Table of Content

28 February 1992, Volume 10 Issue 1

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STUDIES ON THE STRAIN DIFFERENCES OF SCHISTOSOMA JAPONICUM IN THE MAINLAND OF CHINA VII.GENETIC VARIATION AND DIFFERENTIATION OF FIVE ISOLATES

1992, 10(1):  1-4. 

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Electrophoretic techniques have been used to survey five different field-collected isolates (i.e.,Anhui,Hubei,Guangxi,Sichuan and Yunnan) of S.japonicum from the mainland of China for genetic variation at 7 isozymes markers representing 9 loci.Polymorphic loci were observed for LDH-1,LDH-2,MDH and PGM,respectively,the proportion of polymorphic loci being 44.4%.Average heterozygosity varied between 0.223 and 0.425,with a mean of 0.332.Nei's genetic distance (D) among the populations of 5 isolates gave values between 0.001 and 0.039 with an average of 0.023,indicating that these isolates are very closely rehted.

TRANSMISSION ELECTRON MICROSCOPY OF TEGUMENT AND DIGESTIVE TRACT OF SCHISTOSOMA JAPONICUM DURING LONG TERM CULTIVATION IN VITRO

1992, 10(1):  5-7. 

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The article reports the result of the observation on the ultrastructure of the tegument and digestive tract of Schistosoma japonicum schistosomula cultured 130 days in vitro in 841 medium,and the 42-day-old adults from infected animal cultured 23 days in 851 medium by transmission electron microscopy.The results showed that the ultrastructure of the tegument including sensory papillae and flame cells was normal.The matrix seemed to have no vacuolization.The surface of the esophagus was highly rugous.Many laminae and lipid droplets filled the lumen of the digestive tract and the tubules-like depressions from basal lamina were occured.All these structures seemed to have less variation as compared with the prior authors' data from normal worms.It is considered that the above two media which we cesigned were suitable for the growth and development of the schistosomes due to the tegument and digestive tract maintained their metabolic function in the long-term cultivation in vitro.(Figs 1-10).

THE DYNAMICS OF ANTIGEN AND ANTIBODY IN SCHISTOSOMA JAPONICUM EGG GRANULOMA AND ITS RELATIONSHIP WITH THE GRANULOMA RESPONSE

1992, 10(1):  8-10. 

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Synchronous Schistosoma japonicum egg granuloma in the lung of mouse model and PAP (peroxidas-anti-peroxidase) technique were employed to study the dynamics of antigen and antibody in the egg granuloma of S.japouicum and its relationship with the granuloma formation.The granulomatous response began on the 3.5 day after egg injection and increased to the maximal size at the 4th week.Lymphocyte populations and macrophage comprised an important part of lesions during the time of acute granulomatous response (7-28 day).Using PAP staining,the SEA within egg could be detected at high level on the first day after egg injection and then declined gradually.On the contrary,the SEA around egg was minimal at the first week and increased to the peak at the 4th week,then decreased gradually.No antibody could be detected throughout the experimental period (35 days).The results suggested that 1) the antigen of Schistosoma egg is the essential factor for the granuloma formation.2) S.japonicum egg granuloma could be formed in unsensitized mouse.3) The mechanism of egg granuloma formation of S.japonicum is similar to that of S.mansoni.

THE ACTIVITY OF TUMOR NECROSIS FACTOR IN SCHISTOSOMIASIS JAPONICA

1992, 10(1):  11-13. 

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The activity of tumor necrosis factor (TNF) in both the culture supernatant of pheripheral blood mononuclear cells (PBMC) and the serum was examined in 20 patients of advanced schisto-somiasis at 8th week after pyquiton treatment,and 6 rabbits infected with S.japonicum cercariae at 8th and 16th weeks post-infection and using L929 cell cytotoxic assay.The results showed that the capacity of PBMC to produce TNF in response to LPS from patients with late schistosomiasis was significantly lower than that from normal subjects (P0.01),whereas the activity of TNF in serum was significantly elevated (P0.01).The capacity of PBMC to produce TNF and the TNF activity in serum of infected rabbits showed sharp rise at 8th week post-infection (P0.01),and remained high at 16th week,then returned to normal level 8 weeks after treatment.The persistent rise of TNF in the circulation indicate that TNF might take part in the formation of hepatic fibrosis.

STUDY ON TOXICOLOGY OF A NEW ANTHELMINTIC “85012”

1992, 10(1):  14-17. 

