Table of Content

30 October 2010, Volume 28 Issue 5

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A New Species of Culicoides （Beltranmyia） from China （Diptera ∶ Ceratopogonidae）

QUFeng-yi;CAOMin;LIURong-xing_;

2010, 28(5):  1-324. 

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Objective To investigate the species of blood-sucking midges of Shanghai. Methods The specimens were collected by light-trap from Pudong Airport （reed marshes）, Shanghai， during the years of 2007-2009. The captured samples was briefly classified, and mounted slide specimens with phenol-balsam method. The microscopic examination including morphological and numerical characters for species identification. Results A new species of biting midges named Culicoides （Beltranmyia） shanghaiensis sp. nov. was obtained， its diagnostic characters were as follows: ① wing without distinct pale or dark markings， only 0-7 macrotrichia find in basal cell; ② the female eyes separate， with pubescence between facets， proboscis head ratio （P/H） 0.7, while the head proboscis ratio （H/P） 1.45, antennal ratio （AR） 1.18, sensila coeloconica present on segments 3-14 （frequency: 1.0, 0.7, 0.5, 0.5, 0.7, 0.5, 0.8, 0.7, 1.0, 1.0, 1.0, 1.0, respectively, n=17） and absent on segments 4-10 indefinitely， with one developed spermatheca；③ the male′s parameres fused narrowly on base. The new species is allied to Culicoides homochrous Remm，but can be distinguished chiefly by: its wing with numerous macrotrichias distributing in basal cell; female eyes separate, bare, H/P 1.03-1.04; male′s parameres separate. Besides, the Culicoides charadraeus Arnaud and Culicoides rarus Das Gupta also resembling with the new species, it can be differentiated from the two former species by the female eyes bare and contiguous （or narrowly separated）, AR＞1.61, no macrotrichia in basal cell of the wings, and the different male genitalia features. Conclusion A new species of biting midges collected from Shanghai is described, and some diagnostic characters are discussed for distinguishing the closely allied species.

论著

Subcellular Mimical Localization of the ATP Synthase B Subunit from Clonorchis sinensis under Different Conditioins of Cell Cycling

ZHOUHong-juan;YUXin-bing;XuJin;HUFeng-yu;HUANGCan;ZHAOJun-hong;MAChang-ling;ZHENGXiao-ling;HUXu-chu*

2010, 28(5):  2-329. 

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Objective To investigate the subcellular localization of ATP synthase b subunit from Clonorchis sinensis under different conditions of Hela cell cycling, and the effect of this protein on the expression of its encoding-gene and homologous host genes. Methods pEGFP-N1-CsATP-synt_B and the vector pEGFP-N1 were transfected into Hela cells, respectively. Transfected cells were synchronized in G₀/G₁ by serum starvation, G₁/S, S cells by double thymidine block, and G₂/M cells by thymidine-Nocodozale block. After synchronization, the subcellular localization of the expressed fusion protein was observed with a laser confocal microscope. The expression level of this fusion protein in cells was detected by flow cytometry (FCM). The expression of CsATP-synt_B and HomoATP-synt_B in different cell cycle phases accessed by RT-PCR. Results FCM results indicated that in the G₀/G₁ phase the expression of pEGFP-N1 vector was decreased significantly, while pEGFP-N1-CsATP-synt_B expression showed an upward trend. In the other phases of cell cycle, the protein expression was similar in the above two kinds of plasmids. The intact CsATP-synt_B was expressed in mitochondria in the G₀/G₁， S， and G₂/M phases and nucleus during G₁/S phase. After the fusion proteins entered the nucleus, the mRNA expression of CsATP-synt_B and HomoATP-synt_B increased significantly. Conclusion CsATP-synt_B can be expressed in the nucleus during G₁/S phase, and regulated by the cell cycle and energy requirements.

Heterologous expression of attacin gene from Musca domestica in Pichia pastoris

LIXiao-bo;WANGTing-ting;MAYan;ZHUJia-yong*;LIUMan-yu;JINXiao-bao

2010, 28(5):  3-336. 

