Table of Content

31 October 2008, Volume 26 Issue 5

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论著

Effect of Tribendimidine, Artesunate, Artemether and Praziquantel，Administered Intragastrically at Single, Multiple or Combined Doses, to Rats Infected with Clonorchis sinensis

XIAOShu-hua*;XUEJian;MarcelTANNER;ZHANGYong-nian;JenniferKEISER;JürgUTZINGER;QIANGHui-qin;LIUXiao-yun

2008, 26(5):  1-326. 

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Objective To assess the efficacy of single，multiple or combined oral doses of tribendimidine，artesunate，artemether and praziquantel against Clonorchis sinensis in rats. Methods A total of 147 rats，each infected with 50 C. sinensis metacercariae，were used in experimental chemotherapy. All the drugs used were administered intragastrically 42-44 d after infection. ① Sixty infected rats were randomly divided into 11 groups（4-5 rats per group）and the following drug dose-schedules were applied，i.e. under the same total dose tribendimidine or praziquantel was given at a single dose of 150 mg/kg，or given at smaller divided doses of 75 mg/kg（qd for 2 d），50 mg/kg（qd for 3 d），25 mg/kg（tid for 2 d）; artesunate or artemether was given at a single dose of 75 mg/kg，or given a half dose of 37.5 mg/kg daily for 2 days. ②Eighty-seven infected rats were randomly divided into 15 groups（4-6 rats per group） for combined treatment with the following drug administration regimens，i.e. artesunate or artemether 30 mg/kg plus praziquantel 150 mg/kg or tribendimidine 50 mg/kg and 75 mg/kg，respectively; tribendimidine 50 mg/kg plus praziquantel 150 mg/kg；tribendimidine 75 mg/kg plus praziquantel 187.5 mg/kg. A single dose of each drug mentioned above was also involved. Untreated C. sinensis-infected rats served as control. Rats were killed 14 days post-treatment，worms recovered from the bile duct and the liver tissue，mean worm burden reduction calculated and mean worm burden compared between the groups using non-parametric method（Mann-Whitney test）. Results Rats infected with C. sinensis and treated at a single 150 mg/kg dose of either tribendimidine or praziquantel resulted in a worm reduction of 57.2% and 63.8%，respectively. Whilst administration of tribendimidine at smaller but multiple doses given within 2-3 days at the same total dosage resulted in a slightly higher worm reduction（77.1%-79.4%），the opposite trend was observed for praziquantel（50.6%-54.2%）. However，for both tribendimidine and praziquantel，the difference of mean worm burden lacked statistical significance between single and multiple doses. Infected rats administered either artesunate or artemether at a single dose of 75 mg/kg or a daily dose of 37.5 mg/kg for 2 days，the worm reduction was 100% and 90.4%-98.5%，respectively. Combined treatment with low doses of tribendimidine(50 mg/kg or 75 mg/kg)plus praziquantel(150 mg/kg or 187.5 mg/kg)resulted in a worm reduction of 74.9%-100%, which were higher than those of 26.9%-79.6% obtained from a single dose of each drug used. High worm reduction of 74.4%-97.9% was also observed when administering a low dose of artesunate or artemether（30 mg/kg）plus a low dose of tribendimidine（50 mg/kg or 75 mg/kg）or praziquantel（150 mg/kg）. Mean worm reduction of 24.8%-79.6% were seen when drugs used at single doses. Conclusion The investigation confirmed that tribendimidine，artesunate，artemether and praziquantel are all efficacious against C. sinensis，and that drug combination acts synergistically.

Construction and Expression of Plasmodium berghei Chimeric Protein in Pichia pastoris and its Immunogenicity in Mice

CAOYi;ZHANGDong-mei;PANWei-qing*

2008, 26(5):  2-331. 

