Table of Content

30 April 1997, Volume 15 Issue 2

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论著

EVALUATION OF THE DIAGNOSTIC VALUE OF E.m.18k Da AND E.m.16 k Da ANTIGENSIN ECHINOCOCCUS MULTILOCULARIS BY WESTERN BLOTTING

MaLiang;AkiraIto;LiuYuehan;WangXiaogen;YaoYunqing;YuDenggao

1997, 15(2):  1-68. 

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AIM: To study the usefulness of 18 kDa/16 kDa antigens in larval Echinococcus multilocularis, designated E. m. 18 kDa and E. m. 16 kDa, for immunodiagnosis of human alveolar echinococcosis. METHODS: Western blot analysis based on the detection of antibody responses against E. m. 18 kDa and E. m. 16 kDa and commercially available E. m. 2-ELISA were evaluated comparatively using 24 sera from patients with alveolar echinococcosis, 55 sera from patients with cystic echinococcosis, 33 sera from patients with cysticercosis and 30 sera from healthy controls. RESULTS: All 24 (100% ) sera from alvrolar echinoco-ccosis patients recognized both E. m. 18 kDa and E. m. 16 kDa in Western blotting, whereas 23 of them were positive in E. m. 2-ELISA. The cross-reactivity with sera from Cystic echinococcosis was 16.4% for E. m. 18 kDa, 29.1% for E. m. 16 kDa and 23.6% for E. m. 2, while the cross-reactivity with sera from Cysticercosis was 3.0% for E. m. 18 kDa, 33.3% for both E. m. 16 kDa and E. m. 2. None of the sera from healthy controls showed reactivity with E. m. 18 kDa, E. m. 16 kDa or E. m 2. CONCLUSION: Our findings confirmed the high potential of E. m. 18 kDa and E. m. 16 kDa for immunodiagnosis of alveolar echinococcosis, and E. m. 18 kDa in particular.

MOLECULAR CLONING AND SEQUENCE ANALYSIS OF GENES ENCODING REGIONS16—17 IN MSP1 FROM TWO ISOLATES OF PLASMODIUM FALCIPARUM FROM CHINESE PATIENTS WITH CEREBRAL MALARIA

BianZhongqi;GuanWeibin;SongGuanhong;WangGongzhuo;LiFangyin

1997, 15(2):  2-75. 

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AIM: To further provide scientific evidence for designing safe and effective vaccines of human cerebral malaria. METHODS: Genomic DNA samples of two P. falciparum isolates prepared directly from 5 cases of cerebral malaria patients’blood in Mengla County, Yunnan Province (CMH/YN) and in Yingjiang County, Yunnan Province (CYJ/YN) were used for polymerase chain reaction (PCR) amplification and the two pairs of oligonucleotides for the highly conserved genes encoding the regions 12—17 in MAD20 merozoite surface protein 1(MSP1) of Papua New Guinea strain of P. falciparum were used as primers. The PCR products were digested with EcoRI and KpnI, respectively, and the generated fragment regions 16—17 were cloned into M13mp18 and M13mp19 vectors and their DNA were analyzed as the templates for DNA sequencing by the dideoxy chain-termination method. RESULTS: Compared with the three other published-MAD20, K1 and Wellcome sequences, DNA sequences of regions 16—17 in MSP1 from two isolates CMH/YN and CYJ/YN of P. falciparum from Chinese patients with cerebral malaria examined contained identical genes which were composed of 918 bp, encoding 306 amino acid, and containing 12 cysteines, which consisted of 2 epidermal growth factor (EGF) —like domains, respectively, and they were highly homologous up to 98.6% with that of MAD20 strain except that it contained an additional single base deletion at positions 4869 nucleotides and 5 point mutations. CONCLUSION: The results demonstrate for the first time that both DNA sequence determined from two isolates CMH/YN and CYJ/YN of P. falciparum from Chinese patients with cerebral malaria belong to the MAD20 allelic dimorphic family, and the deduced amino acids 1691—1 701 have been found to be TCTEEDSGSSR epitopes defined by monoclonal antibodies in P. falciparum.