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Objective To construct recombinant expression plasmid with an antibacterial peptide （attacin） mature peptide gene from Musca domestica and express an antibacterial peptide attacin in Pichia pastoris. Methods The coding region of attacin mature peptide was cloned into the expression vector pPIC9K. The recombinant plasmid was linearized by digestion with SacⅠand transformed into P. pastoris GS115 by electroporation. The insert clones containing multiple copies were selected with geneticin，pha，enotype and PCR. The expression of P. pastoris GS115/pPIC9K-attacin were induced with methanol. The supernatant of the expressed protein was analyzed by SDS-PAGE and Western blotting. The anti-bacterial activity of expression product was analyzed by agarose diffusion assay. Results The expression plasmid was constructed and transformed into P. pastoris GS115. SDS-PAGE and Western blotting analysis indicated that the recombinant containing recombinant plasmid pPIC9K-attacin expressed a M_r 22 000 protein. The expression product inhibited the growth of E. coli K12D31. Conclusion The attacin gene from Musca domestica has been expressed in P. pastoris.

Cloning，expression and immunologic identification of Eg10 gene of Echinococcus granulosus

DUJuan;ZHANGWei;WANGYa-na;WANGJie;WANGShu-jing;ZHANGYan;GAOPeng;LIJu-yi;ZHAOWei*

2010, 28(5):  4-342. 

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Objective To clone and express Eg10 gene of Echinococcus granulosus, and investigate the immunological characteristic of the recombinant. Methods Eg10 gene was subcloned into pET28a vector. The recombinant plasmid was transformed into E. coli BL21 and induced with IPTG. The recombinant protein was purified with His-bind purification kit. Forty-eight mice were randomly divided into 4 groups. Mice in groups A and B were injected with PBS and PBS+Freund′s adjuvant（100 μl） as control. Mice in groups C and D were immunized with 10 mg and 50 μg purified Eg10 antigen formulated in Freund′s adjuvant, respectively. All the mice received three immunizations at 2-week intervals with the same dose of antigen. Serum samples were collected at pre-immunization and certain time after immunization. The immunological characteristics of recombinant Eg10 was analyzed by Western blotting and ELISA. Results The recombinant Eg10 protein （M_r 31 000） was expressed in E. coli BL21. The recombinant Eg10 and expression product of PET28a/Eg10 were recognized by sera from mice immunized with recombinant Eg10. ELISA showed that the titer of IgG reached a peak at the 8th week in groups C and D, the level of IgG in sera of groups C or D was higher than that of groups A or B （P＜0.05） at the 2nd, 4th, 6th, 8th, and 10th week. There was no significant difference in the level of IgG between group C and group D （P＞0.05）. Conclusion The Eg10 gene has been expressed with immunogenicity.

Application of double antibody sandwich ELISA for detection of nucleoside triphosphate hydrolase-Ⅱ protein of Toxoplasma gondii

HUXin;ZHUGEQing-yun;LIYa-fei;PANChang-wang;TANFeng*

2010, 28(5):  5-347. 

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Objective To establish a double antibody sandwich ELISA method for detection of nucleoside triphosphate hydrolase-Ⅱ （NTPase-Ⅱ） protein of Toxoplasma gondii. Methods BALB/c mice were immunized with recombinant NTPase-Ⅱ （rTgNTPase-Ⅱ） protein of T. gondii. The hybridomas that secreted high titer of monoclonal antibodies （mAbs） with high specificity were screened and used to establish the double antibody sandwich ELISA for the detection of rTgNTPase-Ⅱ. In order to evaluate the sensitivity of the method, the concentration of whole-tachyzoite lysate and rTgNTPase-Ⅱ was detected, respectively. Serum samples from patients with malaria （7 cases）, schistosomiasis （12 cases）, paragonimiasis （14 cases） and cysticercosis （10 cases） were examined by the same method. Results Two hybridoma cell lines, MNT1 and MNT2, were developed for secreting mAbs against rTgNTPase-Ⅱ. Western blotting analysis showed that the two mAbs specifically recognized rTgNTPase-Ⅱ and whole-tachyzoite lysate. The MNT1 was used as coating antibody, and HRP-labeled MNT2 as secondary antibody. The double antibody sandwich ELISA detecting rTgNTPase-Ⅱ was developed with a minimum concentration of 6 μg/ml for whole-tachyzoite lysate and 1.5 μg/ml for rTgNTPase-Ⅱ. An overall specificity of 100% was determined. Conclusion The double antibody sandwich ELISA based on MNT1 as coating antibody and MNT2 as secondary antibody has a high specificity.

