Table of Content

30 December 2008, Volume 26 Issue 6

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述评

Malaria Situation in the People′s Republic of China in 2007

ZHOUShui-sen;WANGYi;FANGWen;TANGLin-hua

2008, 26(6):  1-403. 

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论著

Discovery of a Residual Focus of Bancroftian Filariasis after Declaration of its Transmission Interruption in Guangxi

LIXue-ming;YANGYi-chao;HUANGKeng-ling;XUHong-bo;XIEZu-ying;MAIFu-zhen;SHANGShao-ming;SUAi-rong;LIShu-lin;OUFang-qi;WUQin-hua;WUWei-ping;ZHUXue-hua;JIANGZhi-hua

2008, 26(6):  2-408. 

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Objective To describe the discovery of a residual foci of bancroftian filariasis in Fuchuan County where the disease was announced to have been eliminated， and reveal its epidemiologic feature. Methods The investigation was carried out from August 2007 to March 2008 among residents in Changtang village where the first case of filariasiswas found and the neighboring villages. They were screened with two thick blood smears. Immunochromato-graphic technology （ICT） was conducted for those going out but returned and those in surrounding areas. Vector mosquitoes were collected and dissected to find filaria larvae. Historical documents were reviewed and relevant people were interviewed. Results In Changtang administrative village， 1 052 residents were screened and 19 cases with microfilaremia were found in 2 natural villages， with a Mf-positive rate of 1.8% （5.1% in Gangshang and 1.4% in Yinshan respectively）. No Mf-positive case was found in 4 119 residents screened in other 3 villages. The average microfilaria density in the 19 cases was 17.37/60 μl blood. All the 19 cases belonged to 12 families，and 13 cases were relatives to each other，which showed a feature of spatial clustering and family clustering. More patients were identified in the age groups of 20-29 and 50-59, and 57.9% of them were older than 50 years. No larvae were found in 54 Culex pipiens fatigans dissected. Conclusion The Changtang village is identified as a residual focus of bancroftian filariasis with a low, limited endemicity. More cases have been among the elderly with a low average microfilaremia.

A Multi-gene DNA Vaccine Encoding Non-fusion Membrane-anchoring Antigen against Schistosoma japonicum

TANGCheng-wu;LIUShuo-jie;MAYan-bin;LIANGliang;GUOPing;WANGShu-yu;GAOHong;DUANQiu-hong;CHENGJi-zhong;DAIWu-xing*

2008, 26(6):  3-416. 

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Objective To construct DNA vaccine （pIRES-Sj97-Sj14-Sj26） and study its immunogenicity and protective immunity against Schistosoma japonicum. Methods The plasmid pIRES-Sj97-Sj14-Sj26 containing fatty binding protein （Sj14）， GST （Sj26） and paramyosin （Sj97） was constructed and expressed on the membrane. RT-PCR was used to detect the expression of Sj14 mRNA， Sj26 mRNA and Sj97 mRNA in the Hela cells， and IFA for detecting the ex-pression of trans-membrane Sj14， Sj26 and Sj97. Sixty BALB/c mice were randomly divided into 3 groups. Mice in each group respectively received normal saline， pIRES blank vector and pIRES?-Sj97-Sj14-Sj26 by intramuscular injection. Two weeks after the 3rd immunization, 10 mice from each group were sacrificed and total IgG in serum and the level of IFN-γ were detected by ELISA and lymphocyte stimulating index（SI） by MTT. FCM was used to analyze the subgroups of splenocytes. The level of NO secreted by peritoneal macrophages was determined by nitrate reductase approaches. The left 10 mice in each group were challenged with （40±1） cercariae of S. japonicum by abdominal skin penetration. Forty-five days after challenge， mice were sacrificed，and numbers of recovered worms and hepatic eggs were counted． Results RT-PCR showed the expression of Sj14 mRNA, Sj26 mRNA and Sj97 mRNA. IFA proved the expression of Sj26, Sj14 and Sj97 protein. Level of total IgG in the vaccination group, saline group and pIRES blank vector group was （5.62±0.64）、 （1.22±0.20） and （1.48±0.36） mg/ml respectively, showing a statistical significance （P<0.01， P<0.05）. The NO level in macrophages was （321.19±18.03）、（184.12±11.05） and（213.51±15.93） nmol/ml in the 3 groups respectively （P<0.05）, and the lymphocyte stimulating index in the 3 groups was （2.25±0.29）、（1.18±0.07） and（1.22±0.09） respectively（P<0.01）. The INF-γ level was higher in the vaccination group than others （P<0.01）. The percentage of CD4+ and CD8+ T cells increased considerably （P<0.01）. The vaccination group showed a worm reduction rate of 39.9% （P<0.01） and an egg reduction rate of 43.9% （P<0.01）. Conclusion The vaccine candidate pIRES-Sj97-Sj14-Sj26 induces an immune protection in BALB/c mice against Schistosoma japonicum.

