)(1967)       Save
    To understand the progress of, summarize the lessons learned from and analyze the challenges in the national schistosomiasis elimination program of China in 2024, this article presented the endemic situation of schistosomiasis and national schistosomiasis surveillance results in the People's Republic of China in 2024. By the end of 2024, Shanghai Municipality, Zhejiang Province, Fujian Province, Guangdong Province and Guangxi Zhuang Autonomous Region continued to consolidate schistosomiasis elimination achievements, and 7 provinces of Jiangsu, Sichuan, Yunnan, Hubei, Hunan, Anhui and Jiangxi maintained the criteria of schistosomiasis transmission interruption. A total of 450 counties (cites, districts) were found to be endemic for schistosomiasis in China in 2024, including 26 061 endemic villages covering 73 630 500 residents at risk of infections. Among the 450 counties (cities, districts) endemic for schistosomiasis, 388 (86.22%) achieved the criteria of schistosomiasis elimination and 62 (13.78%) achieved the criteria of transmission interruption. In 2024, a total of 4 102 624 individuals received immunological tests for schistosomiasis in China, with 44 823 sero⁃positives identified (1.09% seroprevalence), and a total of 169 722 individuals received parasitological examinations, with 1 egg⁃positives detected. A total of 27 321 cases with advanced schistosomiasis were documented in China by the end of 2024. In 2024, a total of 575 686 bovines were raised in schistosomiasis⁃endemic villages of China, and 113 842 bovines received immunological tests, with 235 sero⁃positives detected (0.21% seroprevalence), while no egg⁃positives were identified among the 167 475 bovines receiving parasitological examinations. In 2024, snail survey was performed covering an area of 680 498.27 hm2 in China, and 190 778.66 hm2 snail habitats were identified, including 59.09 hm2 emerging snail habitats and 704.23 hm2 reemerging snail habitats. In 2024, a total of 19 665 schistosomiasis patients receiving chemotherapy with praziquantel in China, and expanded chemotherapy was given to humans at 571 722 person⁃times and to bovines at 306 740 herd⁃times. In addition, snail control with chemical treatment covered 117 111.37 hm2 snail habitats across China in 2024, and the actual area of chemical treatment was 66 562.95 hm2, while environmental improvements were performed in snail habitats covering an area of 1 374.26 hm2. The national schistosomiasis surveillance results showed that the mean prevalence rates of Schistosoma japonicum infections were both 0 among humans and bovines in China in 2024, and no S. japonicum infection was detected in snails. These data demonstrated that the prevalence of schistosomiasis remained at a low level in China in 2024; however, the areas of snail habitats remained high and the number of fenced cattle showed a slight increase. To address these risks, it is imperative to maintain the integrated strategy with an emphasis on management of the source of S. japonicum infection and intensified snail control in high⁃risk areas, and to reinforce schistosomiasis surveillance and forecast and snail control in high⁃risk areas.  
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    Advances in detection methods for fish⁃borne parasites in aquatic products
    ZHANG Yaoguang, WANG Zhenyu, CHEN Jian, YU Qing
    Chinese Journal of Schistosomiasis Control    2024, 36 (4): 428-434,438.  
    Abstract510)      PDF(pc) (1021KB)(1944)       Save
    Consumption of raw and semi⁃raw aquatic products is strongly associated with the development of fish⁃borne parasitic diseases. Detection of fish⁃borne parasites in aquatic products is of great significance for the prevention and control of fish⁃borne parasitic diseases. This review describes the advances in the application of etiological, molecular biological and immunological techniques alone and in combinations for detection of fish⁃borne parasites, so as to provide insights into detection of fish⁃borne parasites.
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    How do female mosquitoes determine the most suitable males for mating?
    LI Yitong, LI Dong, LIU Xiaofei, WANG Ying, LIU Tingting, XU Yanqiu, DENG Shengqun
    Chinese Journal of Schistosomiasis Control    2024, 36 (3): 321-328.  
    Abstract316)      PDF(pc) (1001KB)(1684)       Save
    More than 80% of the world’s populations are at risk of vector⁃borne diseases, with mosquito⁃borne diseases as a significant global public health problem. Mosquito populations control is critical to interrupting the transmission of mosquito⁃borne diseases. This review summarizes the physical attributes, smell, vision, touch, and hearing of mosquitoes to unravel the preferences of female mosquitoes, and describes the mechanisms underlying the best male mating by female mosquitoes, so as to provide new insights into management of mosquito⁃borne diseases.
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    Hepatic T cell subtypes and functional analysis among alveolar echinococcosis patients using single⁃cell RNA sequencing
    CHEN Si, WANG Xiangqian, JIA Wanzhong, CAI Qigang, ZHANG Xueyong, ZHANG Qiang, ZHENG Haibo, ZHU Linghong, LI Bing, WANG Wei, HAN Xiumin
    Chinese Journal of Schistosomiasis Control    2024, 36 (5): 481-493.  
    Abstract572)      PDF(pc) (4415KB)(1574)       Save
    Objective To investigate T cell subtypes and their functions in liver immune microenvironments among patients with alveolar echinococcosis (AE) using single⁃cell RNA sequencing (scRNA⁃seq).  Methods Four AE patients that were admitted to Qinghai Provincial People's Hospital in 2023 for hepatic surgery for the first time were enrolled, and liver specimens were sampled 1 cm (peri⁃lesion, PL group) and > 5 cm from AE lesions (distal lesion, DL group) among each patient. Finally, a total of eight liver specimens were sampled from four AE patients for scRNA⁃seq analysis. Genome and transcriptome data of liver specimens were processed using the software Cell Ranger and R package. Differentially expressed genes (DEGs) and their biological functions were analyzed using gene ontology (GO) enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis, and the primary intercellular communication patterns and interaction mechanisms were identified among T cell subtypes in liver specimens using the CellChat package. In addition, the developmental stages of T cells were subjected to trajectory analysis with the monocle package to investigate the expression of genes associated with cell growth and tumor transformation, and to predict the developmental trajectories of T cells. Results All four AE patients were female, with a mean age of (25.00 ± 9.06) years, and there were three cases from Jiuzhi County, Golog Tibetan Autonomous Prefecture and one case from Chengduo County, Yushu Tibetan Autonomous Prefecture, Qinghai Province. The viability of single⁃cell samples from eight liver specimens was 90.41% to 96.33%, and a total of 81 763 cells were analyzed, with 19 cell types annotated. Of these cell types, 13 were immune cells (87.60%), and T cells (33.13%), neutrophils (15.40%), and natural killer cells (11.92%) were the three most common cell types. Re⁃clustering of 27 752 T cells and proliferative T cells identified 10 distinct T cell subtypes, with CD8+ cytotoxic T cells (23.43%), CD8+ naive T cells (12.80%), and CD4+ effector memory T cells (17.73%) as dominant cell types. The proportions of T helper 2 (Th2) cells (5.19% vs. 3.63%; [χ2]  = 38.35, P < 0.01) and CD4+ effector memory T cells (21.59% vs. 13.67%; [χ2]  = 244.70, P < 0.01) were significantly higher in liver specimens in the PL group than in the DL group, and the proportion of CD4+ helper T cells was significantly lower in the PL group than in the DL group (7.50% vs. 14.75%; [χ2]  = 330.52, P < 0.01). KEGG pathway analysis revealed that Th2 cells were significantly enriched in cell apoptosis and multiple cancer⁃associated pathways, and CD4+ effector memory T cells were significantly enriched in the regulation of cytokines and chronic inflammation, while CD4+ helper T cells were significantly enriched in immune responses regulation. Trajectory analysis of T cells showed that CD4+ helper T cells were at an earlier developmental stage relative to Th2 cells and CD4+ effector memory T cells, and the expression of inhibitor of DNA binding 3 (ID3), thioredoxin interacting protein (TXNIP), Bcl2⁃associated athanogene 3 (BAG3) and heat shock protein family B (small) member 1 (HSPB1) genes appeared a tendency towards a decline over time. Conclusions CD4+ effector memory T cells and CD8+ cytotoxic T cells are primary interacting cells in the liver specimens of AE patients. Reduced expression of Th2 cells and CD4+ helper T cells contributes to an inhibitory immune microenvironment, which promotes immune evasion by Echinococcus multilocularis, and Th2 cells are significantly enriched in multiple cancer⁃associated pathways, which may be linked to the invasive growth of E. multilocularis.
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    Evaluation of surgical efficacy in patients with hepatic cystic echinococcosis in Gansu Province from 2006 to 2023
    CHENG Xixi, FENG Yu, WANG Xu, WANG Zhiyi, LEI Jiaxi, JIANG Mingzhe, YANG Guobing, ZHANG Xiaojuan, YANG Shijie, WANG Liying
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 247-254.  
