Table of Content

16 June 2015, Volume 27 Issue 3

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Integration and demonstration of key techniques in surveillance and forecast of schistosomiasis in Jiangsu Province I Layout and effect of the demonstration sites for schistosomiasis surveillance and forecast

SUN Le-Ping, LIANG You-Sheng, DAI Jian-Rong, HONG Qing-Biao, XU Ming, WANG Wei, GAO Yang, XIE Chao-Yong, WANG Lin, ZUO Yin-Ping, WU Feng, YANG Kun

2015, 27(3):  221.  DOI: 10.16250/j.32.1374.2015087

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Objective Objective To explore and integrate the key techniques used in the surveillance and forecast of schistosomiasis in the water regions along the Yangtze River，so as to provide technical support for identifying rapidly the risk of schistosomiasis transmission and implementing control measures targeting the risk. Methods Methods According to the distribution of water systems and water regions along the Yangtze River in Jiangsu Province，the demonstration sites for surveillance and forecast of schistosomiasis were set across the province，where the integration and demonstration of the techniques regarding monitoring of Schistoso? ma japonicum infection in sentinel mice，human and animal activities，release of forecast information，and emergency treat? ment of water regions at risk of infection were performed. The pattern of human and animal activities was compared with the S. ja? ponicum infection in sentinel mice in the demonstration sites，and the operability of the release of information and emergency treatment of the risk of S. japonicum infection was evaluated. Results Results A total of 50 demonstration sites for surveillance and forecast of schistosomiasis were set in fixed anchor points，opening of the navigation lock to the Yangtze River，freight terminal， agritainment places，ferry，large construction places，and places for guaranteeing the Youth Olympic Games in 23 counties（dis? tricts）of 5 cities，Jiangsu Province. During the period between May and September，2014，the infectivity of water body was monitored by using 5 batches of sentinel mice，with a 99.06%（4 954/5 001）gross recovery rate of mice. S. japonicum infection was detected in a demonstration site，and an infected mouse was found，with a 0.02%（1/4 933）gross positive rate of sentinel mice. The field survey showed 2 088 person?times contacting water，including 91.95%（1 920/2 088）contacting water due to the production such as capturing fish，harvesting and cultivating crops，and repairing and building boats，and 8.05%（168/2 088）contacting water due to the life activity，such as fishing，washing vegetables and playing with water. The people contacted water predominantly in August and September（49.57%） . A total of 859 boats containing 1 877 boatmen were observed，68.22% （586/859）of which were fishing boats containing 1 306 fishermen（69.58%） . There were 32 sheep found in 4 demonstration sites，and 3 sheep were seen in the demonstration site with infected sentinel mouse. Four blue forecasts（emergence of water con? tact）and one orange forecast（S. japonicum?infected sentinel mouse detected）were released across the province，with one fore? cast map released which showed 5 sites with fishing and one site with sheep grazing，one emergency response system initiated， mollusciciding implemented in 10 hm2 high?risk regions，120 sheep grazed in fence，and 35 fishermen given health?education materials，schistosomiasis examination and preventive therapy. In addition，no acute schistosomiasis occurred in the demonstra? tion site with S. japonicum?infected sentinel mice. Conclusions Conclusions The integration and demonstration of the techniques regarding monitoring of S. japonicum infection in sentinel mice，human and animal activities，release of forecast information，and emer? gency treatment of water regions at risk of infection provides an effective approach for the large?scale surveillance and forecast of schistosomiasis.

Analysis of endemic changes of schistosomiasis in China from 2002 to 2010

WANG Qiang, XU Jing, ZHANG Li-Juan, ZHENG Hao, RUAN Yao, HAO Yu-Wan, LI Shi-Zhu, ZHOU Xiao-Nong

2015, 27(3):  229.  DOI: 10.16250/j.32.1374.2015080

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Objective Objective To provide the reference for schistosomiasis control through analyzing the changes of endemic status of schistosomiasis in China in recent years. Methods Methods The annual data of schistosomiasis prevention and control from 2002 to 2012 were collected, and the descriptive analysis was conducted to describe the changes of schistosomiasis endemic status. Re Re? ? sults sults Sichuan, Yunan and Jiangxi provinces reached the criteria of transmission controlled from 2002 to 2010 while Hubei, Hu? nan, Jiangxi and Anhui provinces reached the criteria of infection controlled in 2008. The number of counties where the trans? mission of schistosomiasis was controlled decreased from 110 in 2002 to 80 in 2010. The numbers of estimated schistosomiasis cases and reported acute cases fell from 810.4 thousands and 913 in 2002 to 325.8 thousands and 43 in 2010, respectively, and they were reduced by 59.79% and 95.40% respectively. The number of infected bovine went down from 23 199 in 2002 to 7 173 in 2010, with a fall of 69.03%. However, the Oncomelania hupensis snails breeding areas kept in 3.7 to 3.8 billion m2 and among which, 125 million m2 areas were newly discovered in non?endemic areas during the nine years. In 2010, 90% of schistosomiasis cases, acute cases, infected bovine, and snail breeding areas were concentrated in 5 provinces, especially in the Dongting Lake and Poyang Lake areas and the regions along the Yangtze River. Conclusions Conclusions The epidemic of schistosomiasis decreased sig? nificantly from 2002 to 2010. The lake regions are the key and tough areas for schistosomiasis prevention and control. To consoli? date the achievements, the surveillance and case management need to be strengthened as the extension of snail breeding areas and acute cases reported from other places becomes common.