Evaluation on Malaria Situation in Areas along Yellow River and Huaihe River by Indirect Fluorescent Antibody Test

ZHENGXiang;FANGWen;HUANGFang;SHENYi-zu;SUYun?-pu;HUANGGuang-quan;ZHOUShui-sen*

2008, 26(6):  4-421. 

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Objective To evaluate malaria situation by indirect fluorescent antibody test （IFAT） in 12 villages of six counties along Yellow River and Huihe River in Anhui， Henan and Hubei Provinces. Methods Crosssectional seroepidemiologic survey was conducted through IFAT in Anopheles sinensis area and A. anthropophagus plus A. sinensis area respectively in 2006 and 2007. The blood-filter papers collected from residents were tested by IFAT to calculate malaria antibody positive rate （APR） and positive GMRT in different age groups. The predictive infection probability was evaluated by reversible catalytic model based on antibody positive rate in the group of less than 20 year-old and compared to the annual parasite incidence. Results In 2006 and 2007， the average antibody positive rate in the investigated villages was 6.1% and 12.0% respectively. In An. sinensis areas， the APR in 2007（12.0%）was significantly higher than that in 2006 （4.1%） （χ₂=69.9， P<0.01） while in An. anthropophagus plus An. sinensis areas there was no significant difference in 2006 （10.3%） and 2007 （12.1%） （χ₂=0.17， P>0.05）. The average positive GMRT of the 12 villages was 26.2 in 2007， slightly higher than that in 2006. In both An. sinensis area and An. anthropophagus plus An. sinensis area， the predictive infection probabilities were higher than the annual parasite incidence with an average 117.3 and 17.20 times higher than the latter respectively. 87.8% of the antibody positive cases were asymptomatic， indicating that there were potential infection sources with symptomatic parasitaemia in the areas. Conclusion Malaria is still prevalent in areas along Yellow River and Huihe River and the actual number of malaria cases are underestimated by the case report system. Malaria reemergence may occur in An. sinensis area but relatively stable in An. anthropophagus plus An. sinensis area.

Urticaria in Relation to Mite Sensitivity and Immunotherapy with Injectio dermatophagoidei farinae

XINGDao-rong;WENTing-huan;YUYang-lin;WEIZhi-ping;LIYi-ming;HANTian

2008, 26(6):  5-427. 