CLINICAL OBSERVATION ON RED BLOOD CELL IMMUNE FUNCTION AND SERUM CYTOKINESIN PATIENTS WITH AMOEBIC LIVER ABSCESS

ZhuJiayong;WangWeiwei;HuangJing;TangBin;LaiXiuqiu;ChenChuan

1997, 15(2):  3-78. 

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AIM: To observe the changes in the red blood cell immune function (RBC-C3bRR, RBC-ICR) and serum soluble IL-2 receptor (IL-2R), tumor necrosis factor α(TNFα), IL-6, IL-8 in 40 patients with clinically confirmed amoebic liver abscess. METHODS: By sandwich ELISA and flow cytometry. RESULTS: Levels of IL-2R (672.48±291.52, U/ml), TNFα(0.874±0.452, ng/ml), IL-6 (0.198±0.021, ng/ml), IL-8 (0.648±0.075, ng/ml) in patients with amoebic liver abscess were significantly higher than those of healthy subject s, while RBC-C3bRR were significantly lower than that of healthy subjects . There was a negative correlation between the levels of IL-2R and RBC-C3bRR. The CD4/CD8 cell ratio (0.74±0.30) was markedly declined in peripheral blood of patients in comparison with healthy subjects (2.12) as detected by flow cytometry. CONCLUSION: The immune responses of both red blood cells and T cells were considerably suppressed in patients with amoebic liver abscess.

STUDIES ON ANTI-FEMALE FECUNDITY AND ANTI- EMBRYONATION IMMUNITY INDUCED IN MICE WITH 26/28 kDa ANTIGENS OF IMMATURE EGGS OF SCHISTOSOMA JAPONICUM

WangShiping;ZhouMibo;ShenGuoli;ChenYan;LiHua;ZhaoWeixian

1997, 15(2):  4-83. 

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AIM: To observe the effect of antiembryonation and anti-female fecundity immunity in mice induced by the 26/28 kDa antigens purified from soluble immature eggs antigen (SIEA) of Schistosoma japonicum. METHODS: BALB/c mice were immunized with SIEA 26/28 kDa or SIEA-I antigen to examine if these antigens can induce anti-disease immunity as determined by egg burden in liver, intestines and faeces on day 46 after challenge infection with cercariae of S. japo icum. RESULTS: A comparison of the egg burden in tissues and faeces on day 46 after challenge infection showed that the number of mature eggs in the livers, intestines and faeces was significantly reduced in mice immunized with SIEA 26/28kDa by 83.6%, 93.3% and 87.3%, respective-ly, whereas the number of dead eggs was much greater in the tissues of immunized mice than in that of control mice. In addition, the number of eggs in the uteri of female adults was obviously decreased in mice immunized with SIEA 26/28 kDa antigens or with the SIEA. However, these immune responses were not induced by the SEA or SIEA-I. CONCLUSION: The 26/28 kDa antigens were major components in SIEA responsible for both anti-fecundity and antiembryonation immunity.

EFFECT OF INHIBITION OF WORM OVULATION IN SCHISTOSOMA JAPONICUM-INFECTED RABBITS ON THE DETECTION OF CIRCULATING MEMBRANE ANTIGEN

QiuLishu;ZhangYonghong;LiHao;XueHaichou;XuYueqin

1997, 15(2):  6-85. 

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AIM: To understand the effect of inhibiting worm ovulation on the detection of circulating membrane antigen in Schistosoma japonicum infected rabbits. METHODS: Fifteen New Zealand rabbits were each infected with 250 S. japonicum cercariae per cuttem and a chemical, allyl thiourea, which inhibit egg formation was administrated per os at days 19 and 46 after infection. Sera were collected from rabbits before and weekly after infection and subjected to dot-ELISA for circulating antigen detection. RESULTS: Group 1 rabbits were administered allyl thiourea 295- 590 mg/d 19d post infection (p. i.) on 3 consecutive days per wk until 8 wk p. i., no circulating membrane antigen could be detected in the blood stream throuhgout the experiment. Group 2 rabbits received the same dose of allyl thiourea as that in group 1 starting from d46 p. i.. Dot-ELISA positive reaction was obtained 6 wk p. i. and lasted for 2 wk. In Group 3 untreated infected rabbits, dot-ELISA positive reaction was also detected at 6 wk p. i. and lasted for 2 wk. CONCLUSION: The positive circulating membrane antigen reaction is closely related to egg laying by the female worms.