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Objective To produce an erythrocytic stage chimeric protein of Plasmodium berghei in Pichia pastoris and evaluate its immunogenicity. Methods The DNA sequences of AMA1（Ⅲ） and MSP1-19 from P. berghei homologous to the corresponding sequences of P. falciparum chimeric antigen 2.9 (PfCP-2.9) were fused to generate a chimeric gene，designated as PbCP-2.9. The resulting gene was redesigned using Pichia preferential coden usage and expressed in P. pastoris in the secreted form. The recombinant protein was purified by Ni-NTA affinity chromatography. Three groups each with 10 BALB/c mice were immunized subcutaneously with 20 μg of purified PbCP-2.9 antigen formulated in Freund’s adjuvant，Montanide ISA720 and Montanide IMS 1 312，respectively. Three control groups each with 10 mice received only adjuvants emulsified with PBS. All the mice received three immunizations at 2-week intervals with the same dose of antigen. Serum samples were collected at pre-immunization and one week after each immunization，and were analyzed for specific antibodies by ELISA and reaction with natural P. berghei proteins by IFAT. Results The PbCP-2.9 antigen with Mr 26 400 was successfully expressed in P. pastoris in secreted form. The recombinant protein can be recognized by the serum against blood stage parasites of P. berghei. High antibody responses were detected in all three PbCP-2.9-immune groups of mice by ELISA. However，mice immunized with PbCP-2.9 antigen in Freund’s adjuvant produced higher antibody titers than those with PbCP-2.9 antigen in Montanide ISA 206 and Montanide IMS 1312 adjuvants. The mean antibody titer in Freund’s adjuvant was 6.9-fold higher than in Montanide ISA 206 adjuvant and 5.6-fold higher than in Montanide IMS 1312 adjuvant after the second immunization （F＝81.06，P＜0.01). In addition，after the third immunization the mean antibody titer in Freund’s adjuvant was 3.7-fold higher than in Montanide ISA 206 adjuvant and 3.3-fold higher than in Montanide IMS 1312 adjuvant（F＝13.29，P＜0.01). The results from IFAT assay demonstrated that the immune sera recognized the surface proteins of P. berghei parasites. Conclusion The codenoptimized PbCP-2.9 gene has been constructed and expressed in P. pastoris. The chimeric antigen is highly immunogenic in mice and the immune sera can interact with natural proteins of P. berghei parasite.

In vitro Observation on the Apoptosis Induced by H₂O₂ in Protoscolex of Echinococcus granulosus

KANGJin-feng*;HUHan-hua;Baishanbieke;CHENRong;Abulizi

2008, 26(5):  3-337. 

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Objective To explore the apoptosis induced by hydrogen peroxide( H₂O₂) in protoscolex of Echinococcus granulosus. Methods Protoscoleces were cultured in vitro，and used for the experiment in 2 groups：RPMI 1640 medium and RPMI 1640 medium added with glutamine. They were then treated with different concentrations of H₂O₂ to induce apoptosis. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay (TUNEL) was employed to observe the apoptosis. Protein expression of caspase-1, caspase-3 and Fas was detected by SP immunohistochemical technique，stained with DAB re-stained with hematoxylin. A yellow or brown color nucleus revealed positive apoptosis cells in protoscolex，a brown reaction product in cytoplasm showed positive cells of caspase-1 and caspase-3，and brown cell membrane and cytoplasm revealed Fas product；otherwise it was judged as negative. According to the percentage of positive cells in a protoscolex，the expression level was divided as 4 grades. The percentage of less than 5% was regarded as “-”，5%-25% as “+”，26%-50% as “++”，more than 50% as “+++”. The experiments were repeated 2 times with controls. Results In RPMI 1640 group，positive TUNEL was found in the protoscolex induced by 1 mmol/L H₂O₂ inducing for 12 hours. Induced by 1 mmol/L H₂O₂ for 4 h，the “+ - ++”expression rate of caspase-1 and caspase-3 in the protoscoleces was 86.6% and 77.8%，and for 8 h，86.6% and 80.0% respectively，a significant increase in comparison to the control（P＜0.05). Induced by 5 mmol/L H₂O₂ for 4 hours，the “++ - +++”expression rate of caspase-1 was 93.0%，and the “+ - ++”expression rate of caspase-3 was 89.5%；induced for 8 h，the “++ - +++”expression rate of caspase-1 decreased to 53.2%，and the “+ - ++”expression rate of caspase-3 decreased to 48.4% and “-”expression rate increased to 46.8%. Under 5 mmol/L H₂O₂ for 4 h the expression rate of caspase-3 significantly decreased at 8 h（P＜0.01). In the group of RPMI 1640 plus glutamine，induced by 5 mmol/L H₂O₂ for 8 h，the “++ - +++”expression rate of caspase-3 in protoscolex was100%(P＜0.01). However，in RPMI 1640 group，induced by 5 mmol/L H₂O₂ for 8 h，the “++ - +++”expression rate of caspase-3 in protoscolex was 32.2% and 46.8% were negative. The Fas product with positive reaction in protoscolex was found in both control and induced groups：in RPMI 1640 group，under 5 mmol/L H₂O₂ induced for 4 h，“+ - ++” expression rate was 53.0% and control was 20.0%；and in the group of RPMI 1640 plus glutamine，under 5 mmol/L H₂O₂ induced for 8 h，“+ - ++” expression rate was 88.7% and control was 71.4%，increased in both groups after inducion (P＜0.05). Conclusion Apoptosis in the protoscolex of E. granulosus may be induced by H₂O₂.

