Table of Content

30 December 2003, Volume 21 Issue 6

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论著

Content Change of CollagenⅠ, Ⅲ in Liver and Spleen of Patients with Advanced Schistosomiasis Japonica

CHENJia-lin~;TAOJun~;CAIWei-min~;ZHANGYan-ping~

2003, 21(6):  2-325. 

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　Objective To study the change of the content of collagen typeⅠand collagen type Ⅲ (C-Ⅰ, C-Ⅲ) in liver and spleen of patients with advanced schistosomiasis japonica. Methods Sirius red staining was used to examine pathologically the liver and spleen biopsy materials from 55 patients at the advanced stage of schistosomiasis and 5 healthy persons as control. The stained slides were observed under polarizing microscope and results were interpreted by computer imaging analysis system. (Results) The content of C-Ⅰand C-Ⅲ in liver tissue and hepatic sinusoids increased significantly in the patients than that of the control (P<0.01), the ratio of C-Ⅰ/C-Ⅲ decreased considerably (P<0.01). With the progress of fibrosis in the liver, the content of C-Ⅰand C-Ⅲ in hepatic sinusoids decreased, and the ratio of C-Ⅰ/C-Ⅲ increased. The content of C-Ⅰand C-Ⅲ in spleen tissue also increased significantly in schistosomiasis patients than that of the control (P<0.01), the ratio of C-Ⅰ/C-Ⅲ decreased (P<0.01). The C-Ⅰand C-Ⅲ content increased with the progress of spleen fibrosis while the ratio of C-Ⅰ/C-Ⅲ decreased. Conclusion The result shows that the content of C-Ⅰand C-Ⅲ increased in liver and spleen of the patients with advanced schistosomiasis japonica, the increase of C-Ⅲ was more significant than that of C-Ⅰ.


Effect of Mitochondrial Membrane Potential in the Neurotoxicity of Artemether

ZHAOYan-hong;WANGJing-yan

2003, 21(6):  3-329. 

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　Objective To study the effect of mitochondrial membrane potential and cellular membrane permeability in the neurotoxicity of artemether. Methods Mitochondrial membrane potential and cellular membrane permeability in pheochromocytoma cell and primary cultured rat neuronal cell were measured by flow cytometry, using rhodamine 123(Rh123) and PI as fluorescent dye, respectively. Mitochondrial swelling was measured by spectrometer. Results Artemether could decrease the mitochondrial membrane potential and increase the cellular membrane permeability in both cell types in a dose-dependent manner. In addition, artemether led to mitochondrial swelling with dose-effect and time-effect relationships. Conclusion The decrease of mitochondrial membrane potential plays an important role in the neurotoxicity of artemether. The drug can change the mitochondrial membrane potential and mitochondrial swelling by affecting the permeability transition pore complex located in the mitochondrial membrane, increasing cellular membrane permeability, causing an obstruction and finally leading to neurotoxicity.


Immunolocalization of the Signaling Protein l4-3-3 of Schistosoma japonicum

LIUQing-zhong*;SHENJi-long~**;WANGXue-ong

2003, 21(6):  4-332. 

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　Objective To study the localization of the signaling protein 14-3-3 of Schistosoma japonicum (Sj14-3-3)in the parasite. Methods Cercariae were collected from the infected Oncomelania hupensis for the infection of rabbits. Fifteen-day-old schistosomula and adult worms obtained from infected rabbits 15 and 42 days post-infection were used for frozen sections and indirect immunofluorescence staining with monoclonal antibody to rSj14-3-3. Results The results showed that the Sj14-3-3 distributed mainly in the tegument, subtegument, muscle, and parenchyma of both adult worms and 15-day-old schistosomula. Conclusion The wide distribution and large sites of Sj14-3-3 in the parasite were clearly demonstrated, which established a significant clue for further studies of biologic actions and application of 14-3-3 protein.


National Surveillance of Schistosomiasis from 2000 to 2002

ZHAOQi~;ZHAOGen-ming~;CHENXian-yi~;WANGLi-ying~;HENa~;WeiJian-guo~;JIANGQing-wu~

2003, 21(6):  5-337. 

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　Objective To identify the trends of endemic situation at the twenty-one national surveillance sites from 2000 to 2002. Methods According to the protocol of the national surveillance system, longitudinal surveillance of endemic situation of schistosomiasis was carried out at the twenty-one surveillance sites since 2000. Results Relative indexes of endemic situation of schistosomiasis decreased in two surveillance sites annually. The infection rate of Schistosoma japonicum in human population decreased in most surveillance sites. The infection rate of cattle decreased in eight surveillance sites. However, acute cases were found in two sites each year. The density of live snails and infected snails was higher compared with that in 2000 among eight sites and infected snails were found in inner embankment in Junshan surveillance sites, Hunan Province, in 2002 at first time. The snail ridden areas increased significantly in Fengjin sites, Shanghai, where schisotomiasis was eliminated years ago. Conclusion Current control strategies have some impact on the morbidity of schistosomiasis. However, these strategies should be lasted for longer time and surveillance on snails should be enhanced.


