Chinese Journal of Schistosomiasis Control

Recombinant expression Schistosoma japonicum and immunogenicity identification of antigen epitopes inducing T-cell response

Li Jian, Yin Xu-ren, Yu Chuan-xin, Xu Yong-Iiang, Hua Wan-quan, He Wei, Liang Yo

2007, 19(4): 241-246.

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Objective To synthesize and fusion express the predicted T-cell epitopes of Schistoso-
ma japonicum., and analyze their immunogenicities. Methods The plus and minus oligo-nucleic acid
strands of epitopes P7, P17, P18 were synthesized following their DNA sequence, respectively.
The Nco I restriction enzyme sites were added to the 5' end of epitope gene and the Xho I restriction
enzyme sites were added to the 3' end of epitope gene. The plus and minus strand of each epitope
gene was annealed to form double strand DNA fragments. Then the double strand DNA fragments
encoding epitope peptide were cloned into the site between Nco I and Xho I of plasmid pET32c(+)
to construct recombinant plasmid which was transformed into E.coli DH5a. The recombinant plas-
mid containing P7, P17, P18 genes respectively was identified by PCR, restriction digestion and
DNA sequencing, and then transformed into E.coli BL21 (DE3) for expressing the fusion protein.
The fusion protein of peptide-thioredoxin(Trx) was expressed by inducing with IPTG and analyzed
with SDS-PAGE. The fusion proteins were purified with N12+ column affinity chromatography.
Meanwhile, the peptides P7, P17, P18 were synthesized artificially following their amino acid se-

quence. By using the purified epitope peptide fusion proteins and synthesized epitope peptides, the
splenic cells of C57BL/6J mice immunized with ultraviolet-attenuated cercaria of Schistosoma japon-
icum were stimulated respectively. The stimulation activity of fusion proteins and synthesized pep-
tides were assayed by detecting the incorporation rate of 3H-thymidine. Results The double strand
DNA fragments of epitopes P7, P17, P18 were successfully cloned to form recombinant plasmids,
all of which could express a 20 kDa fusion protein. Both the fusion protein and synthesized epitope
peptides of P7 and P17 were able to stimulate the lymphocyte cells to proliferation effectively. Con-
clusion The peptide P7 and peptide P17 are testified as T-cell epitopes of Schistosoma jaPon,icum.

Glucose tolerance test of advanced schistosomiasis patients

GUO Hou-xiang, YANG Jin-hong, ZHAI Sha-hu

2007, 19(4): 246-246.

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Cloning and expression of gene encoding myophilin-like protein of Schistosoma japonicum and study on the antigenicity of recombinant protein

Tong Qun-bo, Liu Shu-xian.,Li Xiao-hong, Xu Yu-xin, Shen Yu-juan, Cao Jian-ping

2007, 19(4): 247-251.

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Objective To clone and express the gene encoding Schistosoma japonicum myophilin-
like protein (SjcMLP) and to study the antigenicity of the recombinant protein. Methods The
SjcMLP gene was amplified by PCR. The PCR product was cloned into T vector, and then sub-
cloned into expression vector pQE30. The recombinant plasmid of pQE30-SjcMLP was transformed
into E. coli M15, and induced with IPTG for expression. The bacteriallysis was conducted by ultra-
sonication and the supernatant was analysed by SDS-PAGE. The recombinant protein (reSjcMLP)
was purified with the Ni-NTA resin, and analysed with SDS-PAGE and Western blot. The titers of
sera from C57BL/6 mice immunized subcutaneously with reSjcMLP were detected by ELISA. Re-
suIts The results of SDS-PAGE and Western blot showed that the molecular weight of expressed
fusion protein was around 24. 8 kDa and was recognized by the sera from the mice infected with
Schistosoma japonicum. The purified protein of reSjcMLP was coated for ELISA test and the IgG
titers in the sera from the mice immunized with reSjcMLP were as high as l : 12 800 reacted with.
However, no significant difference was found in worm reduction rates between the immunized mice
and control mice. Conclusions The fused recombinant protein of reSjcMLP is successfully ex-

Construction of a T7 phage display cDNA library from lung of Microtus |fortis

2007, 19(4): 252-256.

