Table of Content

29 February 2012, Volume 30 Issue 1

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Control Situation and Primary Task of  Key Parasitic Diseases in China

LEI Zheng-Long, WANG Li-Yang

2012, 30(1):  1-1-5. 

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In the“11th Five-year Plan” period, China unveiled a mid-term planning on the control of the key parasitic diseases, including schistosomiasis, malaria, echinococcosis (hydatid disease), as well as some other parasitic diseases. It clarifies the goals on the control of the major parasitic diseases in the national control program during the “12th Five-year Plan” period (2011-2015) , formulates current main tasks based on analyzing the progress of the national control program on key parasitic diseases, in order to meet the challenges appearing in the future implementation.

Assessment of Health Education against Echincococcosis in  Tibetan Region of Sichuan Province，China

XIAO Ning1,5 *，ZHOU Zhang-jun2，CHEN Xing-wang1，ZENG Yun-zhong2，WANG Zai-yue3，

2012, 30(1):  2-6-11. 

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 Objective   To find an effective strategy of implementing health education in Tibetan Regions so as to make echinococcosis control sustainable at a large scale.  Methods   During July to November of 2008, surveys were conducted on health education requirements among various populations in the form of questionnaire and group discussion in endemic areas of echinococcosis in three counties of the Ganzi Tibetan Autonomous Prefecture, Sichuan. Based on the obtained results，health education materials suitable for different populations were designed. The materials were applied for conducting health education in field at Tagong Township, Kangding of Ganzi in October, 2009. By the survey in May, 2010，the improved effect on knowledge and behavior change was compared before and after (6 months later) health education in order to assess the usefulness of these materials. Simultaneously, Xinduqiao Township of Kangding was selected as control. This town is neighboring to the study area with similar natural and demographic conditions, in which no health education activities related to the study was implemented.  Results   Various populations showed their interested requirements for health education materials and ways. Based on the information collected，a series of materials were designed and applied for health education activities in field. The results indicated that，compared with the data before health education，improvement of the knowledge and behaviors against echinococcosis among students and local residents showed significant increase (P<0.05)，but only one knowledge point about the importance of washing hands before meal among students showed no significant difference (P>0.05) in spite of the rates increasing from 88.6% (78/88) before health education to 95.5%(84/88) after education. The local officers and monks also showed to some extent improvement on behavior manners. The rate of correct treatment of livestock viscera increased from 37.1%（13/35）and 30.3%（10/33）before the education to 82.9%(29/35） and 78.8%(26/33）(P<0.01)，while the rate of not feeding stray dogs  among monks was 6.4% (3/47) before and 10.6% (5/47)(P>0.05) after the education. The knowledge-increasing and behavior-improving rates on playing dogs among the students，residents and monks increased by 18.2% and 19.3%，42.8% and 54.3%，6.4% and 14.9%，respectively. There was no considerable change in the control township.     Conclusion   The materials designed for health education are acceptable by most populations. The improvement rate of knowledge and behaviors is considerable among the groups. Use of the materials receives expected effect on health education and health promotion.

Immunological Analysis of Piperaquine-resistant Murine Model of Plasmodium berghei ANKA Strain

HUANG Xian-xi，ZHOU Li-min，YI Guo-hui，WU Jin-yan，PAN Zai-yong，XUE Wei-ling，GUO Hong*

2012, 30(1):  3-12-17. 

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Objective   To analyze the immunological characteristics of murine model of piperaquine sensitive （PQS） line and resistant （PQR） line of Plasmodium berghei （Pb） ANKA strain.  Methods   64 Kunming mice were divided into three groups, 16 in each of groups A and C， 32 in group B（16 of 32 were used for observing suvival days）. Each mouse in groups A and B was infected with 1×10⁷ erythrocytic stage parasites of PbPQS and PbPQR，respectively. Mice in group C were injected with the same volume of normal saline. On days 4，8，12 and 16 after inoculation，4 mice from each group were sacrificed. Blood samples were collected for thin blood smear examination，and parasitemia rate calculated. Spleens were removed and spleen lymphocytes suspension prepared. Spleen lymphocytes were stimulated with ConA，and cell proliferation was measured by Cell Counting Kit-8 （CCK-8） assay. Nitrogen oxide （NO）and IFN-γ level of spleen cell culture supernatants were detected by the Griess reagent and ELISA methods，respectively. Another 10 mice were each inoculated with 1×10⁷ parasites of PbPQR line，and the mice were then challenged with lethal PQS line when the parasites turned into blue stained cells. The parasitemia and survival days were recorded.  Results   The average survival time of group A was （9.0±3.0） d，the parasitemia rate was over 50% at 6-12 days post-infection with severe anemia. On 16th day post-infection，no death was recorded in group B with a parasitemia rate of （26.66±2.54）%. After ConA stimulation，the proliferation of spleen lymphocytes in groups A （0.65±0.08） and B （0.86±0.20） at 12 days after infection was significantly higher than that of group C （0.18±0.03） （P<0.01）. NO level in spleen cell culture supernatant increased with prolonged infection time. On 12th day post-infection，NO level of groups A［（48.80±3.49） μmol/L］ and B［（54.80±2.17） μmol/L］ was higher than that of group C ［（7.80±0.71） μmol/L］ （P<0.01）. IFN-γ concentration in spleen lymphocytes culture supernatant increased with prolonged infection time. The highest IFN-γ level of group A was （752.20±39.49） pg/ml on 12th day post-infection，while in group B it was （855.80±33.65）  pg/ml on 8th day after infection，then decreased on 12th day ［（620.20±27.11） pg/ml］. IFN-γ  level showed a significant difference between groups A and B （P<0.01）. In 10 days after challenge，the parasitemia rate in PQR group was up to （2.44±2.07）%，and gradually disappeared. No parasite was detected on 40th day after challenge and no mice died.   Conclusion   The proliferation of spleen cells，NO and IFN-γ  levels of spleen lymphocytes culture supernatant in PbANKA strain PQR line are significantly higher than that of PQS line. PbPQR line can induce certain protective immunoreaction.

