Table of Content

30 June 1999, Volume 17 Issue 3

Previous Issue    Next Issue

论著

ESTABLISHMENT AND APPLICATION OF DIPSTICK SANDWICH COLLOIDAL DYE IMMUNOASSAY FOR CIRCULATING ANTIGEN DETECTION IN SCHISTOSOMIASIS PATIENTS

XIEYanhui;XUEChunliang;LOUWenxian

1999, 17(3):  1-131. 

Asbtract ( )   PDF (278KB) ( )  

Related Articles | Metrics

　AIM:To set up a simple immunoassay for the detection of circulating antigens in host serum. METHODS: A kind of colloidal pale purple dye (HFRL) used as a staining reagent for immunoassay was first selected from the dyes produced in China and an optimum condition for labelling the dye onto the combined IgM antibodies was explored. A dipstick sandwich colloidal dye immunoassay (DS DIA) for the detection of circulating antigens in host sera was established. RESULTS:The minimal concentration of SEA detected was 5 ng/ml by DS DIA. Serum circulating antigen detection in 14 cases of acute schistosomiasis japonica ,113 chronic cases and in 113 healthy controls revealed that the sensitivity was 100% in acute cases, 52.2% in chronic cases and the specificity was 92.9% for uninfected students.The sensitivity and specificity of DIA were similar to those of dot ELISA . A certain degree of diagnostic complementarity up to 76.1% in sera from these patients was seen when DS DIA and dot ELISA were used in parallel. CONCLUSION:DS DIA is a simple, economical and reliable method for detecting circulating antigens of Schistosoma japonicum ,having a wide potential value especially for field use.


PRELIMINARY STUDY ON THE LEVELS OF NATURAL ANTIBODIES AGAINST SCHISTOSOMA JAPONICUM IN MICROTUS FORTIS IN DONGTING LAKE AREA

HEYongkang;LUOXinsong;YUXinling;LINJinlian;LIUShuxian;ZHANGXinyue;LIYi

1999, 17(3):  2-134. 

Asbtract ( )   PDF (191KB) ( )  

Related Articles | Metrics

　AIM:To explore the detection rate and the levels of natural antibodies to cercarial antigen(CA),juvenile antigen(JA),adult worm antigen (AWA) and soluble egg antigen(SEA),respectively,derived from Schistosoma japoncum in Microtus fortis distributed in Dongting Lake area.METHODS:The antibodies in wild Microtus fortis (WMF),laboratory bred Microtus fortis (BMF)and laboratory bred Microtus fortis 15 days after S.japonicum infection (BMF i15 ) were detected by indirect ELISA and a comparison of the antibody levels between Microtus fortis including WMF,BMF,BMF i15 and albino mice without infection (AM), 15 days after infection(AM i15 ) and 42 days after infection (AM i42 ) was made.RESULTS:The positive rates of detection for anti JA Ab in WMF and BMF were 94.6%and 94.4%,respectively,followed by 85.5%and 83.3 % for anti AWA Ab,58.1% and 59.4% for anti SEA Ab,and 14.3% and 13.9% for anti CA Ab.The levels of antibodies to the above four antigens in BMF i15 increased significantly, the detection rates being 100% for anti JA Ab and anti AWA Ab, 84.4% for anti SEA Ab and 65.6% for anti CA Ab.The levels of antibodies in WME, BMF and BMF i15 had the similar trend(JA>AWA>SEA>CA),and those in BMF i15 to CA,JA,AWA and SEA increased by 1.9,2.2,2.1 and 1.5 folds,respectively,when compared with BMF.CONCLUSION:Natural antibodies to Schistosoma japonicum were found in wild and labortory bred Microtus fortis .The levels of antibodies in BMF i15 to four above antigens were apparently higher than those in AM i15 .


TAXONOMIC STUDIES ON ECHINOCHASMUS FUJIENENSIS AND ITS RELATED SPECIES BY RANDOM AMPLIFIED POLYMORPHIC DNA ANALYSIS AND EXPERIMENTAL INFECTION

CHENGYouzhuZHANGYaojuanLINChenxinZHANZihaoYUXiaozhongLINJinxiangCAIMingjieCHEWeihong

1999, 17(3):  3-139. 

