Table of Content

30 June 2001, Volume 19 Issue 3

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论著

Efficacy of Ablendazole Emulsion in Treatment of 212 Patients with Cystic Echinococcosis

CHAIJun-jie;Menghebat;JIAOWei;SUNDe-yu;LIANGBin;SHIJin-cao;FUCheng;LIXiong;MAOYi-ding;WANGXiu-ling;Dolikun;Guliber;WANGYan-chun;GAOFang-hua;XIAOShu-hua

2001, 19(3):  1-134. 

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　Objective To verify the efficacy of abendazole emulsion, a new formulation of abendazole, in treatment of human cystic echinococcosis. Methods 212 patients with liver cystic echinococcosis were treated orally with albendazole emulsion at a daily dose of 10 mg/kg or 12.5 mg/kg for 3 to 12 months or over one year. The therapeutic efficacy was mainly \{evaluated\} by image feature examined with B utrasound examination, a short-term efficacy at the completion of treatment and a long-term efficacy followed-up for 1-4 years. Results In 212 patients treated with albendazole emulsion at a daily dose of 10 mg/kg and 12.5 mg/kg, the average cure rate, improved rate and the rate of no avail were 74.5%, 99.1% and 0.9% respectively after termination of the treatment, and the average long-term rates were 83.1%, 89.3% and 0.6% respectively. Recurrence occurred in 18 patients(10.2%). The results indicated that the best efficacy was seen in patients treated with albendazole 12.5 mg/kg daily for 9 months. Better response was also found when the recurrent patients were re-treated with albendazole. Conclusion The efficacy of albendazole emulsion on patients with liver cystic echinococcosis is much better than that of albendazole tablet or capsule and mebendazole. Meanwhile, the efficacy of albendazole emulsion is stable with less adverse effects. The results suggest that albendazole emulsion could be the drug of choice for treatment of cystic hydatid disease.


Gene Cloning, Construction and Expression of Single-Chain Fv (scFv) Against the Membrane Protein of Schisotosoma japonicum

YUXiao-cong;JIANGXin;HUANGHao-min;ZHANGZhong;LINQing;GUANXiao-hong;HUANGHua-liang*

2001, 19(3):  2-140. 

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　Objective To construct single chain antibody specific to membrane protein of Schistosoma japonicum by gonetic engineering technique. Methods The V\-H (heavy-chain variable region) and V\-L (light-chain variable region) genes were amplified by PCR from the genomic DNA of NP11-4 cell line, and sequenced by Sanger's method. The ScFv was constructed in pTHA90 vector using V\-H and V\-L genes, then expressed by IPTG. Results The V\-H and V\-L genes were obtained through PCR. The DNA sequences showed that V\-H and V\-L were new variable region genes of antibody. They were registered by GenBank. A ScFv gene with (Gly4Ser) 3 intralinker in the pTHA90 vector was successfully constructed. The ScFv was expressed as thioredoxin-fused proteins about 36.2 kDa. Conclusion A specific ScFv against the membrane protein of Schistosoma japonicum was constructed and expressed.


Efficacy of Albendazole Immunoliposome Against Echinococcosis Granulosus in Mice

NiuRong-li;XueHong-xie;MoHong-mei

2001, 19(3):  3-144. 

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　Objective To evaluate the effect of albendazole immunoliposome (IL-Alb) against Echinococcus granulosus. Methods Mice infected with protoscolices of E.granulosus were divided into five groups. Four groups were treated with albendazole (Alb), albendazole liposome (L-Alb), albendazole sulfoxide liposome (L-Albso), and IL-Alb respectively at a dosage of 100 mg (Alb)/(kg·d)×5 d for 3 courses. The fifth group was established as control. The major criteria for evaluating the effects included a reduction rate of E.granulosus tissue wet weight, histopathological examination of the cysts by both light microscopy and electron-microscopy, and the content of albendazole-sulfoxide in cysts detected by HPLC. Results The efficacy of albendazole immunoliposome was significantly higher than that of albendazole liposome, and much higher than that of albendazole. The reduction rates of cyst tissue weight of IL-Alb group, L-Alb group and Alb group were 91.5%, 80.3%, 61.2% respectively as compared to control group; the concentration of Albso in cyst tissue of the above groups were 5.15 μg/g, 2.18 μg/g, 0.76 μg/g respectively (P<0.01). The histopathological damages of cysts were also found most severely in the group of IL-Alb. \{Conclusion\} Immunoliposome as a targeting carrier may significantly strengthen the therapeutic effect of albendazole on echinococcosis granulosus.