In vitro effect of immune sera on the invasion of Trichinella spiralis infective larvae into intestinal epithelial cells and their development

WANGShu-wei;CUIJing*;WANGZhong-quan;WANGLi

2010, 28(5):  6-352. 

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Objective To observe the effect of sera from mice immunized with excretory-secretory （ES） antigen or surface antigen of Trichinella spiralis larvae on the invasion of intestinal epithelial cells in vitro by the larvae and on their development. Methods HCT-8 cells grown to confluence were overlaid with the larvae suspended in semisolid medium （RPMI 1640 medium +1.75% agarose）， and the larvae were then observed by using an inverted microscope after being incubated at 37 ℃ under 5% CO₂ for 12、24、36、72、96 h. The larval development and its invasion into intestinal epithelial cells were observed under inverted microscope after 15 min when HCT-8 cells were overlaid with the larvae suspended in semisolid medium supplemented with immune sera. Finally， the 1st stage and 2nd~4th stage larvae were observed and enumerated by indirect fluorescent antibody test （IFAT） after 36 h incubation. Results When the larvae were cultured in semisolid medium， they invaded the HCT-8 cell monolayer and molted 1-2 times at 36 h to 72 h of culture. Early adult was observed at 96 h of culture. Cephalic caps on larvae were found at 15 min of culture when the larvae were cultured with medium containing immune sera， but the caps were not observed on those cultured with sera of normal mice or without sera. And the larvae with cephalic caps did not invade the cell monolayer. When the larvae were cultured with immune sera for 36 h， the percentage of 2nd~4th stage larvae （2.25%， 2.20%） were significantly lower than that of those cultured in normal sera （24.7%） （P<0.05）. Conclusion The sera from mice immunized with excretory-secretory antigen or surface antigen of T. spiralis larvae prevent the invasion of the larvae into intestinal epithelial cells in vitro and impede the development （ecdysis） of the larvae.

Multiplex PCR assay for the detection of Angiostrongylus cantonensis larvae in Pomacea canaliculata

WEIFu-rong;LIUHe-xiang;LVShan;HULing;ZHANGYi*

2010, 28(5):  7-358. 

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Objective To establish a multiplex PCR assay for detecting Angiostrongylus cantonensis larvae in Pomacea canaliculata. Methods A pair of specific primers was designed based on the sequences of the small subunit rDNA of A. cantonensis （GenBank jAY295804）， in combination with 16s rDNA specific primers of P. canaliculata, a multiplex PCR was developed. The PCR was performed on positive and negative snails， and the amplified products were analyzed by agarose gel electrophoresis and DNA sequencing. DNA template of 200 Ⅲ stage larvae of A. cantonensis was diluted by negative snail DNA（1 200 ng/μl，120 ng/μl，12 ng/μl，1 200 pg/μl，120 pg/μl and 12 pg/μl），to find the minimum detectable level. Single blind method was used to evaluate the accuracy. After being detected by lung microscopy, 172 snails from field were tested by the multiplex PCR to assess the sensitivity and specificity. Results Agarose gel electrophoresis and DNA sequencing analysis indicated that the target sequences were efficiently amplified by the PCR assay（550 bp for P. canaliculata，405 bp for A. cantonensis）. The minimum detectable level was 120 pg/μl. The coincidence between the two methods stood for 84.3% (145/172), including 45 positives and 100 negatives. 24 snails were PCR positive and microscopy negative, 3 snails were PCR negative and microscopy positive. The sensitivity and specificity of multiplex PCR was 93.8% and 80.6%, respectively. Its positive rate （40.1%, 69/172） was significant higher than that of lung-microscopy （27.9%，48/172）（χ²=14.8，P＜0.01）. Conclusion A multiplex PCR method has been developed for the detection of A. cantonensis larvae in P. canaliculata.

