Table of Content

31 May 1990, Volume 8 Issue 2

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PRELIMINARY STUDY ON THE SURFACE ANTIGEN GENE OF PLASMODIVM MALARIAE

1990, 8(2):  81-83. 

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The genomic Hind Ⅲ λgt1149 library from Plasmodium malariae was screened with two types of oligonucleotides, both of which were from the P190 gene of P. falciparum. One, MAD20 type, was made from spanning nwcleotides 4488-4562 in the P100 gene of MAD20 strain and another, K1 type, was made from spanning nucleotides 3652-3692 in the P190 of K1 strain. Both were end-labelled with 32P-ATP as probes before hybridization. Two clones were selected. One clone, designated XMSA-1, was specifically recognized by the MAD20 type oligo probe; the other,designated XMSA-2,by the K1 type oligo probe. XMSA-1 and AMSA-2 inserts were obtained by Hind Ⅲ digestion of the two-clone phage DNA. The analysis of the two inserts showed that the size of X.MSA-1 is 5.5 kb whilst that of XMSA-2 is 3.5kb. The MSA-1 and MSA-2 inserts recloned into PUC8 were digested with the restriction enzymes Bg1 Ⅱ, EcoR Ⅰ , Xba Ⅰ , Hind Ⅱ and Pst Ⅰ. The results showed that the MSA-1 DNA had one Bgl Ⅱ site, one EcoR Ⅰsite, two Xba Ⅰ sites, one Hind Ⅱ site and one Pst Ⅰ site. The MSA-2 DNA had only one Hindn site.The surface antigen gene of P. malariae was little known. This result also showed that there was probably an analogue of P190 on the surface of P. malariae, and they might fall into two types. This study is informative for further investigation on malariae parasites (Figs. 1-8).

ULTRASTRUCTURAL STUDIES ON EFFECTS OF COLCHICINE IN TREATING HEPATIC FIBROSIS OF SCHISTOSOMIASIS RABBITS

1990, 8(2):  84-87. 

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8 rabbits were infected percutaneously with the cercariae of Schistosoma japonicum; After hepatic fibrosis had developed in these rabbits 4 months after infection, 4 out of the 8 infected rabbits were given colchicine orally at a dosage of 40/ig/kg per day for 7 weeks. Another uninfected rabbits were used as controls. The therapeutic effects of col-chicine on hepatic fibrosis were studied by transmission electron microscopy and mor-phometry, in which the area of collagen microfibrils in the space of disse and liver cells was measured.The results showed that colchicine relieved ultrastructural injury of liver cells and reduced the number of active fibroblasts and collagen microfibrils. The area of collagen microfibrils measured in the liver of the infected rabbits and colchicine-treated rabbits accounted for 42.5% and 0.2%, respectively (P0.01), suggesting that colchicine has therapeutic effect against schistosomal liver fibrosis (Figs. 1-4).

STUDY ON MCABS AGAINST PROTEIN "TARGET ANTIGEN" IN SCHXSTOSOMA JAPONICUM

1990, 8(2):  88-91. 

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In the studies presented here, we demonstrated the feasibility of producing large number of hybridoma cell lines secreting McAbs against S. japonicum including 16 cell lines secreting IgG McAbs (7 for IgG1, 5 for IgG2a, 1 for IgG2b, 3 for IgG3) and 13 McAbs for IgM, on the basis of successfully extracting 24-26kD and 90kD "target an tigen" proteins known as important antigens for inducing schistosome protective immunity All the above cell lines were characterized for localization of the McAbs, mediating in vitro ADCC against schistosomula, epitope recognized by the McAbs on different kinds of schistosome antigens. Studies have shown the evidences that the McAbs can be used to identify the "target antigen" on the surface of schistosome, to isolate and purify "target antigen" of S. japonicum by chromatography column bearing McAbs, as well as to immunize animals as antigens for accumulation of data in the development of vaccine against S. japonicum via anti-idiotype antibodies.

STUDIES ON THE STRAIN DIFFERENCES OF SCHISTOSOMA JAPONICUM IN THE MAINLAND OF CHINA I. COMPATIBILITY BETWEEN SCHISTOSOMES AND THEIR SNAIL HOSTS

1990, 8(2):  92-95. 