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Objective To investigate the prevalence of mite sensitivity in patients with urticaria or other skin rashes，and to observe the clinical efficacy of a specific immunotherapy（SIT）by the Injectio dermatophagoidei farinae for the patients. Methods In 7-year period（1998-2005）, skin prick test（SPT）with a dust mite（Df）allergen was carried out to detect the prevalence of mite sensitivity in OPD patients suffering from skin rashes. Among the patients sensitive to mite with SPT≥++ response, 3 groups were established. In group A, routine SIT with Injectio dermatophagoidei farinae was conducted. In 9-week increasing dose phase, three stepwise increasing volumes（0.3ml, 0.6 ml and 1.0 ml）each case was injected subcutaneously with mite concentration of 1 ∶ 100 000（w/v）, 1 ∶ 10 000（w/v）or 1 ∶ 5 000（w/v）respectively once a week, followed by a maintenance dose phase for an injection with 1 ∶ 5 000（w/v）1.0 ml/wk for 6 weeks. Group B received rush SIT with mite injections. A total of 15 injections in a course of therapy with same concentration and volume was given as those for the routine ones except shortened intervals, namely, 9 initial injections completed in 3 days by three injections of each concentration per day with two 30 min intervals, maintenance doses were then provided in 6 days with 1 ∶ 5 000（w/v）1.0 ml/d. Thereafter，both groups A and B were maintained for one year with a dose of 1 ∶ 5 000（w/v）1.0 ml every 2 wk. Group C received antihistamine treatment as control, the patients received daily oral Ebastine 10 mg in the morning and Cetirizine dihydrochloride 10 mg in the evening for one week course and pro re nata later. Levels of serum tIgE and serum mite sIgE were detected by ELISA in 20 urticaria cases before and after one year mite SIT. Results Altogether, 2 685 cases with skin rashes were detected by Df allergen SPT. The prevalence of urticaria cases sensitive to mite was 70.3%（1 754/2 496），which was higher than that of eczema 63.5%（54/85）and anaphylactoid purpura 60.6%（63/104）（P<0.05）. 248 cases of urticaria sensitive to mite with SPT≥++ response received SIT with Injectio dermatophagoidei farinae for one year, clinical evaluation revealed an overall efficacy of 91.1%（226/248）with 66.1%（164/248）of excellent or good results，significantly higher than that of antihistamine treatment［12.7%（20/158）］（P<0.01）. Faster improvement of clinical symptoms was shown in rush SIT （group B）than that of routine one（group A）, with higher efficacy in group B than group A（excellent and good results being 76.7% and 55.0% respectively）（P＜0.05）. Serum tIgE and mite sIgE in 20 urticaria cases were detected before and after one year mite SIT，showing that tIgE decreased by half in 40%（8）of the patients，while serum mite sIgE level increased significantly（P<0.01）one year later. Conclusion Mite allergen SPT is an etiological diagnostic technique for urticaria patients sensitive to mite. Clinical efficacy of mite allergen SIT has been proved to be good for the patients，and the rush SIT shows quicker effect of relieving symptoms and better efficacy than that of the routine immunotherapy.

Suppression Effect of Different Stage Antigens of Schistosoma japonicum on Airway Inflammation in a Murine Model of Asthma

JIANGZi-wei;MOHong-mei;WANGLei;CHENGYu-li;WANGQi-hong;LEIJia-hui;LIUWen-qi;LIYong-long*

2008, 26(6):  6-431. 

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Objective To investigate the effect of antigens of different stage Schistosoma japonicum on airway inflammation in a murine model of asthma. Methods 48 female BALB/c mice were randomly divided into eight groups. Mice in group A were given normal saline of equal volume as control. Group B was asthma model which was established by intraperitoneal and intranasal challenge with OVA. Mice in groups C，D and E were immunized with soluble egg antigen（SEA）, soluble male worm antigen（SWA），and schistosomulum antigen（SSA）respectively 4 times in a week interval, followed by OVA sensitization as in group B 1 week after the final immunization. Mice in groups F，G，and H were immunized with SEA, SWA, and SSA respectively but sensitized and challenged with saline instead of OVA. 48 hours after asthma was induced, the mice were sacrificed. Leukocytes and eosinophils were counted in bronchoalveolar lavage fluid（BALF）. The level of IL-5，IL-10 and IFN-γ in BALF was detected. Pathologic changes in lung tissues were observed. Results Inflammation cells，especially eosionphils，appeared in airways of mice in groups B，C，D and E，but with much less number in groups C， D and E. No inflammation cells were seen in airways of group A mice. The number of leukocytes，eosinophils and level of IL-5 in BALF of group B ［（98.4±16.1）×10⁴/ml，（17.6±4.3）×10⁴/ml，（197.9±36.5）pg/ml respectively］ were significantly higher than those of group A ［（8.2±1.1）×10⁴/ml，（0.02±0.01）×10⁴/ml，（12.3±7.4）pg/ml］，however the levels of IL-10 and IFN-γ were significantly lower than that of group A （P<0.05）. The number of leukocytes, especially eosinophils, in BALF of groups C，D and E was significantly lower than that of group B. The level of IL-5 in BALF of groups C，D and E was significantly reduced，while that of IL-10 and IFN-γ in BALF of the 3 groups was significantly higher than group B （P<0.05）. Conclusions The immunization with S. japonicum antigens can effectively modulate the level of cytokines and inhibit the eosinophil infiltration and airway inflammation in asthmatic mice.

Expression of Multiepitope Antigenic Gene of Toxoplasma gondii in Transgenic Tomatoes

ZHOUXiao-hong;HUANGXiao-qin;ZHONGRen-jia;ZHUQiao-zhen;ZHAOYing;GUOYu-qi;CHENXiao-guang*

2008, 26(6):  7-437. 