Second Sampled Survey on the Distribution of Human Parasites in Zhejiang Province

TUXing-guo;YAOLi-nong;HUANGXue-min;CHENHua-liang;YUKe-gen;JIANGMiao-gen;ZHUWen-ming;CHENYu-man;LIUBei-dou;LEIChang-qiu

2001, 19(3):  4-148. 

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　Objective Twenty-six species of human parasites were recorded in Zhejiang Province in 1987-1989 as a part of the national investigation on the distribution of human parasites, and the total prevalence was 80.2%. In order to find out possible changes on the composition of parasite species and decrease of prevalence after control intervention particularly mass chemotherapy in the past years and provide evidence for an improved control strategy, the second sampled survey was carried out from 1998 to 1999. Methods Ten counties were identified randomly out of 28 counties where the last survey was conducted following the same sampling method. Same technics were used for case detection and data processing. Results The total \{infection\} rate in a sample of 15 698 was 22.84% in 30 investigation spots in 10 counties, and 17 species of parasites were \{revealed\}. The overall prevalence was reduced by 71.51% in comparison to that of 1989, and the number of parasite species was 17, 9 less than that of the last investigation. Conclusions The prevalence of human parasites has greatly declined in this province due to the socioeconomic development and adoption of comprehensive control measures focusing on mass chemotherapy in the past decade.


Partial Sequence of Sporogony Stage-specific 18S Ribosomal DNA of Plasmodium yoelii and Its Application for Detection of Parasites

XUXiao-chun;QUFeng-yi;SONGGuan-hong

2001, 19(3):  5-152. 

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　Objective To determine sequence of sporogony stage-specific (S type) 18S ribosomal RNA gene of Plasmodium yoelii (P.y) By265 strain, and by using it to detect the malaria parasites within vector mosquito. Methods A pair of conserved DNA primers, universe primer (Pu) and reverse transcription one (Pr), was designed and synthesized according to sequence of the 18S rRNA gene of Plasmodium berghei (P.b). The segment of the S type 18S rDNA of P.y was amplified by reverse transcript-polymerase chain reaction (RT-PCR) from dissected midguts of Anopheles stephensi infected with P.y on the 7th day after infective blood-meal, and its sequence was then determined. One P.y sporogony stage-specific primer (Pys) was selected according to the sequence. Using this primer and Pr, the parasites within mosquitoes were semi-quantitatively detected through RT-PCR between 1-7 d post-infection. Results The length of the amplified segment was 920 bp. Alignment in match region of the 18S rDNA among S type of P.y (PyS), S type of P.b (PbS) and asexual blood stage-specific one of P.y (PyA) revealed that the similarity between the former and the latter two reached 95.3% and 94.0% respectively. The density of amplified band was significantly concordance with the intensity of oocyst in the midgut. Sensitivity of RT-PCR method was higher than that of the traditional dissection and oocyst observation also. The assay could detect the 18S rRNA molecule of the parasites on the third day post-infection while their oocysts were difficult to be recognized under an optical microscope at that time. Conclusion This S type 18S rDNA sequence in P.y species was first reported (AF266261). As a molecular marker, it could be applied to monitoring the parasite development in its vector at an earlier stage semi-quantitatively with an adequate sensitivity and specificity.


Screening and Cloning of Genes Encoding Schistosoma japonicum Antigens Related to the Serum Antibodies in Mirotus Fortis

YANYu-tao;LIUShu-xian;SONGGuang-cheng;XUYu-xin;HEYong-kang

2001, 19(3):  6-156. 

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　Objective To understand and identify the molecules related to the natural resistance to Schistosoma japonicum infection in Mirotus fortis. Methods Sera from Mirotus fortis without schistosome infection were collected. The S.japonicum adult worm cDNA library was immunologically screened with the sera. The positive recombinants were identified, cloned, sequenced and analysed with software and internet. Results Seven genes encoding antigens relevant to sera antibodies in Mirotus fortis were cloned and sequenced. These antigens included glyceraldehyde 3-phosphate dehydrogenase (GAPDH), serine protease inhibitors(SERPIN), 70 kDa heat shock protein(HSP70), 22.6 kDa membrane-associated antigen, paramyosin (Sj97), cytochrome C and cathepsin B. Conclusion Many protein molecules might have been involved in natural resistance to \{S.japonicum\} infection in Mirotus fortis. The above 7 kinds of molecules may be identified as new candidates of vaccine against \{S.japonicum\} infection.