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Objective To construct a T7 phage display cDNA library from the lung of Microtus
fortis for further screening the schistosomiasis-resistence-related genes of Microtus fortis. Methods
mRNA was isolated from total RNA extracted from the lungs of Microtus fortis by TRIzol
reagent, and was used to synthesize double strain cDNA by the reverse transcription. Then the
double strain cDNA was given with EcoR I and Hind I[ adhering ends by ligation with the direc-
tional EcoR I /Hind II linkers and digestion with EcoR i and Hind m. The double strain cDNA
fragments longer than 300 bp in length were fractionated by the Mini Column, and ligated into the
T7 Select l0-3b vector with EcoR I and Hind m adhering ends. After packaging in vitro, the re-
combinant T7 Select l0-3b was transformed into BLT5403 to construct a T7 phage display cDNA li-
brary. Results The library constructed here contained l. 5 Xl06 clones and the titer of the amplied
library was l. 1X101z pfu/ml. The PCR identification results of 100 clones picked at random showed
that 91% clones were recombinant and 90% of recombinant clones contained cDNA fragments
longer than 300 bp in length. Conclusion A T7 phage display cDNA library from the lung of M/-
crotus fortis is successfully constructed.

Epidemiological investigation on new schistosomiasis endemic area in Ouchi River

2007, 19(4): 256-256.

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Surveillance on schistosomiasis in China in 2006

Zhu Rong, Dang Hui, Guo Jia-gang

2007, 19(4): 257-262.

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Objective To understand the endemic situation of schistosomiasis among 80 national
surveillance sites for schistosomiasis in 2006,so as to provide scientific evidence for making out
schistosomiasis prevention and control measures. Methods According to the national surveillance
protocol, the endemic situation of schistosomiasis in residents and livestock, and the status of On-
comelania snails and other factors were investigated in the 80 national surveillance sites for schisto-
somiasis, and all the data were collected and analysed. Results Among the 80 national surveillance
sites,the average positive rate of residents with sera examination was 13. 60% and the average posi-
tive rate with stool examination was l. 59%. The average positive rate of floating population with
sera examination was 5.79%. There were 143 advanced patients among the national surveillance
sites, and 12 acute patients were reported in 2006, but no breaking-out epidemic reported. The pro-
portion of domestic animals stable breeding were about 4. 74% of 5 365 examined animals in 2006,
and the others were not. The infection rate of domestic animals was 5. 93% that declined by 34. 49%
compared with that in 2005 and especially for the infection rate of cattle and sheep. A total of
7 429. 63 hmz of areas were surveyed. The areas of snail habitats, infected snail habitats and the
new snail habitats were 4 994. 01,621. 75 hm2 and 21. 75 hm2,respectively. The density of living
snails and infection rate of snails were 0. 544 8/0. 1 m2 and 0. 37%, which increased compared with
infection rate of 0. 26% in 2005. Conclusion The endemic situation of residents, cattle and snail
habitats declined in 2006 compared with those in 2005 in the national surveillance sites.

Analysis of endemic situation of surveillance sites of schistosomiasis in Jiangsu Province, |2006 |i |Status of residents and livestock infection

Hong Qing-biao, Huang Yi-xin, Sun Le-ping, Zhang Jian- feng, Liang You-sheng,etc

2007, 19(4): 263-267.

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Objective To understand the current endemic status of schistosomiasis of residents
and livestock in Jiangsu Province, in order to provide a scientific basis for decision-making of pre-
vention and control. Methods According to the national and provincial surveillance protocol, 82
representative endemic villages were selected as surveillance sites and the infection of Schistosoma
japonicum in residents and livestock were investigated. Results The positive rates of serological
and stool examination for local permanent residents were l. 63% and 0. 035%, respectively. The
positive rates of serological and stool examination for floating population were 2. 42% and 0. 988% ,
respectively. It was estimated that the infection rates of S. japo挖icum in residents, cattle and goats
were 0. 007 4%, 0. 80% and l. 90%, respectively. Conclusions The endemic status of schistoso-
miasis in Jiangsu Province is low and stable, the infection rate of floating population is much higher
than that of local permanent residents, the livestock with dispersal breeding is seldom, and the
whole infection rate is low, but it is still high in some special areas. Therefore, the floating popula-
tion and livestock are import epidemic factors in some local areas of Jiangsu Province.