Trend analysis of schistosomiasis endemic situation in Anhui Province from 2004 to 2014

ZHANG Shi-Qing, GAO Feng-Hua, HE Jia-Chang, ZHANG Gong-Hua, WANG Hao, WANG Tian-Ping

2015, 27(3):  235.  DOI: 10.16250/j.32.1374.2015026

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Objective Objective To analyze the changes of schistosomiasis endemic situation in Anhui Province from 2004 to 2014， so as to provide the evidence for formulating“The thirteenth five year”schistosomiasis control plan. Methods Methods The information for schistosomiasis control and the data of the endemic surveillance of schistosomiasis were collected and analyzed for the chang? es of schistosomiasis endemic situation in Anhui Province from 2004 to 2014. Results Results The Oncomelania hupensis snail areas presented on a rising trend，and reached to the largest area with about 3.1 billon m2 in 2011. However，the snail areas decreased from 2012，and reduced by 10.55% in 2012 compared with the areas in 2011，and reduced to the lowest level in 2014 in recent 10 years. The density of living snails presented a fluctuation situation from 2004 to 2008，and on a decreasing trend from 2008， and the density of living snails was below 1 snail/0.1 m2 after 2011. The infection rate of snails remained stagnant state from 2004 to 2011，and decreased rapidly in 2012，and no schistosome infected snails were found in 2013 and 2014. The schistosome in? fection rate of residents decreased gradually，and the number of acute schistosomiasis was under 50 cases with scattered distribu? tion after 2006，and no acute cases occurred in 2013 and 2014. The infection rate of livestock was above 1% from 2004 to 2011， and reduced to 0.55% in 2012，and it was the first time that the infection rate of livestock was lower than that of residents in the same year in 2014. The progress for achieving the goal of schistosomiasis transmission control accelerated after 2011，and the number of counties that reached the standard of schistosomiasis transmission controlled from 2012 to 2014 was 4，9 and 14 and the number of townships was 33，76 and 32，respectively. Conclusion Conclusion Schistosomiasis control has achieved remarkable effec? tiveness in Anhui Province，but there still exists hard work to consolidate the achievement and reach schistosomiasis transmis? sion interrupted.

Effect of ditching for drain on control of Oncomelania hupensis snail in beaches of Dongting Lake

WEI Wang-Yuan, BU Kai-Ming, WEI Kai-Lin, LUO Zhi-Hong, REN Guang-Hui, CHEN Xiang-Lin, YI Jian-Min, LIU Yu, XIANG Yang, TANG Ke-Wen, YAN Jian-Hui, XIA Meng, DING Liang, LU Xian-Jiang, NIE Dong-Song, LI Yuan

2015, 27(3):  241.  DOI: 10.16250/j.32.1374.2014133

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Objective Objective To evaluate the effect of ditching for drain on the control of the breed of Oncomelania hupensis snails in beaches of Dongting Lake. Methods Methods From November，2009 to November，2012，an O. hupensis snail infested beach of the Yueyang jail and an O. hupensis snail infested beach of Junshan District were selected as research fields in the eastern Dongting Lake area，and the former，as the intervention field，was performed with the ditching for drain by excavators and the latter，as the control field，was not. Results Results Before the project implemented，the average soil moisture contents on the beaches in dry seasons of the two fields were both about 35.56%. After the project implemented，in the intervention field，the average soil mois? ture content was 26.53% which was significantly lower than that（35.56%）in the control field（F = 6.53，P < 0.05） . The under? ground water levels in different heights in the intervention field were lower than those in the control field（ χ2 = 33.33，P < 0.05） . Before the project implemented，the natural death rates of the snails were 0.98% and 0.89% in the two research fields re? spectively（P > 0.05），and after the project implemented（in 2012），no adult and young snails were found in the interventional field，but in the control field，the average densities of living snails and young snails were 29.37 snails/0.1 m2 and 213±108.45 snails/0.1 m2 respectively. Conclusion Conclusion The intervention of ditching for drain can decrease the soil moisture contents quickly and change the ecological condition， therefore， can control the breed of O. hupensis snails in the beaches of Dongting Lake.

Investigation of theoretical knowledge on schistosomiasis prevention and control among professionals in county level in Hunan and Hubei provinces

ZHANG Li-Juan, ZHU Rong, XU Jing, LI Shi-Zhu

2015, 27(3):  246.  DOI: 10.16250/j.32.1374.2015055

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Objective Objective To understand the knowledge level on schistosomiasis prevention and treatment among professionals of schistosomiasis endemic counties in Hunan and Hubei provinces，so as to provide the basis for the ability construction of schistosomiasis control institution. Methods Methods The theoretical test was applied to investigate the mastering situation on schistoso? miasis prevention and control among professionals of 12 selected schistosomiasis endemic counties in Hunan and Hubei provinc? es，and the results were analyzed statistically. Results Results Ninety?six professionals were surveyed. The average score was 66.94± 11.53，in the range of 34?91，and the pass rate was 75.00%. The scoring rates of the knowledge points of the test and treatment of schistosomiasis，snail survey and killing as well as basic knowledge and laws and regulations about schistosome were 68.69%，70.54% and 73.19%，respectively. On the knowledge points of the test and treatment of schistosomiasis and basic knowledge and laws and regulations about schistosome，the differences among different education backgrounds were significant （F = 3.337，4.793，both P < 0.05），and the scores were higher in professionals with higher diploma. In the scores，there were no statistical differences between or among different genders，age groups，professional titles or specialties（all P > 0.05） . Con Con? ? clusion clusion The overall knowledge level on schistosomiasis prevention and treatment of the professionals from 12 schistosomiasis endemic counties in Hunan and Hubei provinces is low. Therefore， the learning of relative knowledge should be strengthened.