    Abstract201)      PDF(pc) (1150KB)(1574)       Save
    Objective To evaluate the therapeutic efficacy for surgical treatments among patients with hepatic cystic echinococcosis in Gansu Province, so as to provide insights into optimization of the diagnosis and treatment strategies against hepatic cystic echinococcosis. Methods The demographic and clinical data of all echinococcosis cases included in central government fiscal transfer payment program for echinococcosis control and undergoing surgical treatments in Gansu Province from 2006 to 2023 were captured. Hepatic cystic echinococcosis patients with complete medical records and follow⁃up data were included in the study, and patients' characteristics, including hospital where patients received diagnosis and treatment, methods of case identification, year of surgery, classification of lesions, number of lesions, size of lesions, course of disease, surgical methods, and post⁃surgical follow⁃up data. The cure and recurrence of hepatic cystic echinococcosis were evaluated according to the Guidelines for Management of Echinococcosis Patients in the Central Government Fiscal Transfer Payment Program, and the cure and recurrent rates were calculated. Results Data were collected from 1 686 surgical patients with hepatic cystic echinococcosis. According to the inclusion and exclusion criteria, 1 222 hepatic cystic echinococcosis patients undergoing surgical treatments were included during the period from 2006 to 2022, including 1 166 cured patients (95.42%) and 88 patients with post⁃surgical recurrence (7.20%), and the cure rate of surgical treatments appeared a tendency towards a rise among patients with hepatic cystic echinococcosis from 2008 to 2022 ([χ2]trend = 19.39, P < 0.05). The cure rates of hepatic cystic echinococcosis were 100% (177/177), 94.81% (128/135) and 94.62% (861/910) among patients detected through regular physical examinations, screened by the central government fiscal transfer payment program for echinococcosis control, and those who passively sought healthcare services, respectively ([χ2] = 9.95, P < 0.05). The cure rates of hepatic cystic echinococcosis were 95.96% (1 046/1 090) among patients with a disease course of 2 years and less and 90.90% (120/132) among patients with a disease course of over 2 years ([χ2] = 6.87, P < 0.05), and there were significant differences in the cure rates among patients with hepatic cystic echinococcosis in terms of number of lesions ([χ2] = 24.44, P < 0.05) and surgical methods (P < 0.05). The cure rate of hepatic cystic echinococcosis patients was significantly higher following initiation of the central government fiscal transfer payment program for echinococcosis control (96.06%, 1 096/1 141) than before the program (86.42%, 70/81) ([χ2] = 16.06, P < 0.05), and the cure rate of hepatic cystic echinococcosis patients was significantly higher in designated hospitals (96.48%, 741/768) than in non⁃designated hospitals (93.37%, 366/392) ([χ2] = 5.78, P < 0.05). The median follow⁃up period was 4 (interquartile range, 7) years among 1 222 hepatic cystic echinococcosis patients undergoing surgical treatments. The recurrent rate of hepatic cystic echinococcosis appeared a tendency towards a decline from 2008 to 2022 ([χ2]trend = 36.86, P < 0.05), with a reduction from 23.08% (9/39) in 2008 to 1.85% (1/54) in 2021, and the post⁃surgical recurrence rate of hepatic cystic echinococcosis was lower following initiation of the central government fiscal transfer payment program for echinococcosis control (5.87%, 67/1 141) than before the program (25.93%, 21/81) ([χ2] = 45.51, P < 0.05). In addition, the post⁃surgical recurrence rate of hepatic cystic echinococcosis was higher in non⁃designated hospitals (10.46%, 41/392) than in designated hospitals (5.60%, 43/768) ([χ2] = 9.12, P < 0.05), and there was a significant difference in the post⁃surgical recurrence rate among patients with hepatic cystic echinococcosis in terms of surgical methods (P < 0.05), with the highest recurrence rate (11.54%) seen among patients undergoing percutaneous fine⁃needle aspiration of cyst fluids⁃based surgical procedures (P < 0.05). Conclusion Since the initiation of the central government fiscal transfer payment program for echinococcosis control in Gansu Province in 2006, an increase in the surgical cure rate and a reduction in the recurrence of hepatic cystic echinococcosis had been found among patients with hepatic cystic echinococcosis, indicating a high overall therapeutic efficacy.
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    Feasibility of the spatiotemporal filtering model for analyzing the spatiotemporal distribution of reported schistosomiasis cases 
    XU Jiayao, WANG Zengliang, GAO Fenghua, ZHANG Zhijie
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 232-238, 275.  
    Abstract268)      PDF(pc) (1718KB)(1564)       Save
    Objective To investigate the feasibility of the spatiotemporal filtering model in analysis of reported schistosomiasis cases, so as to provide insights into analysis of complicated data pertaining to schistosomiasis control. Methods Demographic and epidemiological data of reported schistosomiasis cases in Anhui Province from 1997 to 2010 were collected from Anhui Provincial Center for Disease Control and Prevention, and the annual prevalence of Schistosoma japonicum human infections was calculated. The meteorological data were captured from meteorological stations in counties (cities, districts) of Anhui Province where schistosomiasis cases were reported from 1997 to 2010 at the National Meteorological Information Center, including monthly average air temperature and precipitation. Meteorological data were interpolated using the inverse⁃distance weighting method, and the annual average air temperature and annual precipitation were calculated in each county (city, district). The centroid of the county (city, district) where schistosomiasis cases were reported was extracted using the software ArcGIS 10.0, and the Euclidean distance from each centroid to the Yangtze River was calculated as the distance between that county (city, district) and the Yangtze River. The global Moran's I of the prevalence of S. japonicum human infections in Anhui Province for each year from 1997 to 2010 were calculated to analyze the spatial autocorrelation. A spatial weight matrix was constructed using Rook adjacency, and a first⁃order temporal weight matrix was built to quantify the relationship between disease changes over time. Subsequently, a spatiotemporal structure matrix was constructed. A negative binomial model was built based on the spatiotemporal structure matrix and data pertaining to reported schistosomiasis cases, and a linear model was created between the residual of the model and candidate set feature vectors to determine the optimal subset composition of the spatiotemporal filter through stepwise regression. Then, a spatio⁃temporal filtering model was constructed using the negative binomial model. Negative binomial models, Bayesian spatial models, and Bayesian spatiotemporal models were constructed and compared with the spatiotemporal filtering model to validate the performance of the spatiotemporal filtering model, and cross⁃validation was conducted for each model. The goodness of fit was evaluated using the deviance information criterion (DIC) and Watanabe⁃Akaike information criterion (WAIC), and the effectiveness of model validation was assessed using mean squared error (MSE), while the accuracy of assessment results was assessed using coefficients and their 95% confidence intervals (CI), and the computational efficiency was assessed based on the running time of the model. The four feature vectors with the largest Moran's I values were selected to identify regions with autocorrelation through their schematic diagrams to investigate the differences in spatiotemporal patterns of specific regions.       Results Of all models created, the spatiotemporal filtering model exhibited the highest goodness of fit (DIC = 3 240.70, WAIC = 3 257.80), the best model validation effectiveness (MSE = 42 617.52), and the runtime was 3.18 s, exhibiting the optimal performance. Across all modeling results, the distance from the Yangtze River showed a negative correlation with the number of reported schistosomiasis cases (coefficient values = -4.93 to -3.78, none of the 95% CIs included 0), and annual average air temperature or average precipitation posed no significant effects on numbers of reported schistosomiasis cases (both of the 95% CIs included 0). Schematic diagrams of feature vectors showed that the transmission of schistosomiasis might be associated with water systems in Anhui Province, and localized clustering patterns were primarily concentrated in the northern and western parts of schistosomiasis⁃endemic areas in the province. Conclusion The spatiotemporal filtering model is an effective spatiotemporal analysis characterized by simple modeling, user⁃friendly operation, accurate results and good flexibility, which may serve as an efficient alternative to conventional complex spatiotemporal models for data analysis in schistosomiasis researches.
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    Impact of the number of microsatellite markers on the analysis of population genetic diversity of Schistosoma japonicum
    LONG Juan, MA Lang, ZONG Hongying, ZHOU Zhipeng, YAN Hao, ZHAO Qinping
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 239-246.  