In vitro Potentiation of Chloroquine Activity in Plasmodium falciparum by Ketotifen and Cyproheptadine

QUANHong;TANGLin-hua*

2008, 26(5):  4-342. 

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Objective To study the potentiation of chloroquine activity and mechanism by ketotifen and cyproheptadine in in vitro cultured Plasmodium falciparum Fcc SM1/yN strain. Methods In vitro cultured Fcc SM1/yN strain was added to pre-prepared drug plates at 50 μl/well after synchronization to make final concentration of 0.312 5-2 560 nmol/L for chloroqine and of 9.80-5 000 nmol/L for ketotifen or cyproheptadine. After 34 hours’ culture in 37 ℃，the number of schizonts with 3 or more nuclei was calculated among 200 parasites under microscope. Calculated half inhibitive concentration (IC₅₀) of chloroquine and every drug combination to parasite as well as chloroquine activity enhancement index (AEI) of ketotifen (or cyproheptadin). Time dependency of potentiation was studied. All data were analyzed statistically with SPSS 13.0. After 20 hours’ action of one optimal combination dose of chloroquine/ketotifen or chloroquine/cyproheptadine，RNA of the Fcc SM1/yN strain was extracted and real-time PCR was used to determine the expression level of pfcrt and pfmdr1 genes. Results The best potentiation effect was observed with ketotifen or cyproheptadine of 625 nmol/L，with IC₅₀ of 74.53 nmol/L for chloroquine/ketotifen and 89.7 nmol/L for chloroquine/cyproheptadine respectively，and activity enhancement index (AEI) of 0.42 for chloroquine/ketotifen and 0.30 for chloroquine/cyproheptadine respectively. Combination of 625 nmol/L ketotifen or cyproheptadine with 5 nmol/L chloroquine showed the highest potentiation potency. 6-7 hours during which ketotifen or cyproheptadine was added after chloroquine showed the highest effect，with IC₅₀ of 67.70 nmol/L for chloroquine/ketotifen and 81.53 nmol/L for chloroquine/cyproheptadine respectively，and the AEI was 0.47 for chloroquine/ketotifen and 0.37 for chloroquine/cyproheptadine respectively. After action of chloroquine/ketotifen or chloroquine/cyproheptadine at one optimal combination dose，expression level of pfcrt gene increased by 91% and that of pfmdr1 gene decreased by 14% respectively. Conclusion Appropriate combination of chloroquine/ketotiphen or chloroquine/cyproheptadine potentiates chloroquine against in vitro cultured P. falciparum. 6-7 hour period is an optimal time when ketotifen or cyproheptadine was added after chloroquine. Potentiating activity of ketotifen and cyproheptadine may be related to the expression level of pfcrt and pfmdr1 genes.

Epidemiological Survey on Malaria Situation in Motuo County of Tibet，China

ZHANGZai-xing;ZHOUHong-ning;ZHAOXiao-tao;CHANGFu-xing;WANGHong-ju;LIXiao-ju;ZHUOMAYang-jin;CIRENQuzhen;BIANMAZuoma;SANGDANLamu;ZHANGWei;YONGJian;XUHui-mei;BIANJiang;WANGLi-ying

2008, 26(5):  5-348. 