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"85012",a new anthelmintic containing amidine synthesized in our laboratory,was ex perimeatally effective against Nippostrongylus braziliensis an.d Ancylostoma caninum.In acute and subacute toxicity test,it was proved lowly toxic.There was no evidence that the drug induced any damages to main organs and tissues of animals.In order to evaluate its potential mutagenicity and teratogenicity,micronucleus test,bone marrow metaphase analysis,CHL cells chromosomal aberration assay,spermatic aberration test as well as teratogenicity test were performed.No mutagenic and teratogenic effects were observed.

STUDIES ON THE EFFECT OF IMMUNOENHANCING REAGENT “425” ON EGG GRANULOMA FORMATION IN MICE INFECTED WITH SCHISTOSOMA JAPONICUM

1992, 10(1):  18-21. 

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Fourteen-34 days after Schistosoma japonzcum-infected mice were injected intraperitoneally with immunoenhancing reagent,Chinese angelica root extract ("425"),the specific antibody (IgG) levels were significantly higher in Group Ⅲ (362.67±162.21-480.00±289.45) than the control group (66.67±39.68-245.33±101.56) in experiment one,and in Gronp Ⅱ(488.00±320.38-768.00±267.38) than the control group (256.00±189.07-394.67±141.06) in experiment two.The egg granulomatous formation were markedly diminished in the injected mice.The ratios of granulomas vs.eggs' mean diameters of Group in of mice were 5.24±1.04-3.95±0.77 and those of the control group were 6.59±1.19-5.29±0.94 in experiment one,those of the group Ⅱ were 3.75±0.71-4.15±0.73 and those of the control group were 4.94±0.81-5.36±0.97 in experiment two.Meantime the egg antigen levels in the injected mice might be lower.This study shows that the control of immunomodulation of granulomatous formation in the hosts injected with immunoenhance reagent "425" can be induced.

PRODUCTION AND CHARACTERIZATION OF A MURINE PROTECTIVE MONOCLONAL ANTIBODY AGAINST SCHISTOSOMA JAPONICUM SCHISTOSOMULA

1992, 10(1):  22-25. 

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Eight murine monoclonal antibodies against surface determinants of Schistosoma japoni-cum (Chinese mainland strain) schistosomula were generated,of which only one monoclonal IgM antibody (N15D9) gave protection at level ranging from 14 to 39% in experiments of passive transfer or inhibition of infectivity while the others did not exhibit significant levels of passive protection.Further characterization of N15D9 antigen specificity showed that 96 and 14 kDa antigen molecules in cercaria,and 132 and 10 kDa in schistosomula could be recognized by N15D9 in Western blot assay.Furthermore,the 96 and 132 kDa molecules could also be recognized by pooled infected human sera while 14 kDa and 10 kDa only by sera from mice vaccinated with 3-hour schistosomula.The molecules recognizable by N15D9 were surface epitopes repeatedly expressed on cercaria,in vitro 3-hour mechanically transformed schistosomula and 5 day lung-stage schistosomula,as demonstrated by indirect immuno-fluorecence surface binding assay.

GENERATION AND IMMUNOCHEMICAL CHARACTERIZATION OF AN ANTI-EGG MONOCLONAL ANTIBODY SPECIFIC TO SCHISTOSOMA JAPONICUM

1992, 10(1):  26-29. 

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Monoclonal antibodies (McAbs) were generated from mice immunized with soluble egg antigen (SEA) of Schistosoma japonicum.Five of which were specific to SEA of S.ja-ponicum.The isotype of these McAbs was IgG1.Immunoblotting showed that the approximate molecule weight of the antigens recognized by the McAbs were 22kDa,116 kDa and 200 kDa respectively.At least 3 isomorphs of the 22 kDa antigen recognized by 1E1 were found at pI 4.6-6 with 2-D Western blot.Moreover,immunoprecipitation using 125I-labeled S.japonicum SEA demonstrated that only one band corresponding to 30 kDa was recognized by 1E1

LOCALIZATION OF THE ANTIGEN IN ERYTHROCYTIC STAGES OF PLASMODIVM YOELII BY IMMUNO-ELECTRON MICROSCOPY

1992, 10(1):  30-32. 

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In this study,the antigen recognized by the protective McAb M26-32 in erythrocytic stages of P.yoelii was localized by immuno-electron microscopy with LR Whithe resin embedding and colloidal gold probe cytochemical techniques.The results indicated that the antigen which reacts specifically to McAb M26-32 was mainly localized within the cytoplasm of early and late trophozoites,schizonts and merozoites,being the common antigen of asexual blood stages of the plasmodium.The amount of the antigen was on the increase during the deve-lopment of trophozoite,while a portion of the antigen might be transported outward by cxocytosis of the parasites and then be localized in the cytoplasm of the infected erythrocytes adjacent to the parasites.

ROLES OF T CELL SUBSETS IN THE PROTECTIVE IMMUNITY OF MICE AGAINST PLASMODIUM YOELII

1992, 10(1):  33-36. 