Asbtract ( )   PDF (318KB) ( )  

Related Articles | Metrics

　AIM:To explore the identification and differentiation of Echinochasmus fujianensis,Echinochasmus japonicus(Jiangxi strain and Fujian strain), Echinochasmus liliputanus Anhui and Echinochasmus perfoliatus Hubei.METHODS:Random amplified polymorphic DNA analysis (RAPD) and experimental animal infection were performed.RESULTS:469 polymorphic DNA fragments were obtained by 28 primers from 4 Echinochasmus species and strains in Fujian,Anhui and Jiangxi.20.8% and 97.6% of the fragments in Echinochasmus fujianensis were the same as those in Echinochasmus japonicus Fujian strain and in Echinochasmus liliputanus Anhui, respectively,99.7% of the fragments were the same between Echinochasmus japonicus Jiangxi strain and Fujian strain. CONCLUSION: Echinochasmus fujianensis and Echinochasmus liliputanus Anhui are the same species.Echinochasmus fujienensis is an independent species different from Echinochasmus japonicus.Polyinfection of Echinochasmus fujianensis,Echinochasmus japonicus and Echinochasmus perfoliatus exist in all the 3 provinces,Hubei,Anbui and Fujian,of which Echinochasmus fujienensis is a dominant species.


CHANGES IN TRANSFORMING GROWTH FACTOR β1 mRNA AND ITS CLINICAL VALUE IN PATIENTS WITH SCHISTOSOMAL LIVER FIBROSIS

CHENFeng;CAIWeimin;CHENZhi;LIURonghua

1999, 17(3):  4-142. 

Asbtract ( )   PDF (252KB) ( )  

Related Articles | Metrics

　AIM:To explore the changes in TGF β1 mRNA in patients with schistosomal liver fibrosis.METHODS:Reverse transcription polymerase chain reaction and dot blot analysis were used to determine the level of TGF β1 mRNA in PBMC. The serum levels of hyaluronic acid, type IV collagen, laminine and procollagen Ⅲ peptide were also measured simultaneously. RESULTS: The mean level (m ±s) of PBMC TGF β1 mRNA was significantly higher in late stage schistosomiasis group (LSS)(1.26±0.14), liver cirrhosis group (LC)(2.05±0.81) and hepatocellular carcinoma group (HCC) than that in control group (0 62 ±0 40) (P<0.05). PBMC TGF β1 mRNA level was higher in LC and HCC than that in LSS (P<0.05). No significant difference was found in TGF β1 mRNA between PBMC and liver tissue in patients with HCC. All patients with increased fibrogenic activity (i.e., abnormal levels of serum HA, Col IV and LN ) had increased levels of TGF β1 mRNA. CONCLUSION:The level of TGF β1 mRNA is correlated with the fibrotic activity of liver,the increasement of TGF β1 mRNA in schistosomiasis patients after praziquantel treatment might induce liver fibrosis.


CLONING AND SEQUENCE ANALYSIS OF HISTIDINE RICH PROTEIN Ⅱ GENE FRAGMENT OF PLASMODIUM FALCIPARUM YUNNAN STRAIN

XIAOJianhua*;LIMing;BIHuixiang;WANGPing;LIYingjie

1999, 17(3):  5-145. 

Asbtract ( )   PDF (197KB) ( )  

Related Articles | Metrics

　AIM:To determine the nucleotide sequence of the partial gene of the histidine rich proteinⅡ (HRPⅡ) the Plasmodium falciparum PFD 3/YN and find out the differences of the HRPⅡ gene sequence between this isolate and other isolates.METHODS:The histidine rich protein Ⅱ gene fragment was amplified by polymerase chain reaction and cloned into M13 bacteriophage.M13 HRPⅡ single strand DNAs of three positive clones were extracted, respectively.The nucleotide sequence of the HRPⅡ gene fragment was determined by the dideoxy chain termination method.Using PCGENE software to compare and analyze the HRPⅡ gene sequence among the different isolates.RESULTS: Different degrees of diversity of the HRPⅡgene sequences were found among Plasmodium falciparum PFD 3/YN(from China)and two other isolates(7G8 from Brazil and D10 from Gambia).The HRPⅡ in the three isolates exhibited 70.3% homology in amino acid sequences and 68.6% homology in the nucleic acid sequences.CONCLUSION:There were differences in HRPⅡ gene sequence among the Plasmodium falciparum PFD 3/YN and two other isolates(7G8 and D10).


DETECTION OF CIRCULATING ANTIGEN IN SERUM AND CEREBROSPINAL FLUID FOR DIAGNOSIS OF CEREBRAL CYSTICERCOSIS

LINXinjing;LIGuiping;HUOHaiying;XUFengquan;LIQingshanZHAOZhongpinInstructor:GELingyun

1999, 17(3):  5-148. 