Analysis of epidemic situation of malaria in Jiangsu Province from 2005 to 2014

LUO En-Pei, WANG Wei-Ming, LIU Yao-Bao, CAO Yuan-Yuan, ZHOU Hua-Yun, XU Tan

2015, 27(3):  251.  DOI: 10.16250/j.32.1374.2015046

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Objective Objective To understand the epidemic situation and epidemiological characteristics of malaria in Jiangsu Prov? ince from 2005 to 2014，so as to provide the evidence for formulating and adjusting the strategies and measures on malaria elimi? nation in this province. Method Method The epidemiological data of malaria in Jiangsu province from 2005 to 2014 were collected， and the prevalence situation，infection sources as well as the temporal，regional and population distribution of the cases were an? alyzed. Result Result A total of 5 069 malaria cases were reported in Jiangsu Province form 2005 to 2014，there were 3 422 cases （67.51%）of vivax malaria，1 497 cases（29.53%）of falciparum malaria，and 150 cases（2.96%）that were unclassified. Among all the reported cases，2 139（42.20%） were local cases，1 131 were （22.31%） imported cases from other places in Chi? na ，and 1 799 （35.49%） were imported cases from abroad. There were 3 809 male cases and 1 260 female cases，with a sex ratio of 3.02 ∶1. The patients were mainly aged from 25 to ＜60 years（accounting for 67.15%），and were mainly young adult farmers （40.26%）and migrant workers（23.63%），who mainly distributed（accounting for 60.58%）in Xuzhou，Suqian，Huai’ an，Su? zhou and Nantong cities. The predominant malaria type from 2005 to 2009 were vivax malaria，while after 2010，it changed to falciparum malaria. Conclusions Conclusions The malaria epidemic situation has been effectively controlled in Jiangsu Province，which re? flects the initiative achievements of malaria elimination. However，there are still many imported malaria cases from abroad，and the species of the Plasmodium are diverse. Therefore，the imported malaria from abroad still remains the key of malaria control in Jiangsu Province.

Establishment and application of monitoring and evaluation indicator framework for malaria elimination at province and county levels in China

QIAN Ying-Jun, WANG Duo-Quan, DENG Yao, XIAO Ning, LI Shi-Zhu

2015, 27(3):  255.  DOI: 10.16250/j.32.1374.2015067

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Objective Objective To establish a monitoring and evaluation indicator framework in the context of malaria elimination in pilot areas in China，so as to further assess the malaria elimination work in the future. Methods Methods Based on the consultancy， and according to the government documents such as Action Plan for Malaria Elimination in China（2010-2020），Technical Guidance for Malaria Elimination，Programme Evaluation on Malaria Elimination，the indicators were set up at both provincial and county levels and their application was conducted in Yunnan and Anhui provinces to assess the accountability，accessibility and applicability. Results Results An indicator framework was built up including four key components，which were input，process， output and impact indicators，in which 95 were provincial indicators and 98 were county indicators. The completion rate of the indicators of Yunnan and Anhui provinces were 94.7%（90/95）and 95.8%（91/95）respectively；and the completion rate of in? dicators of Tengchong and Feidong counties were 93.9%（92/98）and 92.9%（91/98）respectively. Conclusion Conclusion The estab? lished indicator framework is reliable and applicable，which could effectively help to track the malaria elimination progress and identify gaps.

Investigation on human Cryptosporidium infection in local area of Anhui Province

WANG Qing-Quan, GUO Jian-Duo, CAO Zhi-Guo, WANG Qi-Zhi, LIU Dao-Hua, WANG Tian-Ping

2015, 27(3):  263.  DOI: 10.16250/j.32.1374.2015049

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Objective Objective To investigate the status of Cryptosporidium infection in the population in the local area of Anhui Prov? ince，and discuss the risk factors of the infection，so as to provide the evidence for the prevention and treatment of cryptosporidi? osis. Methods Methods Qianshan County and Lingbi County of Anhui Province were selected as investigation spots，and the oocysts of Cryptosporidium in the feces of the investigation objects and the specific IgG antibody against Cryptosporidium in the serum were checked by using the pathogenic modified acid fast staining method and ELISA，respectively，so as to determine the status of Cryptosporidium infection in these investigation objects. At the same time，the questionnaire surveys were conducted in the inves? tigation objects so as to know about the risk factors of Cryptosporidium infection. Results Results A total of 668 people were investigat? ed in the two counties，635 people received etiological examinations，and 15 people were positive with the positive rate of 2.36%；642 people received serological examinations，and 140 people were positive with the positive rate of 21.81%；628 peo? ple received pathogenic and serological examinations at the same time，and the examination results of the both methods showed that 12 people were positive（there were 4 people in Qianshan County and 8 people in Lingbi County），and the positive rate was 1.94%. The rates of Cryptosporidium infection in the population of Qianshan County and Lingbi County were 1.24%（4/322）and 2.71%（8/295）respectively，and the difference had no statistical significance（P > 0.05） . The single factor analysis found that the rate of Cryptosporidium infection was higher in the children and diarrhea patients；the multivariate logistics regression analy? sis indicated that the rate of Cryptosporidium infection was higher in the people who bred poultry and the diarrhea patients. Con Con? ? clusions clusions The positive rate of serum antibody of Cryptosporidium in the population of the local area of Anhui Province is higher， which indicates that the previous infection is serious，and the rate of Cryptosporidium infection in human is relative to the age， diarrhea and whether there are poultries to be bred in the family，which is worthy of attention in the future prevention and treat? ment.