    Abstract220)      PDF(pc) (1112KB)(1512)       Save
    Objective To examine the impact of different numbers of microsatellite markers on the analysis of population genetic diversity of Schistosoma japonicum, so as to provide insights into studies on the population genetic diversity of S. japonicum. Methods Oncomelania hupensis snails were collected from a wasteland in Gong'an County, Hubei Province, and 37 S. japonicum⁃infected O. hupensis snails were identified using the cercarial shedding method. A single cercaria released from each S. japonicum⁃infected O. hupensis snail was collected, and 10 cercariae were randomly collected from DNA extraction. Nine previously validated microsatellite loci and 15 additional microsatellite loci screened from literature review and the GenBank database and confirmed with stable amplification efficiency were selected as molecular markers. Genomic DNA from cercariae was subjected to three multiplex PCR amplifications of microsatellite markers with the Type⁃it Microsatellite PCR kit, and genotyped using capillary electrophoresis. The population genetic diversity of S. japonicum cercariae DNA was analyzed with observed number of alleles (Na), effective number of alleles (Ae), observed heterozygosity (Ho), expected heterozygosity (He), and polymorphism information content (PIC), and tested for Hardy⁃Weinberg equilibrium (HWE) and linkage disequilibrium (LD). To further investigate the impact of the number of microsatellite loci on the population genetic diversity of S. japonicum, the number of microsatellite markers was sequentially assigned from 1 to 24, and the mean and standard deviation of Na were calculated for S. japonicum populations at different locus numbers. In addition, the coefficient of variation (CV) of allelic number (defined as the ratio of the standard deviation to the mean) was determined, and the variation in Na with increasing microsatellite locus numbers was analyzed. Results Genomic DNA from 345 S. japonicum cercariae was selected for genotyping of 24 microsatellite markers, and all 24 microsatellite loci met linkage equilibrium (standardized linkage disequilibrium coefficient D′ < 0.7, r2 < 0.3) and deviated from Hardy⁃Weinberg equilibrium (P < 0.001). The mean Na, Ae, Ho and He were 27.46 ± 2.18, 12.46 ± 0.95, 0.46 ± 0.03, and 0.91 ± 0.01 for 24 microsatellite loci in S. japonicum cercarial populations, respectively, and PIC ranged from 0.85 to 0.96, indicating high genome⁃wide representativeness of 24 microsatellite loci. The mean value of Na-Ae was higher in genotyping with 9 previously validated microsatellite loci (19.88 ± 8.43) than with all 24 loci (14.99 ± 8.09). As the number of microsatellite loci increased, the mean Na showed no significant variation; however, the standard deviation gradually decreased. Notably, if the locus number reached 18 or more, the variation in the standard deviation of Na remarkably reduced. In addition, the standard deviation of Na at 18 loci was less than 5% of the mean Na at 24 loci, with a CV of 4.6%. Conclusions The number of microsatellite loci significantly affects the population genetic diversity analysis of S. japonicum. Eighteen or more microsatellite loci are recommended for analysis of the population genetic diversity of S. japonicum under the current conditions of low⁃prevalence infection and unbalanced genetic distribution of S. japonicum.
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    Spatiotemporal distribution of Aedes albopictus and its influencing factors in China from 2000 to 2019
    JIAO Zerui, QU Lei, WANG Duoquan, ZHANG Yi, LÜ Shan
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 268-275.  
    Abstract800)      PDF(pc) (1564KB)(1472)       Save
    Objective To investigate the spatial distribution of Aedes albopictus in China at different time periods from 2000 to 2019, so as to provide insights into precise management of Ae. albopictus in China. Methods  Data pertaining to the distribution of Ae. albopictus in China from 2000 to 2019 were collected through literature retrieval with terms of "Aedes albopictus", "monitoring", "survey", "density", "distribution", and "outbreak" in national and international databases. The title and time of the publication, sampling sites, sampling time, mosquito capture methods, and mosquito species and density were extracted, and the longitude and latitude of sampling sites were obtained through Baidu Map. Meteorological element data at meteorological observation stations within China were obtained from the National Climatic Data Center of the United States, and the annual maximum temperature, annual minimum temperature, average temperature in January, average temperature in July, annual temperature range, daily temperature range and relative humidity were calculated and subjected to Kriging interpolation. Monthly cumulative precipitation grid data and monthly average temperature grid data with a resolution of 1 km for China from 2000 to 2019 were obtained from the National Tibetan Plateau Scientific Data Center, and the annual precipitation and annual average temperature were calculated cumulatively. Population density data in China from 2000 to 2019 were obtained from the WorldPop Hub, and the gross domestic product (GDP) in China was obtained from the Institute of Geographic Sciences and Natural Resources Research, Chinese Academy of Sciences. The above data were divided into 5⁃year intervals to calculate data during the periods from 2000 to 2004, from 2005 to 2009, from 2010 to 2014, and from 2015 to 2019. Ae. albopictus distribution data were modeled in China from 2000 to 2019 and during each period with the classification random forest (RF) model, to predict the distribution of Ae. albopictus across the country and analyze the distribution of Ae. albopictus based on the seven major climate zones in China. The performance of RF models was evaluated by accuracy, precision, recall, and area under the receiver operating characteristic curve (AUC), and the importance of each feature in the RF model was evaluated with mean decrease accuracy (MDA). Results  A total of 1 191 Chinese publictions and 391 English publications were retrieved, among which 580 articles provided detailed data on the sampling sites of Ae. albopictus and specific sampling years, meeting the inclusion criteria. A total of 2 234 Ae. albopictus sampling sites were included in China from 2000 to 2019, and RF modeling results showed that the overall Ae. albopictus distribution area was mainly found in southeastern and southwestern provinces of China from 2000 to 2019, with scattered distribution in coastal areas of northeastern provinces, such as Liaoning Province. The accuracy, precision, recall and AUC of the RF model were 0.915 to 0.947, 0.933 to 0.975, 0.898 to 0.978, and 0.902 to 0.932 for the distribution of Ae. albopictus at different time periods from 2000 to 2019. Among all features in the RF models, population density was the most contributing factor to the distribution of Ae. albopictus in China, followed by GDP, and all meteorological variables contributed relatively less to the predictive power of the RF model. In China's seven major climate zones, Ae. albopictus was almost entirely distributed in the marginal tropical humid region, the north subtropical humid region, and the warm temperate semi⁃humid region. The combined distribution area of these three zones accounted for 100.0% of the national distribution area from 2000 to 2004, from 2005 to 2009, and from 2010 to 2014, and 99.9% from 2015 to 2019, and the proportion of Ae. albopictus distribution area in the warm temperate semi⁃humid region increased gradually from 20.2% to 30.2%. Conclusions Ae. albopictus is mainly distributed in the southeastern and southwestern provinces of China and is greatly influenced by population and economic factors. The warm temperate semi⁃humid region in China is gradually becoming a hot spot for the distribution of Ae. albopictus.
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    Biological threats to global malaria eliminationⅠAntimalarial drug resistance
    LU Feng
    Chinese Journal of Schistosomiasis Control    2024, 36 (3): 233-238.  
    Abstract442)      PDF(pc) (1470KB)(1315)       Save
    Malaria is an infectious disease that seriously threatens human health. Currently, malaria control mainly depends on antimalarial chemotherapy. However, antimalarial drug resistance is becoming increasingly severe, which poses a great challenge to malaria control, notably treatment of Plasmodium falciparum malaria. To address this challenge, there is a need to facilitate development of novel antimalarial drugs and innovation of treatment strategies, as well as reinforce surveillance and research on antimalarial drug resistance. This article reviews the main categories and use guidelines of current antimalarial agents, summarizes the current status and monitoring methods of antimalarial drug resistance, and proposes the response to antimalarial drug resistance, so as to provide insights into the use of antimalarial drugs and response to antimalarial drug resistance, and contribute to global malaria elimination.
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    Sequencing and analysis of the complete mitochondrial genome of Bulinus globosus
    QIAN Peijun, Masceline Jenipher Mutsaka⁃Makuvaza, LÜ Chao, QIAN Yingjun, WANG Wenya, CHEN Shenglin, XU Andong, XUE Jingbo, XU Jing, ZHOU Xiaonong, Nicholas Midzi, LI Shizhu
    Chinese Journal of Schistosomiasis Control    2025, 37 (2): 116-126, 135.  
    Abstract363)      PDF(pc) (3009KB)(1272)       Save
    Objective To analyze the structural and phylogenetic characteristics of the mitochondrial genome from Bulinus globosus, so as to provide a theoretical basis for classification and identification of species within the Bulinus genus, and to provide insights into understanding of Bulinus⁃schistosomes interactions and the mechanisms of parasite transmission.  Methods        B. globosus samples were collected from the Ruya River basin in Zimbabwe. Mitochondrial DNA was extracted from B. globosus samples and the corresponding libraries were constructed for high⁃throughput sequencing on the Illumina NovaSeq 6000 platform. After raw sequencing data were subjected to quality control using the fastp software, genome assembly was performed using the A5⁃miseq and SPAdes tools, and genome annotation was conducted using the MITOS online server. Circular maps and sequence plots of the mitochondrial genome were generated using the CGView and OGDRAW software, and the protein conservation motifs and structures were analyzed using the TBtools software. Base composition and codon usage bias were analyzed and visualized using the software MEGA X and the ggplot2 package in the R software. In addition, a phylogenetic tree was created in the software MEGA X after sequence alignment with the software MAFFT 7, and visualized using the software iTOL.  Results          The mitochondrial genome of B. globosus was a 13 730 bp double⁃stranded circular molecule, containing 2 ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and 13 protein⁃coding genes, with a marked AT preference. The mitochondrial genome composition of B. globosus was similar to that of other species within the Bulinus genus. Phylogenetic analysis revealed that the complete mitochondrial genome sequence of B. globosus was clustered with B. truncatus, B. nasutus, and B. ugandae into the same evolutionary clade, and gene superfamily analysis showed that the metabolism⁃related proteins of B. globosus were highly conserved, notably the cytochrome c oxidase family, which showed a significant consistency.  Conclusions This is the first whole mitochondrial genome sequencing to decode the compositional features of the mitochondrial genome of B. globosus from Zimbabwe and its evolutionary relationship within the Bulinus genus, which provides important insights for further understanding of the phylogeny and mitochondrial genome characteristics of the Bulinus genus.