Asbtract ( )   PDF (213KB) ( )  

Related Articles | Metrics

　AIM:To detect circulating antigen(CAg)in serum and cerebrospinal fluid(CSF)from patients with cerebral cysticercosis with specific antiserum by sandwich ELISA.METHODS:Antisera were prepared from rabbits immunized respectively with 3 antigens with molecule weights of 64 kDa,53 kDa,32 kDa～30 kDa extracted from Cysticercus cellulosae and purified by SDS PAGE.RESULTS:When samples from patients were tested with antiserum against 53 kDa antigen,the CAg positive rate was 93.8% in serum and 91.7% in CSF of 32 patients with active cysticercosis,whereas only one positive was found in CSF in 16 patients with inactive cysticercosis.The detection rate of CAg was significantly higher with anti 53 kDa antiserum than with anti 64 kDa and anti 32 kDa～30 kDa antiserum.CONCLUSION:Sandwich ELISA using antiserum against 53 kDa antigen to detect CAg was found to be a promising assay for diagnosis and evaluation of treatment efficacy of active cerebral cysticercosis in terms of its high sensitivity and specificity.


STUDY ON FAMILY AGGREGATION OF CASES OF ADVANCED SCHISTOSOMIASIS JAPONICA

LIUYing;YUANHongchang;LINDandan;HUFei;LIUYuemin;ZHANGShaoji;ZHAOGenming;JIANGQingwu

1999, 17(3):  7-151. 

Asbtract ( )   PDF (246KB) ( )  

Related Articles | Metrics

　AIM:To explore the family aggregation of advanced schistosomiasis japonica.METHODS:Eighty one cases of advanced schistosomiasis(AS) and 67 cases of non advanced schistosomiasis with history of infections in Yushan County,Jiangxi Province were chosen as proband groups and control groups respectively,then grades 1 and 2 relatives of them were investigated on AS.Family aggregation of AS was analyzed through comparing the prevalence rate between the close and distant relatives of probands and controls and fitting the observed distribution of AS cases among the population by zero truncated Poisson distribution and zero truncated negative binomial distribution. RESULTS:The prevalence rate was higher in the close relatives (Group Ⅰ relatives) of the probands than in the distant relatives(Group Ⅱ relatives)of the probands and in the controls relatives.The observed distribution of AS was beyond the probability of the zero truncated Poisson distribution,but consistent with the zero truncated negative binomial distribution.CONCLUSION:Family aggregation of advanced schistosomiasis does exist.


MORPHOLGICAL,HISTOLOGICAL AND HISTOCHEMICAL OBSERVATIONS ON THE EFFECT OF ALBENDAZOLE ON ENCYSTED LARVAE OF TRICHINELLA SPIRALIS IN MICE

CHENXiaoningCHENPeihuiJIFengqingWANGYixingWANGFengWANGFengyun

1999, 17(3):  8-154. 

Asbtract ( )   PDF (352KB) ( )  

Related Articles | Metrics

　AIM: To study the mechanism of action of albendazole on encysted larvae of Trichinella spiralis in mice. METHODS: Twelve Kunming strain mice each infected with 200 T. spiralis larvae were equally divided into two groups. Six mice of the treatment group were treated with 30 mg of albendazole/kg daily for 5 days. Morphological, histological and histochemical methods were used. RESULTS: The encysted larvae of albendazole treated group became significantly damaged,most worms were shrunken and surrounded by inflammatory cells.Histochemical study demonstrated that the glycogen and RNA content of the larvae in the treated group was decreased,the activities of SDH,ATPase, ACP were lower than those of the control group. CONCLUSION: Albendazole can affect the physiological function of Trichinella spiralis .


GENOTYPE AND SEQUENCE ANALYSIS OF MEROZOITE SURFACE PROTEIN 1 OF PLASMODIUM FALCIPARUM ISOLATES IN YUNNAN PROVINCE

ZHUXinping;ZHOULei;LIUQiang;GAOXin

1999, 17(3):  9-158. 