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Objective To make an epidemiological investigation on malaria in Motuo County，Linzhi Prefecture of Tibet. Methods In July of the year 2006，the following activities were conducted in 2 selected villages from each of the three townships，i.e.，Motuo，Dexing and Beibeng：malaria history survey among inhabitants in recent 2 years; collection of blood samples of inhabitants for examining malaria parasites，IFAT and detecting G6PD，respectively; mosquito collection in human dwellings and cattle shelters at night and various resting sites at day-time; mosquito collection by outdoor human baiting capture; classification and composition calculation of mosquito species and manbiting rates; ELISA for detecting sporozoite infection of Ａnopheles. Results The mean rate of two-year malaria history was 8.98%（118/1 314） and the parasite rate was 3.13%（38/1 216， all P. vivax） in the inhabitants. The parasite positive rate among the feverish patients was 7.14%（3/42）. IFAT revealed a malaria antibody rate of 40.24%（472/1 173）. The G6PD deficiency rate was 1.74%（21/1 208）. Five hundred and thirteen anopheline mosquitoes were caught. They were An. maculatus（474） which occupied 92.4%（474/513）， An. peditaeniatus（35），An. kochi（3）and An. sinensis（1）. The mean indoor density of An. maculatus was 4.75/night in human houses，and 69.5/night in cattle shelters. The outdoor human biting rate was 22.75/half-night/person，and the sporozoite rate of An. maculatus in anopheline saliva glands was 0 by ELISA. Conclusion Motuo County is an endemic area of vivax malaria with An. maculatus as the potential vector.

Artesunate in Interrupting the Transmission of Plasmodium falciparum

CHENPei-quan*;XUYing;CHENDong;HEKun-rong;OUFeng-zhen;FUChong-wei;FULin-chun;LIGuo-qiao

2008, 26(5):  6-352. 

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Objective To observe the effect of artesunate（ATS） on the infectivity of Plasmodium falciparum gametocytes（PFG）. Methods 31 volunteers with falciparum malaria and gametocytemia were randomly divided into 3 groups：artesunate (ATS) group (15 cases)， quinine (QN) group (10 cases) and placebo group (6 cases). Each case in ATS group received 6-day course of oral artesunate (200 mg at 0，6 and 24 hours then 100 mg daily for 4 days). Cases in QN group each received 21-dose course of quinine sulfate (500 mg/time) over seven days. Cases in placebo group took 2 tablets of vitamin B composites，three times per day for seven days. Peripheral PFG were counted daily in all cases until the clearance of PFG. Mosquitoes (Anopheles dirus)were fed with venous blood of patients on the 1st，7th，14th，21st and 28th day，respectively. Results All cases in placebo group were PFG positive at the whole course by blood smear examinations. The PFG relative density in ATS group were （12.5±3.3）%，（1.2± 0.4）%，（0.3± 0.1）% on 7th，14th，21st day respectively，and the mean PFG clearance time was（22.0±1.4）d. The PFG relative density in QN group were（173.9±47.0）%，（112.5±45.4）%，（32.5±17.8）% at 7th，14th，21st day respectively，and the mean clearance time of PFG was （32.5±2.1）d （t=4.731，P＜0.01). PFG remained positive on the 28th day in placebo group. The infectivity test to mosquitoes showed on 14th day the positive rate in ATS group，QN group and placebo group were 0，35.0% and 48.7% respectively. In ATS group，the sporozoite rate of anopheline mosquitoes were 14.8% and 0 at 7th，14th day，while in QN group，142.0%，98.6% and 20.3% at 7th，14th，21st day respectively. In placebo group，the infection rate of sporozoites remained stable. Conclusion Oral administration of artesunate with a total dosage of 1 000 mg in 6 days inhibits the infectivity of PFG.

Pathological Change in the Brain of Mice Infected with Angiostrongylus cantonensis

GUOPeng-juan;ZHANXi-mei;GANMing;PANZhi-hua;YUYan-jie;ZHANGMei-chun;QUZhen-yu;LIZhuo-ya;HEAi*

2008, 26(5):  7-355. 