Species identification of freshwater snail Planorbella trivolvis and analysis of its potential distribution

LI Xiao-Heng, GAO Shi-Tong, GU Wen-Biao, ZHANG Yi, GUO Yun-Hai

2015, 27(3):  268.  DOI: 10.16250/j.32.1374.2015069

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Objective Objective To identify the species classification of an ornamental Planorbidae from a flower market in Shanghai and analyze its potential distribution in China. Methods Methods In August 2013，six freshwater snail specimens were collected from the Wanshang flower market. The species was identified by morphology and molecular biology. An ecological niche model was constructed based on the native geographic presence occurrence data，and projected onto the whole of China to predict the poten? tial distribution. Results Results Their shell external morphology suggested that the specimens belonged to Planorbella trivolvis（Say 1817）of Planorbidae，which is native in North America. The sequence data of a fragment of the mitochondrial cytochrome c oxi? dase subunit I（COI）confirmed its identification. A total of 2 294 georeferenced occurrence points in North America were car? ried out from the Global Biodiversity Information Facility databases and 614 records with coordinates were used to produce a North American native niche model by a maximum entropy method（Maxent） . The projection on China results suggested high probabilities of occurrence mostly in Henan Province and its borderland with nearby provinces. Conclusions Conclusions P. trivolvis is sim? ilarly with Biomphalaria species from shell morphology. It is the first records of the species in China，and the field dispersal is not clear.

Molecular characteristics and RNA interference efficiency of Schistosoma japonicum Sj79 gene

JIANG Yan-Yan, YUAN Zhong-Ying, XU Yu-Xin, ZANG Wei, CAO Jian-Ping, WANG Ying, YIN Jian-Hai, SHEN Yu-Juan

2015, 27(3):  273.  DOI: 10.16250/j.32.1374.2015050

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Objective Objective To study the structural features and characteristics of a novel gene Schistosoma japonicum 79（Sj79） ， and observe its effect of RNA interference（RNAi），so as to provide the experimental basis for its further function study and mechanism study of anti reproductive development of schistosome. Methods Methods The gene structure and characteristics of Sj79 were analyzed by bioinformatics methods. Then the expressions of Sj79 messenger RNA（mRNA）during the different develop? mental stages of schistosome were analyzed and the effects of RNAi silencing were observed by the soaking method. The tran? scriptional levels of Sj79 after RNAi were detected by real time PCR. Results Results The open reading frame of Sj79 contained 696 base pairs with an exon structure. The gene had obvious stage specificity，and its transcriptional level in mature female worms was the highest. After soaking for 3 d， the Sj79 mRNA level［（41.0 ± 12.3） %］in the siRNA?1 group with low dosage（20 nmol/L） was lower than that in the siRNA?NC group［（103.2 ± 14.4） %］，the difference was statistically significant（t = 3.28， P < 0.05） . When with high dosage（200 nmol/L），both the Sj79 mRNA levels in the siRNA?1 group ［（15.8 ± 10.9） %］and siRNA?2 group ［（11.1 ± 8.8） %］were significantly lower than that in the siRNA?NC group［（100.1 ± 6.3） %］ （t = 13.44，27.84，both P < 0.01） . After soaking for 7 d，only the Sj79 mRNA levels in the siRNA?1group［（43.4 ± 4.5） %］and siRNA?2 group［（62.5 ± 5.4） %］with low dosage were lower than that in the siRNA?NC group［（100.4 ± 5.2） %］，and the differences had statistical sig? nificance （t = 8.33，5.07，both P < 0.01） . Conclusion Conclusion Through this study，we have improved the mRNA sequence and genom? ic information of Sj79 gene，and understood its structural features，as well as selected out two effect fragments siRNA?1and siRNA?2，which will provide the basic evidences for the further study on egg laying interference of the female adult worm of schisto? some in vitro.

Recombinant expression of Schistosoma japonicum fructose-1, 6-bisphos-phate aldolase and its expression in different developmental stages of S. japonicum

YAN Ke, ZHONG Zheng-Rong, XU Yun-Xia, DING Shu-Qin, HU Jian-Guo, XU Yuan-Hong, LUO Qing-Li, SHEN Ji-Long