Asbtract ( )   PDF (198KB) ( )  

Related Articles | Metrics

　AIM: To identify the genotype of merozoite surface protein 1 of Plasmodium falciparum in Yunnan Province and explore the polymorphism of MSP1 genes in geographical characteristics and genetics. METHODS: Nested polymerase chain reaction was applied to genotyping of P. falciparum isolated in Yunnan Province. Some of parasite alleles were sequenced by dye primer cycle sequencing. RESULTS: In 30 P. falciparum infections, 38 different alleles were found. Of them, the dominant allele was a variant of MAD20, while was K1 less and the RO33 was few. In addition, incidences of mixed allele infections were observed. Sequence analysis showed that DNA sequences of MAD20 , K1 and RO33 alleles from Yunnan were highly homologous with those of international standard strains, respectively. CONCLUSION:Composition and sequence characteristics of P.falciparum arasite population in the endemic area can be detected by genotyping with MSP1 as genetic marker, which would be useful for the prevention and treatment of malaria.


PREPARATION OF MONOCLONAL ANTIBODIES BY INTRASPLENIC IMMUNIZATION OF MICE WITH URINE CIRCULATING ANTIGEN FROM PATIENTS INFECTED WITH SCHISTOSOMA JAPONICUM

WUChenyun;LOUWenxian;XUEChunliang

1999, 17(3):  10-161. 

Asbtract ( )   PDF (272KB) ( )  

Related Articles | Metrics

　AIM: To explore an efficient method for preparing monoclonal antibodies against circulating antigen of S.japonicum in the urine of schistosomiasis patients. METHODS: The urine of schistosomasis patients were precipitated with 5% trichloroacetic acid,and then used to immunize the BALB/c mice by intrasplenic immunization to prepare McAb. RESULTS: Two monoclonal cell lines against the CAg in the urine of schistosomiasis patients have been established.The immunoglobulin subclass of the 2 McAbs (2E6 and 2B11) were identified as IgM.The titres of 2E6 and 2B11 were 2.56×10 5 and 6.4×10 4. On immunodiffusion , McAb 2E6 showed one precipitation line with IHU CAg, however, both McAbs did not react with NHU-CAg.Using ascitic fluid of 2B11 and 2E6 as capture antibody separately, and HRP H11 as labelled antibody to detect the urine of 12 acute schistosomasis patients, 2B11 2E6 gave 6 and 3 positive, respectively, whereas 8 normal urine were all negative. CONCLUSION: It is feasible to use CAg from schistosomasis patientsurine to prepare anti CAg McAb by intrasplenic immunization in mice.


OBSERVATION ON THE DYNAMICS OF SPECIFIC ANTI GST ANTIBODIES IN RABBITS IMMUNIZED WITH RECOMBINANT 26 kDa GST OF SCHISTOSOMA JAPONICUM

XUYuxin;SONGGuangcheng;LIUShuxian

1999, 17(3):  11-164. 

Asbtract ( )   PDF (248KB) ( )  

Related Articles | Metrics

　AIM: To investigate the dynamics of the anti GST antibodies in rabbits immunized with recombinant 26 kDa GST of Schistosoma japonicum . METHODS: The specific antibodies were detected weekly by GST ELISA in rabbits vaccinated with purified recombinant 26 kDa GST antigen of S.japonicum plus FCA/IFCA;rabbits immunized with 0.85% saline plus FCA/IFCA served as control group. RESULTS: The specific anti GST antibodies began to increase at week 4 post immunization,the mean OD value in rabbits vaccinated with recombinant GST could reach 1.03±0.46(adjuvant control group only 0.42±0.04).The experiment was completed at week 65 post immunization when the mean OD values for specific anti GST antibody in immunized rabbits were 0.94±0.26(0.29±0.16 in the control group). CONCLUSION:Strong anti GST antibody responses could be induced in rabbits vaccinated with recombinant 26 kDa GST of S.japonicum.


LONGITUDINAL OBSERVATION ON A SCHISTOSOME INFECTED SNAIL SPOT AS A SOURCE OF INFECTION IN A MOUNTAINOUS VILLAGE OF DALI CITY

LIFei;XIADaiguang;MACanhua;JIAXuemei;

1999, 17(3):  12-166. 

Asbtract ( )   PDF (119KB) ( )  

Related Articles | Metrics

　AIM: To observe longitudinally the changes in survival and infection rate of Oncomelania snails in a positive snail spot in Yunnan.METHODS: A positive snail spot in a ditch in Shalimuzhuang Village,Dali City,was continually observed on the changes in the density of snail population and natural infection rate of the snails by random environmental sampling for 8 times in 4 successive years. RESULTS: During the 8 times of snail survey,living snails were found relatively stable in the ditch,in which positive snails were obtained in 6 occasions,the average infection rate being 3.99%. CONCLUSION: The density and the positive rate of snails in the mountain region remains relatively stable as a source of human infection. Elimination of the positive snail spots is necessary for controlling schistosomiasis.


