Table of Content

30 November 1996, Volume 14 Issue 4

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DETECTION OF EXCRETED CIRCULATING ANTIGEN(CCA) IN URINE OF INFECTED HOSTS BY MEANS OF SANDWICH STRIP ELISA

1996, 14(4):  253-256. 

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AIM:To set up a simple immunoassay for detection of the excreted antigens in host urine.METHOD:A modified double anitbody sandwich ELISA in the form of a strip reagent which combined the advantages of both dot ELISA and plate ELISA for the detection of the gut associated circulating cathodic antigen(CCA) in the urine of Schistosoma japonicum infected rabbits and patients was developed.RESULTS:All the 9 reconstituted lyophilized urine samples collected from heavily infected rabbits,stored for over 10 years,gave strong positive reactions up to an end point titre of 512 -1 ,while only one out of 11 urine samples from normal rabbit stored and reconstituted in the same manner showed a weak reaction which faded at a dilution of 8 -1 .Urine CCA detections in 58 schistosomiasis patients collected from Sichuan Province,China and in 60 healthy students from the Shanghai Second Medical University revealed an assay sensitivity of 43.1% and a specificity of 96.7%. The detectability has been found similar to the parallelly performed plate ELISA for urine CCA.A certain degree of diagnostic complementarity in parallel up to 58.6% in these patient samples was seen with no increase of false positive cases.For identification of the diagnostic specificity,an appropriate amount of the capture antibody was added to the infected rabbit urine,a dose dependent competitive inhibition of the reaction intensity was observed,indicating the specific recognition of the target molecules in the urine by the anti CCA McAb system.The possible diagnostic complementarity via different methods using the same probe system was discussed.CONCLUSION:Results of this study have revealed the possibility as well as various advantages of the sandwich reagent strip assay to be used in the field for an non invasive detection of the excreted antigens in urine of infected individuals.

DYNAMIC STUDY ON PHENOTYPE OF LYMPHOCYTES IN MICE AFTER VACCINATION WITH RADIATION ATTENUATED CERCARIAE OF SCHISTOSOMA JAPONICUM AND S.MANSONI

1996, 14(4):  262-265. 

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AIM:Vaccination with radiation attenuated cercariae of schistosomes induced partial resistance in several hosts against challenge with normal cercariae.In the study,phenotype of lymphocytes in mice after vaccination with uv attenuated cercariae were investigated.METHOD:Female NMRI mice were used, each received either 200 untreated or 500 attenuated cercariae of S.japonicum or S.mansoni 6-7 mice of each group were killed at 3 d,5 d,7 d,10 d,14 d and 21 d after infection or vaccination.The axillary lymphnodes and the spleens were removed.Cells were prepared separately and were analysed by double colour immunofluorescence with conjugated monoclonal antibodies in a FACScan cytofluorimeter at 488 and 560 nm. RESULTS:Immunization with 500 attenuated cercariae was followed by a three to four fold increase in cell number in axillary lymph nodes,irrespective of schistosome species.In both cases,CD + 4 as well as B cell numbers increased,reaching peak levels at around 10 days;while CD + 8 cells did not appreciably vary over the three weeks of observation.CONCLUSION:On the basis of this study it is suspected that CD + 4 cells might play a significant role in the protective immunity elicited by uv attenuated cercarial vaccine.

ANALYSIS ON HOMOLOGY IN SEVERAL ISOLATES OF LEISHMANIA FROM

1996, 14(4):  266-269. 

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AIM: To identify Leishmania isolates in Phlebotomus (P.major wui) and cutaneous leishmaniasis (CL) patients from,Karamay, Xinjiang . METHOD:Study on the characterization and genotype of Leishmania isolates from the sandflies, CL patients and the reference strain by analysing simultaneouly kinetoplast and chromosonal DNA. RESULTS: The nDNA genotype analysis showed that isolates from the sandflies, CL patients and Leishmania infantum are highly homologous. CONCLUSION: Leishmania infantum is the pathogen of cutaneous leishmaniasis in Karamay, Xinjiang, and P.major wui, the vector of this disease.

CHARACTERIZATOIN OF cDNA OF THE MEMBRANE ANTIGEN Pc90 ON ERYTHROCYTE INFECTED WITH PLASMODIUM CHABAUDI II.CHARACTERIZATION OF THE IMMUNE POSITIVE CLONE Pc90 1 AND ITS EXPERSSION IN E. COLI

1996, 14(4):  274-279. 