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    Cross⁃border joint prevention and control of tropical diseases in countries along the "Belt and Road" Initiative: a framework and roadmap
    QIAN Yingjun, DING Wei, LI Hongmei, WANG Duoquan, LÜ Shan, LI Shizhu, ZHOU Xiaonong
    Chinese Journal of Schistosomiasis Control    2025, 37 (1): 8-13.  
    Abstract376)      PDF(pc) (1292KB)(1203)       Save
    Recently, there has been an increasing risk of importation of tropical diseases into China and the resultant retransmission in the country with the in⁃depth implementation of the "Belt and Road" Initiative, which poses a serious threat to the national public health security. To effectively respond to the cross⁃border transmission risk of tropical diseases and facilitate the process towards tropical disease control and elimination in China and the countries along the "Belt and Road" Initiative, this article analyzes the current status and governance risks of major imported tropical diseases, cross⁃border joint prevention and control polices implemented for tropical diseases and challenges in the establishment of the joint prevention and control system for tropical diseases in China, and discusses the establishment and implementation path of the joint prevention and control system for tropical diseases in countries along the "Belt and Road" Initiative. This path covers the establishment of cross⁃border cooperation mechanisms, research and development and pilot production of Chinese public health products, and implementation of key cross⁃border tropical disease prevention and control projects. The establishment of this system will further improve Chinese prevention and control capabilities for key cross⁃border tropical diseases, build a demonstrative prevention and control model for tropical diseases, and promote international technical exchanges and cooperation of tropical diseases. 
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    Generation of a dense granule protein 3 gene⁃deficient strain of Toxoplasma gondii and its virulence testing
    WANG Peihao, WU Minmin, DU Jian
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 304-309.  
    Abstract159)      PDF(pc) (1729KB)(1202)       Save
    Objective To generate a dense granule protein 3 (GRA3) gene⁃deficient mutant of the Toxoplasma gondii ME49 strain and to test the virulence of the mutant. Methods Gene⁃deficient parasites were generated with the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR⁃associated protein 9 (CRISPR/Cas9) system. Guide RNA (gRNA) was designed using the E⁃CRISPR software, and mutated on the pSAG1::Cas9⁃U6::sgUPRT plasmid using the Q5 site⁃directed mutagenesis kit to generate the pSAG1::Cas9⁃U6::sgGRA3 plasmid. A GRA3 donor plasmid containing GRA3 gene upstream sequences, pyrimethamine resistant gene dihydrofolate reductase⁃thymidylate synthase (DHFR⁃TS) and GRA3 gene downstream sequence was generated, and GRA3 donor DNA was amplified using PCR assay. The pSAG1::Cas9⁃U6::sgGRA3 plasmid and GRA3 donor DNA were electroporated into tachyzoites of the wild⁃type T. gondii ME49 strain. Then, parasite suspensions were inoculated into human foreskin fibroblast (HFF) cells and screened with pyrimethamine to yield pyrimethamine⁃resistant parasites for monoclonal screening. The GRA3 gene deficient monoclonal strain (ME49Δgra3) of T. gondii was identified using PCR and Western blotting assays, and the expression of GRA3 protein was determined in the T. gondii ME49Δgra3 strain using Western blotting. Subsequently, 1 000 freshly lysed tachyzoites of T. gondii ME49 and ME49Δgra3 strains were transferred to 12⁃well plates seeded with HFF cells, and incubated at 37 ℃ containing 5% CO2 for 7 days, and the number of plaques was counted by staining with crystal violet solutions. HFF cells infected with tachyzoites of T. gondii ME49 and ME49Δgra3 strains were stained using Giemsa solutions, and the numbers of cells containing 1, 2, 4, and > 4 T. gondii parasitophorous vacuoles were counted. In addition, the survival rates of C57BL/6 mice infected with T. gondii ME49 and ME49Δgra3 strains were compared 35 days post⁃infection. Results PCR assay revealed successful amplification of both the upstream and downstream homologous arm bands of the DHFR⁃TS gene in the T. gondii ME49Δgra3 strain, and no corresponding bands were amplified in the ME49 strain. The GRA3 band was amplified in the ME49 strain, and the DHFR⁃TS band, rather than GRA3 band, was amplified in the ME49Δgra3 strain. Western blotting determined absence of GRA3 protein expression in the ME49Δgra3 strain. Crystal violet staining showed that the T. gondii ME49 strain produced more plaques than the ME49Δgra3 strain [(352.67 ± 26.39) plaques vs. (235.00 ± 26.29) plaques; t = 5.472, P < 0.01], and Giemsa staining revealed that the proportion of T. gondii parasitophorous vacuoles containing at least four T. gondii tachyzoites was higher in HFF cells infected with the ME49 strain than in those infected with the T. gondii ME49Δgra3 strain [(75.67 ± 2.52)% vs. (59.67 ± 2.31)%; t = 8.113, P < 0.01], and the proportion of T. gondii parasitophorous vacuoles containing at least 1 or 2 T. gondii tachyzoites was higher in HFF cells infected with the T. gondii ME49 strain than in those infected with the T. gondii ME49Δgra3 strain [(24.33 ± 2.52)% vs. (40.33 ± 2.31)%; t = -8.113, P < 0.01]. In addition, mice infected with the T. gondii ME49 and ME49Δgra3 strains started to die 8 and 9 days post⁃infection, and the 35⁃day mortality rates of mice infected with T. gondii ME49 and ME49Δgra3 strains were 10.00% and 70.00% post⁃infection ([χ2] = 6.762, P < 0.01). Conclusions The T. gondii ME49Δgra3 strain has been successfully generated, and GRA3 protein may increase the virulence of the T. gondii ME49 strain.
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    Strategic thinking on management of cross⁃boundary imported schistosomiasis
    XU Jing, LI Shizhen, LI Qin, GUO Suying, LI Shizhu, ZHOU Xiaonong
    Chinese Journal of Schistosomiasis Control    2025, 37 (2): 107-111.  
    Abstract359)      PDF(pc) (1187KB)(1177)       Save
    Schistosomiasis is prevalent in 78 countries and territories worldwide, while the eastern and western parts of sub⁃Sahara Africa bear the highest disease burden due to schistosomiasis. Recently, climate change, international trade and travel, urbanization and war have increased the risk of cross⁃boundary importation and transmission of schistosomiasis, and schistosomiasis has increasingly become a public health concern in non⁃endemic countries and territories. Biomphalaria straminea, the intermediate host of Schistosoma mansoni, has colonized in southern China and its habitats continue to move northward. In addition, cross⁃boundary imported cases of schistosomiasis have been reported occasionally in China. However, the real number of cases may be underestimated greatly due to insufficient diagnostic capacity and weak awareness of case reporting for overseas imported schistosomiasis in healthcare facilities. It is necessary to establish a multi⁃party collaborative mechanism, improve corresponding systems and technical specifications, reinforce surveillance and early warning, and border management, enhance technical reserves and capability building, and improve the awareness of schistosomiasis prevention and healthcare⁃seeking among entry⁃exit personnel, in order to effectively address the threat of cross⁃boundary imported schistosomiasis.
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    Preliminary study on the biological characteristics of heat shock cognate protein 20 of Schistosoma japonicum
    YU Xingang, YUAN Kaijian, LI Yilong, MU Xuanru, XU Hui, LI Qiaoyu, ZENG Wenjing, FU Zhiqiang, HONG Yang
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 294-303.  
