Table of Content

30 November 1993, Volume 11 Issue 4

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PURIFICATION OF 31/32 kDa PROTEINS OF ADULT SCHISTOSOMA JAPONICUM AND STUDIES ON THEIR PROTECTIVE IMMUNITY

1993, 11(4):  241-243. 

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Schistosoma japonicum adult worm 31/32 kDa proteins (Sj 31/32) were separated on polyacrylamide slab gels and purified by electrophoretic elution. These purified proteins were used to immunize mice in order to observe their protective immunity against challenge. The results of SDS-PAGE,EITB and ELISA indicated that the 31/32 kDa proteins separated and purified by means of these methods were pure and active. It was assumed that Sj 31/32 proteins could reduce worm burden and inhibit the fecundity of schistosome and formation of egg granuloma. The results suggested that 31/32 kDa 5. japonicum proteins might be an important component of a multivalent vaccine against schistosomiasis japonica (Figs. 1 - 4).

STUDIES ON THE ESTABLISHMENT OF MALARIAL ANIMAL MODEL OF SHORT-TERM RELAPSE IV. SHORT-TERM RELAPSE IN RHESUS MONKEYS INFECTED WITH SPOROZOITES OF PLASMODIUM CYNOMOLGI

1993, 11(4):  244-247. 

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The present paper reports that the short-term relapse could be artificially made by the application of the experimental method, and thus we established the monkey model of the short-term relapse. According to the experimental design,when the parasitemia was detected in rhesus monkeys infected with sporozoites of Plasmodium cynomolgi, a combined therapy of pyronaridine 6 mg/kg body weight, artemether 10 mg/kg and chloroquine 20 mg/kg once daily for 3 days was carried out to clear the erythrocytic parasite and then the short-term relapse was observed in the animal follow-up for 100 days. The combined therapy was given again when relapse occurred.One onset of relapse occurred on 47 days after therapy in monkey Ml94 infected with sporozoites 11×103. In M195 infected with sporozoites 55×104, relapes occurred for 3 times on 30,37 and 51 days respectively after medication,but during a follow-up period of 200 days,no relapse was shown in M192 and M193 infected with sporozoites 50 and 55×102, respectively.The results showed that the frequency of the short-term relapse was relative to the sporozoites inoculation,and sporozoites less than 11×103 were not suitable for making short-term relapse in animal model. No long term relapse could be seen in all the 4 monkeys until 400 days.Under existing conditions that the relapse was generally explained with hypnozoite assumption, the establishment of animal model of short-term relapse could be tenable. The reasons were:(l) P. cynomolgi from Vietnam,a parasite with the biological characteristic of short-term relapse, was recognized; (2) A series of studies for the relationship between the parasite and monkey host was carried out and the experimental basis of the model establishment was put forward by the authors; (3) This study did not exclude the hypnozoite assumption.The significance of this animal model study at least included : (1) to innovate the treatment countermeasure of the short-term relapse of malaria; (2) to study the biology of the short-term relapse; and (3) to test the antimalarial drugs for the radical cure of vivax malaria. As for the animal model of the long-term relapse,it will be necessary for study to select the monkey parasite which has the biological characteristic of long-term relapse. The verification of the relative problem,hypnozoite assumption,will also be necessary.

THE RESIDUAL CORRECTING PREDICTION OF GREY MODEL FOR MALARIA CASES IN CHINA

1993, 11(4):  248-250. 

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In this study a grey model GM(1,1) was developed on the basis of malaria cases reported for the period of 1986-1990 in China. In order to further enhance the predicting power of the model,residual correction is performed and the residual correcting GM(1,1)model was established. The result showed that the predicting performance of the latter model was much improved. The number of predicted cases for 1991(101 770) was conformed to the number of actual malaria cases reported (101 636).

EVALUATION OF THE EFFICACY OF PRAZIQUANTEL-LIPOSOME AGAINST ALVEOCOCCOSIS

1993, 11(4):  251-254. 