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AIM: To characterize the immunopositive clone expressing Pc90 of Plasmodium chabaudi and then express it in E. coli. METHOD: A cDNA clone (designated Pc90 1) was deleted with ExoⅢ and sequence analysis was performed with chain terminating, then transferred to E. coli for fushion expression. RESULTS:The clone Pc90 1 with a length of 2 581 nucleotides was identified. Its deduced coding capacity was approximately 61 kDa (532 amino acids). The open reading frame extended to position 1 569, which was followed by two consecutive stop codons. Surprisingly, the 3 putative untranslated regions were relatively long. Pc90 1 showed a remarkable repeat structure composed of four distinct types of repetitive elements (between 100 and 355 amino acids). Five sequencing subclones were subjected to expression in pEX32 vectors,but only subclone SK5 expressed a 35 kDa protein. The Pc90 1 has no homology with all known sequences by a search of EMBL and Swiss Prot data bases. The characterization of Pc90 1 deduced protein was identical with Pc90, an phosphoprotein with an isoelectric point of 4.6. CONCLUSION: Pc90 1 was a possible candidate expressing Pc90 protein.

EPIDEMIOLOGICAL SURVEY BY RANDOM SAMPLING ON PATIENTS WITH CHRONIC FILARIASIS IN HUNAN PROVINCE WHERE THE DISEASE HAD BEEN BASICALLY ELIMINATED

1996, 14(4):  280-284. 

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AIM: To survey the morbidity of chronic filariasis in Hunan Province where the disease had been basically eliminated during 1991 1993. METHOD: 108 villages of 12 counties/cities were randomly selected and investigated on fliariasis endemicity. Local residents above ten years of age were inquired about illness and susbjected to physical examination. RESULTS: 470 chronic filariasis cases were discovered, the prevalence being 0.4% in a population of 111 127, of which elephantiasis accounted for 0.1%, hydrocele 0.2% and chyluria 0.1%. A total of 386 cases were detected before, and 84 after, the basic elimination of filariasis, the prevalence being 0.4% and 0.07%, respectively.CONCLUSION: The results indicated that after basic elimination of filariasis, new elephantiasis cases have not been detected in endemic areas of malayan filariasis, while few new cases of chyluria and hydrocele continued to delevop in areas endemic for bancroftian and mixed filariasis.

CALCULATION OF INFECTION RATE AND NUMBER OF INFECTED PERSONS IN NATION WIDE SAMPLING INVESTIGATIONS ON SCHISTOSOMIASIS JAPONICA

1996, 14(4):  290-293. 

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AIM:To offer the method of calculating infection rate and number of infected persons in nation wide sampling investigations on schistosomiasis japonica. METHOD: Calculation formulae were established on the basis of schistosomiasis epidemiology and biomedical statistics, adopting multiple stratified cluster sampling. RESULTS: Thirteen formulae were derived for calculation of infection rate and number of infected persons in different endemic provinces with different types of endemic areas. CONCLUSION: The presented method is simple, rational and available.

HISTOPATHOLOGICAL STUDIES ON ROSACEA BY LIGHT AND TRANSMISSION ELECTRON MICROSCOPY

1996, 14(4):  299-302. 

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AIM: To understand different histopathological and ultrastructural changes in three forms of rosacea as well as the parasitism of Demodex species in situ . METHOD: Fourteen biopsy and dissected materials of three forms of rosacea were collected and observed under light and transmission electron microscope. RESULTS: Demodex folliculorum and Demodex brevis were found parasitizing the dilated follicles and proliferative sebaceous glands. Chronic inflammatory cells of various types were present in the dermis. In acne rosacea and rinophyma rosacea, many dilated or proliferative capillaries were viewed with thinning or stratified thickenning of the basement membrane, and degranulation of the mast cells. CONCLUSION: pathological changes of rosacea of dermatitis type is due to the parasitism of Demodex species, while the histopathological changes of other types of rosacea are caused by bacterial infection together with hair follicle mite infestation, possibly being not only related to cellular and humoral immunity, but also to types III and IV allergy.

OBSERVATION ON THE HISTOLOGICAL CHANGES OF MOSQUITO LARVAE INFECTED BY LAGENIDIUM GIGANTEUM *

1996, 14(4):  303-306. 