    Abstract214)      PDF(pc) (1631KB)(1171)       Save
    Objective To clone and express the heat shock cognate protein 20 (SjHsc20) of Schistosoma japonicum, and to preliminarily investigate its biological characteristics. Methods The target fragment of the SjHsc20 gene was amplified using PCR assay and cloned into the pET⁃28a(+) expression plasmid to generate the recombinant expression vector pET⁃28a(+)⁃SjHsc20, which was then transformed into Escherichia coli BL21 (DE3) competent cells. The recombinant SjHsc20 (rSjHsc20) protein was induced with isopropyl β⁃D⁃thiogalactopyranoside (IPTG) and purified, and the expression of the rSjHsc20 protein was checked with sodium dodecyl sulfate⁃polyacrylamide gel electrophoresis (SDS⁃PAGE). The immunogenicity of the rSjHsc20 protein was detected using Western blotting, and the transcriptional levels of SjHsc20 were quantified in S. japonicum worms at different developmental stages and in male and female adult worms using real⁃time quantitative PCR (RT⁃qPCR) assay. Thirty female BALB/c mice at ages 6 to 8 weeks were divided into three groups, including the rSjHsc20 immunization group, the PBS control group, and the ISA 206 adjuvant group, of 10 mice in each group. Mice in the rSjHsc20 immunization group were subcutaneously immunized with 20 μg rSjHsc20 on days 1, 15 and 31, and animals in the PBS control group were subcutaneously injected with the same volume of PBS on days 1, 15 and 31, while mice in the ISA 206 adjuvant group were subcutaneously immunized with the same volume of ISA 206 adjuvant on days 1, 15 and 31, respectively. All mice in each group were infected with (40 ± 2) S. japonicum cercariae via the abdomen 14 day following the last immunization. Levels of serum specific IgG and its subtypes IgG1 and IgG2 antibodies against rSjHsc20, and the serum titers of anti⁃rSjHsc20 antibody were detected in mice using indirect enzyme⁃linked immunosorbent assay (ELISA). All mice were sacrifice 42 days post⁃infection, and S. japonicum worms were collected from the hepatic portal vein and counted. The eggs per gram (EPG) , worm burden reductions and egg burden reductions were estimated to evaluate the protective efficacy of the rSjHsc20 protein. Results The SjHsc20 gene had an open reading frame (ORF) with 756 bp in length and encoded 252 amino acids, and the rSjHsc20 protein had a relative molecular mass of approximately 29 kDa. The rSjHsc20 protein was recognized by the serum of mice infected with S. japonicum and the serum of mice immunized with the rSjHsc20 protein, indicating that rSjHsc20 had a good immunogenicity. There was a significant difference in the transcriptional levels of the SjHsc20 gene among the 7⁃day (1.001 4 ± 0.065 7), 12⁃day (2.268 3 ± 0.129 2), 21⁃day (1.378 5 ± 0.160 4), 28⁃day (1.196 4 ± 0.244 0), 35⁃day (1.646 3 ± 0.226 1), 42⁃day worms of S. japonicum (1.758 0 ± 0.611 1) (F = 38.45, P < 0.000 1), and the transcriptional level of the SjHsc20 gene was higher in the 12⁃day worms than in worms at other developmental stages (all P values < 0.000 1). The serum levels of anti⁃rSjHsc20 IgG antibody were 0.106 6 ± 0.010 7, 0.108 3 ± 0.010 4, and 0.553 2 ± 0.069 1 in the PBS control group, ISA 206 adjuvant group, and rSjHsc20 immunization group following the last immunization, respectively, and the serum levels of IgG1 antibody were 0.137 3 ± 0.054 0, 0.181 1 ± 0.096 8, and 1.765 8 ± 0.221 1, while the levels of IgG2a antibody were 0.280 3 ± 0.197 6, 0.274 0 ± 0.146 3, and 1.560 4 ± 0.106 0, respectively. There were significant differences in the serum levels of anti⁃rSjHsc20 IgG (F = 397.70, P < 0.000 1), IgG1 (F = 401.00, P < 0.000 1) and IgG2a antibodies (F = 229.70, P < 0.000 1) among the three groups, and the serum levels of anti⁃rSjHsc20 IgG, IgG1 and IgG2a antibodies were higher in the rSjHsc20 immunization group than in the PBS control group and the ISA 206 adjuvant group (all P values < 0.000 1). There was a significant difference in the IgG1/IgG2a ratio among the rSjHsc20 immunization group (1.177 2 ± 0.143 6), the PBS control group (0.428 4 ± 0.199 8) and the ISA 206 adjuvant group (0.559 9 ± 0.181 1) (F = 43.97, P < 0.000 1), and the IgG1/IgG2a ratio was > 1 in the rSjHsc20 immunization group, which was higher than in the PBS control group and the ISA 206 adjuvant group (both P values < 0.000 1). The titers of serum anti⁃rSjHsc20 antibody were all above 1∶16 384 in the rSjHsc20 immunization group following immunizations on days 1, 15 and 31, indicating that the rSjHsc20 protein had a strong immunogenicity. The mean worm burdens were (16.60 ± 5.75), (15.80 ± 5.58) worms per mouse and (14.40 ± 5.75) worms per mouse in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group 42 days post⁃infection with S. japonicum cercariae (F = 0.50, P > 0.05), and the EPG  were 68 370 ± 22 690, 67 972 ± 19 502, and 41 075 ± 13 251 in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group (F = 4.55, P < 0.05), with lower EPG in the PBS control group and the ISA 206 adjuvant group than in the rSjHsc20 immunization group (both P values < 0.05). Immunization with the rSjHsc20 protein resulted in a worm burden reduction of 13.25% and an egg burden reduction of 39.92% relative to the PBS control group.  Conclusions SjHsc20 is successfully cloned and expressed, and the rSjHsc20 protein induces partial immunoprotective effects in mice, which provides a basis for deciphering the biological functions of SjHsc20 and assessing the potential of SjHsc20 as a vaccine candidate.
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    Application of eco⁃immunology in parasitology and parasitic diseases research: a review
    WANG Xu, YIN Jianhai, LIU Hua, HAN Shuai, HU Yuan, SHEN Yujuan, CAO Jianping
    Chinese Journal of Schistosomiasis Control    2025, 37 (4): 428-433, 446.  
    Abstract212)      PDF(pc) (1122KB)(1136)       Save
    As an emerging interdisciplinary field bridging ecology and immunology, eco⁃immunology focuses on the co⁃evolutionary dynamics between hosts and parasites within natural environments, and aims to unravel the ecological mechanisms underlying the formation of host immune strategies, so as to provide new insights into parasitology and parasitic diseases research. Based on case studies of diverse host⁃parasite systems, including insects⁃protozoans, fish⁃cestodes, amphibians⁃nematodes, reptiles⁃arthropods, birds⁃ectoparasites, and mammals⁃helminths, this review summarizes critical eco⁃immunological principles, including host tolerance trade⁃offs under resource constraints, transgenerational epigenetic adaptation, nutrition⁃immunity interactions, and immune conflicts triggered by multiparasite co⁃infections. In addition, the article discusses the feasibility and practical pathways of ecological management and interventions to achieve biodiversity conservation and disease control based on the eco⁃immunological theory, so as to provide innovative insights into responses to address ecological conservation and public health challenges in the context of global changes.
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    Effectiveness of the integrated schistosomiasis control programme in Sichuan Province from 2015 to 2023
    PU Chen, ZHANG Yu, WAN Jiajia, WANG Nannan, SHANG Jingye, XU Liang, CHEN Ling, CHEN Lin, WU Zisong, ZHONG Bo, LIU Yang
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 284-288.  