Anaesthesia selection of advanced schistosomiasis patients with surgical therapy

Cao Xiao-lin, Jiang Hui-wen, Zhang Bao-long

2007, 19(4): 267-288.

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Wu Ming-can, Chen Shi-jie, Luo Guo~cai, Liu Jian-xiong, Yu Hui, Li Mao-kun, etc

2007, 19(4): 268-273.

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Objective To explore the clinical characteristics and diagnostic methods of cerebral
schistosomiasis. Methods The data of CT, MRI and the cerebral spine fluid (CSF) immunoassay
in 42 patients with cerebral schistosomiasis were analyzed retrospectively. Results Forty cerebral
schistosomiasis patients were diagnosed by the iconography and immunoassay, 1 by the postopera-
tive pathological examination and l by diagnostic treatment with praziquantel. Conclusions CT and
MRI are of great value to determination of the position and nature of the cerebral schistosomiasis.
The sensitivity and accuracy of MRI are better for indicating the main pathological changes of the
disease than those of CT. The CSF immunoassay plays an important role in the differential diagno-
sis of cerebral schistosomiasis.

Systemic skin anaphylaxis induced by praziquantel chemotherapy: one case report

2007, 19(4): 273-273.

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Growth difference of Plasmodium vivax in Anopheles sinensis and An. anthropophagus during the periods of clinic attack and diapause of vivax malaria patients

2007, 19(4): 274-277.

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Objective To compare the growth of Plasm,odium vivax in A规opheles sinensis(An., s)
and Anopheles anthropop矗agus (An. a) during the periods of malaria clinic attack and diapause.
Methods The blood samples of patients during the clinic attack and diapause of vivax malaria pa-
tients in the vivax epidemic area in China were collected, feeding the mosquitoes of An. s and An. a
by using the artificial membrane feeding system /咒 vivo in the lab, and the mosquitoes were dissect-
ed during the day 7-9th and 14th after the infection and the oocysts and sporozoites in the stomach
and gland of the mosquitoes were counted, respectively. Results The oocyst positive rate in An.s
fed by Plasmodium vivar during the fever stage was lower than that in non-fever stage, the sporo-
zoite positive rate in An. a fed by Plasmodium vivax during the fever stage was lower than that in
non-fever stage. The positive mosquito rate with oocyst and sporozoite infected by Plasmodiu,m v/-
var in the fever stage to An. s and An., a were lower than those in non-fever stage. The infective
sporozoite intensity of An.s fed by Plasmodium vivar in the fever stage was lower than that in non-
fever stage, but the reverse result was found to An. a. Conclusion There is a significant difference
between the periods of malaria clinic attack and diapause of the oocyst and sporozoite infection to An. s and An,. a.

Wu Cheng-guo,Xu Xing-jian,Wang Ru-bo,Xiao Bang-zhong, Wei Feng-hua,etc

2007, 19(4): 278-280.

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Objective To study the risk factors of schistosomiasis transmission in the Three
    Gorges Reservoir areas (TGRA) and provide the technique project for dynamic monitoring schisto-
    somiasis in TGRA. Methods The analogue experiment of Oncomelania snail survival was done in
    TGRA. The potential risk factors of schistosomiasis were investigated and the potential risk factors
    included the Oncomelania snail imported into TGRA and the imported infection sources of schistoso-
    miasis in the floating population and livestock. The potential infected factors of schistosomiasis
    transmission were investigated with the change of social-economy development after the dam con-
    struction. At last, the emphases of schistosomiasis monitoring were found out. Results   On.comela-
    nia snails could survive and breed in the appropriate environment of TGRA. The infection sources
    of schistosomlasis were mainly the floating population from endemic areas. With the plants and live
    stock imported, the snails and infection sources of livestock might be imported to TGRA. The so-
    cial-economy development might increase the risk of schistosomiasis imported. The TGRA hasoeen
    a potential prevalence area of schistosomiasis. Conclusion The emphases of monitoring schistoso-
    miasis in TGRA should be the infection sources of schistosomiasis such as floating population, im-
    ported livestock and the risk factors of importing Oncomelania snails.

