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Table of Content

    31 May 1993, Volume 11 Issue 2
    AMPLIFICATION, CLONING AND SEQUENCE ANALYSIS OF A SSUrRNA GENE FRAGMENT OF PLASMODIUM VIVAX ISOLATES FROM YUNNAN PROVINCE
    1993, 11(2):  81-85. 
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    According to known SSUrUNA sequences ot Plasmodium vivax , correlated protozoa and human being, sequences of oligonucleotide primers were defined with computer programming. Specific SSUrDNA fragment of P. vivax , about 640 bp in length, was directly amplified by two temperature point polymerase chain reaction from extracted genomic DNA of two blood samples of vivax malaria patients from Yunnan Province. Using dideoxynu-cleotide terminator method, the sequences of amplified DNA fragments were determined separately and showed no difference between the two samples. However, comparison of the sequence reported by Waters AP and McCutchan TF(1989)and that of amplified fragment of Yunnan P. vivax isolates revealed the existence of nucleotide substitution and deletion which occured respectively in the sites 269 and 630,and resulted in the change of restriction map.
    FURTHER REPORT ON APPLICATION OF DNA PROBE IN DIAGNOSIS OF VIVAX MALARIA INFECTION
    1993, 11(2):  86-88. 
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    Southern blotting with a labeled and linearized pUC 19 DNA containing a specific fragment of 0. 24 kb DNA of Plasmodium vivax asexual blood stages (kindly offered by Dr. C. Kidson )was used for further identification of blood samples showing positive reaction by dot-blot hybridization. The results showed that those with positive reaction from patients with P. vivax, with P. falciparum or with fever but with negative microscopic findings were also positive by Southern blotting. It was confirmed that some of those positive with P. falciparum were likely to infect P. vivax at the same time. So did a part of those with fever but negative in the blood films (Figs. 1,2).
    ROLE OF MACROPHAGE IN PRESENTING ANTIGENS OF SCHISTOSOMA JAPONICUM
    1993, 11(2):  89-92. 
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    Indirect ELISA was employed to monitor the serum anti-UEA(urea soluble egg antigen of Schistosoma japonicum )antibody level of mice immunized by a. UEA pulsed macrophage (M + ); b. Cultural supernatant of M+; c. paraformaldehyde fixed M(P -M)pulsed with UEA; d. Ammonium chloride treated M0 (NH4Cl-M0) pulsed with UEA, e. P -M0 pulsed with trypsin digested UEA (T-UEA ); f. NH4C1-M pulsed with T-UEA. The normal M its supernatant and the culture media RPMI 1640 acted as the negative control.The results showed : 1. Serum anti-UEA antibody levels of mice immunized by a and b raised markedly, indicating that the immunogenicity of UEA might be kept up after M processing and the antigenic message could be transferred either by the M+ or by its supernatant; 2. Mice immunized by c and d gave similar results, but the anti-UEA antibody level of the former was higher than that of the latter, suggesting that polyformaldehyde could not alter the UEA binding site on the surface of M; 3. In the case of mice immunized by e and f , the antibody levels were much lower than that of mice immunized by c and d, suggesting that UEA binding sites on M surface as well as UEA immunogenicity could be changed by trypsin.
    STUDIES ON THE STRAIN DIFFERENCES OF SCHISTOSOMA JAPONJCUM IN THE MAINLAND OF CHINA XIII. CONCLUSION
    1993, 11(2):  93-97. 