Cloning and Identification of an Unknown Gene Encoding 10.6 kDa Protein of Schistosoma japonicum

SHENJi-jia;JIANGZuo-jun;YUXin-bing;WANGXue-long;WANGWei

2001, 19(3):  7-159. 

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　Objective To screen a new schistosome vaccine candidate. \ Methods\ Schistosoma japonicum adult cDNA library was screened using sera from immune rabbits vaccinated with irradiated cercariae and monoclonal antibodies against membrane antigen of S.japonicum schistosomula. Three different fragments of S.japonicum cDNA genes were cloned into pGEM-T vector. The sequences of the inserts were determined using an automatic DNA sequencer and were analysed using Blast program. One of the unknown genes (B8) was selected and its ORF sequence (291 bp) was subcloned into eukaryotic expression vector. The recombinant plasmids were identified by restrictive enzymes and PCR amplification. The positive recombinant plasmids (pBK/SjB8) were transformed into host bacteria XL1-blue, and were then induced by IPTG for expression. SDS-PAGE and Western blotting analysis of total cellular protein from the bacteria were performed to detect the gene products. Results The results demonstrated that ORF of SjB8 gene was subcloned into the plasmid pBK-CMV and could express as fusion protein in XL1-blue. The results of SDS-PAGE and Western-blot also showed that the molecular weight of the fusion protein with 3 kDa β-galactosidase was approximately 13.6 kDa and the actual molecular weights of the SjB8 was 10.6 kDa. The expressed fusion product of pBK/Sj-B8 could be recognized by immune serum and McAb. Conclusion A new gene of S.japonicum vaccine candidate (SjB8) was cloned into eukaryotic expression vector pBK-CMV and could express 10.6 kDa schistosome protein. The results provide foundation for further study of the protein for its posibility as candidate vaccine.


Effect of Dibutyl Phthalate on Demodicidosis

YUANFang-shu;GUOShu-ling;QIUZhen-xu;DENGShu-hai;HUANGGui-hua

2001, 19(3):  8-162. 

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　Objective To evaluate the curative effect and safety of dibutyl phthalate on demodicidosis. Methods A single blinded and controlled study of human demodicidosis treated with dibutyl phthalate was conducted. One hundred and forty three patients with demodicidosis, including 81 acne and 62 rosacea, randomly divided into trial and control groups. The trial group was treated with dibutyl phthalate and control group with "new fumanling" cream twice a day in the early morning and evening respectively for six weeks consecutively. Results The rates of excellent, good, and fair efficacy and total effective rate in the trial group with acne were 53.7%, 41.5%, 4.9% and 100% respectively, with a significant difference to the control group (P<0.05). The rates in the trial group with rosacea were 40.6%, 40.6%, 18.8% and 100% respectively, with no statistical difference to the control group (P>0.05). No complaint of side effects in the trial group was recorded. Conclusion Dibutyl phthalate is highly effective to demodicidosis without prominent adverse reactions.


Preliminary Study on Isolation, Purification and Hydrolytic Activity of Cysteine Proteinases in Entamoeba histolytica

YANZhe;CHENSheng-liang;MAOSun-zhong

2001, 19(3):  9-165. 

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　Objective To explore the invading mechanism of amebae in lamina porpria and observe the interaction between the cysteine proteinase (CP) of Entamoeba histolytica and laminin. Methods CP was identified by laminin-sepharose affinity chromatography, followed by isolation, purification and inhibitor experiment. The hydrolytic activity was measured by gelatin electrophoresis. Results Purified CP of E.histolytica showed a strong affinity with laminin. The molecular weight of CP is 27 kDa. It can be inhibited by EC-64 and exhibited a protein hydrolytic activity. Conclusion The specific affinity and hydrolytic activity of CP might play an important role in its invasion to the basement membrane of intestinal mucosa.


实验报道

Studies on Immunological Reaction of the Antiserum of Recombinant Secreted Protein from Ancylostoma Caninum

WENLi-yong;HotezPJ

2001, 19(3):  10-168. 