Immune response elicited by the recombinant protein and plasmid DNA of complex antigen ROP2-SAG1 from Toxoplasma gondii

LIWen-shu;XIEZi-xin;CHENQing-xin;CHENShao;ZHANGLi-fang*

2010, 28(5):  8-363. 

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Objective To study the immune response elicited by the recombinant protein vaccine and DNA vaccine of the complex antigen ROP2-SAG1 from Toxoplasma gondii. Methods Sixty female BALB/c mice were randomly divided into 4 groups （15 per group）. Mice in rROP2-SAG1 group were immunized subcutaneously with 2.5 μg rROP2-SAG1 protein formulated in Freund′s adjuvant. Mice in control group received only adjuvant emulsified with normal saline. Mice in recombinant plasmid pcROP2-SAG1 and control plasmid pcDNA3.1 groups were each injected intramuscularly with 100 μg of pcROP2-SAG1 and pcDNA3.1, respectively. All the mice received three immunizations at 2-week intervals. Serum samples were collected at 25, 45, and 70 days after immunization for determining antibody IgG, and at 2 weeks after the last immunization IgG1 and IgG2a were detected all by ELISA. Cell counting kit-8 （CCK-8） was used to determine the splenocyte proliferation, and the supernatant of cultured splenocytes was collected for the detection of IFN-γ by ELISA. Results The level of IgG continued to rise in rROP2-SAG1 group after immunization, and similarly in pcROP2-SAG1 group. At 2 weeks after the last immunization, level of IgG1 （1.538±0.183） was higher than that of IgG2a （0.618±0.122）（P＜0.05） in rROP2-SAG1 group. Whereas no significant difference between IgG1 （1.107±0.137） and IgG2a （0.830±0.185） was observed in pcROP2-SAG1 group （P＞0.05）. Compared with the pcROP2-SAG1 group （A₄₅₀=0.123±0.018）, more significant proliferation response of splenocytes was observed in rROP2?-SAG1 group （0.348±0.042） （P＜0.05）. There was no significant difference（P＞0.05） of IFN-γ and IL-2 in the supernatant of cultured splenocytes between the groups of rROP2-SAG1 and pcROP2-SAG1. Conclusion The antibody level and splenocyte proliferation have been significantly higher in mice immunized with recombinant protein rROP2-SAG1 than those with recombinant plasmid pcROP2-SAG1.

Acute infection of Toxoplasma gondii affects the level of oxygen free radicals in serum and testes of mice

YANGJun-feng;YUEHui-ping;HOUYu-ying*;LIUZhi-shen;RAOHua-xiang;HEYan-xia;ZHANGTing-ting;HUANGChun-yan;GUOLi-na

2010, 28(5):  9-367. 

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Objective To observe the impact of Toxoplasma gondii infection on the level of oxygen free radicals and antioxidant enzyme in serum and testes of mice. Methods 40 BALB/c male mice were randomly divided into four groups. Five mice from each group were injected intraperitoneally with 2.5×10³ tachyzoites of T. gondii, the others received PBS. Mice were sacrificed on the 1st, 3rd, 5th, and 7th day after inoculation. Samples of serum and testes were collected to determine the content of oxygen free radicals and superoxide dismutase（SOD）. Results The concentration of the oxygen free radicals （NO, ·OH, O^2-） in serum and testes of the mice increased along with the days of infection. The concentration of SOD reached a peak on the 3rd day after the injection and then decreased. Both of oxygen free radicals and SOD showed a statistical difference with the control （P＜0.01）. Conclusion Acute infection of T. gondii leads to an increase of oxygen free radicals and SOD in the serum and testes of mice.