DETECTION OF ANTI-TOXOPLASMA ANTIBODIES FROM TUMOR PATIENTS BY DIFA

NiuAn'ou;FuXiaoyu;H.Hlobil

1997, 15(2):  7-88. 

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AIM: To survey the prevalence of toxoplasmosis in tumor patients in Hubei and to examine the significance of direct immunofixation agglutination (DIFA), IgG-IFAT, and ISAGA for dignosing Toxoplasma infection. METHODS: Sera from 100 tumor cases were tested by DIFA for detecting anti-Toxoplasma IgG antibodies. The positive sera were re-examined by IgG-IFAT and then detected by Toxo-IgG/IgA/IgM-ISAGA. RESULTS: Using DIFA, the total anti-Toxoplasma antibody positive rate was 12% (12/100). Using IgG-IFAT, the corresponding rate was 100%, with one case suspected to be IgA positive. Four cases were IgM positive, two of them were with active infection and one was dubious. The rates of Toxoplasma infection in patients with lung cancer, non-Hodgkin lymphoma and breast cancer were 22.2% (6/27), 19.3% (5/26) and 7.1% (1/14), respectively. CONCLUSION: DIFA is more sensitive and specific for diagnosing Toxoplasma infection and that cancer patients are more susceptible to Toxoplasma infection.

DETECTION OF SPECIFIC ANTIBODY IN SALIVA OF PATIENTS WITH HYDATID DISEASE

JiangLi;TangXu;YuZhihong

1997, 15(2):  8-92. 

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AIM: To explore the diagnostic value of the detection of specific antibody in the saliva of patients with hydatid disease. METHODS: SPA sandwich-ELISA method was used to determine IgG in saliva of patients with hydatidosis and normal subjects. Antigen neutralization test was performed to identify the specificity of the salivary IgG. Serum and saliva antibodies were assayed by ELISA for a comparison. RESULTS: The salivary IgG levels in 32 normal subjects and 32 hydatidosis patients were 94.2±55.5 μg/ml and 105.6±65.4 μg/ml, respectively, being significantly different (P< 0.01). The antigen neutrali-zation test revealed that the inhibition rate of salivary IgG antibody responses in 40 hydatidosis patients were 30%-82.1%, and no inhibition effect was found in 32 normal subjects, demonstrating that the IgG antibody in saliva was specific antibody against echinococcus antigen. The IgG-ELISA tests of serum and saliva specimens from 89 hydatidosis patients and 129 normal subjects showed positive rate of 92.1% and 78.7% in serum and saliva, respectively. The coincidence rate for positives of saliva and serum were 80.5% while that for negatives were 98.3%. CONCLUSION: Salivary specific IgG antibody can be used as a parameter for the diagnosis of hydatid disease.

IN VITRO ASSAY INCORPORATED WITH METABOLISM FOR SCREENING ANTIMALARIALS

NiYichang;XuYueqin;ZhuZiyang

1997, 15(2):  10-95. 

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AIM: To develop an in vitro assay incorporated with metabolism for screening antimalarials. METHODS: Antimalarial activities of chloroquine, pyronaridine, cycloproguanil and proguanil were tested with an in vitro assay based on the routine in vitro culture of P. falciparum incorporated with a metabolic system comprising rat liver microsomes and NADP cofactors. RESULTS: Except proguanil, the three drugs perse all showed antimalarial activities, but proguanil must be metabolized into its active metabolite for the antimalarial effect. CONCLUSION: The in vitro assay incorporated with metabolism could overcome the shortcoming of miss-screening the potential antimalarials which must be metabolically biotransformed into the active metabolite.