2015, 27(3):  277.  DOI: 10.16250/j.32.1374.2015010

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Objective Objective To clone，express and purify Schistosoma japonicum fructose?1， 6?bisphosphate aldolase（SjFBPA） in E. coli and observe its expression in different developmental stages of S. japonicum. Methods Methods FBPA gene was amplified from S. japonicum adult worm cDNA by using PCR. The amplified product was recombined into pET28a plasmid，and inducibly expressed with IPTG in E. coli BL21. SDS?PAGE and Western blotting were employed to analyze and identify the recombinant protein SjFBPA（rSjFBPA） . Then，rSjFBPA was purified by chromatographic purification and its purity was analyzed by SDS? PAGE. The protein concentration of rSjFBPA purified was measured by the BCA method. Furthermore，SjFBPA mRNA was ana? lyzed in different developmental stages of S. japonicum by RT?PCR. Results Results SjFBPA was successfully amplified by using PCR and identified by restriction enzyme digestion and sequencing. The Western blotting analysis confirmed that the recombinant pro? tein could specifically reactive to the anti ?His? tag monoclonal antibody. The concentration of the purified recombinant protein was about 4 mg/ml. The result of RT?PCR showed that SjFBPA mRNA was expressed in cercaria，schistosomulum，adult worm and egg of S. japonicum. Conclusion Conclusion SjFBPA is successfully recombined and expressed in a prokaryotic system，and SjFBPA mRNA is expressed in cercaria， schistosomulum，adult worm and egg of S. japonicum.

Isolation and identification of macrophages from liver of Microtus fortis

HU Yuan, SUN Lei, XU Yu-Xin, CAO Jian-Ping

2015, 27(3):  282.  DOI: 10.16250/j.32.1374.2015060

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Objective Objective To separate and purify intrahepatic macrophages from Microtus fortis（Mf）and identify its phagocy? tosis. Methods Methods The intrahepatic macrophages from Mf were separated and purified by perfusion，collagenase digestion and density gradient centrifugation. The function of the cells was identified by FACS analysis and ink phagocytosis activity. Results Results The macrophage cells from the liver of Mf were obtained. These cells were bright and circular，and grew adhering to the wall. The proportion of the living cells was 95%. The binding rate of these cells from Mf with anti?mouse CD14 antibody（Clone，Sa2? 8）was about 50% of the rate of macrophage from C57BL/6 mice with this monoclonal antibody. The result of ink?phagocytosis ex? periment of macrophage cells from the liver of Mf was positive. Conclusion Conclusion The method above mentioned is useful to separate and purify macrophage from the liver of Mf. The study builds the foundation for further research on macrophages of Mf against Schistosoma japonicum.

Cloning, prokaryotic expression and immunological identification of Toxoplasma surface antigen IMP1

KOU Jing-Xuan, ZHAO Gui-Hua, WEI Qing-Kuan, XU Chao, ZHU Song, YIN Kun

2015, 27(3):  285.  DOI: 10.16250/j.32.1374.2015025

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Objective Objective To subclone，express and identify the immune mapped protein 1（IMP1）which encodes a surface an? tigen of Toxoplasma gondii. Methods Methods The cDNA of T. gondii RH strain was synthesized by reverse transcription PCR，the IMP1 open reading frame（ORF）was amplified by PCR using the T. gondii RH strain cDNA as template，the PCR products were identified by TA?cloning and sequencing，then the IMP1 ORF was subcloned into the NdeⅠand Xho I sites of the vector pET28b，and the positive recombinant pET28b?IMP1 was identified by double?digesting and sequencing. The protein of 6 × His tagged IMP1 was inducibly expressed in E. coli strain BL21（DE3）with isopropyl β?D?1?thiogalactopyranoside（IPTG），and the induction time，concentration of IPTG and temperature gradients to optimize protein expression conditions were determined. After the cells carried IMP1 were induced by the optimized conditions and harvested，the resulting bacteria were suspended in resuspension buffer and lysed by sonication，and the supernatants were loaded onto the Ni 2+ Chelating Sepharose Fast Flow col? umn for affinity chromatography of the N?terminal 6 × His tagged IMP1 protein. Finally，the fusion IMP1 proteins were identified by Western blotting. Results Results The ORF sequence of IMP1 was successfully subcloned from the cDNA of Toxoplasma Gondii RH strain，and the amplified product was sequenced and identified，based on which the IMP1 ORF gene was inserted into the prokaryotic expression vector pET28b，and the recombinant pET28b?IMP1 was constructed successfully. The double?digesting and sequencing results indicated the validity of the recombinant vector. And the optimized conditions for the expression of IMP1 was determined，namely 0.3 mmol/L IPTG induction for 9 h at 20 ℃. Furthermore，IMP1 protein was expressed solubly and che? lated on Ni 2 + sepharose beads with high affinity，thus this protein could be purified efficiently by affinity chromatography. The pure fusion protein was confirmed with fine immunocompetence by SDS?PAGE and Western blotting. Conclusions Conclusions IMP1 protein can be high efficiently expressed by the E. coli prokaryotic expression systems，the protein of IMP1 is soluble and has stable characters. The study may lay a useful foundation for the following works including in vivo expression of IMP1，crystal structure study of IMP1 and anti?toxoplasmosis subunit vaccine development.