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In this paper, praziquantel, an anti-hydatidosis drug, was encapsulated with lipsome by REV (Reverse -phase -evaporation) method, the encapsulated percentage was 60%. The drug distribution dynamics in mice of both praziquantel (PZQ) and praziquantel liposome (PZQ-Lip) were determined using RP-HPLC method. The results indicated that the drug concentration of blood, liver and spleen in the PZQ-Lip group was higher than those in the PZQ group from 1/2 to 16 hours post ip. the t 1/2 of the former was considerably prolonged. The toxicity of PZQ -Lip tested by LD50 was decreased about one fourth as compared with PZQ (3372 vs. 2 454 mg/kg). There was a significant difference in the cyst inhibition rate between PZQ-Lip group (68. 7%) and PZQ group (14. 3%)(P0. 01) as shown by the results of secondary alveococcosis mice treated with either drug at 500 mg/kg· d × 12 d for four consecutive courses. Histological observation of the germinal layer after treatment showed that the damage of PZQ-Lip group was more severe than that of PZQ group. The ul-trastructural observation showed that both drugs had marked effects on the organelles of cells. The above experiments indicated that the efficacy of PZQ-Lip was more effective than PZQ for treatment of alveococcosis.

DIAGNOSIS AND TREATMENT OF CYSTICERCOSIS OF CEREBRAL VENTRICLES

1993, 11(4):  255-257. 

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31 patients with cysticercosis of cerebral ventricles verified by operation or pathological investigation were reported. All patients were between 7 and 64 years of age and 14 were females. All had a single cyst. Since 29 patients (94%) were without a history of intestinal taeniasis, it was proposed that most patients of cysticercosis of cerebral ventricles were caused by hetero-infection and the entrance of Cysticercus into brain ventricle was through choroid plexus along the cerebro-spinal fluid. This is probably the reason why it occurs mostly in the 4th ventricle. The clinical manifestation of cysticercosis of cerebral ventricles were paroxysmal headache and vomiting caused by increased intracranial pressure. Ventricu-lography and CT scanning have considerable diagnostic value. Removal of Cysticercus by surgical operation is successful (Figs. 1 - 8).

GENOMIC CLONING OF FASCIOLA HEPATICA SECRETORY ANTIGENS IN ESCHERICHIA COLI

1993, 11(4):  258-261. 

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This paper reported the immunoscreening of three recombinants expressing secretory antigens from the genomic DNA library of F. hepatica and the primary study of expression of the three recombinants. To construct the genomic DNA library, DNA from the adult F. hep-atica was cut with Sau3AI to an average length of about 2. 0 kb and inserted into the BamHI site of the expression vector pUC18, the size of the genomic DNA library of F. hepatica being 1. 5×105 recombinants.Three recombinants expressing antigenic determinants of F. hepatica pFHl6, pFH23, pFH48 were detected through primary screening and rescreenging with F. hepatica infected rabbit serum(1 : 50) preabsorbed to remove antibodies to E. colt and SPA-HRP(1 : 40). The test of the ability of expressing fusion proteins showed pFH16pFH48pFH23. These studies provide the possibility of further research on the expression of recombinant antigenic genes, the immunity of their expressed products and the protection of animals.

PURIFICATION AND ISOLATION OF DIFFERENT STAGES OF PLASMODIUM VIVAX, P. FALCIPARUM AND P. BERGHEI

1993, 11(4):  262-266. 

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The present paper reported the results of purification and isolation of different stages of P. vivax, P. falciparum and P. berghei by Percoll gradient centrifugation after removal of leukocytes (WBCs) by passing the blood suspension through CF -11 cellulose columns. 94. 7% and 75. 4% of WBCs from the patients* blood of vivax malaria and the mice blood infected with P. berghei respectively were removed with no alteration of the parasite density and the malaria stage ratio after the cellulose filtration.The densities of the uninfected and the infected erythrocytes with different stages of 3 species of malaria parasites were determined by isopycnic centrifugation on multi continuous gradients of Percoll. The mean densities (kg/L) of schizonts, trophozoites, ring form and uninfected cells were approximately 1. 059,1. 072,1. 096 and 1. 099 in P. vivax) 1. 070,1. 079,1.10 and 1. 10, gametocytes 1. 085 in P. falciparum; 1. 059,1. 062,1. 103 and 1. 104 in P. berghei, respectively. Based on these values, 3 different density gradients of Percoll were devised to separate erythrocytes infected with the more mature stages (trophozoites and schizonts) from ring forms and uninfected cells. By these method, it is possible to obtain high purified parasites of 93. 5% from blood with starting parasitaemias of 0. 8% in P. vivax, 100% from 2. 3% in P. falciparum and 93. 0% from 30. 6% in P. berghei, being raised bv 113. 8, 43. 4 and 2. 0 times respectively.