Study on method of determination of appropriate sample size of Oncom,elania hupensis 'in marshland and lake regions

Ni Ying, Cui Dao-yong| Zhang 2hi-jie, Peng Wen-xiang

2007, 19(4): 281-284.

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Objective To explore a possible method for determining the appropriate sample size
of Oncomelania hupensis in the marshland and lake regions. Methods Four bottomlands were se-
lected randomly along the Qiupu River in Guichi District, Anhui Province. Oncomelania snails were
surveyed carefully in these bottomlands and brought back to the laboratory to judge whether they
were alive or not with mild water. All the data of Oncomelania hupen,sis were doubly input into the
EpiData software and checked for the errors. Repeated sampling was performed in the four databas-
es according to different sample sizes by using SAS software and the corresponding snail density was
also calculated. According to the values of sample size and corresponding snail density, sample size-
snail density absolutely changing curve and relatively changing curve were drawn to determine the
appropriate sample size of snail survey. Results The appropriate sample sizes of snail survey were
40,200,100 and 200 frames in Gutan bottomland, Caizi bottomland, Qiupu bottomland and Ouyang
bottomland, respectively. Conclusion Sample size-snail density absolutely changing curve and rel-
atively changing curve could provide some instructions for choosing the appropriate sample size dur-
ing the process of snail survey in the marshland and lake regions.

Analysis of acute schistosomiasis:a report of 88 cases

Rong Xian-bing, Yuan Mei-zhi, Dong Juan| Peng Xiao-wu, Fu Zheng-yin

2007, 19(4): 284-295.

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Molluscicidal experiment of endophytes from Pseudolarix kaempferi Gord

Guo Shang-bin, He Jia, Cheng Xi, Zhong Nan

2007, 19(4): 285-288.

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Objective To screen 18 endophytes from Pseudolarir kaempferi Gord for molluscici-
dal effect and identify them by morphology. Methods Molluscicidal tests were performed according
to the immersion test suggested by WHO and the strain screened was identified by the slide culture.
Results The mortality rates of snails immersed by JJ18 broth salified (pH = 7) were 26.7%,
76. 7% and l00. 0% for 24,48 h and 72 h, respectively, and 53. 3% and 86. 7% in 5% and 10%
concentrations of JJ18 broth, respectively. The active components were extracellular moiety of the
broth which had no acute toxicity to fish, and JJ18 strain belonged to Aspergillus. Conclusion Ex-
tracellular moiety of endophyte JJ18 from Pseudolarir kaempferi Gord is a new resource of mollusci-
cide.

Sensitivity analysis of the yearly extreme low temperature to predict the distribution of On.comelania hupensis

Cui Dao-yong,Zhang zhi-jie, Ni Ying', Pen.g Wen.-xiang, Zhuang Jian-lin, etc

2007, 19(4): 289-292.

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Objective To explore the sensitive temperature index in order to predict the distribu-
tion of Oncomelania hupensis. Methods Sixty-one weather stations, distributed in Shanghai, Jiang-
su. Zhejiang, Anhui, Jiangxi, Hubei, Hunan, Hebei, Henan: Shandong, Shanxi, Shaanxi,
Sichuan provinces (municipality) , were selected as research stations. North latitude 34^。 was set as
the boundary of Oncomelan,ia hupensis's distribution. The differences of yearly extreme low temper-
ature and mean temperature in a year between snail areas and no snail areas were tested respectively
by using t test to show their respective significance on the prediction of Oncomelania hupensis's dis-
tribution. Finally, unconditional logistic regression analysis was used to determine which tempera-
ture index had the better effect to predict the distribution of Oncomelan,ia hupensis. Results Both
the yearly extreme low temperature and mean temperature in a year had significant effect on the dis-
tribution of Oncomelania hupensis under the t test (t= - 6. 49, P<0.01; t= - 3. 93, P<0. 01).
The differences of the yearly extreme low temperature and mean temperature in a year between snail
areas and no snail areas were 6. 72 'C and 3. 02 'C respectivcly, and the degree of their differences
was about 2. 23 times. The impact of yearly extreme low temperature was more important than that
of mean temperature in a year because mean temperature in a year became no longer statistically sig-
nificant if they were tested simultaneously by using unconditional logistic regression analysis. Con-
clusion Yearly extreme low temperature may be more sensitive on predicting the distribution of Oncomelania hupensis than mean temperature in a year.