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AIM:To investigate the mechanism of Lagenidium giganteum for invading and killing mosquito larvae, the interaction between the fungus and mosquitoes,and histological changes of mosquito larvae infected by this fungus were observed in the laboratory. METHOD: The larvae of Culex quinquefasciatus and Aedes albopictus of different infection periods by L. giganteum were fixed.Their paraffin sections were made and stained with H. E. method, observed with Olympus microscope and photoed with PM 10 auto camera. RESULTS: During the early stageⅠof infection, many fine branchy vegetative mycelia of L. giganteum were found in tissue gap of the head and thorax; later, the mycelia entered the hemocoel and developed to a large number; subsequently, they invaded the muscles,stomach,middle gut, and fat bodies of the host. Some hyphae even grew in cells of these organs or tissues and destroyed them. In stage Ⅱ of infection,remarkable destruction and disappearance of tissues were observed. The vegetative mycelia developed into reproductive form which were septated and were shorter and wider. Then, they became sporangia in round,ovoid or bead string forms. By the end of infection,most of the normal tissues of larvae disappeared, and the cytoplasm and nuclei of mycelia and sporangia of the fungus also disappeared. Only empty tubes derived from the cell wall of the fungus were found in larval bodies. CONCLUSION: It was considered as one of the lethal causes of mosquito larvae that L. giganteum invaded and destroyed cells of tissues and organs, exerting damage consequence on the host.

EFFECT OF INOCULATED SPOROZOITES OF PLASMODIUM YOELII ON THE ULTRASTRUCTURE OF RAT LIVER

1996, 14(4):  307-309. 

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AIM: To examine the effect of inoculated sporozoites of Plasmodium yoelii on the early cellular responses of rat liver. METHOD: Ultrastructure of rat liver was observed by transmission electron microscopy 30 minutes after inoculating with nine million sporozoites. RESULTS: Phagocytic activity of Kupffer cells(Kc) was increased. There were a lot of phagolysosomes in the cytoplasm of Kc. Blood platelets were swollen and adhered to Kc or endothelial cells and to form microthrombi. Some hepatic cells were damaged. The mitochondria of hepatic cells were vacuolated and the mitochondrial cristae were dissolved.There were cytolytic focal necrosis in the cytoplasm. CONCLUSION: Acute cellular responses of rat liver occurred 30 minutes after the inoculation of nine million of P. yoelii sporozoites. Some sporozoites were damaged by Kupffer cells. The ultratructural changes of mitochondria of hepatic cells contributed at least partly to the degeneration of exo erythrocytic stage of the parasite.

STUDIES ON EFFICACY OF PRAZIQUANTEL AND MEBENDAZOLE-MEDICATED SALT IN TREATMENT OF ECHINOCHASMUS FUJIANENSIS INFECTION

1996, 14(4):  310-313. 

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AIM: To study the effect of praziquantel and mebendazole medicated salt for treating Echinochasmus fujianensis infection. METHOD: 109 cases with E. fujianensis infection were randomly divided into four groups, and administrated respectively with a single dose of praziquantel: 5 mg/kg (A), 2.5 mg/kg (B); or daily dose of mebendazole medicated salt contai ning the drug 80 mg × 10 d (C), 40 mg × 10 d (D). The parameters for evaluating efficacy were egg negative conversion rates and egg reduction rates. RESULTS: Four weeks after treatment, the negative conversion rates were 100%, 92.3%, 85.2% and 71.4% in groups A, B, C and D, respectively; the egg reduction rates were 84.8%-100%. The curative effects among groups A, B and C are not statistically different. CONCLUSION: Praziquantel is the drug of choice for treatment of E. fujianensis, while mebendazole is also effective,especially for those with mixed infections of other intestinal namtodes.

COMPARATIVE STUDIES ON THE TREATMENT OF DRUG RESISTANT FALCIPARUM MALARIA WITH SINGLE DOSE OR TWO DAY REGIMENS OF PYRONARIDINE/SULFADOXINE PYRIMETHAMINE PLUS PRIMAQUINE

1996, 14(4):  314-317. 

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AIM: To observe the comparative effects of pyronaridine/sulfadoxine pyrimethamine with single dose or two day regimens in combination with primaquine in 2 endemic areas of multidrug resistant falciparum malaria in Hainan Island. METHOD: A total of 100 patients with acute falciparum malaria were randomly divided into 2 groups. Group A was orally administered with a single dose of pyronaridine(PND) 500 mg/sulfadoxine(S)1 000 mg and pyrimethamine(P) 50 mg composite plus primaquine(Pq)22.5 mg, 24 hours later Pq 22.5 mg was given again; and Group B received a dose of PND 400 mg/S 500 mg and P 25 mg composite plus Pq 22.5 mg daily for 2 days. RESULTS: The average fever subsidence time of groups A and B was 39.8±27.2 and 42.1±20.5, the asexual parasite clearance time was 46.9±13.3 and 51±14.1 ( P 0.05), the rates of asexual parasite eliminated within 24 hours were 80% and 78%, respectively; the asexual parasite clearance rate within 28 days of both groups was 100%. Gametocytes were cleared within 21 days. The side effects were mild, but hemolysis reaction occurred in 3 cases of each group. CONCLUSION: Both presented regimens could cure multidrug resistant falciparum malaria.