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    Up to the present, Schistosoma japonicum endemic in the mainland of China has used to be considered being of only one strain, Chinese mainland strain. It is well known that the endemic areas of schistosomiasis japonica in the Chinese mainland are mainly confined to regions south of Yangtze River, but they are discontinuous, showing conspicuous geographic and topographic isolation. So far, little is known about the characteristics of schistosomes isolated from various topographic areas. We have tackled the problem by comparing the biological, pathological and immunological characteristics of 5. japonicum isolated from the following five localities; 1. Guichi in Anhui, at the lower reaches of the Yangtze River in the east; 2. Jianli in Hubei, at the middle reaches of the Yangtze River in the middle; 3. Guiping in Guangxi, a karst land in the south; 4. Tianquan in Sichuan, a mountainous region in the west; 5. Eryuan in Yunnan, a high mountainous region in the southwest. The following items were observed and compared including morphometric data, susceptibility of six mammalian hosts, prepatent period, compatibility between larvae and snail hosts, size of hepatic granuloma produced by eggs,immunoreactions in experimental animals, sensitivity to praziquantel, SDS-PAGE protein pattern and its antigenicity analysis, DNA hybridization and genetic variation and differentiation by analysis with multilocus enzyme electrophoresis. By means of these multidisciplinary methods, from morphological to molecular level, the following conclusions may be drawn from our results. The evidences indicate firstly that the Chinese mainland strain of S. japonicum in the mainland of China comprises a strain complex with several components of geographically distributed strains. At least four distinct strains, i. e. Yunnan, Guangxi, Sichuan and Anhui-Hubei, are present. The Chinese An-hui-Hubei strain occupies territories of the Yangtze valley within Hubei and Anhui provinces.
    EFFECT OF SOME FACTORS ON THE DEVELOPMENT OF EXOERYTHROCYTIC STAGE OF PLASMODIUM YOELII
    1993, 11(2):  98-101. 
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    Using orthogonal design, the effect of environmental temperature, photoperiod , splenec-tomy and oestrogen levels upon the development of exoerythrocytic forms (EEF) of P. yoelii was observed. The results indicated that a significant number of viable sporozoites were cleared in the spleen, since the EEF density was much higher in the livers of splenectomized rats (1. 73/mm3)than in sham-operated counterparts(0. 55/mm3). The effect of high level oestrogen on EEF density was also evident since there was a significant difference between experimental(0. 86) and control rats(1. 42). Low environmental temperature caused the development of EEF stunted and asynchronous,but no significant effect on the density of EEF in this group was found. The density and average diameter (AD) of EEF between rats illuminated 8 and 16 hours per day were identical. The interaction between splenectomy and elevated oestrogen level offset each other, whereas the interactions between other two factors showed no difference by analysis of variance.
    ULTRASTRUCTURAL LOCALIZATION OF 145/102 kDa ANTIGENS IN ERYTHROCYTIC STAGES OF PLASMODIVM FALCIPARUM
    1993, 11(2):  102-104. 
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    The ultrastructural localization of the 145/102 kDa antigens recognized by the possible protective monoclonal antibody (MeAb) M26-32 in erythrocytic stages of Plasmodium falci-parum, FCC1/HN, in vitro, was investigated by immuno-electron microscopy with LR White resin embedding and colloidal gold probe cytochemistry techniques. The results showed that the gold particles were mainly localized within the cytoplasm of ring forms, trophozoites,schizonts and merozoites of the Plasmodium. Some gold particles were found to locate on the pellicular complex of the plasmodium surface or in the cytoplasm of the infected erythrocytes. The results indicated that 145/102 kDa antigens were the common cytoplasmic antigens of asexual blood stages of Plasmodium falciparum, FCC1/HN, while a portion of the antigens could be transported to the cytoplasm of the infected erythrocytes via the pellicular complex of the plasmodium surface (Figs. 1-4).
    IMMUNOELECTRON-MICROSCOPIC LOCALIZATION OF A 54-kDa PROTEIN OVEREXPRESSED BY CHLOROQUINE-RESISTANT PLASMODIUM BERGHEI ANKA STRAIN
    1993, 11(2):  105-107. 
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    A 54-kDa protein overexpressed by chloroquine-resistant Plasmodium berghei ANKA strain was first reported by us. In this paper, the localization of this protein by immunoelec-tron microscopy is presented. The results showed that the protein was mainly scattered inside the cytoplasm of the early date trophozoites and schizonts of erythrocytic stage of P. berghei ANKA strain , and some of it was also found in cytoplasm of erythrocytes infected with parasites. The protein content was much higher in chloroquine-resistant P. berghei ANKA strain than in chloroquine-sensitive P. berghei ANKA strain, suggesting the importance of this protein in understanding mechanism of chloroquine resistance in malaria parasites .
    ANALYSIS OF RELATIVE DEGREE OF THE GREY SYSTEM ON SOCIOECONOMIC FACTORS CONTRIBUTING TO MALARIA TRANSMISSION
    1993, 11(2):  108-111. 