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　Objective To analyze the immunological reaction of the antiserum of recombinant secreted protein from Ancylostoma caninum with antigens of various species hookworms at different developmental stages. Methods SDS-PAGE and ELIB technique were employed in the study. Results and Conclusion The protein component of Ac-rAsp-1 was 45 kDa, its immune serum can recognize the antigens of Ac-L\-3 and Ac-rAsp-1 protein, but not react to the antigens of Ad-A?Ad-L\-3?Na-A?Ac-A?Nb-A and Ac-rAsp-2 protein. The protein component of Ac-rAsp-2 was 24 kDa, its immune serum can recognize the antigens of Ad-A?Ad-L\-3?Na-A?Ac-A?Ac-L\-3 and Ac-rAsp-2 protein, but not react to the antigens of Nb-A and Ac-rAsp-1 protein.


Morphological and Ultrastructural Observation of Blastocystis hominis

HENi;ZHANGYue-qing;HONGMing-li;CONGMin

2001, 19(3):  11-172. 

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　Objective To observe the morphology and ultrastructure of Blastocystis hominis. Methods Morphological observation was made with 4-5 days cultured B.hominis by light microscopy, and similar material fixed with 4% glutaraldehyde was used for transmission electron microscopy. Results Several forms of B.hominis were observed including vacuolar, \{granular\}, amebic, multifission and cystic forms. The multiplication patterns of B.hominis included both binary fission and sporogony. Under transmission electron microscope, the nuclei, mitochondria, rough endoplasmic reticula and lysomes were observed in addition to lipid droplets in its cytoplasm, and glycogen in the central vacuole. Conclusion The central vacuole of vacuolar form may be related to the storage of the excreta. The amebic form of B.hominis might be pathogenic.


Detection of DNA of Toxoplasma gondii in Rat by Using Polymerase Chain Reaction

GENGZhi-hui;HECheng-yan;ZHANGYong-sheng;LIShu-hong;DUJun;LIULi;FANGYan-qiu;ZHUGang;LIJia-he

2001, 19(3):  12-175. 

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　Objective To study the significance of DNA of Toxoplasma gondii in peripheral blood. Methods DNA of T.gondii in peripheral blood of 50 infected rats was detected by polymerase chain reaction. A pair of primers was designed, according to the sequence P30 gene specific to T.gondii, to amplify DNA from T.gondii by PCR. Results The primers amplified DNA specifically from T.gondii and could not amplify DNA from humans, uninfected rat and mouse and from Trichomonas vaginalis and Entamoeba histolytica. DNA of two Toxoplasma parasites was detected by 35 cycles of amplification, indicating a fair sensitivity of the PCR system. Conclusion PCR may have a value for early diagnosis of T.gondii infection in rat.


Analysis on Karyotypes of ANKA strain of Plasmodium berghei

CHENYing-dan;ZHANGJia-xun;LINGBao-ying

2001, 19(3):  13-178. 

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　Objective To analyze the molecular karyotypes of ANKA strain of Plasmodium berghei and demonstrate the size and number of chromosomes. Methods To isolate the genome DNA of P.berghei ANKA strain and analyze molecular karyotypes through CHEF-Ⅲ pulsed field gel electrophoresis (PFGE). Results The number of chromosomes was found to be 14, and their size ranged from 0.6 Mb to 3 Mb. Chromosomes number 5 to 7 and 9 to 12 appeared co-migrated in the gel. Conclusion PFGE technique is useful for analyzing the molecular karyotypes and may be also useful for further study to locate the special gene on chromosomes and carry out the genetic characters and mechanism of drug resistance.


Observation on Eggs of Oncomelania Hupensis Hupensis with Scanning Electron Microscope

XIAQuan-bin;YUANYou-bing;LIUBing;TANPei-ping

2001, 19(3):  14-181. 

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　Objective To observe the structure of the mud hull packed Oncomelania eggs and the surface structure of colloid membrane called the third grade membrane of eggs. \ Methods\ Scanning electron microscopy was used to observe Oncomelania snail eggs with integral mud hull collected from eastern Dongting Lake. \ Results and Conclusion\ The mud hull of eggs was made of unshapen small humification combined with earth granules with a diameter of 2.6-9.2 μm. The mud hull in 60um thickness was honeycomb-like in shape with many small holes and small folds on the wall. There were many round or irregularly round hollownesses on the inner layer of mud hull that contacts colloid membrane but no hole through mud hull. There were some protein fiber networks covering on the colloid membrane and apophysis. The structure of the mud hull showed that the exchange of matter was maintained between eggs and outside, and the mud hull is of great importance to regulating temperature and moisture for the growth of eggs by preventing hydrosoluble substances from penetrating into eggs. The protein fiber networks act on gluing mud hull and buffering outside power. The dense glue membrane might be a main barricade to prevent \{pharmaceutical\} molecules from penetrating into eggs.