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Objective To observe the pathological change in the brain of Angiostrongylus cantonensis-infected mouse. Methods Forty-eight mice were orally infected each with 40 third stage larvae of A. cantonensis，3 mice were sacrificed at 7、10、13、16、19、22、25、28 days postinfection respectively for worm recovery，and another 3 mice were for observing the histopathological change in tissue sections of the brain. Results Ten days postinfection，worms were found in the brain of the infected mice with a mean worm number of （7.0±1.7） per mouse. Ｔｈｅ ｈighest number of worms was found at 16 days postinfection，with a mean of（23.7±4.9）per mouse. Notable symptoms of nervous system were seen on 15 days postinfection. Most mice died around 22 days postinfection. Histological examination revealed mechanical damages. Cavitis and inflammation were observed in the brain parenchyma. Worms were seen in the subarachoid space. Meningitis-like signs started at 13 days and aggravated then. Conclusions Infection of A. cantonensis causes pathological change in mouse brain and the process is aggravating with postinfection time.

Cloning，Expression and Purification of Allergen Arginine Kinase from Periplaneta americana and its Allergic Activity

CHENJia-jie;XIALi-xin;LIUZhi-gang*;LIUWen;JIKun-mei

2008, 26(5):  8-360. 

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Objective To clone the gene of arginine kinase（AK） from Periplaneta americana，produce its recombinant protein and investigate its allergenicity. Method The cDNA of AK was cloned using specific primers from the total RNA of P. americana. The cloned gene was inserted into pMD18-T vector and digested by BamHI and HindⅢ. The cDNA was sequenced and subcloned into pET-28a expression vector. The cloned AK cDNA gene was expressed in Escherichia coli BL21（DE3） by IPTG induction. The recombinant AK(rAK) was purified by metal(Ni²⁺) chelating affinity chromatography. Its allergenicity was examined by both Western blotting and enzyme-linked immunosorbent assay (ELISA). Result The cloned cDNA ORF sequence (Accession no. EU429466) contained 1 068 bp and encoded 365 amino acids. Its sequence homology with the published one (Accession no. AY563004) was 99.9% at nucleotide level. The allergen rAK was highly expressed in E. coli BL21 (DE3) as a soluble protein mainly with the molecular weight of about Mr 45 000 under induction of IPTG and purified by 6-His-tag purification system. Both in the non-denaturalization and denaturalization conditions，the recombinant allergen was identified as its affinity to IgE antibodies from the cockroach-allergic patient sera by Western blotting and ELISA. Conclusion The recombinant cockroach arginine kinase has been obtained with proper allergenicity.

实验研究

Effect of the Liposome Albendazole and Huai-Er Fungus Extract on HepaticInfection of Echinococcus granulosus in Mice

LVHai-long;PENGXing-yu*;ZHANGShi-jie;Aduwayi;YANGHong-qiang;SUNHong;YANG-Jun;LIBin-jun;LIUYuan-kun

2008, 26(5):  9-365. 

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Objective To explore the effect of Huai-Er fungus extract (HEF) and liposome albendazole (L-ABZ)on the hepatic infection of Echinococcus granulosus(Eg) in mice，and understand the influence of improved host immunity on postoperative recurrence. Methods Female Kunming mice were immuned by intraperitoneal injection Eg cyst fluid from sheep，IgG positive mice were divided into drug treatment groups(A，B and C) and control group(D). Mice in group A were administered by gavage with single L-ABZ(75 mg/kg)，group B with single HEF(15 000 mg/kg)，and group C with combination therapy (L-ABZ 75 mg/kg plus HEF 15 000 mg/kg)，once every two days for one week. The prostoscoleces were treated with balanced solution for 20 minute and inoculated in the liver by open abdomen of the anesthetized mice. Imitating to an “open sub-adventitial total exocystectomy”， prostoscoleces outside the cyst led to reinfection. 72 hours later，groups A，B and C were re-treated for a month. Mice in group D were given with distilled water (0.3 ml per mouse). Group E was set as blank control (n=8). Another 120 IgG positive mice were divided as groups F，G，H and I，and treated in parallel with the groups A，B，C and D respectively；these mice were inoculated with prostoscoleces which were treated in advance with 75% alcohol， 20% hypertonic saline，L-ABZ and PBS for 20 min respectively. In 3 months after infection，all the mice were sacrificed to evaluate the efficacy，which covered infection rate，pathological change，spleen index，level of IgG and IgE，CD4+ and CD8⁺ cells. Results The recurrence rate in group C (5.7%) was lower than that of groups A (17.1%) and B (24.2%)，with hydatid cysts in white colour and nodular，showing degeneration of the germinal and laminated layers. Groups A，B and C showed lower spleen index(A：3.84±0.86，B：3.95±1.01，and C：3.27±0.52)，and lower IgE level (A：0.06±0.08，B：0.07±0.08 and C：0.03±0.03) than group D (5.46±0.52 and 0.20±0.02，respectively)(P＜0.05)，especially in group C；and also lower CD8⁺ in groups A (16.61±3.89)，B(18.18±3.90) and C(15.38±2.63) than group D (32.90±4.71)(P＜0.05)，but higher CD4⁺/CD8⁺ in groups A(3.21±0.70)，B(3.05±0.66) and C (3.53±0.57) than group D (1.57±0.26)(P<0.05)，especially in group C. The infection rate of mice in group F，G and H was 0，0 and 23.1% respectively，and that of group I，31.2%（P＜0.01）. Conclusion The combination of HEF and L-ABZ considerably improves the immunity of the hosts and HEF may have a synergetic action to L-ABZ in reducing the recurrence of “sub-adventitial total exocystectomy”. 75% alcohol and 20% hypertonic saline show better effect of inactivating prostoscoleces than L-ABZ in surgical operation.