SCANNING ELECTRON MICROSCOPIC STUDY ON THE INFLUENCE OF MONOCLONAL ANTIBODY ON THE INVASION OF MACROPHAGE BY LEISHMANIA PARASITES

1993, 11(4):  267-269. 

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The interaction of promastigotes of Leishmania with mouse peritoneal macrophage in vitro was studied by scanning electron microscopy. The results showed that when promastigotes had been incubated with 1/1 000 McAb for 1 h, morphological changes could be seen in most promastigotes, some promastigotes were lysed and some promastigotes although adhered to the macrophage, but no penetration could be seen. When 1/1 000 McAb had been inactivated at 56℃ for 1 h and then incubated with promastigotes, scanning electron microscopic examination showed the promastigotes closely adhered to the surface of macrophage with their anterior end "buried" within macrophage.The results of the present study suggested that the mechanism of promastigote-macrophage adhesion was ligand-receptor binding interaction (Figs. 1-11).

STUDIES ON IMMUNOENZYMATIC STAINING TECHNIQUE USING ULTRASONICATED DRY EGG SECTION ON PVC MEMBRANE FOR DIAGNOSIS OF SCHISTOSOMIASIS JAPONICA

1993, 11(4):  270-272. 

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Immunoenzymatic staining technique (TEST) using PVC-membrane coated with Schisto-soma japonicurn ultrasonicated dry egg(UDE) and liver egg PVC-IEST was carried out to detect specific antibodies in sera from 159 cases with schistosomiasis japonica, the positive rates being 98. l%(156/159)and 97. 5% (155/159) .respectively. The sera from 63 healthy individuals showed false positive reaction in 2 cases and 1 case,respectively,suggesting that the sensitivity and specificity of the two tests were similar. However,the positive intensity was higher in UDE section than in liver egg section(F0. 05). As the UDE section could be stained more obvious and the PVC membranes are cheap and could be easily brought so that this method might be aplicable for practical purposes.

COMPARATIVE STUDIES ON THE DIAGNOSIS OF CLONORCHIASIS BY IEST AND IFAT

1993, 11(4):  273-275. 

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Both immunoenzymatic staining technique (TEST) and indirect fluorescent antibody test (IFAT) with frozen sections of adult Clonorchis sinensis as antigen were employed for detecting 51 cases with clonorchiasis and 50 healthy persons. The positive rate was 92% with I-EST and 88% with IF AT. The results showed no significant statistic difference (P0. 05). The false positive rates were 2% with the former and 4% with the latter. When sera from 22 cases with acute schistosomiasis, 20 cases with chronic schistosomiasis and 15 cases with paragonimiasis were examined by IEST and IF AT, cross-reactions were 14%, 5%and 0% with IEST, and 14%,10%and 0% with IFAT, respectively. The results showed that both IEST and IFAT are useful methods for serological diagnosis of clonorchiasis and the antigen on the gut was well demonstrated, while IEST might be more suitable in field surveys.

DETECTION OF CAg IN CSF OF 231 CEREBRAL CYSTICERCOSIS PATIENTS

1993, 11(4):  276-278. 

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The results of detection of circulating antigen(CAg) in cerebrospinal fluids(CSF) of 231 cerebral cysticercosis patients with McAb(4F8)-based ELISA were reported and compared with the case histories , clinical manifestations and CT scans. No relationship was found between CAg detected in CSF of these cases and history of taeniasis. However, the positive rate of CAg in cerebral cysticercosis patients with subcutaneous nodules was found significantly higher than those in cases with simple cerebral cysticercosis and in cases with subcutaneous nodules disappeared after anti-Cysticercus therapy before CSF collection. The results of CAg detection in CSF were related with the CT findings as well as the stage of the disease. It is thus indicated that MeAb(4F8)-based ELISA might be useful not only for diagnosis of neurocysticercosis, but also for the evaluation of efficacy of anti-Cysticercus therapy.

COMPARISON OF THE REPETITIVE DNA SEQUENCES BETWEEN PARAGONIMUS WESTERMANI AND PAGUMOGONIMUS SKRJABINI FROM SIX AREAS

1993, 11(4):  279-281. 