Cloning and epitope prediction of AgB antigen gene family of Echinococcus granulosus and Echinococcus multilocularis

JIANGLi*;FENGZheng;HUWei;ZHANGYao-guang

2010, 28(5):  10-371. 

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Objective To clone the subunits of AgB antigen gene family of Echinococcus granulosus（Eg） and Echinococcus multilocularis （Em）, analyze the gene sequences by bioinformatics methods, and predict the potential antigen epitopes. Methods Specific primers were designed based on the reference sequences from GenBank database. Total DNA was extracted from E. granulosus and E. multilocularis, which collected from the endemic areas of Xinjiang, Gansu and Sichuan. The PCR products were cloned into TA vector for sequencing. The online bioinformatics systems such as Bioedit software, Blastn, NPS@ and IEDB were used for the analysis. Results A total of five AgB gene subunits were cloned from E. granulosus and E. multilocularis, respectively, and identified by sequencing. Sequence homology analysis indicated that EmAgB subunits were highly conserved, while the EgAgB subunits were variable. Sequence alignment of AgB subunit genes from E. granulosus and E. multilocularis showed that the identity ranged from 87.69% to 100%. Three major types of secondary structure in the AgB subunits were alpha helix, extended strand, and random coil. Compared with other subunits, the higher percentages of random coil were found in AgB1, AgB2 and AgB4. Ten epitope areas were predicted from the five AgB subunits which were located in AgB1: 1-7 and 21-27, AgB2: 1-7 and 29-36, AgB3: 1-11 and 18-28, AgB4: 1-13, 27-37 and 39-60, AgB5: 1-11, repectively. Conclusion The majority of epitopes in EgAgB and EmAgB subunits have the same or similar sequences. Ten predicted epitopes are mainly located at the N-terminal sequences. Among the five subunits, AgB1, AgB2 and AgB4 display higher antigenicity.

综述

Research progress on midgut bacteria of anopheline mosquitoes and their application in malaria control

LIMei;TANGLin-hua*

2010, 28(5):  11-376. 

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Midgut bacteria in anopheline mosquitoes could control the mosquito populations and block Plasmodium development. The article summarizes the role of bacteria in the mosquito nutrition, the midgut bacterial flora in anopheline mosquitoes and their application in disease control, transstadial transfer of bacteria within mosquitoes, the molecular immune responses of the mosquito to bacteria and the feasibility of applying bacteria to control malaria.

Effect of toll-like receptors on helminth infection

YUYan-rong*;QIYong-fen;

2010, 28(5):  12-381. 

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Toll-like receptors（TLRs） as a family of pattern-recognition receptors are vital for immune and inflammatory responses, and also play an important role in helminth infection. By regulating TLRs expression and function， helminth can alter host immune response, interfere with disease progression， and change the immune microenvironment of host. This review summarizes the biological characteristics of TLRs and recent advances of TLRs in immune modulation on helminth infection.

Formation mechanism and the function of parasitophorous vacuole of Toxoplasma gondii

PENGHong-juan

2010, 28(5):  13-386. 

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Toxoplasma gondii is a ubiquitous, obligate intracellular protozoan parasite in human and animals. Chronic infection with this parasite is likely one of the most common infection in human. Following invasion of host cells T. gondii resides within membrane-bound vacuoles known as parasitophorous vacuole （PV）, which can protect the parasite from the endosomal acidification and lysosomal fusion of host cells. It plays an essential role in the whole parasitic process of T. gondii. This review summarizes the mechanism of PV formation and its function in the host cells.

Progress on the application of Cryptoporidium infected animal models and in vitro cultivation

YINJian-hai;SHENYu-juan;CAOJian-ping*

2010, 28(5):  14-392. 