    Abstract245)      PDF(pc) (1106KB)(1129)       Save
    Objective To investigate the effectiveness of the integrated schistosomiasis control programme in Sichuan Province during the stage moving from transmission interruption to elimination (2015—2023), so as to provide insights into formulation of the schistosomiasis control measures during the post⁃elimination stage. Methods Schistosomiasis control data were retrospectively collected from departments of health, agriculture and rural affairs, forestry and grassland, water resources, and natural resources in Sichuan Province from 2015 to 2023, and a database was created to document examinations and treatments of human and livestock schistosomiasis, and snail survey and control, conversion of paddy fields to dry fields, ditch hardening, rivers and lakes management and building of forests for snail control and schistosomiasis prevention. The completion of schistosomiasis control measures was investigated, and the effectiveness was evaluated. Results A total of 20 545 155 person⁃times received human schistosomiasis examinations in Sichuan Province during the period from 2015 to 2023, and 232 157 person⁃times were seropositive, with a reduction in the seroprevalence from 2.10% (44 299/2 107 003) in 2015 to 1.12% (9 361/837 896) in 2023 ([χ2] = 7.68, P < 0.001). The seroprevalence of human schistosomiasis appeared a tendency towards a decline in Sichuan Province over years from 2015 to 2023 (b = -8.375,t = -10.052,P < 0.001); however, no egg positive individuals were identified during the period from 2018 to 2023, with the prevalence of human Schistosoma japonicum infections maintained at 0. Expanded chemotherapy was administered to 2 754 515 person⁃times, and medical assistance of advanced schistosomiasis was given to 6 436 person⁃times, with the treatment coverage increasing from 46.80% (827/1 767) in 2015 to 64.87% (868/1 338) in 2023. Parasitological tests for livestock schistosomiasis were performed in 35 113 herd⁃times, and expanded chemotherapy was administered to 513 043 herd⁃times, while the number of fenced livestock decreased from 121 631 in 2015 to 103 489 in 2023, with a reduction of 14.92%. Snail survey covered 433 621.80 hm2 in Sichuan Province from 2015 to 2023, with 204 602.81 hm2 treated by chemical control and 4 637.74 hm2 by environmental modifications. The area of snail habitats decreased from the peak of 5 029.80 hm2 in 2016 to 3 709.72 hm2 in 2023, and the actual area of snail habitats decreased from the peak of 8 585.48 hm2 in 2016 to 473.09 hm2 in 2023. The mean density of living snails remained low across the study period except in 2017 (0.62 snails/0.1 m2). Schistosomiasis control efforts by departments of agriculture and rural affairs in Sichuan Province included conversion of paddy fields to dry fields covering 153 346.93 hm2, hardening of 6 110.31 km ditches, building of 70 356 biogas digesters, replacement of cattle with 227 161 sets of machines, and captive breeding of 21 161 070 livestock from 2015 to 2023, and the control efforts by departments of water resources included rivers and lakes management measuring 5 676.92 km and renovation of 2 331 irrigation areas, while the control efforts by departments of forestry and grassland included building of forests for snail control and schistosomiasis prevention covering 23 913.33 hm2, renovation of snail control forests covering 8 720 hm2 and newly building of shelterbelts covering 764 686.67 hm2. All 63 endemic counties (cities and districts) had achieved the criterion for schistosomiasis elimination criteria in Sichuan Province by the end of 2023. Conclusion Following the integrated control efforts from 2015 to 2023, remarkable achievements have been obtained in the schistosomiasis control programme in Sichuan Province, with all endemic counties successfully attaining the schistosomiasis elimination target at the county level.
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    Human infection with Gongylonema pulchrum: a case report and review of relevant literature during the recent 10 years
    TANG Feng, SUN Xiaofeng, XU Xiangzhen, MAO Fanzhen, LIU Yaobao
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 332-336.  
    Abstract178)      PDF(pc) (1359KB)(1106)       Save
    This article presents the diagnosis and treatment processes, and morphological and genetic testing of Gongylonema pulchrum in a case with G. pulchrum found in the oral mucosa. In addition, this article reviews publications pertaining to G. pulchrum human infections by Chinese scientists during the recent 10 years and summarizes the demographic and clinical characteristics, location and number of parasites, diagnosis and treatment processes, and epidemiological surveys of cases infected with G. pulchrum, so as to provide insights into improving the diagnostic capability among clinicians. 
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    Prediction of suitable habitats of Phlebotomus chinensis in Gansu Province based on the Biomod2 ensemble model
    YU Dawei, HOU Yandong, HE Aiwei, FENG Yu, YANG Guobing, YANG Chengming, LIANG Hong, ZHANG Hailiang, LI Fan
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 276-283.  
    Abstract323)      PDF(pc) (1586KB)(1074)       Save
    Objective To investigate the suitable habitats of Phlebotomus chinensis in Gansu Province, so as provide insights into effective management of mountain⁃type zoonotic visceral leishmaniasis (MT⁃ZVL).  Methods The geographical coordinates of locations where MT⁃ZVL cases were reported were retrieved in Gansu Province from 2015 to 2023, and data pertaining to 26 environmental variables were captured, including 19 climatic variables (annual mean temperature, mean diurnal range, isothermality, temperature seasonality, maximum temperature of the warmest month, minimum temperature of the coldest month, temperature annual range, mean temperature of the wettest quarter, mean temperature of the driest quarter, mean temperature of the warmest quarter, mean temperature of the coldest quarter, annual precipitation, precipitation of the wettest month, precipitation of the driest month, precipitation seasonality, precipitation of the wettest quarter, precipitation of the driest quarter, precipitation of the warmest quarter, and precipitation of the coldest quarter), five geographical variables (elevation, annual normalized difference vegetation index, vegetation type, landform type and land use type), and two population and economic variables (population distribution and gross domestic product). Twelve species distribution models were built using the biomod2 package in R project, including surface range envelope (SRE) model, generalized linear model (GLM), generalized additive model (GAM), multivariate adaptive regression splines (MARS) model, generalized boosted model (GBM), classification tree analysis (CTA) model, flexible discriminant analysis (FDA) model, maximum entropy (MaxEnt) model, optimized maximum entropy (MAXNET) model, artificial neural network (ANN) model, random forest (RF) model, and extreme gradient boosting (XGBOOST) model. The performance of 12 models was evaluated using the area under the receiver operating characteristic curve (AUC), true skill statistics (TSS), and Kappa coefficient, and single models with high performance was selected to build the optimal ensemble models. Factors affecting the survival of Ph. chinensis were identified based on climatic, geographical, population and economic variables. In addition, the suitable distribution areas of Ph. chinensis were predicted in Gansu Province under shared socioeconomic pathway 126 (SSP126), SSP370 and SSP585 scenarios based on climatic data during the period from 1991 to 2020, from 2041 to 2060 (2050s), and from 2081 to 2100 (2090s) . Results A total of 11 species distribution models were successfully built for prediction of potential distribution areas of Ph. chinensis in Gansu Province, and the RF model had the highest predictive accuracy (AUC = 0.998). The ensemble model built based on the RF model, XGBOOST model, GLM, and MARS model had an increased predictive accuracy (AUC = 0.999) relative to single models. Among the 26 environmental factors, precipitation of the wettest quarter (12.00%), maximum temperature of the warmest month (11.58%), and annual normalized difference vegetation index (11.29%) had the greatest contributions to suitable habitats distribution of Ph. sinensis. Under the climatic conditions from 1991 to 2020, the potential suitable habitat area for Ph. chinensis in Gansu Province was approximately 5.80 × 104 km2, of which the highly suitable area was 1.42 × 104 km2, and primarily concentrated in the southernmost region of Gansu Province. By the 2050s, the unsuitable and lowly suitable areas for Ph. chinensis in Gansu Province had decreased by varying degrees compared to that of 1991 to 2020 period, while the moderately and highly suitable areas exhibited expansion and migration. By the 2090s, under the SSP126 scenario, the suitable habitat area for Ph. chinensis increased significantly, and under the SSP585 scenario, the highly suitable areas transformed into extremely suitable areas, also showing substantial growth. Future global warming is conducive to the survival and reproduction of Ph. chinensis. From the 2050s to the 2090s, the highly suitable areas for Ph. chinensis in Gansu Province will be projected to expand northward. Under the SSP126 scenario, the suitable habitat area for Ph. chinensis in Gansu Province is expected to increase by 194.75% and 204.79% in the 2050s and 2090s, respectively, compared to that of the 1991 to 2020 period. Under the SSP370 scenario, the moderately and highly suitable areas will be projected to increase by 164.40% and 209.03% in the 2050s and 2090s, respectively, while under the SSP585 scenario, they are expected to increase by 195.98% and 211.66%, respectively. Conclusions The distribution of potential suitable habitats of Ph. sinensis gradually shifts with climatic changes. Intensified surveillance and management of Ph. sinensis is recommended in central and eastern parts of Gansu Province to support early warning of MT⁃ZVL.
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    Risk of and response to cross⁃border importation and secondary transmission of malaria
    ZHOU Yaowu, ZHANG Li, XIA Zhigui
    Chinese Journal of Schistosomiasis Control    2025, 37 (1): 14-18.  
    Abstract291)      PDF(pc) (1058KB)(1038)       Save
    Currently, the global malaria epidemic is still severe. China was certified malaria⁃free by WHO in 2021; however, there is a risk of cross⁃border importation and secondary transmission of malaria via aircrafts and through land bordering countries in China because of the complex environments in border areas between China and neighboring malaria⁃endemic countries, notably the explosive growth in the malaria epidemic in Myanmar in China⁃Myanmar border areas. This article summarizes typical cases of secondary transmission caused by imported malaria across the world, and proposes responses to cross⁃border importation and secondary transmission of malaria in non⁃border and border areas of China, in order to effectively reduce the risk of malaria importation and secondary transmission and consolidate the hard earned malaria elimination achievements.
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    Application of health education during the progress towards malaria elimination in China: a review
    ZHOU Meijin, LIN Kangming
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 321-324.  