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The genomic DNA of the two lung flukes (P. westermani and P. skrjabini)from six areas was digested separately by three restriction endonucleases BamHI,Hae Ⅲ and Hind Ⅲ (followed by agarose gel electrophoresis. Repetitive DNA were visusalized as distinct bands in stained gels. Restriction fragment length differences were detected between the two species. The majority of bands of repetitive DNA were similar with minor differences within the same species collected from different areas,upon digestion with certain restriction endonucleases.The results demonstrated that the technique for demonstrating the band pattern of repetitive DNA with three restriction endonucleases can be used for the identification of P. westermani and P. skrjabini and the geographic relationship of the same species from different areas (Figs. 1 - 3).

STUDIES ON THE CHANGES IN INFECTION RATES OF WHIPWORM AND ASCARIS IN POPULATION AFTER CONTINUOUS CHEMOTHERAPY

1993, 11(4):  282-285. 

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The inhabitants with positive eggs of whipworm and ascaris in a village of Jiangdou County were examined and treated with different antihelmintic drugs twice a year. The un-cured population were treated by drug administration once again. On an average the efficiency of drug treatment (cure ratex treatment rate) was 56. 7% in whipworm and 64. 5% in ascaris. The program had been carried out for five years. The positive rate of eggs of whipworm and ascaris in the population decreased from 64. 7% and 52. 1% to 2. 7% and 4. 1%,respectively, the mean increase in infection rates among population at 5 or 7 months after treatment was 2.68(0.86-5.74)% and 6. 94(1. 72-17. 27)% .respectively.

DETECTION OF SERUM ANTIBODIES IN RATS INFECTED WITH ANGIOSTRONGYLUS CANTONENSIS BY INDIRECT FLUORESCENT ANTIBODY TEST

1993, 11(4):  286-287. 

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Frozen section specimens with a thickness of 4 μm were made from adult worms of An-giostrongylus cantonensis in order to detect the serum antibodies in rats infected with An-giostrongylus cantonensis by indirect fluorescent antibody test (IFAT). The results showed that the positive rates of the serum samples 2 and 4 weeks after infection were 92. 0%(46/ 50) and l00% (19/19), respectively. The results of serum samples in healthy rats and normal saline (as control) were negative. This technique proved to have a highly detectable rate and the positive reaction might occur in early infection. It might be used for the early supplementary diagnosis of angiostrongyliasis.

EFFECT OF SPLENECTOMY ON DISORDER OF CELLULAR IMMUNOREGULATION IN PATIENTS WITH ADVANCED SCHISTOSOMIASIS

1993, 11(4):  288-290. 

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The peripheral blood natural killer(NK)cell activity,the percentage of E-C,bR rosettes, phagocytosis of neutrophil and lymphocyte blastogenesis were studied to assess the cellular immunoregulation in patients with advanced schistosomiasis.The results showed that the NK cell activity and the percentage of E-C,bR rosettes were significantly lower in patients with splenomegaly than in patients who had undergone splenectomy and in normal subjects.The phagocytic index of neutrophil and lymphocyte blastogenesis was reduced in patients with advanced schistosomiasis,but no significant difference was found between the patients with and without splenectomy.These results suggested that splenectomy not only reduce the portal hypertension but also improve the regulatory functions of cellular immunity which may enhance the patient's resistance against pathogens.

EXPERIMENTAL AND CLINICAL STUDY ON CRYPTOSPORIDIASIS III. ULTRASTRUCTURAL OBSERVATION ON CRYPTOSPORIDIUM IN IMMUNOSUPPRESSED MICE

1993, 11(4):  291-293. 

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The ultrastructure of Cryptosporidium parvum, which parasitized on the surface of the intestine of the immunosuppressed mice,was studied by transmission electron microscopy. The parasite was enveloped by the parasitophorous vacuole developed from the host epithelial cell. An electron dense band was found at the attachment site between the Cryptoporidium and the host cell. At the attachment site,the cytoplasma membrane of Cryptosporidium appeared in the form of many membranous folds,and contacted closely with the plasma membrane of the epithelial cell. Rhoptry-like structure and small electron dense granules were found in the merozoite,but their effects on the development of Cryptosporidium ar?still unknown (Figs. 1 - 5).