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The genus Cryptosporidium is composed of protozoan parasites that infect epithelial cells in the microvillus border of the gastrointestinal tract of all classes of vertebrates, and cause severe diarrheal disease in a variety of neonatal animals, children and immunocompromised persons. Establishment of Cryptosporidium infected animal models and its in vitro cultivation system have established a good foundation for characterizing life cycle stage, exploring immunological mechanism, developing vaccines, screening and evaluating potential drugs, as well as assessing oocyst inactivation techniques. This paper reviews recent development and application of the Cryptosporidium infected animal models and its in vitro cultivation.

研究简报

Construction and identification of a full-length cDNA library from Spirometra erinaceieuropaei

FANGZheng-ming;WANGSheng;SUBin-tao;GONGNian-qiao;LIUWen-qi;MINGChang-sheng*

2010, 28(5):  15-331. 

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20 ml peritoneal lavage fluid of mice infected with Toxoplasma gondii RH strain was diluted to 250 ml with sterilized physiological saline, and filtered through cellulose membrane filters （pore size：5 μm）. The filtrate was centrifuged at 1 512×g for 15 min， and the sediment was pure T. gondii tachyzoites which were then sonicated. The soluble antigen was prepared by centrifugation at 11 200×g for 30 min. Sera of T. gondii infected SD rat and normal SD rats were collected for immunodetection of soluble antigen. The specificity and valence of soluble antigen were detected with indirect ELISA. The mean removal rates of mouse leukocytes and erythrocytes were 99.9% and 80.3%, respectively, and recovery rate of tachyzoites was 71%. The soluble antigen was extracted from purified T. gondii （1.38 mg per mouse）. Indirect ELISA showed that the lowest effective antigen concentration was 5 μg/ml.

Real-time fluorescence quantitative polymerase chain reaction detection for Toxoplasma gondii B1 gene in mice urine

LUOFang-jun;YAOHong-feng;WANGYong-ming;TANFeng*

2010, 28(5):  16-338. 

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A pair of specific primers and a TaqMan probe were designed based on the sequence of Toxoplasma gondii B1 gene from GenBank database. Total DNA of T. gondii was extracted from fresh mice urine. DNA fragment of B1 gene was amplified by PCR. The PCR product was cloned into pMD18-T vector. Following identification, the positive recombinant plasmid was used as reference template to generate standard curve and melt curve. Sensitivity, reproducibility, linear range and stability of reference plasmids were determined. The sensitivity of this method was 10⁴ copies/ml. The coefficient of variation （cv） of intra-assay and inter-assay were 2.42% and 4.18%, respectively. Linear range was （10³-10⁷） copies/ml. The specificity was 100%. The reference materials were stable. Real-time FQ-PCR of T. gondii DNA in mice urine has been constructed, which is a convenient, sensitive and reliable method for quantifying T. gondii DNA in mice urine.

Screening of mimic antigen epitopes of Echinococcus granulosus by phage-displayed peptide library

XUANYan;ZHENGHong*;LIANGChao

2010, 28(5):  17-354. 

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The 7-mer phage random peptide libraries was screened with the specific anti-EgB polyclonal antibody. Sixty clones picked randomly were deduced by DNA sequencing and their amino acid sequences were compared with EgB. After 5 rounds of panning, the phage recovery rate increased from 4.15×10^-5 to 4.30×10^-2. 45 out of 60 selected clones were ELISA positive by anti?鄄EgB polyclonal antibody with a positive rate of 75%, 45 positive clones were sequenced and the predicted amino acid sequences showed low homology to EgB. The reactivity of selected peptide and sera from patients of anaphylactic shock induced by Echinococcus granulosus was detected by ELISA. The result showed that the absorbance value（A₄₁₀） of 45 postive clones detected by using serum samples from patients was twice higher than that by using 7 peptide library. The 45 positive clones exhibited specific binding to anti-EgB antibody. The peptides screening from 7-mer phage random peptide libraries may be the mimic epitopes of anaphylactic shock induced by E. granulosus.

Construction and identification of a full-length cDNA library from Spirometra erinaceieuropaei

LVGang*;LUYa-jun;FANZhi-gang;SHIDa-zhong;GANXiu-feng;ZHONGSai-feng

2010, 28(5):  18-394. 