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Objective To obtain the transgenic tomato plant expressed multiepitope antigenic gene of Toxoplasma gondii（Tg-MAG）. Methods Tg-MAG under the control of E35S promoter， E35S-E81.1 chimeric promoter and E8 2.2 promoter in the plant expression vectors was transferred into the tomato cotyledons and hypocotyls via Agrobacterium-mediated T-DNA transformation. The candidate buds were obtained and elongated by kanamycin resistant screening. After further rooting selection， the selected tomato lines were trained and transplanted to soil and grown in a greenhouse till blossoming and bearing fruit.The transgenic tomato plants were identified using PCR， RT-PCR and Western blotting. Results Tg-MAG was introduced into the transgenic plant genomic DNA and transcribed correctly at expected size 360 bp identified by PCR and RT-PCR. Western blotting results indicated that the recombinant Tg-MAG from 8 transgenic lines， expressed in tomato fruits at expected size 11 900， 2 of them showed strong reaction band with HRP labeled McAb K7H3 against the major surface antigen 1（ SAG1） of T. gondii tachyzoites. Conclusion Tg-MAG transgenic tomato lines were obtained.Tg-MAG recombinant protein with good immune activity was expressed successfully in transgenic tomato fruits.

In vitro Effect of （+）-Usnic Acid on Toxoplasm gondii Tachyzoites

WEILin-lin;CHENGYan-bin*;SIKai-wei;GUNing;LIXiao-qi;LIChen;YUANYu-kang

2008, 26(6):  8-441. 

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Objective To explore the effect of usnic acid on Toxoplasma gondii tachyzoites in vitro. Methods There are four groups named as （＋）-usnic acid group, acetylespiramycin group, DMSO group and normal saline group. Groups of （＋）-usnic acid and acetylespiramycin were further divided into 4 subgroups with final concentration of 5, 10, 25, 50 μg/ml respectively. Normal saline group and DMSO group were respectively given equal volume normal saline and 1% DMSO. Each group have 15 parallel tubes with 1 ml （1×10⁶/ml） T. gondii tachyzoites aqueous suspension. At 1 h, 2 h and 4 h after drug treatment, tachyzoites were counted by light microscope with 0.4% Trypan blue staining. Tachyzoites in aqueous suspension was collected, and washed 3 times by PBS solution. Normal mice were inoculated intraperitoneally and observed for three generations. The cultivated rat cardiofibroblasts were then infected in vitro with T. gondii tachyzoites. At the same time, rat cardiomycytes invasion by T. gondii tachyzoites was investigated. Results At 4 h treated by 10, 25 and 50 μg/ml （＋）-usnic acid, 100% T. gondii tachyzoites were stained. Some tachyzoites were swelling, blunt or round in the two ends; and granules appeared in the cytoplasm, the nuclei were deep stained. The changes of tachyzoites in acetylespiramycin group were similar to （＋）-usnic acid group, 100% T. gondii tachyzoites were stained in 50 μg/ml acetylespiramycin subgroup. In inoculation tests, mice died at 8th to 9th days in 5 μg/ml （＋）-usnic acid subgroup and numerous tachyzoites were detected in ascites. However, most mice survived to be killed in the other （＋）-usnic acid subgroups and the tachyzoites were not found in ascites. All mice in acetylespirmycin groups died at 6th to 8th days after inoculation and many tachyzoites or psudocysts were observed in mice ascites. In infecting cell tests, the cultivated rat cardiofibroblasts were infected in vitro by the tachyzoites after treated with 5 μg/ml （＋）-usnic acid for 4 h， and psudocysts were formed in infected cells. It was negative in the other subgroups of （＋）-usnic acid. But the cultivated rat cardiofibroblasts were infected to varying degree in acetylespiramycin groups, normal saline group and DMSO group. Conclusion （＋）-Usnic acid has a remarkable effect on T. gondii tachyzoites.

Therapeutic Efficacy of Naphthoquine Phosphate Combined with Artemisinine against Plasmodium knowlesi

WANGJing-yan;DINGDe-ben;LIGuo-fu;ZHAOJing-hua

2008, 26(6):  9-444. 