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    This paper reports relative degree of the grey system analysis on socioeconomic factors contributing to malaria transmission in Hainan province. The results indicated that the prior factors related to malaria transmission are economic conditions, primary health care, human behavior and culture. It is suggested that rebuilding primary health care and strengthening health propaganda would be advantageous to malaria control in these areas.
    STUDIES ON THE SUSCEPTIBILITY OF ANOPHELES ANTHROPOPHAGVS TO EXPERIMENTAL INFECTION WITH WUCHERERIA BANCROFTl
    1993, 11(2):  112-115. 
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    This paper reports the susceptibility of Anopheles anthropophagus (Xu et Feng, 1975) to experimental infection with Wuchereria bancrofti(Cobbold, 1877), comparing with that of An. sinensis Wiedemann, 1828, Culex pipiens quinquefasciatus Say, 1823, and Aedes togoi (Theobald, 1907). Of 188 An. anthropophagus ,202 An. sinensis, 280 Cx. pipiens quinquefasciatus and 129 Ae. togoi infected by the blood of a bancroftian microfilaremia with mff density of 190 mff/20cmm,the infective rates were 35. 64%,9. 41%,30. 00% and 65. 89%,respectively ; while of 188 An. lesteri anthropophagus , 134 An. sinensis, 289 Cx, pipiens quin-quefasciatus and 176 Ae. togoi infected by the blood of another bancroftian microfilaremia with mff density of 83 mff/20cmm, the infective rates were 18. 09%, 3. 73%. 13. 84% and 39. 77%,respectively. It is concluded that the susceptibility of An. anthropophagus to experimental infection with W. bancrofti is significantly higher than that of An. sinensis, significantly lower than that of Ae. togoi and at the same level with that of Cx. pipiens quinque-fasciatus(Figs. 1-7).
    THE ESTABLISHMENT OF GENOMIC DNA LIBRARIES FOR THE HUMAN MALARIA PARASITE PLASMOD1VM FALC1PARVM
    1993, 11(2):  116-119. 
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    The DNA of Plasmodium falciparum has been purified and fragmented with restriction endonuclease BamHI. The fragments have been incorporated in vitro into derivatives of bac-teriophage lambda EMBL4 digested with BamHI and Sal I. The recombinant mixture has been ligated and packaged in vitro. The recombinant phages have been identified in E. coli L95host cell and the libraries have been established in which most of the parasite DNA is represented. The ligation proportion of vector to insert is 3 : 1. The recombinant phages of 4×105 have been obtained. By plaque hybridization, we have been able to recover from these libraries specific clones containing repetitive DNA sequences.
    STUDIES ON DISTRIBUTION AND BEHAVIOR OF ANOPHELES MINIMUS AND ITS ROLE OF MALARIA TRANSMISSION IN HAINAN PROVINCE AT PRESENT
    1993, 11(2):  120-123. 
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    Anopheles minimus was once a main malaria vector in Hainan Island and had been e-liminated basically through the campaign of indoor residual spraying launched in 1959. It again became an incriminated vector of some focal malaria outbreaks in recent years. The present study was conducted in a selected county-Danxian and a typical hilly area-Feibar in the west part of Danxian county in 1989-1990.An. minimus was found in 50% and 62. 5 % of the surveyed sites at mountainous and hilly area of Danxian county,but not found in coastal region. An. minimus was found in all 18 sites surveyed in Feibar district constituting 52% of anopheline composition. Man-biting rate made by human-baited collection was 3. 2 before midniaght and 38. 2 when collected through whole night in some sites. However, the behaviour characteristics of An. minimus has changed. It has become exophilic,exophagic, and has an equal preference for man and cattle. The vectorial capacity of An. minimus estimated by quantitative data was in accord with malaria infection rate in Feibar district ,and the malaria infection rate among the inhabitants in three types of residential quarter with different socioeconomic conditions. Malaria infection rates of residential quarter of land-reclamation outcomers, villagers and state farm residents were 10%,2. 9% and 0. 5% respectively during 40 days from July to August,1990.Owing to the fact that An. minimus has become a secondary vector only next to An. dirus, with a wide range of distribution and a considerable different characteristics in behaviour compared to that before spraying campaign , it is suggested that a malaria control programme must be seriously planned to adjust the new problem of malaria epidemiology in Hainan Province.