    Abstract149)      PDF(pc) (986KB)(980)       Save
    Elimination of malaria is one of important global public health targets. Malaria was once highly prevalent in China; however, China was certified malaria⁃free by WHO in 2021 following decades of integrated control efforts. As an effective intervention, health education plays a critical role during the progress towards elimination of malaria in China, which remarkably increases the public awareness and action capability of malaria prevention and control knowledge. In addition, health education is of great significance to reduce the risk of re⁃establishment of imported malaria following disease elimination in the country. This article reviews the application of community⁃based and school⁃based health education, health education activities targeting entry⁃exit personnel and healthcare workers, and diversified media propagation in the progress towards elimination of malaria in China, so as to provide insights into formulation of malaria control strategy during the post⁃elimination stage in the country. 
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    Analysis and prediction of global burden due to cystic echinococcosis from 1990 to 2035
    LAI Zhen, LIU Gang, ZHAO Haili, QIU Miaomiao, CHEN Jian, LUO En, XIN Junguo, YANG Xiaohong
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 255-267, 283.  
    Abstract233)      PDF(pc) (2761KB)(971)       Save
    Objective To investigate the trends in the global burden due to cystic echinococcosis from 1990 to 2021, and to predict the global burden of cystic echinococcosis from 2022 to 2035, so as to provide insights into formulation of the cystic echinococcosis control strategy. Methods The global age⁃standardized prevalence, mortality, disability⁃adjusted life years (DALYs) rates and their 95% uncertainty intervals (UI) of cystic echinococcosis from 1990 to 2021 were captured from the Global Burden of Disease Study 2021 (GBD 2021) database, and the trends in the global burden of cystic echinococcosis from 1990 to 2021 were analyzed using the Joinpoint regression model. The associations between the global burden of cystic echinococcosis and socio⁃demographic index (SDI) were examined using a smoothing spline model and frontier analysis, and the global burden of cystic echinococcosis was projected from 2022 to 2035 using the Bayesian age⁃period⁃cohort (BAPC) model.  Results The global age⁃standardized prevalence, mortality and DALYs rates of cystic echinococcosis were 7.69/105 [95% UI: (6.27/105, 9.51/105)], 0.02/105 [95% UI: (0.01/105, 0.02/105)], and 1.32/105 [95% UI: (0.99/105, 1.69/105)] in 2021. The global age⁃standardized prevalence of cystic echinococcosis appeared a tendency towards a rise by 0.14% per year from 1990 to 2021, and the global age⁃standardized mortality and DALYs rates of cystic echinococcosis appeared a tendency towards a decline by 4.68% and 4.01% per year from 1990 to 2021, respectively. Joinpoint regression analysis showed that global age⁃standardized prevalence of cystic echinococcosis appeared a tendency towards a decline from 1990 to 2000 [annual percent change (APC) = -0.66%, 95% confidence interval (CI): (-0.70%, -0.61%)] and from 2005 to 2015 [APC = -0.88%, 95% CI: (-0.93%, -0.82%)], and towards a rise from 2000 to 2005 [APC = 3.68%, 95% CI: (3.49%, 3.87%)] and from 2015 to 2021 [APC = 0.30%, 95% CI: (0.19%, 0.40%)]. The age⁃standardized prevalence (r = -0.17, P < 0.05), mortality (r = -0.67, P < 0.05) and DALYs rates of cystic echinococcosis (r = -0.60, P < 0.05) all correlated negatively with SDI across 21 geographical regions from 1990 to 2021, and the age⁃standardized mortality (r = -0.61, P < 0.05) and DALYs rates (r = -0.44, P < 0.05) both correlated negatively with SDI across 204 countries and territories in 2021. Frontier analysis revealed that the age⁃standardized DALYs rate of cystic echinococcosis was still not in line with the frontier in some high⁃SDI countries or territories. In addition, the global age⁃standardized prevalence was projected with the BAPC model to appear a tendency towards a rise among both men [estimated annual percent change (EAPC) = 0.18%, 95% CI: (0.13%, 0.23%)] and women [EAPC = 0.29%, 95% CI: (0.24%, 0.34%)] from 2022 to 2035, and the global age⁃standardized mortality [men: EAPC = -4.71%, 95% CI: (-4.37%, -4.71%); women: EAPC = -4.74%, 95% CI: (-4.74%, -4.74%)] and DALYs rates [men: EAPC = -3.35%, 95% CI: (-3.36%, -3.34%); women: EAPC = -3.17%, 95% CI: (-3.18%, -3.16%)] were projected to appear a tendency towards a decline among both men and women.  Conclusions The global burden of cystic echinococcosis appeared an overall tendency towards a decline from 1990 to 2021; however, the global prevalence of cystic echinococcosis is projected to appear a tendency towards a rise from 2022 to 2035. Intensified cystic echinococcosis control programmes are recommended. 
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    Progress of researches on mechanisms underlying immune escape of Plasmodium
    WU Yuhuang, HE Jing, CAO Xinghang, LI Juntong, SHEN Shuchu, DU Youqin, TAN Chao
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 325-331.  
    Abstract270)      PDF(pc) (1041KB)(963)       Save
    Malaria, a parasitic disease caused by infection with the species of Plasmodium and transmitted by Anopheles mosquito bites, is one of the major public health challenges that seriously threaten human health. Malaria parasites present diverse immune escape strategies to escape from the recognition and clearance of the host immune system, which poses a great challenge to the malaria control programme. This review presents the advances in the mechanisms underlying the immune escape of Plasmodium, including antigenic variation, epigenetic regulation, antagonism against IgM antibody, activation of the cyclic guanosine monophosphate⁃adenosine monophosphate (GMP⁃AMP) synthase⁃stimulator of interferon genes (cGAS⁃STING) signaling, suppression of splenic immune functions, and molecular camouflage, so as to provide insights into development of malaria vaccines and antimalarial agents and formulation of the malaria control strategy. 
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    The future is already here and the wave is coming: progress and challenges in the research and application of artificial intelligence-empowered malaria control programmes
    ZHU Guoding, CAO Jun
    Chinese Journal of Schistosomiasis Control    2025, 37 (2): 112-115.  
    Abstract282)      PDF(pc) (1090KB)(938)       Save
    The rapid rise and fast development of artificial intelligence (AI) has brought unprecedented opportunities and challenges to all sectors, including disease prevention control. Malaria is one of the world's most devastating infectious diseases. This article summarizes the advances in the research and application of AI⁃empowered malaria control programmes, analyzes key challenges during the implementation of malaria control programmes, and proposes future development directions and research proprieties, so as to provide insights into facilitating the translation of AI⁃driven strategies in global infectious disease control efforts.
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    Practice and effectiveness of the optional parasitology curriculum Two Sides of the Same Coin in Parasitic Diseases among international students for the Bachelor of Medicine and Bachelor of Surgery program
    ZHANG Donghui, CHEN Lu, XU Zhipeng, JI Minjun, CHEN Lin
    Chinese Journal of Schistosomiasis Control    2025, 37 (3): 310-315.  
    Abstract176)      PDF(pc) (1026KB)(937)       Save
    To promote convergence education among national and international students, Nanjing Medical University designed an optional curriculum Two Sides of the Same Coin in Parasitic Diseases among international students for the Bachelor of Medicine and Bachelor of Surgery (MBBS) program, and a research⁃based teaching method was employed to teach 31 international students. This curriculum contained three parts, including parasitology⁃related knowledge, basic knowledge and skills of scientific research, and frontier topics in scientific research advances, and the assessment contents included course sign⁃in, classroom activity participation and scientific research presentation. Curriculum evaluation showed 100.0% (8/8) of students in batch 2018—2021, 81.8% (9/11) in batch 2022, and 83.3% (10/12) in batch 2023 with a total score of 80 points and higher, with median scores (interquartile range) of 91.1 (7.0), 90.8 (5.7) points and 90.8 (5.7) points, respectively (H = 0.04, P > 0.05). In addition, a questionnaire survey was performed with a Likert scale to assess the interests in curriculum learning, and the curriculum practicality and importance among 31 international students, and a total of 27 valid questionnaires were recovered, with a recovery rate of 87.1%. Questionnaire survey showed that the median total scores (interquartile range) were 7.0 (0.8) points among batch 2018—2021 students, 6.4 (1.4) points among batch 2022 students and 6.0 (1.1) points among batch 2023 students (H = 2.64, P > 0.05). Collectively, these data demonstrate that this optional curriculum improves the interests in parasitology learning of the international students for MBBS program, as well as their capability of self⁃directed learning, teamwork and innovative. 
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    Analysis and forecast of the disease burden of schistosomiasis in China from 1992 to 2030
    LIN Kai, ZHANG Chenhuan, XU Zhendong, LI Xuemei, HUANG Renzhan, LIU Yawen, YU Haihang, GU Lisi
    Chinese Journal of Schistosomiasis Control    2025, 37 (1): 24-34.  