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Objective To study the antimalarial activity of naphthoquine phosphate combined with artemisinine against Plasmodium knowlesi in rhesus monkey. Methods Monkeys were randomly divided into 9 groups （3/group）. The monkeys in groups A and B were treated i.g. once daily for 3 days with 6 or 10 mg/kg of naphthoquine phosphate respectively. Those in groups C and D were treated i.g. twice for the 1st day and once for the 2nd and 3rd day with 31.6 or 100 mg/kg of artemisinine respectively. In groups E, F and G, they were treated i.g. only once with the combination of naphthoquine phosphate 10 mg/kg and artemisinine 10，20 or 25 mg/kg respectively. Groups H and I served as controls which were treated i.g. only once with 10 mg/kg of naphthoquine phosphate and 30 mg/kg of artemisinine respectively. Parasitemia was examined beginning 24 h after drug administration. The observation lasted 105 days when no more parasite was found. Results At 24 h after drug administration, the parasite reduction rate in all groups was higher than 90%. The parasite clearance time for groups E, F and G was （56.0±16.0）, （53.3±4.6），and （56.0±8.0） h respectively, more rapid than that of Group H ［（69.3±4.6） h］. There were 1, 3, 3, 2, 2，and 3 monkeys in groups A, B, D, E, F，and G respectively which were cured. No monkeys were cured in groups C, H and I. Conclusion The combination of naphthoquine phosphate and artemisinine is superior to the single component and the optimum proportion in the combination is 1:2.5 in treating P. knowlesi infection in monkeys.

实验研究

Effect of Maternal Anti-Trichinella Antibodies on Intestinal Worm Burden in Sucking Mice

WANGYan-juan;XUDong-mei;CUIJing;WANGZhong-quan*

2008, 26(6):  10-449. 

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Objective To observe the transfer route of maternal anti-Trichinella antibodies and its effect on intestinal worm expulsion after the sucking mice was infected with T. spiralis. Methods Ninety-eight sucking mice （Kunming strain） were divided into 4 groups： mice born and nursed by infected mothers （group A）， mice born of normal mothers and fostered by infected mothers （group B）， mice born of infected mothers and fostered by normal mothers （group C），mice born of and nursed by normal mothers （group D）. Blood was taken from tail veins of 4 groups of sucking mice when they were 14， 21 and 42 days old，respectively. Serum anti-Trichinella antibody level was detected by ELISA using T. spiralis muscle larvae excretory-secretory （ES） antigens. Each sucking mouse were then orally challenged with 200 T. spiralis muscle larvae， and intestinal worm burden was observed 18 h after challenge infection. Results At 18 h after challenge infection， the mean intestinal worm burden of groups A， B， C and D of sucking mice aged 14 days was 5， 5， 19 and 18 larvae respectively. The worm burden of 4 groups of little mice aged 21 days was 18， 19， 75 and 73 larvae， respectively. Groups A and B of 14 and 21 day old mice fostered by infected mothers were shown to harbor obviously fewer worms than groups C and D of mice fostered by normal mothers （F₁₄=10.056， F₂₁=35.062， P<0.01）. Serum absorbance （A₄₉₂） of groups A （0.177， 0.235） and B （0.183， 0.250） of 14 and 21 day old mice was significantly higher than groups C （0.108， 0.105） and D （0.067， 0.065） （F₁₄=75.326， F₂₁=60.867， P<0.01）. The intestinal worm burden in 4 groups of sucking mice aged 14 and 21 days showed significant negative correlation with their serum absorbance （r14=-0.621， r21=-0.756， P<0.01）. The intestinal worm burden in 4 groups of sucking mice aged 42 days was 55， 51， 46，and 60 larvae respectively， with no significant difference （F42=0.916， P>0.05）， their serum anti-Trichinella antibodies were negative. The intestinal worm burden in 4 groups of offspring mice aged 42 days showed no correlation with their serum absorbance （r42=-0.291， P>0.05）. Anti-Trichinella antibodies in sera of sucking mice were detected 6 hours after anti-serum Ig to T. spiralis was intravenously injected into the lactating mothers. Conclusion The maternal anti-Trichinella antibodies have been transferred from mother to filial mice mostly through milk，the antibodies can enhance the worm expulsion from intestine when the sucking mice aged 14-21 days was infected with T. spiralis larvae.

In vitro Culture of Tachyzoites of Toxoplasma gondii RH Strain in HeLa Cells

WULiang;CHENSheng-xia*;LILin-jie;CHEFei-hu;DENGHong-yan;JIANGXu-gan

2008, 26(6):  11-456. 