    FLUIDITY OF RED BLOOD CELL MEMBRANE FROM MOUSE INFECTED WITH MALARIA PARASITE
    1993, 11(2):  124-126. 
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    The fluidity of membrane lipid regions of Plasmodium berghei- or Plasmodium yoelii- infected red blood cells has been determined by the fluorescence polarization technique using 1, 6-diphenyl-1, 3, 5-hexatriene (DPH) as a probe. The results showed that the fluidity of Plasmodium(berghei or yoelii)-infected red blood cell membranes was increased significantly as compared with that of normal controls judging from the degree of polarization and the microviscosity. Its mechanism was discussed briefly.
    ANALYSIS ON THE PROTEIN/POL YPEPTIDE IN CULEX PIPIENS PALIENS (DIPTERA:CULICIDAE) DURING HIBERNATION BY TWO-DIMENSIONAL GEL ELECTROPHORESIS
    1993, 11(2):  127-131. 
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    Analysis on the protein / polypeptide in Culex pipiens pallens ( Diptera : Culicidae) caught from the resting or overwintering places during the 3rd ten-day of September,the 2nd ten-day of December and the 1st ten-day of March in next year in Zhengzhou(34°43'N ,113° 39' E) during hibernation by two-dimensional gel electrophoresis indicated that there were 153, 118 and 169 protein/polypeptide spots in unoverwintering, overwintering and recovering mosquitoes respectively. The molecular weight (MW) of most of the spots was less than 43. 0 kDa, with isoelectric points (pI)between 5. 65-7. 34. The comparisons between each two types of mosquitoes during hibernation revealed that the variant rate of protein / polypeptide spots between overwintering and recovering mosquitoes (31. 01 % ) was significantly higher than that between unoverwintering and recovering females (21.12%) (P0. 05). The variant spots were 13 in unoverwintering, 7 in overwintering and 27 in recovering mosquitoes respectively. The comparisons among three types of mosquitoes during hibernation showed that the variant rate was significantly higher in recovering females(15. 98%) than in unoverwintering (8. 50%)(P0. 05)and overwintering females(5. 93%)(P0. 01). The total variant rate of protein/polypeptide spots of all these three types of mosquitoes was 10. 68%.Because the blood meal of all the mosquitoes used for homogenization was at Sella 1 (empty), and the ovary development was under Christophers Ⅱ(resting stage), the experimental results were not affected by blood meal and the later ovary development (Christophers Ⅱ-Ⅴ).
    SCANNING ELECTRON MICROSCOPIC OBSERVATION ON THE ENCYSTED AND EXCYSTED METACERCARIAE OF PARAGONIMUS HETEROTREMUS
    1993, 11(2):  132-134. 
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    The surface ultrastructure of metacercariae of Paragonimus heterotremus was studied by scanning electron microscope. Encysted metacercaria was egg-shapped with a button-like structure on its end. The oral sucker of the newly excysted metacercaria was larger than the ventral sucker. Single-pointed spines were covered densely on all of the tegumental surface. There were two and three rings sensory papillae on the oral and ventral suckers, respec- lively. Six papillae of the most inner ring were distributed on the ventral sucker inner-pore symmetrically. A few sensory papillae were dispersed on each side of the anterior part of the worms. These papillae of some excysted metacercariae were arranged irregularly in two rows with 5 to 6 papillae in each row (Figs. 1-8).
    SCHISTOSOMA JAPONICUM ADULT WORM RNA ISOLATION WITH CHOMCZYNSKI'S METHOD
    1993, 11(2):  135-137. 
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    A rapid method of total RNA of Schistosoma japonicum adult worm isolation by single step extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described. The method provides a pure preparation of intact RNA and can be completed within several hours. It is particularly useful for isolation of RNA from minute quantity of parasite material. Analysis on agarose gels electrophoresis reveals that the RNA preparations show a distinct band comigrating with 18S rat brain RNA and without large rRNA species. The large rRNA is in vivo nicked. The absence of a band at approximately 26S presumably reflects post transcriptional processing of the large rRNA.