    Abstract773)      PDF(pc) (2297KB)(921)       Save
    Objective To analyze the trends in the disease burden of schistosomiasis in China from 1992 to 2021, and to project the disease burden of schistosomiasis in China from 2022 to 2030, so as to provide insights into the elimination of schistosomiasis in China.  Methods The prevalence, age⁃standardized prevalence, disability⁃adjusted life year (DALYs) rate and age⁃standardized DALYs rate of schistosomiasis, as well as the years lost due to disability (YLDs) rate and age⁃standardized YLDs rate of anemia attributable to Schistosoma infections in China, the world and different socio⁃demographic index (SDI) regions were captured from the Global Burden of Disease Study 2021 (GBD 2021) data resources, and the trends in the disease burden due to schistosomiasis were evaluated with estimated annual percentage change (EAPC) and its 95% confidence interval (CI). In addition, the age, period and cohort effects on the prevalence of schistosomiasis were examined in China using an age⁃period⁃cohort (APC) model, and the disease burden of schistosomiasis was predicted in China from 2022 to 2030 using a Bayesian age⁃period⁃cohort (BAPC) model.  Results The age⁃standardized prevalence and DALYs rate of schistosomiasis, and the age⁃standardized YLDs rate of anemia attributable to Schistosoma infections were 761.32/105, 5.55/105 and 0.38/105 in China in 2021. These rates were all lower than the global levels (1 914.30/105, 21.90/105 and 3.36/105, respectively), as well as those in the medium SDI regions (1 413.61/105, 12.10/105 and 1.93/105, respectively), low⁃medium SDI regions (2 461.03/105, 26.81/105 and 4.48/105, respectively), and low SDI regions (5 832.77/105, 94.48/105 and 10.65/105, respectively), but higher than those in the high SDI regions (59.47/105, 0.49/105 and 0.05/105, respectively) and high⁃medium SDI regions (123.11/105, 1.20/105 and 0.12/105, respectively). The prevalence and DALYs rate of schistosomiasis were higher among men (820.79/105 and 5.86/105, respectively) than among women (697.96/105 and 5.23/105, respectively) in China in 2021, while the YLDs rate of anemia attributable to Schistosoma infections was higher among women (0.66/105) than among men (0.12/105). The prevalence of schistosomiasis peaked at ages of 30 to 34 years among both men and women, while the DALYs rate of schistosomiasis peaked among men at ages of 15 to 19 years and among women at ages of 20 to 24 years. The age⁃standardized prevalence of schistosomiasis showed a moderate decline in China from 1992 to 2021 relative to different SDI regions [EAPC = -1.51%, 95% CI: (-1.65%, -1.38%)], while the age⁃standardized DALYs rate [EAPC = -3.61%, 95% CI: (-3.90%, -3.33%)] and age⁃standardized YLDs rate of anemia attributable to Schistosoma infections [EAPC = -4.16%, 95% CI: (-4.38%, -3.94%)] appeared the fastest decline in China from1992 to 2021 relative to different SDI regions. APC modeling showed age, period, and cohort effects on the trends in the prevalence of schistosomiasis in China from 1992 to 2021, and the prevalence of schistosomiasis appeared a rise followed by decline with age, and reduced with period and cohort. BAPC modeling revealed that the age⁃standardized prevalence and age⁃standardized DALYs rate of schistosomiasis, and age⁃standardized YLDs rate of anemia attributable to Schistosoma infections all appeared a tendency towards a decline in China from 2022 to 2030, which reduced to 722.72/105 [95% CI: (538.74/105, 906.68/105)], 5.19/105 [95% CI: (3.54/105, 6.84/105)] and 0.30/105 [95% CI: (0.21/105, 0.39/105)] in 2030, respectively. Conclusions The disease burden of schistosomiasis appeared a tendency towards a decline in China from 1992 to 2021, and is projected to appear a tendency towards a decline from 2022 to 2030. There are age, period and cohort effects on the prevalence of schistosomiasis in China. Precision schistosomiasis control is required with adaptations to current prevalence and elimination needs.  
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    Impact of Toxoplasma gondii type I rhoptry protein 16 on programmed cell death ligand 1 expression and its binding to programmed cell death 1 in lung adenocarcinoma cells
    LI Guangqi, ZHOU Yuning, MA Shaohan, TIAN Mei, DANG Tiantian, ZHAO Zhijun
    Chinese Journal of Schistosomiasis Control    2025, 37 (1): 44-54.  
    Abstract228)      PDF(pc) (2105KB)(914)       Save
    Objective To investigate the impact of Toxoplasma gondii type I, II and II rhoptry protein 16 (ROP16) on programmed cell death ligand 1 (PD⁃L1) expression in lung adenocarcinoma cells, and to examine the effects of T. gondii type IROP16 protein on the relative PD⁃L1 expression, the relative PD⁃L1 distribution on the cell membrane surface, and the binding of programmed cell death 1 (PD⁃1) to PD⁃L1 in lung adenocarcinoma cells. Methods Lentiviral vectors overexpressing T. gondii type I, II and III ROP16 proteins were generated, and transfected into the human lung adenocarcinoma A549 cell line. A549 cells were used as a blank control group, and A549 cells transfected with an empty lentiviral expression vector were used as a negative control group, while A549 cells transfected with lentiviral vectors overexpressing T. gondii type I, II and III ROP16 proteins served as experimental groups. Stably transfected cells were selected with puromycin and verified using Western blotting, quantitative real⁃time PCR (RT⁃qPCR), and immunofluorescence assays. The PD⁃L1 expression was quantified at translational and transcriptional levels using Western blotting and RT⁃qPCR assays in A549 cells in the five groups, and the relative PD⁃L1 distribution was detected on the A549 cell membrane surface using flow cytometry. In addition, the effect of T. gondii type I ROP16 protein on the PD⁃1/PD⁃L1 binding was measured in A549 cells enzyme⁃linked immunosorbent assay (ELISA).   Results The relative ROP16 protein expression was 0, 0, (1.546 ± 0.091), (1.822 ± 0.047) and (2.334 ± 0.089) in the blank control group, negative control group, and the T. gondii type I, II and III ROP16 protein overexpression groups (F = 1 339.00, P < 0.001), and the relative ROP16 mRNA expression was (2.153 ± 0.949), (2.436 ± 1.614), (14.343 ± 0.020), (12.577 ± 0.285) and (15.090 ± 0.420) in the blank control group, negative control group and the T. gondii type I, II and III ROP16 protein overexpression groups, respectively (F = 483.50, P < 0.001). The ROP16 expression was higher in the T. gondii type I, II and III ROP16 protein overexpression groups than in the blank control group at both translational and transcriptional levels (all P  values < 0.001). Immunofluorescence assay revealed that T. gondii type I, II and III ROP16 proteins were predominantly localized in A549 cell nuclei. Western blotting showed that the relative PD⁃L1 protein expression was (0.685 ± 0.109), (0.589 ± 0.114), (1.007 ± 0.117), (0.572 ± 0.151), and (0.426 ± 0.116) in the blank control group, negative control group, and the T. gondii type I, II and III ROP16 protein overexpression groups (F = 9.46, P < 0.05), and RT⁃qPCR assay quantified that the relative PD⁃L1 mRNA expression was (1.012 ± 0.190), (1.281 ± 0.465), (1.950 ± 0.175), (0.889 ± 0.251), and (0.230 ± 0.192) in the blank control group, negative control group, and the T. gondii type I, II and III ROP16 protein overexpression groups (F = 14.18, P < 0.05). The PD⁃L1 expression was higher in the T. gondii type I ROP16 protein overexpression group than in the blank control group at both translational and transcriptional levels (both P values < 0.05). Flow cytometry detected that the relative distributions of PD⁃L1 protein were (10.83 ± 0.60)%, (11.23 ± 0.20)%, and (14.61 ± 0.50)% on the A549 cell membrane surface (F = 28.31, P < 0.05), and the relative distribution of PD⁃L1 protein was higher in the T. gondii type I ROP16 protein overexpression group than in the blank control group and negative control group (both P values < 0.001). ELISA measured significant differences in the absorbance (A value) among the T. gondii type I ROP16 protein overexpression group, the blank control group and the negative control group if the concentrations of the recombinant PD⁃1 protein were 0.04 (F = 10.45, P < 0.05), 0.08 μg/mL (F = 11.68, P < 0.05) and 0.12 μg/mL (F = 52.68, P < 0.05), and the A value was higher in the T. gondii type I ROP16 protein overexpression group than in the blank control group and the negative control group (both P values < 0.05), indicating that T. gondii type I ROP16 protein promoted the PD⁃L1/PD⁃1 binding in A549 cells in a concentration⁃dose manner.   Conclusions T. gondii type I ROP16 protein overexpression may up⁃regulate PD⁃L1 expression in A549 cells at both transcriptional and translational levels and the relative PD⁃L1 distribution on the A549 cell membrane surface, and affect the PD⁃1/PD⁃L1 binding in a concentration⁃dependent manner.  
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