Biological toxicity of heavy metals to Caenorhabditis elegans

HUANG Yue-E, ZHANG Nan, JIANG Yu-Xin, GUO Wei, LI Chao-Pin

2015, 27(3):  290.  DOI: 10.16250/j.32.1374.2015053

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Objective Objective To evaluate the biological toxicity of heavy metals by using Caenorhabditis elegans. Methods Methods The C. elegans at L4 stage were exposed to CdCl2，CrCl3，As2O3，PbCl2，HgCl2 with low concentrations and M9 buffer（the control group）for 72 h，respectively，and the effects of heavy metals with different concentrations on the survival time and reproduction of C. elegans were evaluated. Results Results After exposure to 2.5，10 μmol/L HgCl2 and PbCl2，10 μmol/L CdCl2，and 50 μmol/L CrCl3 for 72 h，respectively，the life spans and survival curves of the C. elegans were different from those in the control group， the differences were statistically significant（all P < 0.05） . After exposure to CdCl2，CrCl3，As2O3，PbCl2 and HgCl2 with the con? centrations of 2.5，50，100 μmol/L for 72 h，respectively，the generational time and brood size of C. elegans were all different from those in the control group（all P < 0.01） . Among the 5 heavy metals at low concentrations，the reproduction toxicity of Hg was bigger than Pb，Cd，Cr，and the toxicity of As was the weakest. Conclusion Conclusion Heavy metal exposure can affect the life span and reproductive toxicity of C. elegans.

Composition and diversity of Acaroid mites' communities in indoor environments of kindergartens inWuhu City

LIU Ting, WANG Shao-Sheng, ZHAN Xiao-Dong, SUN En-Tao, ZHOU Shu-Lin

2015, 27(3):  295.  DOI: 10.16250/j.32.1374.2015058

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Objective Objective To understand the breeding situation of Acaroid mites in indoor environments of kindergartens in Wuhu City，so as to provide the evidence for its prevention and control. Methods Methods From March to June and September to De? cember in 2014，dust samples were collected from 15 kindergartens of 3 ranks every month. Acaroid mites in the samples were isolated，identified and counted. Results Results Totally 360 samples were selected，and 169 samples（46.94%）had mite infestation. Pyroglyphidae accounted for the most in the population，with the breeding rate of 45.00%. Totally 18 504 mites were found，and Dermatophagoides farina and Dermatophagoides pteronyssinus were the dominant species，with the composition ratios of 30.21% （5 590 mites）and 17.83%（3 300 mites），respectively. The third?ranked kindergartens were in the most serious situation con? cerning with 81.67%（98/120）of the breeding rate of Acaroid mites. The differences were statistically significant among the kin? dergartens with different ranks（F = 6.048，χ2 = 73.523，both P < 0.05） . There were no significant differences about the Aca? roid mite population among different grades in kindergartens（F = 0.132，χ2 = 2.377，both P > 0.05） . Conclusion Conclusion Acaroid mites in the kindergartens in Wuhu City were in serious condition，and the population and species composition of Acaroid mite community is influenced by the human behavior and indoors environments.

Metorchis orientalis founded in basin of Wuhu City, Anhui Province

ZHAN Xiao-Dong, LI Chao-Pin, ZHAO Jin-Hong, WANG Shao-Sheng, GU Sheng-Li

2015, 27(3):  299.  DOI: 10.16250/j.32.1374.2015052

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Objective Objective To investigate the epidemic situation of Metorchis orientalis in Wuhu City，Anhui Province. Meth Meth? ? ods ods Three lakes（Fengming，Longwo and Kui lakes）were selected in Wuhu City，and the poultries around the lakes and fresh? water fishes in the lakes were captured to test the infection of M. orientalis，and the infection rates were calculated. Results Results The adult worms of M. orientalis were found in gall bladder and hepatic duct of domestic ducks with infection rate of 16.7%. The metacercariae of M. orientalis were discovered in the muscle of Pseudorasbora parva and Pseudogobio rivularis with infection rates of 7.13% and 3.38% respectively. Conclusion Conclusion M. orientalis is endemic in Wuhu City， ，which should be paid enough at? tention to the animal husbandry， aquaculture and medicine

Analysis of relationship between natural death of Oncomelania hupensis snails and water level in Eastern Dongting Lake district

GAO Jian-Chuan, ZHOU Yi-Biao, LI Lin-Han, WU Jin-Yi, ZHENG Sheng-Bang, SONG Xiu-Xia, HE Zhong, CAI Bin, YOU Jia-Bian, JIANG Qing-Wu

2015, 27(3):  302.  DOI: 10.16250/j.32.1374.2015023

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Objective Objective To study the reasons of natural death of Oncomelania hupensis snails by comparing the differences of the indicator days covered with water（DCW）in snail marshland and non?snail marshland around the build of Three Gorges Dam in Eastern Dongting Lake areas. Methods Methods Two marshlands were selected，one was a non?snail marshland（Qianliang Lake） and another was a snail marshland（Junshan Park） . The measuring points were set through the mechanical sampling. The snails and elevation of the points were surveyed，and the data of the water levels from the hydrological station were collected，and then DCWs were calculated. Results Results From 1995 to 2013，DCWs of the marshland of natural death of snails were all more than that of the snail marshland（P < 0.01） . In Qianliang Lake marshland，the difference between DCW before natural death and DCW from natural death until the dam was not significant （P = 0.23），while DCWs of the two stages both were more than that after the dam（P1 = 0.045，P2 = 0.002） . Before the build of the dam，DCW of the Qianliang Lake marshland of natural death of snails was more than that after the build of the dam（P = 0.013），and there was the same situation in Junshan Park marshland（P = 0.005） . The relationship between snail density and DCW was not significant in Junshan Park marshland（rs = 0.008， P = 0.914），and the reference range of DCW of all the measuring points was 76-251 days. Conclusion Conclusion In the eastern Dongting Lake district，the build of Three Gorges Dam and DCW may be not the direct factors affecting the natural death of snails and the latter may change the distribution of snails.