INFLUENCE OF FENDONA ON THE CONTENTS OF CHEMICAL ELEMENTS AND AMINO ACIDS IN HEMOLYMPH OF AEDES ALBOPICTUS

ChenPeihui;LiFengwu;BianYinghua;ZengZhaohui

1997, 15(2):  12-99. 

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AIM: To explore the mechanism of the mosquitocidal effect of Fendona. METHODS: The fourth instar larvae of Aedes albopictus were treated with different concent rations of Fendona (alphamethrin). The hemolymph was collected from early emerged female mosquitoes and then subjected to the detection of chemical elements and amino acids. The contents of elements including Fe, Zn, Cu, Cr, K, Na, Ca, Mg, S, P in three different concentrations (0.70, 1.39 and 2.79 μmol/L) of Fendona-treated groups and control group (0.2% acetone-treated) were compared. RESULTS: Compared to the control group, there was a significant increase in the contents of 8 kinds of elements except for Zn, Cr and Ca in the 2.79 μmol/L group; while in 1.39 μmol/L and 0.70 μmol/L groups, the contents of Fe, Mn, K and Mg were higher than those in the control group. Comparing the contents of 15 kinds of amino acids in hemolymph among 1.39 μmol/L, 0.70 μmol/L Fendona and the control group, the levels of 12 kinds of amino acids in Fendona groups were decreased. CONCLUSION: Fendona does influence the contents of many kinds of elements and amino acids in the hemolymph of adult Aedes albopictus.

EXPLORATION ON ANALYTICAL METHOD FOR SAMPLING ERROR OF INFECTION RATE OF RESIDENTS IN SAMPLING SURVEY OF SCHISTOSOMIASIS

WangYanan;HuaZhenghui;WuWeiping;GuoJiagang;ZhengJiang

1997, 15(2):  14-103. 

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AIM: To establish a set of statistic formulas on sampling error of infection rate of residents in sampling survey of schistosomiasis. METHODS: According to the requirement of multiple-stratifying and clustering random sampling, the sampling error of schistosome infection rate of residents was analysed through introducing the theory of error transferring and combining the calcu-lation formula of standard error in clustering random sampling and stratified random sampling. RESULTS: Four standard error formula of prevalence were introduced in different provinces and different types in endemic areas. CONCLUSION: The calculation formulas of multiple-stratifying and clustering random sampling are theoretically set up.

CLONING OF A PARTIAL PARAMYOSIN GENE FROM SCHISTOSOMA JAPONICUM

QiuChiping;TongXiaomei;LiuShuxian

1997, 15(2):  16-106. 

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AIM: To clone apartial paramyosin gene from Schistosoma japonicum of Chinese mainland origin. METHODS: The gene was amplified by PCR according to the published partial paramyosin sequence of S. japonicum of Philippine origin. RESULTS: A gene of about 720bp was cloned from S. japonicum of Chinese mainland origin. CONCLUSION: The analysis of the sequence of this gene revealed 99.03% identity with that of S. japonicum of Philippine origin.

DETECTION OF CIRCULATING ANTIGENSIN ACUTE SCHISTOSOMIASIS PATIENTS AFTER TREATMENT

WuGuilan;WuXiaolan

1997, 15(2):  18-110. 

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AIM: To assess the therapeutic effect of praziquantel in treating acute schistosomiasis and provide evidence for further treatment. METHODS: Circulating antigens (CAg) detection method and etiological detection method. RESULTS: One hundred and ninety acute schistosomiasis patients were treated with praziquntel. After 0.5, 1.5, 3 and 4 years, their negative rate was 83.7%, 87.4%, 99.5% and 98.0%, respectively by fecal hatching method. Five years after treatment, the therapeutic efficacy was evaluated by CAg detection method. Of 164 schistosomiasis patients, 22 were positive (13.4%). As compared with etiological detection method, the coincident positive rate was 68.2%. CONCLUSION: The CAg detection method can reduce the diagnosis-missing rate and save time and labor. Especially, during the late period of the schistosomiasis prevention and treatment, this method can be used to evaluate the therapeutic effect of praziquantel and provide evidence for further treatment of patients.