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BALB/c mice which had developed protective immunity against Plasmodium yoelii (P.y.) challenge were injected with anti-CD4 or anti-CD8 monoclonal antibody,and were then challenged again with P.yoelii.No impairment of protection was observed.CD8+T cells obtained from spleens of these mice were transferred to BALB/c nude mice and induced partial protection,while transfer of CD4+T cells did not so.In P.yoelli-mouse model,the parasites invaded reticulocytes in the early infection,and the infected reticulocytes could be recognized and attacked by sensitized CD8+T cells.In late infection when P.yoelii also invaded mature erythrocytes,the protective effect of CD8+T cells decreased and the main role of protection was played by antibodies.

IN VITRO DEVELOPMENT OF SODIUM ARTESUNATE RESISTANCE IN PLASMODIUM FALCIPARVM

1992, 10(1):  37-39. 

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Plasmodium falciparum(Lab.culture FCR3 isolate) developed resistance to sodium artesunate after exposure to the drug in vitro.The drug effective concentration which resulted in 50 per cent schizont maturation inhibition (IC50) was 1.6ng/ml (4.1nmol/L) before exposure to the drug.After 130 days of discontiguous exposure to sodium artesunate in a stepwise fashion,the sensitivity of the isolate to the drug decreased,with its IC50 3-fold higher than that of the parent isolate.The resistance to artesunate decreased significantly after the resistant line was grown in drug-free medium.

STUDY ON LOCALIZATION OF CYSTIC FLUID ANTIGEN OF CYSTICERCUS CELLULOSAE USING McAb-IGSS

1992, 10(1):  40-42. 

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A study was undertaken on the localization of specific combination of McAb against cystic fluid antigen of Cysticercus cellulosae by immunogold silver staining method (IGSS).It was shown that the combination sites of McAb F3 were distinct in the frozen and paraffin sections of Cysticercus cellulosae.The black granules in the bladder walls presented double-layer distribution and those in the scolex folded sites concentrated into pieces.The combination sites of McAb labeled by IGSS against cystic fluid antigen of Cysticercus cellulosae were consistent with the distribution of the cell layers of Cysticercus cellulosae.The sections in the control group had scarce and irregular distribution,which could be differentiated distindtly from those in the experimental group.

ON ELECTROPHORETIC KARYOTYPE OF ACANTHAMOEBA POLYPHAGA

1992, 10(1):  43-44. 

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An electrophoretic karyotype of Acanthamoeba polyphaga has been preliminarily analysed by means of pulse field gel electrophoresis (PFGE).Ten chromosomal DNA bands are distinguishable on the gel.Using yeast (Saccharomyces cerevisiae) chromosomal DNA as size standard,we estimate the size of the chromosomes to be between about 200 kilobase pairs (kb) and 2 megabase pairs (mb).

COMPARATIVE ANALYSIS OF AMINO ACIDS OF METACERCARIA AND LARVAE OF DIPLOID AND TRIPLOID OF PARAGONIMUS WESTERMANI

1992, 10(1):  45-47. 

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The diploid type of Paragonimus westermani in Heilongjiang Province and triploid type of P.westermani in Liaoning Province were examined for amino acids by high performance liquid chromatography (HPLC).Eleven kinds of amino-acids of diploid type and 15 kinds of triploid type were found in metacercaria; 15 kinds amino acids of diploid type and 16 kinds of triploid type were detected in larvae.The amino acids were more abundant in larvae than in metacercaria and more in the triploid than in the diploid type.The predominant amino acids were cystine and Rlutamic acid.

A STUDY ON THE ECOLOGICAL HABIT OF ARMIGERES SVBALBATUS IN DAWA AREA OF THE MENGSHAN MOUNTAIN IN SHANDONG PROVINCE

1992, 10(1):  48-51. 

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Armigeres subalbatus (A.s.) was reported in Shandong Province for the first time in 1965 and found in five counties of south Shandong including Pingyi,Linyi etc.in 1986.In Dawa area of the Mengshan mountain A.s.alults could be found in the first ten days of May,which increasied in number in July,and become the dominant species in mosquito colonies in Aug.and Sept.,then decreased gradually in number in Oct.and disappeared in Nov.There were two peaks of activity and blood-sucking behavior during the 24 hours of a day,one at dusk and the other at dawn.When the temperature dropped to 16℃ and below in the last ten days of Oct.,the wigglers began their diapause period.The survival ratio reached 90.5% after 12 h freezing at-5℃ and none survived after 60 hours freezing.When the temperature rose to 17℃ and above the over-winterting larvae developed into adults,which could suck blood only at the temperature above 17.5℃.