Killing Effect of Polymorphonuclear Neutrophils on Trichomonas vaginalis

ZHAOJian-ling*;GAOXing-zheng;QUMing

2008, 26(5):  10-369. 

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Objective To study the killing effect of polymorphonuclear neutrophils（PMNs） on Trichomonas vaginalis. Methods The vaginal secretion from a patient with vaginitis was incubated in the liver infusion liquid medium to get T. vaginalis. One ml serum was collected from the patient and heated for 30 min at 56 ℃ to inactivate complement in serum, and was absorbed three times with the parasites at 0 ℃ to make the serum free of antibodies. PMNs were separated from the patient’s blood and purified with density gradient centrifugation and polymer accelerating sedimentation. NBT and safranin O were used to stain the sample. The interaction between PMNs and the parasites was observed under microscope. 300 trichomonads and 3×10⁴ PMNs were incubated for 10，20，30，40，50，60 minutes under the conditions of aerobic or anaerobic, with superoxide dismutase （SOD） and catalase （CAT） or without SOD and CAT, and with complement or without complement. They were then inoculated in solid medium for another five days under the anaerobic condition, and surviving organisms were enumerated. Results PMNs were observed to surround and kill a single trichomonad. In the petri-dish containing PMNs, the surviving rate of the parasites in anaerobic condition was 85%， only 3% in aerobic condition （P<0.01）. SOD and CAT reduced the killing effect of PMNs， with a surviving rate of 98% and 94% respectively after 60 min incubation. Without SOD and CAT, the surviving rate is only 2% （P<0.05）. PMNs in the serum without antibodies killed all the parasites，while the complement-inactivated serum fail to kill them. Conclusion The trichomonacidal activity of PMNs relies on the presence of oxygen and complement in the serum of patient.

现场研究

Epidemiological Investigation of Angiostrongylus cantonensis in Jiangmen of Guangdong Province

ZHANGYun;HUANGDi;TANQi-ming;CHENDai-xiong*;ZHANXi-mei

2008, 26(5):  11-373. 

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Objective To make an epidemiological survey on Angiostrongylus cantonensis in Jiangmen city of Guangdong Province. Methods From October 2006 to November 2007, the characteristics of A. cantonensis infection were investigated in Jiangmen district in various hosts, including the third stage larva infection in the snails Achatina fulica and Pomocea canaliculata by digestion method, and the adult A. cantonensis in rats by the dissection of heart and lungs. Relevant symptoms and dietary habits in Jiangmen residents who were randomly recruited were also investigated by questionnaire, and the specific IgG and IgM antibodies against A. cantonensis in their sera were detected by ELISA. Results　 695 A. fulica and 720 P. canaliculata were examined. The infection rate of third stage larva of A. cantonensis were 45.0% and 1.8% respectively, with an infectiosity of 53.74±147.30 and 5.23±8.51 respectively. Natural infection rate of A. cantonensis in all 229 rats was 4.4%. Among the 300 people surveyed, 11.3% had a history of eating raw or undercooked fish and shrimp, 5.3% directly or indirectly exposed to A. fulica or P. canaliculata. The positive rate of specific IgG antibody against A. cantonensis for serum samples among residents was 14.0% (42/300), and 5 serum samples in the 42 positive samples showed specific IgM antibody, with a positive rate of 1.7%. Conclusion Jiangmen district is an endemic area of A. cantonensis, and the local residents are under the risk of infection.