Research of water contact and labor practice of residents in plateau mountain schistosomiasis endemic areas in Yunnan Province

LI Bing-Gui, CHEN Feng, LI Wen-Bao, YANG Hui, DUAN Yu-Chun, WANG Hai-Ying, MU Liang-Xian, TIAN Shu-Hui, LI Ping, CHEN Shao-Rong, LUO Jia-Jun

2015, 27(3):  306.  DOI: 10.16250/j.32.1374.2014260

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Objective Objective To understand the status of water contact and labor practice of residents in plateau mountain schisto? somiasis endemic areas in Yunnan Province，so as to provide the evidence for formulating the strategy of prevention and control of schistosomiasis in the next step of work. Methods Methods A total of 412 residents aged 6-65 years old in 2 villages were randomly sampled and investigated with questionnaires for their water contacts and labor practices in plateau valley schistosomiasis endem? ic areas. Then the schistosome infection status of the residents was surveyed with the indirect hemagglutination assay（IHA） . Re Re? ? sults sults Among the 412 residents investigated，the rate of water contact was 88.35%，and the main causes of water contact were watering，and swimming and playing. The percentages of residents who had schistosomiasis history were 1.16%，0.00%， 30.80%，3.85%，and 0 in them who had swimming and playing water?contacts，bathing，watering，washing hands，and others， respectively（χ2 = 38.96，P < 0.01） . The positives of IHA in the above?mentioned residents were 18.60%，0.00%，37.60%， 23.08%，and 0，respectively（χ2 = 12.61，P < 0.05） . Conclusions Conclusions The productive infested water contact is the main way of schistosome infection. Therefore，the changes of labor practices and water contact of the residents are very important for the ef? fective prevention and control of schistosomiasis transmission.

Observation on comprehensive control strategy for schistosomiasis with emphasis on infectious source control in Poyang Lake areas from 2009 to 2014

TU Yong-Hong, HONG Xian-Lin, ZENG Ai-Guo, HU Zhuo-Hui, GE Jun, CHEN Hong-Gen, ZENG Xiao-Jun

2015, 27(3):  311.  DOI: 10.16250/j.32.1374.2014266

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Objective Objective To evaluate the long?term effect of the comprehensive control strategy for schistosomiasis with empha? sis on infectious source control in marshland?type endemic areas of Poyang Lake. Methods Methods Three heavy endemic villages with schistosomiasis in the lake areas were selected as pilots for the comprehensive control strategy which included replacing bovine with tractors and forbidding grazing on marshlands. The data of schistosome infections of residents and Oncomelania hupensis snails were collected and analyzed statistically from 2009 to 2014. Results Results There were no patients with schistosomiasis through feces examinations in the 3 pilot villages. The average density of live snails in Niuzhou marshland showed a decline trend and the density was stable in Liulingwei marshland. There were no infected snails in the 2 marshlands，and no schistosome adult worms were found in the sentinel rats Conclusion Conclusion The implantation of the comprehensive control strategy for schistosomiasis with em? phasis on infectious source control can control the prevalence of schistosomiasis during a long period.

Evaluation of implementing effects of Global Fund Malaria Program Round 1 in Junlian County, Sichuan Province

LI Li, XIAO Ning, XU Guo-Jun, LEI Yang, ZHANG Ren-Jie, YAN Da-Pei

2015, 27(3):  313.  DOI: 10.16250/j.32.1374.2015022

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Objective Objective To evaluate the implementing effects of the first round malaria control project supported by the Global Fund on malaria control in Sichuan Province，so as to provide the evidence for intensifying the control strategy of malaria. Meth Meth? ? ods ods Based on the requirements of the malaria project of Globe Fund，the comprehensive malaria control measures including case identification，health education and training for doctors were carried out. Results Results In 2007，the malaria incidence was 0.12/10 000，while it was 1.66/10 000 in 2002，with a reduction rate of 92.77%. The number of malaria endemic towns reduced from 13 to 2. The awareness rate of knowledge about malaria of the local residents rose from 26.86% to 79.83%（χ2 = 403.00， P < 0.01），with an increase rate of 197.21%. The awareness rate of knowledge about malaria of the students rose from 36.87% to 99.22%（χ2 = 359.62，P < 0.01），with an increase rate of 169.11%. Conclusions Conclusions With the support of Globe Fund，the trans? mission of malaria has been effectively controlled. The capacity of malaria control and the malaria control knowledge of the inhab? itants have been enhanced. The implementation of the project has significant effects on malaria control in Sichuan Province.

A bite of parasite, experiences of application of documentaries into parasitology class teaching

GUO Yue, ZHOU Hong-Chang, YAO Yun-Liang, SHAO Sheng-Wen, RU Xiao-Chen, DONG Hai-Yan, ZHANG Ting

2015, 27(3):  316.  DOI: 10.16250/j.32.1374.2015015

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］ ］Medical parasitology education has been facing some difficulties，because it is a course of wide range，lacking clini? cal cases and concerned specimens of parasites currently. In addition，its relationship with life is not closely enough. All these reasons may impact the effect of class education negatively. Therefore，it is important to increase the vitality of parasitology edu? cation and diversify the instructional mode by using the resources from Internet. In recent years，the Discovery Channel has up? loaded a documentary Monsters Inside Me online. This documentary is high professional and closely linked with parasitology. It maintains numbers of clinical cases about parasitic diseases. Each episode is about 3 minutes and shortly enough to be intro? duced into class teaching. However，this resource has not been fully used in domestic temporally. We found that direct introduc? tion of the documentary into class teaching can enrich teaching forms to attract learning interest of students，and finally improve the teaching effect of class. Above that，another popular documentary A Bite of China involves many related knowledge points of parasitology. The appropriate usage of the knowledge can build up close linkage between book and life，which is extremely help? ful to give students a deep impression of parasitology. In brief，it is our strong recommendation to introduce the documentary Monsters Inside Me into class.