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Objective To establish a stable and efficient in vitro culture model for tachyzoites of Toxoplasma gondii RH strain. Methods Tachyzoites were inoculated into HeLa cells to establish an in vitro culture system. The proliferation of tachyzoites was observed under microscope by the method of Giemsa stain. At the same time， the longterm tachyzoites maintenance in HeLa cells was established，and the effect of different temperature and time on the yield and motility of tachyzoites were observed. Results The RH strain tachyzoites were cultured and maintained in HeLa cells. Most HeLa cells were destroyed 96 h after inoculation. In the long-term culture system， the proliferation of tachyzoites was stable and its virulence to mouse showed no decrease. Furthermore， tachyzoites in this system proliferated by 5-20 times and （1-5）×10⁷ tachyzoites were harvested. When cultured in HeLa cells at 37 ℃ for 72 h then at 25 ℃ for another 120 h， the tachyzoites proliferated by more than 40 times with a motility rate of over 90%. However， rare HeLa cells left in the medium were found. Conclusion Tachyzoites of T. gondii RH strain can be subcultured in HeLa cells for a long time， and high proliferation rate of tachyzoites can be obtained from this in vitro culture system.

In Vitro Observation on Albendazole Sulfoxide and its Enantiomers against Echinococcus granulosus Protoscolex

BAOGen-shu*;ZHANGHong;JINGTao;QIAOHua;WANGJing

2008, 26(6):  12-461. 

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Objective To investigate in vitro anti-hydatid efficacy on Echinococcus granulosus protoscolex （EgPSC） by using albendazole sulfoxide （ASOX） and its two enantiomeric antipodes， L-ASOX and D-ASOX. Methods Eg protoscoleces were divided into eight groups and cultured in the DMEM culture media under two concentrations（50 μg/ml and 100 μg/ml） of ASOX， L-ASOX and D-ASOX respectively. The appropriate controls included （i） a culture containing an equal amount of DMSO and （ii） a culture medium alone. The mortality of EgPSC in each group was daily counted until 100% EgPSC death in some groups. Results Significant difference of EgPSC mortality was found among the three drugs with various concentrations compared to control group （P<0.01）, and a significant difference between L-ASOX group and D-ASOX group （P<0.05）. There were no statistical difference between ASOX group and D-ASOX group （P>0.05）， but between ASOX group and L-ASOX group （P<0.05）. On the 9th day of culture, the mortality of protoscleces with the concentration of 50 μg/ml was 93.6%, 56.2% and 99.0% in ASOX, L-ASOX and D-ASOX groups respectively, and those under the concentration of 100 μg/ml were 100%, 74.5% and 100% respectively. The mortality was 19.1% and 22.5% respectively in the control and solvent groups. Conclusion ASOX，L-ASOX and D?-ASOX demonstrate significant effect of anti-Eg protoscolex in vitro. D-ASOX shows stronger effect than L-ASOX.

Anti-fecundity Effect of Soluble Immature Egg Antigen of Schistosoma japonicum in Mice

LICui-ying;LIFei;JIAXue-mei;WANGHong;CHENLian-yong

2008, 26(6):  13-465. 

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Objective To study anti-female fecundity effect in mice immunized with soluble immature egg antigen（SIEA） of Schistosoma japonicum， and observe possible mechanism. Methods Immature eggs were collected from the liver of a rabbit 34 days after being infected with 3000 cercariae of S. japonicum， and soluble antigen was prepared routinely. 20 ICR mice were divided randomly to 2 groups. 11 mice were immunized with SIEA （100 μg per mouse） and Freund adjuvant and the other 9 were injected with equal volume of sterile normal saline and Freund adjuvant as control. All mice received injection 5 times in 2 weeks interval. One week following the last vaccination， mice were challenged with 30±2 cercariae of S. japonicum Yunnan isolate. Feces were collected on day 45. All mice were sacrificed on day 46. The number of worms collected， number of eggs in feces， female uterus and liver， and the number of egg granulomas on the liver surface were compared between the 2 groups. The vitelline gland of female and testes of male worms were examined by transmission electron microscopy. Results There was no significant difference on the worm number between the immunized group and control （P＞0.05）. The eggs per gram feces and liver， eggs in uterus per female，and egg granulomas on the liver surface were （56.68±24.78）， （5 826±437）， （49.94±12.53） and （10.04±1.13）/0.25 cm2， respectively in immunized group， while in control group these were （89.93±32.18）， （10 016±3 541）， （76.54±19.77） and （19.22±2.45）/0.25 cm2 respectively， all with significant difference （P<0.05）. Ultrastructure of the reproductive organs of paired adult worms showed that mature vitelline cells and lipid droplets in the cytoplasm of the cells decreased in the vitelline glands of immunized mice. More sustentacular cells and fewer spermatids were seen in testes of immunized mice. Vacuoles were seen in the cytoplasm of vitelline cells and sustentacular cells. Conclusion The results indicated that SIEA may have an anti-fecundity effect possibly through inhibiting the maturation of germinal cells.