防治研究

Analysis on 2 175 Admitted Cases of Clonorchiasis Sinensis in Guigang City

LIANGShu-de*;WUYan-rong;PANYu-lan

2008, 26(5):  12-375. 

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Objective To understand the epidemiological features of the hospitalized cases with Clonorchis sinensis infection in Guigang City of Guangxi Region. Methods Fecal samples were collected from patients each with 30-50 g and examined by sedimentation technique. Number of eggs was counted in randomly selected cases by Stoll's method. With the number of eggs per gram（EPG）feces, light, medium and heavy intensity of infection were identified as EPG≤500, 501～5 000 and >5 000 respectively. Results Among 2 175 cases with clonorchis eggs, 1 244 cases showed clinical complications（57.2%）. The average EPG in 61 cases counted was 3 055，34（55.7%）showed medium or heavy intensity of infection. The cases distributed in 80% of the townships, more concentrated in 2 towns and 1 farm. The patients aged between 3 to 76 years old with a ratio of male to female 3.6 ∶ 1. 45.5% of the cases were farmers. 1 308 cases（60.1%） were Zhuang Minority. The number of cases seeking medical treatment increases year by year since mid-1980s. 66.0% of the cases had a habit of raw fish-eating. Conclusion Clonorchiasis sinensis widely distributes in Guigang City, number of cases admitted in hospitals increases yearly.

综述

The Neglected Intestinal Parasite Co-infection with AIDS

TIANLi-guang;ZHOUXiao-nong*

2008, 26(5):  13-381. 

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In order to increase the recognition of intestinal parasitic infections in AIDS，this article reviews the common intestinal parasites co-infected in HIV carriers and AIDS patients，namely，Cryptosporidium spp.，microsporidia，Giardia lamblia，and Strongyloides stercoralis，referring to aetiology，epidemiology，clinical features，diagnosis and prevention.

Th17 in the Immunity Against Parasitic Infection

LIUFan;YINHui-bin;SUChuan*

2008, 26(5):  14-386. 

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Th17 lymphocytes have been recently identified as a novel subset of CD4⁺ cells. It has been defined that IL-17，the main product of Th17， plays an important role in immunity against parasitic infection. There is a two-way infl-uence between Th17 and cytokine network： on one hand Th17 consummates cytokine network， on the other hand many cytokines regulate Th17’s activity in parasitic infection. In the anti-parasitic infection process, Th17 cells protect host or promote inflammation， even cause immune pathogenesis in different cases， which comprise host’s immune state， the burden of parasitic infection， as well as the treatment.

研究简报

In vitro Effect of Medicinal Scorpion on the Larvae of Ancylostoma caninum

XUZheng-min*;LIZhi-shan;WENMao-xing;PENGRong-yue;SUNLi;WUXiao-ying;ZHOULe-xiang;TAOYong-ping;YANGLei

2008, 26(5):  15-388. 

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Five gram stool sample from dog infected with Ancylostoma caninum was smeared on filter paper for routine cultivation under 35 ℃ with moist condition for 24 hours. Hookworm larvae were separated and divided into 3 bottles through precipitation-washing method. Traditional Chinese medicine scorpion(2 ml)，albendazole(10 mg) and normal saline (2 ml)(control) were added respectively into the 3 bottles， which were cultured for another 24 hours. The growth and development of the larvae were observed under microscope. In scorpion group，the larva body shrank，development stopped, and with an unclear internal structure. Treated with albendazole，the larvae became rigid and shriveled with rough body surface and vague internal structure. The results indicate that both medicinal scorpion and albendazole have considerable effect in inhibiting the development of hookworm larvae and the effect of albendazole is stronger.

Construction of cDNA Expression Library of Salivary Gland from Boophilus microplus

TIANZhan-cheng;LIUGuang-yuan*;XIEJun-ren;GONGZhen-li

2008, 26(5):  16-391. 