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Oncomelania hupensis hupensis from six localities were used in this study, i.e.Gui-chi of Anhui in the east, Jianli of Hubei in the middle, Guiping of Guangxi in the south, Tianquan of Sichuan and Eryuan of Yunnan in the southwest and Fuqing of Fujian in the southeast. Snails from each locality were individually cross-exposed to 20 miracidia of the different isolates of S. japonicum from the above-named places, with the exception of Fujian Province where no snail could be found naturally infected with S. japonicum.The results showed that snails from one locality were readily infected with the local isolate of S. japonicum. Besides, cross infection also took place readily between the snails and the schistosomes from Hubei and Anhui with snail infection rates of 43.8% and 40.9% respectively.Snails from Sichuan and Yunnan were refractory to infection with schistosome isolates from Hubei and Anhui, but the isolate from Sichuan was able to develop in Oncomelania snails from Hubei and Anhui, resulting in infection rates of 10.2% and 4.5% white that from Yunnan, in snails from Hubei and Anhui in infection rates of 33.6% and 10.8% respectively.Though the Guangxi isolate of S. japonicum developed readily in both Anhui (30.7%) and Guangxi snails (9.4%), the average precercarial period was 100.9 days in the for-mer which was significantly longer than 76.9 days in the latter. None of the other snails from Sichuan, Yunnan and Fujian became infected. On the other hand, snails from Guangxi infected with Anhui parasites also had a longer precercarial period of 92.7 days compared with that of 81.6 days in Anhui snail. The difference between the precercarial periods was statistically significant.Snails from Fujian were readily infected with the isolates from Anhui and Yunnan, and the precercarial period was 76.0 and 63.0 days post infection respectively.

EFFECT OF ALBENDAZOLE AND PYRANTEL IN TREATING INTESTINAL HELMINTHIASIS AND CONTROLLING THE RECURRENCE OF HOOKWORM INFECTIONS

1990, 8(2):  96-99. 

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A total of 720 human intestinal helminthic infections were divided into 4 groups and treated with albendazole 400mg/d × 3d, 400mg/d × 5d, pyrantel 1 500mg/d × 3d, or 1 500mg/d×5d. Half a month after treatment, the negative rates of hookworm egg were 98.6, 98.6, 86.2 and 93.5%, those of ascaris egg were 96.5, 98.2, 92.9 and 96.3%, and those of whipworm egg were 86.4, 89.0, 68.9 and 67.0% respectively. Reduction rate of hookworm egg reached more than 98% in all the 4 groups. Six months after treatment, however, the positive rates of all the 4 groups rose again in varying degrees. The predominant species of hookworm infections was Necator americanus before the treatment and Ancylostoma duodenale after the treatment. It was demonstrated that the recurrence of hookworm infection resulted from A. duodenale infections, while a single dose of 400mg albendazole per day for 3 or 5 days showed good effect in controlling the recurrence of, hookworm infections in a certain area.

CLINICAL OBSERVATIONS ON THE TREATMENT OF HOOKWORM, ASCARIS AND TRICHURIS INFECTION WITH OXIBENDAZOLE

1990, 8(2):  100-103. 

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340 cases of hookworm infection, 196 cases of ascariasis and 178 cases of trichuriasis were treated with an anthelmintic, oxibendazole, 15mg/kg·d × 3d; 102, 70 and 66 cases of respective infections were treated with pyrantel 10mg/kg·d×3d and 108, 74 and 63 cases took placebo for comparison and as control under double-blind observations. Re-examinations of the stool were performed after the treatment. Among the cases treated with oxibendazole, the egg negative conversion rates of hookworm, Ascaris and Trichuris were 70.3-80.6%, 92.5-97.8 and 67.0-71.0% respectively. The egg reduction rates of hookworm were 98.1-98.6%. The larval negative conversion rates of Ancylostoma duodenale and Necator americanus were 77.7 and 83.2% respectively. Among the cases treated with pyrantel, the egg negative conversion rates of the three above mentioned parasites were 73.5, 90.0 and 28.8% respectively. The egg reduction rate of hookworm was 98.8%. Among the cases treated with placebo, the egg negative conversion rates of the three above mentioned parasites were 6.5, 29.7 and 7.9%. No marked adverse reactions were observed by clinical and laboratory examinations.

OBSERVATION ON THE INFECTIVITY OF LEISHMANIA DONOVANI FROM DIFFERENT AREAS TO PHLEBOTOMUS ALEKANDRIFROM XINJIANG

1990, 8(2):  104-107. 