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The full-length pBluescript Ⅱ SK cDNA library of adult Spirometra erinaceieuropaei was constructed by using the SMART method. Data showed that 95.5% of the library was recombinant and the titer of the library was 1.06×106. The average insert size of the library was about 1.4 kb. Forty-eight randomly selected clones were sequenced. A set of 36 effective expressed sequence tags （ESTs） with the average size of 674 bp was obtained after excluding clones shorter than 450 bp. The unigenes occupied 58.3% of the 36 ESTs. The rate of full-length cDNAs were 57.7% （15/26）. The high-quality of full-length cDNA library could be used for large scale EST sequencing.

Cryopreservation of Trichomonas vaginalis and its influence factors

YUANYu-qing;XUEChang-gui*

2010, 28(5):  19-396. 

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Different concentrations of dimethylsulfoxide （DMSO） and glycerol， various densities of Trichomonas vaginalis trophozoites and duration of storage were used to study the effects on the cryopreservation of T. vaginalis at -78 ℃. The optimal densities of trichomonads for cryopreservation were （1-2） ×10⁶/ml. Glycerol and DMSO showed a high protective effect， with a maximum effect at a concentration of 10%， and the survival rates were 38.0% and 31.7%， respectively. There was no significant difference between them（P＞0.05）. The results showed that the above optimized factors can provide a good protection in short-term （1-16 weeks） cryopreservation， and the trichomonads can be incubated to a logarithmic growth phase at 37 ℃.

Clinical analysis of 6 cases with cerebral sparganosis mansoni

CHENXian-han;SHIGuang-feng*

2010, 28(5):  20-398. 

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A retrospective study was carried out to analyze 6 cases of cerebral sparganosis mansoni in Huashan Hospital from August 2005 to August 2009. The epidemiological features, clinical characteristics, therapeutic approaches and outcome of this disease were investigated. Among the patients, 4 cases had a history of eating raw frogs or snakes. 5 showed eosinophilia in peripheral blood, all with positive anti-Sparganum mansoni antibody in serum and cerebrospinal fluid. Cerebral MRI showed placeholder in all patients. Diagnosis was confirmed by pathological examination of operation and species identification. All patients were cured by operation removal and praziquantel treatment.

In Vitro effect of huyinling extract on Trichomonas tenax

LIANGYu-fen;YUNChen-xia;WEIJun-bin;LIQiang;WANGZhi-ping*

2010, 28(5):  21-400. 

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The active ingredient of Huyinling, a combination of Chinese traditional medicine, was extracted by five different ethanol concentrations（40%-80%）. There were seven groups named as five Huyinling ethanol extract groups （40%, 50%, 60%, 70%, and 80%）, metronidazole group and blank control. Each Huyinling ethanol extract group was further divided into five subgroups with final concentration of 6.25, 12.5, 25, 50, and 100 mg/ml, respectively. Metronidazole group was given 10 μg/ml of the drug. Each group had 4 wells with 125 μl T. tenax（2×105/ml）. At 12 h, 24 h and 48 h after drug treatment, the anti-T. tenax effect of Huyinling ethanol extract was tested by microscope counting method. At 24 h the effect of Huyinling on T. tenax was examined with methyl thiazolyl tetrazolium （MTT） assay. The results showed that the higher concentration of Huyinling ethanol extract, the better effect on anti-T. tenax. 60% Huyinling ethanol extract group with concentrations of 6.25 mg/ml and 12.5 mg/ml showed higher anti-T. tenax effect than other groups （P<0.01）. The ethanol extract of Huyinling granules has a remarkable effect on T. tenax, and among the groups, 60% ethanol extract shows the best anti-T. tenax activity.

病例报告

A case of bilateral optic atrophy caused by neurocysticercosis

LVZhong-ping;MENGDan

2010, 28(5):  22-封二. 

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Dipylidium canninum infection in an infant

LILin;ZHANGShi-hai;SHENJie;WEIXiao-lu;WANGZen-xian;LUOQing-li

2010, 28(5):  23-392. 

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