Studies on the Mimic Short Peptide-based Vaccine:Immunoprotection in Mice against Schistosoma japonicum Infection

WANGMin;YIXin-yuan*;ZENGXian-fang;LIXian-ping;ZHOUDong-ming

2003, 21(6):  6-341. 

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　Objective To evaluate the immunoprotective effect of the mimic short peptide-based vaccine of Schistosoma japonicum(S.j) in mice. Methods Phage random peptide library of 12 amino acids was immunoscreened with purified IgG from sera of rats. Three rounds of biopanning were carried out. The positive clones were randomly selected, detected and sequenced. Kunming mice were immunized with positive clones three times (0, 2, 4 weeks). Each mouse was infected with 40 cercariae after 2 weeks of the 3rd immunization. After 42 days of infection, the mice were killed. Adult worms and the liver eggs were counted. Results The specific phages binding to IgG were enriched after 3 rounds of biopanning. Two short peptides were obtained. Compared with the control groups, the mixture of two mimic peptides induced 34.9%(P<0.05) worm reduction and 67.6% (P<(0.001)) total liver egg reduction in mice. Two different mimic peptides induced 31.0% (P<0.05), 14.5% (P>05) worm reduction and 61.2%(P<0.001), 35.7%(P<0.05) total liver egg reduction respectively. The specific antibody could be induced by the mimic peptides and detected by ELISA in immunized mice, and the antibody titer reached more than 1∶6 400. Conclusion The 2 different mimic peptides obtained by immunoscreening phage random peptide library show partial immunoprotection against S.j infection.


Application of Dot-immunogold Filtration Assay in Detection of IgG in Sera of Trichinellosis Patients

LIUYing-jie~;LIUYun~;QUYao-hua~;ZHENGYong~;LIUYan-hong~;ZHANGYong-li~;CHENJuan~

2003, 21(6):  7-344. 

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　Objective To study a new quick method for detecting serum IgG against Trichinella spiralis (T.s). Methods Membrane antigen of muscle larva of T.s was isolated and combined with the nitrocellulose membrane. Goat anti-human IgG was conjugated with the golden pellets of colloidal state which was used as marked antibody for the experiment of dot-immunogold filtration assay to examine specific IgG. Results T.s IgG and its corresponding antigen reacted on the membrane by filtration, and the result could be observed with naked eyes within 10 min. In the examination of 76 clinical serum samples, the positive rate was (94.7%). It was not significantly higher than that of ELISA(χ~2=2.83,P>0.05). Conclusion The dot-immunogold filtration assay is rapid and simple for performing and reading in the detection of IgG against T.s with high sensitivity and specificity.


Preliminary Identification of T Cell Epitopes on 22.6kDa Antigen of Schistosoma japonicum

WANGXin-jun;ZHANGZhao-song;LIGuang-fu;WANGYong;LIUFeng;JIMin-jun;ZHUXiang;CAIXiao-ping;WUGuan-ling

2003, 21(6):  8-348. 

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　Objective To identify the T cell epitopes on 22.6kDa antigen of Schistosoma japonicum (Sj22.6). Methods The primary structure of Sj22.6 molecule was analysed using various predictive algorithms and a panel of 4 peptides were acquired. Their oligonucleotides were designed, synthesized and inserted into the multiple cloning site of plasmid pET-32c(+). The recombinant plasmids were transformed into E.coli BL21 and identified by endonuclease digestion and sequencing. The positive clones containing the recombinant plasmids could express specific fusion proteins (trx-epitope, MW≈20kDa) induced by IPTG. The fusion protein with 6×His could be coupled with NTA resin specifically, and purified by elusion of the column with buffer containing imidazole. The purified fusion proteins were incubated with splenocytes of C3H mice and then, the proliferation of splenocytes was determined by~3H-TdR incorporation assay. Results The recombinant plasmids were constructed successfully and the positive clones containing the recombinant plasmids expressed specific fusion proteins. Three of the purified fusion proteins (P4、P5、P6) could stimulate the lymphocyte proliferation. Conclusion Three T cell epitopes on Sj22.6 antigen were identified.


Study on the Intention of the Administrative Model of Anti\|Schistosomiasis Institutions at County Level

JIANGQing-wu~;WANGWei-bing~;XUBiao~;CHENXian-yi~

2003, 21(6):  9-352. 