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Total RNA were isolated from salivary gland dissected from partially engorged Boophilus microplus. The mRNA was purified. A library of oligo(dT)-primed cDNA with added directional EcoR I/HindⅢ linkers was constructed from the purified mRNA. The constructed cDNA was ligated to the EcoR I/HindⅢarms of the λSCREEN vector. The recombinant phage DNA was packaged by phage-marker packaging extracts，resulting in a primary cDNA library with a size of 1.38×10⁶ PFU. Data showed 100% of the library were recombinant and the titer of the amplified library was 2×10⁹ PFU/ml. A partial cDNA encoding cytochrome oxidase C subunit II of B. microplus was screened from the expression library with rabbit serum against B. microplus salivary gland proteins. The results is suggested that the cDNA expression library has been constructed.

Morphological Observation on the Reproductive System of Adult Schistosoma japonicum under Confocal Laser Scanning Microscopy

ZHANGWei-na;HUANGDa-ke;ZHANGPeng;RENCui-ping;WANGXiao-nan;ZHANGYun-xia;SHENJi-jia*

2008, 26(5):  17-394. 

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BABL/c mice were infected with Schistosoma japonicum cercariae (40±2 per mouse) through abdominal skin. Mice were sacrificed after 35 days to acquire the adult worms which were then fixed，stained，clarified，dehydrated and mounted. The specimens were observed under the confocal laser scanning microscope． The overall morphology of the adult worms was displayed distinctly，especially the testicular lobes，seminal vesicle and genital pore of the male，reproductive system，and the ovary，vitelline glands，oviduct，vitelline duct，seminal receptacle，ootype，mehlis gland，uterus，genital pore and eggs of female reproductive system． The confocal laser scanning microscopy is an alternative method to research organs，tissues and cell structure of schistosome worm.

Preliminary Investigation on Phylogenetic Relationship AmongThree Common Species of House Dust Mites

CUIYu-bao;*;GAOCui-xiang;ZHOUYing;PENGJiang-long;LIULiang

2008, 26(5):  18-396. 

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Dermatophagoides farinae，D. pteronyssinus and Euroglyphus maynei were used for the investigation. The cDNA fragment coding for Der f 1 and Der f 2 were amplified by PCR，cloned and sequenced. By bioinformatics softwares，the amino acid sequences for Der f 1 and Der f 2 were deduced and compared with those for the groups 1 and 2 allergens of D. pteronyssinus and E. maynei available in GenBank. Amino acid sequence similarity analysis showed that Der p 1 shared 84% identical residues with Eur m 1 and 83% with Der f 1. Similarly，Der p 2 shared 87% identical residues with Eur m 2 and 68% with Der f 2. In the two phylogenetic trees constructed with group 1 and 2 allergens，D. pteronyssinus was clustered with E. maynei but not with D. farinae，although D. pteronyssinus and D. farinae belong to the same genus. D. pteronyssinus should be more similar to E. maynei than to D. farinae at evolutional level，which was not consistent with the conventional taxonomical relationship based on their morphological characteristics.

Observation on Mitosis of Trichomonas vaginalis Cultivated in vitro Using Modified Giemsa Staining

FANGZheng-ming*;DENGWei-wen;ZHUHong-gang;YANGZhen-de;LIUJian-jun

2008, 26(5):  19-398. 

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The observation showed that the percentage of Trichomonas vaginalis trophozoites at the stages of interphase，binary fission and multiple fission was 66.5%，24.1% and 9.4% respectively. Cells in binary fission could be classified as premitosis phase，prophase，metaphase，anaphase and telophase. 3 to 8 microcosms were seen in one trophozoite under multiple fission and the percentage of trophozoites with 3 and 4 microcosms occupied 69% and 24.5% respectively. Cells with abnormal morphs were also observed.

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Two imported cases of vivax malaria

XIONGBin;WANGFang

2008, 26(5):  20-373. 

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A case of encephalic paragonimiasis

HUYun-yun;ZENGLi-li;WANGYing

2008, 26(5):  21-399. 

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Case treatment of hydatid disease in pelvic cavity complicated with urethral fistula

ZHANGLin;FUZhen-chao;LIANGDong;LINHai;NURLAN;XUZun-li;DOUWei-long;DUMa-ling

2008, 26(5):  22-400. 

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Two cases infected by Psychoda larvae in Yunnan Province

ZHOUBen-jiang;YANGLi-jun;XIANGZheng

2008, 26(5):  23-封三. 

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