现场研究

Follow-up Survey on the Imported Cases of LymphaticFilariasis in Guangdong Province

ZHANGXian-chang;HUANGShao-yu;DENGZhuo-hui;OUZuo-yan;WUWei-ping;LUOXiong-cai;CHENXi-xin;ZHANGQi-ming;LINRong-xing;RUANCai-wen;WANGJin-long;CUIHui-er

2008, 26(6):  14-411. 

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Objective To make a survey on the possible imported cases with microfilaremia from a recently identified outbreak focus in Fuchuan County of Guangxi Region. Methods Moving people as temporary laborers from Guangxi and periphery residents were screened by thick blood smears for microfilariae and ELISA for specific IgG4 in Sept 2007. Results 989 persons in cities/districts of Nansha, Panyu, Shantou and Zhongshan were investigated. Among 150 persons from Changtang administrative village of Guangxi, 47 were from two outbreak focuses, Ganshang and Yinshan natural villages in Changtang. Three cases with microfilaremia were found, with a microfilariemia density of 24, 20 and 2 per 120 μl blood, respectively. The 3 cases were all from the 2 natural villages, with a microfilariemia positive rate of 6.4% （3/47）. Meanwhile, anti-filaria IgG4 was detected for other 1 501 persons, including those from the outbreak focuses, from surrounding areas and from non-endemic area as control, fifteen cases were found with IgG4 positive. Of the 15 positive cases, 12 were from the 2 outbreak natural villages, with a positive rate of 25.5%, higher than those of others （x₂=295.83, P＜0.01）. The cases with IgG4 positive but microfilaremia negative were reexamined by ICT kit and all showed negative. Conclusion It is suggested that no further transmission occurs although microfilaremia cases have beeen imported from the outbreak focus of Guangxi.

综述

The Structure and Function of Schistosome Tegument and Related Proteomic Study

QIANMen-bao;HUWei*

2008, 26(6):  15-471. 

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Schistosome is covered by a living syncytium， called tegument， which plays important role in nutrient uptake and immune evasion. Recently， tegumental proteomic research identified lots of proteins， of which there may be potential targets for diagnosis， drugs and vaccines.

Field Trial of Malaria Vaccine

CAOJun;GAOQi

2008, 26(6):  16-475. 

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Vaccine is expected to be a promising tool for malaria prevention and control. However, safe and effective malaria vaccine is not yet available for field use so far. They can be pre-erythrocytic stage vaccine, blood stage vaccine and transmission-blocking vaccine. This review summarizes the progress of the vaccine development in the recent field trials.

Limitations of the Application of RNA Interference in the Study of Animal Parasitic Nematodes

WANGZi-jian;WUXiu-ping;DENGHong-kuan;LIUMing-yuan*

2008, 26(6):  17-479. 

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RNA interference is a powerful tool for investigating gene function which has been used extensively in model organisms such as Caenorhabditis elegans and has been adapted as a high-throughput screening method to identify genes. However， recent research indicates that the application of RNAi to animal parasitic nematodes has some problems. For example, the variability of RNAi to different parasitic nematodes or the same nematode in differert stages is great. Reasons to explain why RNAi does not work well in animal parasitic nematodes include： ① the efficacy of RNAi delivery methods in different nematodes is extremely variable；② RNAi mechanism has been applied to related nematodes and most of them are gene functional defect；③ different lifestyles of nematodes may influence on the efficacy of RNAi. If we can find out the methods which can solve these questions， there are still application prospects for using RNAi in parasitic nematodes with improved RNAi effect.

研究简报

Staining Improvement for Blastocystis hominis Specimen

ZHANGRui-qi;CHENHou-biao;WEIHui-ping;SUShui-lian*

2008, 26(6):  18-445. 