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　Objective To investigate current status of schistosomiasis control system at the county level and make recommendations to improve the infrastructure adapting to the prevalent condition of the disease. Methods Through pertinently sampling and semi-quantitative questionnaire among the professional workers, experts and administrators in schistosomiasis control, the primary weaknesses involved in the management system and human resources were analyzed. Results 380 questionnaires were distributed, 33 out of 40 were returned from experts/administrators and 285 out of 340 were received from the professionals. In general, the participants acknowledged the effectiveness of the mechanism on anti-schistosomiasis campaign during the past 40 years. It was found that lack of funds for schistosomiasis control and the poor administrative model of anti-schistosomiasis activities were the main barriers to the control. About half of the participants agreed that the policy about merging the county level anti-schistosomiasis facilities into the Center for Disease Prevention and Control (CDPC) system. The study also showed an inbalanced distribution of the manpower for schistosomiasis control across the endemic regions. The dropout rate of human resources was high due to the poor management and welfare. Conclusion Reforms on the management model should be performed and multiform compensation system should be built in accordance with the prevalence and trends of schistosomiasis.


Study on the Subcloning, Expression and Immunoprotection with Schistosoma japonicum CAI Gene

LUOXiu-ju;YUANShi-shan;YIXin-yuan*;ZENGXian-fang;ZHANGShun-ke;TANGLian-fei;CAIChun;ZHANGJie;LarryMcReynolds

2003, 21(6):  10-356. 

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　Objective To subclone and express the new gene of Schistosoma japonicum (Sj) CAI and evaluate the immunoprotective effect of the recombinant molecule. Methods The cDNA of SjCAI gene was subcloned into expression vector pGEX-5X-3 to form recombinants which were then used to transform to E.coli strain ER 2566. Expression was induced by IPTG. The mice were vaccinated with the expressed protein and the immunoprotective effect was tested. Results Fusion protein of SjGST-CAI was highly expressed in E. coli as inclusion bodies. The worm reduction rate and the liver egg reduction rate in vaccination group of SjGST-CAI were 29.87% and 63.71%, respectively. Conclution SjCAI gene can be highly expressed in E .coli after subcloning into pGEX-5X-3 vector and the expressed fusion protein can induce immunoprotective effect against Sj in mice.


Kinetic Analysis of Cytokines and Immunoglobulin G Subclass in BALB/c Mice Infected with Echinococcus alveolaris

LIFu-rong~;SHIYou-en~;SHIDa-zhong~;DAVuitton~;PSCraig~

2003, 21(6):  11-360. 

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　Objective To observe the dynamic change of immune response in mice infected with Echinococcus alveolaris(AE) at difference period of time, and to explore hostalveo’s immune regulation. Methods The infection lasted and was followed up for 25 weeks. The spleen cells from BALB/c mice infected with AE stimulated with EmAg and ConA or PHA in vitro . IL-2R, IFN-γ, TNF-α, IL-1 and specific IgG subclasses were determined by ELISA. NO was tested by chemical assay. Results NO level sharply rised in 16 weeks after BALB/c mice were infected with AE. The levels of IgG1 and IgG3 significantly increased 8 weeks after infection, and remained elevating throughout the period of observation. IgG3 showed slight increase, IgG2a and IgG2b appeared low level following infection. The production of IL-2R and TNFα increased significantly 8 weeks of infection, while IL-2R sharply decreased in 12 weeks of infection. During the period of 2-12 weeks of infection there was an increase in IL-1 secreting. The level of IL-1 and TNFα rapidly increased since 16 weeks post infection. High level of IFN-γ was detected during the period of observation, and showed a peak at 12 weeks. Conclusion Th1 is the major response in the early stage of infection,which is replaced by Th2 response in later period of infection.


实验报道

Morphological Observation of Haplorchis pumilio

LINJin-xiang;CHENBao-jian;ZHUKai;CHENGYou-zhu;LIYou-song;XULong-shan;ZHANGRong-yan

2003, 21(6):  12-362. 

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　Objective To provide the basis for the differentiation with similar species of intestinal flukes through observing the figure of Haplorchis pumilio. Methods Adults of H. pumilio were collected from the intestine of the cat which was infected with the encysted cercariae of H.pumilio for 45 days. The worms were observed after staining. Eggs and metacercariae of H.pumilio were collected and examined for their shape, size and morphological characteristics. Pseudorasbora parva, the fish host, was examined for the parasitized sites of metacercariae. Results The principal characteristics of the adults is the acetabulum degradation. There are only the genital sucker with 44-48 hamuli. The average measurement of eggs is 31.2×16.7 μm with a smooth shell. Its aceromion is not evident. The average diameter of metacercariae is 168.5 μm. There are squamous spines on metacercaria. The metacercariae only parasitize in the muscle between the basis of the fin and the fish body. The average measurement of metacercaria cyst is 445×95μm, with squamous spines on the body surface. Hamuli are found on the genital sucker of metacercaria cyst. Conclusion The morphological figures and parasitic sites of metacercaria, the genital sucker of the adult, and the number and form of the hamulus on the genital sucker provide basis for distinguishing H. pumilio from other intestinal flukes.