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Hamsters or cotton rats-were infected with 3 stocks of L. donovani, one of which had been isolated from an infected Ph. major wui in Xinjiang desert area and 2 from kala azar patients in a mountain area of Gansu and in a plain area of Henan respectively. The purpose of the study was to compare the infectivity of the parasites from different geographic areas through artificial infection of Ph. alexandri. Sandflies were dissected in 4, 6, 8 and 10 days after feeding on the infected animals to observe the growth and development of promastigotes of different isolates in their alimentary tracts. Evaluation was made on 3 aspects, i. e. infection rates, infectiosity and migration of promastigotes in sandflies. It was demonstrated that Xinjiang isolate of L. donovani appeared to be particulary adapted to Ph. alexandri followed by Gansu isolate, the Henan isolate did not show good adaptation to this sandfly, because the number of promastigotes declined and the infection rate dropped after the digestion of the blood meal in the stomach of the sandflies. It seems that the phenomenon was related to the biological characteristics of L. donovani isolates. These results coincided with those of McAb dot ELISA(Qu,1987) and K-DNA dot hybridridization (Lu Hu 1988) for identification of L. donovani from different areas in China, and were also in parallel to the diverse epidemiological characteristics of different kala azar endemic areas (Guan, 1976) .It is reasonable to infer that there may exist different geographical strains of L. donovani in China.

ULTRASTRUCTURE OF METACERCARIAL MEMBRANE AND IMMUNOHISTOCHEMICAL LOCALIZATION OF METACERCARIAL MEMBRANE REACTION OF PARAGONIMUS WESTERMANI

1990, 8(2):  108-109. 

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The ultrastructure of the tegument of the early excysted P. westermani metacercariae Was observed with transmission electron microscope. The tegument was fine and dense in texture, containing a lot of disk-like bodies and a few mitochondria. The outer-plasma membrane was clear and intact.After the metacercariae were incubated in rabbit anti-serum for 3 hours, metacereanal membrane reaction (MMR) appeared closely to the outer plasma membrane and the tegument was damaged.Indirect peroxidase-labelled immunohistochemical technique was used to identify the location of the MMR at ultrastructural level. The result showed that the antibody-antigen complex was localized on the tegumental surface of the worm. The data constituted the evidence that the MMR resulted from the antibody-antigen reaction and the tegumental surface was the reaction site for the MMR (Figs. 1-4).

PRELIMINARY STUDIES ON TRANSAMINASE OF ONCOMELANIA SNAIL

1990, 8(2):  110-112. 

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By paper chromatography, the tissue homogenate of Oncomelania snails was shown to form glutamic acid at the expense of a-ketogiutarate pms asparuc acia,aialmine otarginine respectively. The existence of alanine-glutamate, aspartate-glutamate and arginine-glutamate transaminase in Oncomelania snail was demonstrated.By using colorimetric method, the activity of aspartate-glutamate transaminase(GOT) and alanine-glutamate transaminase (GPT) of Oncomelania snail was 1.64±0.01 and 0.99 ± 0.01μmol/h·mg protein respectively. GOT and GPT were not inhibited by 2ppm bromoacetamide, but the activity of GPT was suppressed (40 %) by 2ppm nicotinanilide. A combination of 0.5ppm bromoacetamide and 0.5ppm nicotinanilide had no synergetic molluscicidal effect (Fig. 1).

SOLUBLE PROTEIN AND ESTERASE ISOZYME ANALYSIS OF ADULT FEMALE ANOPHELES STEPHEN SI AFTER PLASMODIUM YOELII YOELII-INFECTED BLOOD MEAL

1990, 8(2):  113-116. 

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In this paper, the system of Anopheles stephensi and Plasmodium yoelii yoelii was used as animal model. The soluble protein and esterasc isozymes of adult female mosr quitoes at various times after taking noninfected and plasmodia-infected blood meal were investigated.The result shows that the amount of soluble protein in plasmodia-infected mosquitoes was lower than that in noninfected ones (P0.01); the activity and electrophoretic patterns of EST in infected mosquitoes being also different from those of the noninfected ones.

OBSERVATION ON POLYTENE CHROMOSOMES OF THE OVARIAN NURSE CELL OF ANOPHELES MINIMUS FROM GUANGXI

1990, 8(2):  117-120. 

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The ovarian nurse cell's polytene chromosomes of An. minimus exist as 5 arms, the telocentric X chromosome, the submetacentric chromosome 2 and the metacentric chromosome 3. The X chromosome is easily recognized by its length and shuttle-shaped zone 6, The most important characteristics is that each autosome arm has one to three big puffs. Among the 46 zones, 7A, B and 19C in 2R which is the longest arm in the complement, 28A and 20A, B in 2L, 30A, B and 37D in 3R and 46D and 38A, B in 3L are considered as characteristic zones. It is proposed that this map might be eonsn dered as the "standard" map of the ovarian nurse cell's polytene chromosomes for An, minimus from Guangxi (Fig. 1).

IN VITRO CULTIVATION AND SCANNING ELECTRON MICROSCOPIC OBSERVATION OF PLASMODIUM BERGHEI

1990, 8(2):  121-123. 