STUDY ON FREE AM1NO ACIDS AND PROTEIN IN HEMOLYMPH OF ANOPHELES STEPHEN SI

1992, 10(1):  52-55. 

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The changes in the contents of free amino acids in hemolymph of Anopheles stephenst were determined by automatic amino acid analyzer.The changes in hemolymph protein were determined by ultraviolet absorption method.Free amino acids in hemolymph of infected mosquitoes were compared with those in noninfected mosquitoes.At 4 days after blood meal,6 kinds of amino acids decreased markedly,and 5 kinds of amino acids increased markedly; at 7 days after blood meal,4 kinds of amino decreased markedly,while 7 kinds of amino accids increased markedly; at 11 days after cids blood meal,9 kinds of amino acids decreased markedly,and 4 kinds of amino acids increased remarkably.The protein concentration of infected mosquitoes was higher than that of noninfected ones.

STUDIES ON NEGATIVE CONVERSION OF SCHISTOSOME-INFECTED SNAILS IN MOUNTAINOUS REGION OF YUNNAN

1992, 10(1):  56-58. 

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The monthly cercaria shedding of 90 artificially infected Oncomelania snails was observed under a field condition in mountainous region,Shitoudi Village,Weishan County,Yunnan Province.The results showed that 20% of the snails shed cercaria monthly,42.2% could shed irregularly and 37.8% stopped releasing cercariae after several times of shedding.Final disvection of the snails showed that none cercaria or sporocyst could be found in part of the snails.A total of 304 naturally infected snails were observed individually every month.Samples were taken each month from the snails which did not shed cercaria,then dissecled and examined.The negative conversion rate was calculated in the second,third,fourth,fifth and sixth month after the first shedding,which were 36.1,50.0,41.0,39.8 and 2.6% respectively.

AN IMPROVEMENT OF LIVER EGG-IMMUNOENZYMATIC STAINING TECHNIQUE FOR DIAGNOSIS OF SCHISTOSOMIASIS JAPONICA

1992, 10(1):  59-61. 

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Using PVC membrane instead of common slide in liver egg-immunoenzymatic staining technique (IEST) for diagnosis of schistosomiasis japonica to detect specific antibodies in sera from 179 cases with schistosomiasis japonica,the positive rate was 97.2% (174/ 179).Sera from 63 healthy individuals revealed in one positive (1.5%).No cross reaction was observed in sera from 6 cases of paragonimiasis.PVC-IEST and SLIDE-IEST were comparatively performed on sera from 78 cases with schistosomiasis japonica,the positive rates were 96.2% (75/78) and 97.4% (76/78),respectively (P0.05).As PVC-IEST is simpler to perform than SLIDE-IEST,it is suggested that the former is more suitable for application in the field for diagnostic purposes.

CHANGES IN T LYMPHOCYTES AND THEIR SUBPOPULATIONS AND IgG IN PERIPHERAL BLOOD FROM MICE INFECTED WITH TRICHINELLA SPIRALIS

1992, 10(1):  62-65. 

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Acid α-mphthyl esiere.se (ANAE),a cytoplasmic marker,was used to identify the T lymphocytes and their subpopulations in peripheral blood of mice infected with Trichinella spiralis,and Dot-ELISA was used to identify the serum IgG antibody of the infected mice. The results showed that T lymphocytes increased on the d3 after infection,reaching the peak on d14,and then remainel in number greater than normal up to d77.The spotted granular ANAE positive cells (Help T cells,Th) deereased and the scattered granular ANAE positive cells (Suppresor T cells,Ts) increased,leading to a drop of the Th/Ts ratio.The reduction in host immune function during T.spiralis infection might be related to the drop of Th/Ts ratio.At the same time,the serum IgG antibody increased on d7 after infection,reaching the peak on d28,and remained higher up to d140.Tl:e results indicated that the host's cellular immunological level was lower while the host's IgG antibody level was higher during T.spiralis infection.

ANALYSIS OF NEWBORN LARVA OF TRICHINELLA SPIRALIS BY IMMUNOBLOT

1992, 10(1):  66-68. 

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Newborn larva (NBL) antigens of Trichinella spiralis were analysed by Immuroblot,and were comparied with the adult and muscle larva antigens.The SDS-PAGE patterns of NBL somatic constituents consisted of about 40 polypeptide bands,which were obviously different from those of adult and muscle larva.Immunoblot analysis indicated that immunization with NBL could induce a stage specific immune response.The molecular weight of specific NBL antigens were 129,120,89,87,79,74,72,64,58,43,40,38,34,32,and 20kDa.But during the natural course of the infection,we could not detect the antibodies of anti-NBL in the host.