Analysis of schistosomiasis epidemic in Taizhou City in 2006

Guo Jia-hong, Zhou Hong-wei|Chen Jian-feng

2007, 19(4): 292-292.

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Grading of schistosomiasis patients' hepatic lesions by ultrasonography and analysis of portal vein and splenic vein haemodynamics

Yi Song-tao, Huang Wei-heng, Peng Yi-xin, Guo Jin.-zhi

2007, 19(4): 293-295.

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Objective To study the relationship between the grading of schistosomiasis japonica
hepatic lesions by B-ultrasonography and the portal vein and splenic vein haemodynamics. Methods
The grades of 125 schistosomiasis patients' hepatic lesions were determined by Doppler ultra-
sonography according to the method recommended by WHO/TDR. The portal vein and splenic vein
inner diameters and blood flow velocity were examined and the portal vein and splenic vein volumes
of blood flow were calculated in the patients above mentioned and 51 normal persons. Results The
inner diameters, blood flow velocity and volumes of blood flow of p.ortal vein and splenic vein in the
patients were significantly different from those in the normal persons, and also there were signifi-
cant differences among the patients with different degrees of fibrosis. Conclusions The changes of
schistosomiasis patients' portal vein haemodynamics is connect with the liver parenchyma character-
istics. The step up of the ratio of blood flow volume of splenic vein with the portal vein is break-
down with the aggravate of the hepatic lesion.

Development and application of positioning Oncomelania snail Information Management System based on GIS

Liu Gang, Huang Qiong-yao, Peng Fei, Liu Nian-meng, Sun Hui, Liu Ming, Yang Lin

2007, 19(4): 296-298.

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Objective To develop and apply the Oncomelania snaillnformation Management Sys-
tem based on GIS. Methods With calculating the path by some GPS points and snail information
database, the visualized snail information profiles were drawn automatically. Results The Posi-
tioning Oncomelania snail Information Management System has been developed and running in
Changsha, and it made snail information standardized, visualized and digital, not only supplied
many precise queries, but also formed a link of snailinformation based on years. Conclusions The
positioning Oncomelania snail Information Management System based on GIS raises the efficiency of
information management and the veracity of information, it makes the snail information more inte-
grated and systemic, which provides the basis for developing the On.comelania snail Information
Management System of Hunan Province.

Epidemiological survey of chronic lymphatic filariasis after itsm elimination in Shanghai City

Zhang Xiao-ping, Zhu Min,Cai Li, Wang Long-ying, Wang Zhen-yu, Jin Yan- jun

2007, 19(4): 299-303.

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Objective To understand the status of chronic lymphatic filariasis cases after its e-
limination in Shanghai City. Methods All the chronic filarial cases historically registered were in-
cluded in this survey and some persons with significant signs were also included. The data of clinical
examination and results of serotests and microfilaria examination were recorded and analysed, and
the database was set up. Results A total of 826 chronic filariasis patients were surveyed, who dis-
tributed in 102 townships of 9 districts, Shanghai City. About 87. 17% of the patients were over 70
years old, and 96. 73% of them had the disease over 30 years. The main three chronic signs were
lymphedema/elephantiasis, chyluria, and hydrocele, accounting for 96. 73% , 2. 06% and l. 21% ,
respectively. The cases with Ⅰ-Ⅳstage of limb lymphedema/elephantiasis were for 94. 63%,
ones with V , VI stage of elephantiasis for 5. 37%, and 16. 71% of them were presented with acute
onset of lymphadenitis. In terms of healthy status, 91.77% of the patients were healthy, and
2. 18% of them were unable to care for themselves. The positive rate for filarial specific IgG4 anti-
body was 15. 01%, and no any microfilaria was found in the blood samples among the antibody posi-
tive cases. Conclusions The status of elimination of lymphatic filariasis in Shanghai is maintained.
More attention should be put in the chronic filariasis patients because of their older age and onsets
of the disease.