PILOT STUDY OF SCHISTOSOMIASIS CONTROL IN POYANG LAKE REGION

LINDandan;HidenoriMurakami;ZHANGShaoji;WUZhongdao;NINGAn;TomokoMurakami;TokaoTotsuya;GUXiaonan;HUGuanghan;GAOZulu;LIUYueming;HUFei;CHENTaihui

1999, 17(3):  13-171. 

Asbtract ( )   PDF (166KB) ( )  

Related Articles | Metrics

　AIM:To perform a longitudinal survey of the control of schistosomiasis in Fanhu illage hyperendemic for schistosomiasis, Poyang Lake region of Jiangxi Province from 1992 to 1996. METHODS: Annual mass chemotherapy and health education were implemented in all investigated indiviuals, in 1996, artesunate was used in high risk population in transmission season as a prophylactic measure. RESULTS: The prevalence of S. japonicum infection reduced from 26.0% in 1992 to 10.7% in 1994, the intensity of infection (geometric mean of EPG of feces) in residents decreased from 1.92 in 1992 to 0.55 in 1994, and the morbidity indicators such as hepatomegaly and splenomegaly in the cases also have been improved significantly after chemotherapy. In 1995, both the infection rate and intensity of infection of schistosomiasis were recurred from 10.7% and 0.55 in 1994 to 18.4% and 1.04 respectively due to the floods. In 1996, the infection rate reduced to 5.0% and the intensity of infection also decreased to 0 25 due to the additional measure. CONCLUSION:A mass chemotherapy in conjunction with health education and oral artesunate for preventing reinfection in transmission season is effective for the control of schistosomiasis in lake regions.


实验研究

CONSTRUCTION OF DNA VACCINE INCLUDING A CHIMERIC GENE ENCODING CYSTICERCUS CELLULOSAE ANTIGEN AND PORCINE INTERLEUKIN 4

WUDan;GUOYingjun;SUNShuhan

1999, 17(3):  14-174. 

Asbtract ( )   PDF (227KB) ( )  

Related Articles | Metrics

　AIM: To construct a fusion expression vector for DNA vaccine including porcine interleukin 4(IL 4)and antigen cC1 to enhance the protective immunity of Cysticercus cellulosae antigen cC1. METHODS: The cDNA fragments encoding porcine IL 4 and cC1 were amplified respectively by PCR and then fused.The obtained chimeric gene IL 4cC1 contained a synthetic linker of ten amino acids and the sequence surrounding its 5’ AUG initiatory codon was changed to optimized translational initiation. RESULTS: Identified by restriction enzyme analysis,an insert fragment of 1.5 kb was demonstrated. It had the same sequence as reported and designed by DNA sequencing analysis.CONCLUSION: A fusion expression plasmid containing porcine IL 4 and cC1 was constructed.


临床报道

OBSERVATION ON EFFICACY OF ARTEMETHER COMPOUND AGAINST VIVAX MALARIA

LIXingliang;LIChongzhen;CHELigang;LIUXingzhiLIZonghui;HUANGDeshun;HEWei;DAOQingyang

1999, 17(3):  15-177. 

Asbtract ( )   PDF (185KB) ( )  

Related Articles | Metrics

　AIM: To observe the efficacy of artemether compound against vivax malaria. METHODS: Each artemether compound tablet contains 120 mg benflumetolum and 20 mg artemether.132 patients with vivax malaria were divided into 3 groups.Group A,36 patients received 8 tablets as an initial dose,followed by 4 tablets daily for 2 days;group B,41 patients received 8 tablets as an initial dose, followed by 3 tablets daily for 4 days;group C,55 patients receiving chloroguine primaquine served as control.Two patients of group A were voluntarily to be bitten by Anopheles dirus before and after medication to observe the influence of artemether compound on the sporogony. RESULTS: The average defervescence times for groups A,B and C were 22.3 h、23.2 h and 25.0 h( P >0.05),respectively,the average parasite clearance times were 33.5 h、30.5 h and 44.9 h,respectively,the average parasite clearance times of groups A and B were all significantly shorter than that of group C ( P <0.01).The replase rates of groups A,B and C were 84.9%、78.8% and 22.9%, respectively,followed up at nine months,the relapse rates of groups A and B were higer than that of group C( P <0.01). CONCLUSION: Two regimens of artemether compound have the advantage of high efficacy against vivax malaria.