Effect of clinical pathway on advanced schistosomiasis patients with acites: a report of 220 cases

LIAO Hong-Bao

2015, 27(3):  319.  DOI: 10.16250/j.32.1374.2015007

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Objective Objective To evaluate the effect of the clinical pathway on advanced schistosomiasis patients with acites. Meth Meth? ? ods ods According to the Handbook of Schistosomiasis Control，and the Clinical Pathway on Advanced Schistosomiasis in Hubei Province，the therapeutic effects，disease awareness，and the satisfaction of patients of 220 advanced schistosomiasis patients （ascitic type）who were treated by the traditional pathway in 2013 and 220 patients who treated by the clinical pathway in 2014， respectively，were evaluated and compared with the statistical method. Results Results As the outcomes of the patients had no signifi? cant difference between the traditional pathway therapy and the clinical pathway therapy，the average hospitalization days and medical expense were reduced when the patients were treated by the clinical pathway. The average hospitalization days were re? duced to 12 days from 14 days. The average medical expense was reduced to RMB 5 800 form RMB 6 400. The average rate of awareness about the disease and the satisfaction rate of the patients were raised to 95% in the patients treated by the clinical pathway. The results of t test for the three indicators above mentioned showed significant differences between the two therapies （all P < 0.05） . Conclusion Conclusion The clinical pathway can standardize the medical behavior and avoid the waste of medical resourc? es，so as to improve the treatment of advanced schistosomiasis.

Measures and effect of malaria prevention and control in Libo County

ZHOU Xue-Mei, MO Hai-Liang

2015, 27(3):  321.  DOI: 10.16250/j.32.1374.2014219

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Objective Objective To understand the implementation status and effect of prevention and control of malaria in Libo Coun? ty，so as to provide the evidence for improving the malaria elimination working. Methods Methods The data about malaria from the county CDC and county hospital were collected and 16 villages from 8 townships were randomly sampled and 10 villagers of each village were investigated. Other information about the prevention and control of malaria was also investigated. Results Results The inci? dence of malaria was decreasing annually，from 5.75 per 10 000 in 2008 to zero in 2012. The malaria monitoring could be well conducted in the county and township levels. The infection source could be controlled in time. The utilization rate of anti?mosqui? to facilities in the residents was 93.25% and the awareness rate of knowledge about malaria prevention and control was 40.13%. Conclusions Conclusions The implementation and effect of prevention and control of malaria are satisfactory in Libo County，but the medi? um control is limited and the active protection consciousness of the residents is not strong. Therefore，the task of malaria elimina? tion is still very arduous.

Progress in researches on molecular markers of Plasmodium falciparum drug resistance

ZHANG Mei-Hua, LU Feng, CAO Jun, GAO Qi

2015, 27(3):  323.  DOI: 10.16250/j.32.1374.2014227

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Effective chemotherapy is the mainstay of malaria control. However，it is undergoing the serious threat by resis? tance of falciparum malaria to antimalarial drugs. In recent years，with the development of molecular biology technology，molec? ular markers have been widely used to monitor antimalarial drug resistance. This paper reviews the researches on the common molecular markers related to Plasmodium falciparum drug resistance.

Comparative analysis of epidemiological situation and trategies to control schistosomiasis between China and African countries

YANG Ya, ZHOU Yi-Biao, PAN Xiang, SONG Xiu-Xia, JIANG Qing-Wu

2015, 27(3):  328.  DOI: 10.16250/j.32.1374.2015009

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Schistosomiasis is a neglected tropical disease which is socioeconomically devastating and a significant cause of morbidity in endemic countries or regions. Some countries and regions have brought down the prevalence of schistosomiasis through positive prevention and control programs. However，in the past few years，with the social and economic development and globalization，re?emergence and spread of schistosomiasis led to a growing concern that new endemic areas may occur. This article analyzes the epidemiological situation and the strategies to control schistosomiasis in China and African countries.

One case of overseas imported quartan malaria

SUI Miao-Miao, ZHAO Shuai, LI Jin, XU Yan

2015, 27(3):  332.  DOI: 10.16250/j.32.1374.2014263

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A case of overseas imported quartan malaria was reported in Weihai City. The patient worked in Africa for many years，had no blood transfusion history，and had not been to malaria endemic regions of China. In approximately half a month af? ter returning from Africa，the patient appeared suspected malaria symptoms，such as irregular fever，sweating，and headache. The patient was diagnosed as quartan malaria by a blood test in basic hospital，reviewed with a microscope by Weihai Centre for Disease Control and Prevention，and checked through the microscopic examination of malaria diagnosis and reference laboratory and PCR amplification by Shandong Institute of Parasitic Diseases. The patient was cured after the treatment with chloroquine/ primaquine for 8 days，and did not recur in the 3?month following up.