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Schaudinn solution was used to fix the Blastocystis hominis specimen and an improved Harris hematoxylin staining was applied to stain it. The method shows clearer internal structure of the parasite, simpler and less time-consuming than the traditional iron hematoxylin solution.

Epidemiological Survey on Paragonimiasis in Ningbo City，Zhejiang Province

XUGuo-zhang*;QIANBao-zhen;YELi-ping;ZHANGJie-nan;LUFeng;SUNYa-wei

2008, 26(6):  19-451. 

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Serum samples were collected from 2 643 suspected cases of paragonimiasis in 2000-2007 from the outpatient departments of the city hospitals and surrounding areas， and the infection rate in the inhabitants， the first and second intermediate hosts， and animal reservoir hosts were investigated in the historical endemic areas. Serum samples were detected and 417 were found antibody positive （15.8%）. Among residents in the historical endemic areas，the sero-positive rate was 3.1%（46/1 462），2.8%（18/649） and 3.2% （26/813） in males and females respectively （x₂=0.183 3， P>0.05）. The infection rate in first intermediate host （snails）， second intermediate host （crabs） and animal reservoir hosts was 0.05%（9/19 368）， 31.1% （15 627/50 313） and 11.9% （52/438） respectively. Evidently， natural nidi for Paragonimus spp. still exist in Ningbo City.

Scan Statistic Method with Moving Window and its Application in the Temporal Clustering Detection of Acute Schistosomiasis

XULi-li;CHENGeng-xin;CUIDao-yong;HEZong-gui;ZHOUXiao-nong*

2008, 26(6):  20-458. 

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Acute schistosomiasis cases from 2001 to 2006 in Guichi District, Chizhou City, Anhui Province, were collected and analyzed by scan statistic method with moving window, central degree approach and circular distribution method in order to detect the temporal clustering of schistosomiasis. The analyzed data provided by the method showed that the temporal clustering of acute schistosomiasis from 2001 to 2003 in Guichi District became more concentrative, and then disperse, resulting in a final disappearance of the clustering in 2006 （LLR=4.14, P>0.05）. The temporal clustering for all cases in the six years detected by scan statistic method was between August to September in 2002 （LLR=18.5， P＜0.01）, which was consistent with the real condition. Whereas the results from the central degree and the circular distribution methods could only provide limited information and were biased in the analysis in 2002, indicating that the scan statistic method with moving window can provide ample information with more robust and credible results.

An Epidemiological Survey on Hydatid Disease in Yushu Prefecture of Qinghai Province

LIUHai-qing;HEDuo-long;WUXian-hong;WANGHu;MAXiao;ZHAOYan-mei

2008, 26(6):  21-481. 

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Local inhabitants in 8 towns/townships of the counties Yushu, Zhiduo and Chengduo were examined with serology and ultrasound in 2006. Among 2 251 people tested by indirect hemagglutination （IHA）, 207 showed anti-hydatid IgG positive （9.2%）. Ultrasound examination found 106 cases out of 2 581 people, with a morbidity of 4.1%. Females showed higher sero-positive rate （11.3%） and morbidity rate （5.0%） than males （6.6% and 3.0% respectively）. The highest sero-positive rate was in the group of 40-49 years old （16.4%）. The morbidity rate increased with age, with the highest rate （15.5%） in the group of 60 years old and above. Occupationally, those involved in semi-agriculture and semi-animal husbandry showed highest sero-positive rate and morbidity rate, 21.5% and 11.4% respectively. The survey demonstrated that the prevalence of hydatid disease in human population is at a high level in Yushu Tibetan Autonomous Prefecture.

Cloning and Eukaryotic Expression of the Gene Encoding Myosin from Brugia malayi

XIEDong-fang;FANGZheng;HUANGWei-qun;SHENQin;TONGHai-yan;XUBang-sheng

2008, 26(6):  22-484. 

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Total RNA was extracted from periodic microfilariae of Brugia malayi and its myosin partial gene （Bm-M55） was amplified by RT-PCR. The PCR product was cloned and then subcloned into pcDNA3.1（+）vector. The recombinant eukaryotic plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and was transfected into COS-7 cells subsequently. The expressed protein was identified by SDS-PAGE. Bm-M55 mRNA was highly expressed in transfected COS-7 cells. The deduced amino acid sequence showed to be identical with that of Bm-M55, and the recombinant protein was about Mr 55 000.