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To compare the effects of sera on the growth of Plasmodium berghei, the erythrocytic stages were cultured with rat serum, human umbilical cord serum, human B-type serum, rabbit serum, calf serum and calf serum with hypoxanthine respectively. The topography of the erythrocyte and parasite cultured with calf serum were observed before and 12, 20 and 28 hours after cultivation. All of the sera used did not effectively improve the long-term culture of P. berghei, regardless of some differences in short-term cultures. Under SEM, erythrocytes agglutinated with each other and the surface adhesive materials on the erythrocytes and parasites agglomerated gradually with the cultivation time. The results suggested that the surface adhesive materials were correlated with the destruction of erythrocytes, malnutrition of the parasites and blockade of merozoite reinvasion (Figs. 1-16).

STUDIES ON THE KARYOTYPES OF FASCIOLA SPP.

1990, 8(2):  124-126. 

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The karyotypes of diploid and triploid forms of the common liver fluke were analysed by air-drying method.The chromosome number of diploid form was 2n = 20,n=10, and the karyotype consists of the chromosomes with 2M, 4Sm, 3St, 1T. There were a number of similarities between the karyotypes of diploid form and triploid iorm except that the chromosome number of the triploid was 3n = 30 and n = 10. Judging from the shape of adult worms and their life cycle, the worm of the diploid form is similar to F, gigontica while the worm of the triploid form is similar to F. hepatita.In several specimens of the triploid worms there were some cells having 20 chromosomes that were composed of 10 monovalent chromosomes asd 10 divalent chromosomes. This kind of cell, in a group of 8 cells, was primary spermatocyte. According to the measurement, these chromosomes could be classified into 10 pairs and each pair was made up by one monovalent chromosome and one divalent chromosome. The karyotype of these chromosomes was similar to that of the triploid form. It is a rare phenomenon that one set of monovalent chromosomes and one set of divalent chromosomes co-exist during the same metaphasc (Figs. 1-4).

DETECTION OF SERUM ANTIBODIES IN JIRDS AGAINST ANTIGENS OF BRUGIA MALAYI

1990, 8(2):  127-130. 

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The indirect fluorescent antibody test (1FA) using the infective larva (L3), adult worm (A) and microfilaria (Mf) antigens was applied to detect anti-filarial antibodies in jirds (Meriones unguiculatus) infected with Brugia malayi. The antibodies against L3 appeared within 3 weeks after infection, followed by A and Mf. The serum antibody levels in the jirds with positive infection were compared with those in the jirds with negative infection. Before appearance of microfilaria, there was insignificant difference between these two groups. However, significant difference was observed after the appearance of microfilaria. The method of sonicating L3 and Mf fragment antigens used in this study is simple, and its results are easy to observe.

DETECTION OF SERUM IMMUNOGLOBULINS IN PULMONARY ACARIASIS PATIENTS

1990, 8(2):  131-133. 

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The IgG, IgA, IgM levels in sera of 51 patients suffered from pulmonary acariasis and the serum IgE level in 16 of them were determined using single immunodiffussion technique or biotin-avidin enzyme immunosorbent assay (ABC-ELISA). The results showed that the IgM level of the patients was approaching to that of the normals while IgG (169.77±46.91U/ml), IgA (308.39±91.83U/ml) and IgE (458.90±273.64U/ml) levels of the patients increased significantly. This suggests that some components of mites may have stimulated the humoral immunoreactions of the host.

EPIDEMIOLOGICAL CHARACTERISTICS AND CONTROL OF FILARIASIS IN HUNAN PROVINCE

1990, 8(2):  134-137. 

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Of 98 counties or cities in Hunan Province, 55 were endemic areas of filariasis. The average microfilaria rate was 5.64% (180046/3194102), and the incidence of advanced filariasis, including elephantiasis and hydrocele was 3.29 %. The number of filariasis patients in the whole province was estimated to be 1.63 million, comprising 1.25 million of microfilaretnia cases, Culex fatigans and Anopheles hyrcanus sinensis were the major vectors of bancroftian and malayan filariasis respectively in the province.Control strategies concentrated on the elimination of infection source were implemented on the basis of extensive investigations, and the ensuing examination and treatment of filariasis cases in pilot areas prior to the implementation of province-wide filariasis control. Repeated blood examinations and medications for 3-4 times were carried out in hypo-endemic areas of malayan filariasis, whereas mass treatment with hetrazan-medicated salt containing 0.2% to 0.5% DEC was carried out in meso-and hyper-endemic areas of bancroftian filariasis as well as those situated in remote mountainous regions for six months.Subsequent evaluation and clearance checking showed that microfilaria rate of the whole province has already dropped to 1%. That filariasis was basically eliminated in Hunan was recognized by the Evaluation Mission Group sent by the Ministry of Public Health in 1986.