Expression of Transforming Growth Factor-β1 and Connective Tissue  Growing Factor in Schistosomal Hepatic Fibrosis of Mice

HUANG Jia-quan1，ZHU Jun2，LI Lan1，JIAO Yun-tao1，XU Lei1，TAO Ran1，FAN Xiang-xue1，MA Ke1，GUO Wei1，NING Qin1*

2012, 30(1):  4-20-26. 

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 Objective   To establish the model of hepatic fibrosis in mice infected with Schistosoma japonicum and observe the expression of transforming growth factor-β1 （TGF-β1） and connective tissue growing factor （CTGF） in mice model.  Methods   Thirty BALB/c mice were randomly assigned to control group and model group. Each mouse of model group was infected with (30±1) S. japonicum cercariae through the abdominal skin. Serum samples were collected at 4，6，8，10, and 12 weeks after infection, and were analyzed for the levels of ALT and AST. Pathological changes and proliferation of hepatic collagen fibers in liver tissue were observed after HE staining and Masson staining. Immunohis-tochemistry and real-time PCR were used to detect the protein and mRNA expression of CTGF and TGF-β1.  Results   6-12 weeks after infection, there was significant difference in ALT and AST between model group and control group（P＜0.05）. At 12th week, ALT [（173.53±31.12）U/L ] and AST  [（301.00±34.87）U/L]  in model group were higher than those in control group [(42.00±3.53)  and  (96.58±11.26)U/L]. In model group, egg granulomas formed in the liver, and the formation of hepatic fibrosis was significant in portal areas, and there was tree-like hepatic fibrosis around the portal vein branch. 8 weeks after infection, hepatic fibrosis area in mice of model group increased considerably，and there was significant difference in percentage of positive area of collagen between 12th week [（23.83±1.68）%]  and control group［（1.23±0.14）%］（P<0.05）. 10 and 12 weeks after infection, the percentage of positive area of TGF-β1  [（22.34±2.58）% and （25.82±3.01）%]  and CTGF[（11.32±2.44）% and （14.51±2.05）%]was higher respectively than that of the control ［（2.56±0.87）%，and （1.09±0.73）% （P<0.05）. 6，8，10，and 12 weeks after infection，both TGF-β1 and CTGF mRNA increased gradually, higher than that in control group（P<0.05）. 10 weeks after infection, TGF-β1 mRNA relative transcription level was the highest ( 0.0721±0.0187), and it was 0.0089±0.0037 in control group. CTGF mRNA relative transcription level reached the highest value（0.1136±0.0365）in 12 weeks after infection, while it was 0.0293±0.0184 in control group. CTGF mRNA expression was positively correlated with the duration of infection （r=0.927，P＜0.05）. Conclusion    The area and cell types of TGF-β1 positive expression is the same as that of CTGF in liver tissue of schistosome-infected mice (BALB/c). CTGF mRNA expression is significantly related to the duration of infection, but it is not the case for TGF-β1.

Gene-cloning， Expression and Antigenicity Analysis of Rhoptry Protein 17 of Toxoplasma gondii

WANG Hai-long1，YIN Li-tian2，MENG Xiao-li1，SHEN Jin-yan1，LIU Hong-li1，YIN Guo-rong1 *

2012, 30(1):  5-27-31. 

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Objective  To clone and express the rhoptry protein 17 （ROP17） gene of RH strain of Toxoplasma gondii，analyze the antigenicity of recombinant protein.  Methods   Total RNA was extracted from tachyzoites of RH strain of T. gondii. The open reading frame of TgROP17 gene was amplified with a pair of specific primers which was designed according to the coding sequence of TgROP17 gene （GenBank accession No. AM075203.1），the product of RT-PCR was digested with double restriction enzyme and ligated into a pGEX-6P-1 vector. The recombinant pGEX-6P-1-TgROP17 plasmid was transferred into E. coli DH5α and the positive clones were selected through the colony-PCR and confirmed by the double restriction enzyme digestion and sequencing. The constructed pGEX-6P-1-TgROP17 was transformed into E. coli Rosetta （DE3） and induced with IPTG for expression. The expression products were analyzed through SDS-PAGE followed by Coomassie blue staining. Western blotting assay with GST primary antibody and rabbit anti-T. gondii serum was used to confirm the expression of GST-ROP17 and analyze its antigenic properties.   Results   The product of RT-PCR was with 1 850 bp.  The recombinant plasmid pGEX-6P-1-TgROP17 was confirmed by colony-PCR，double restriction enzyme digestion and sequencing. A soluble recombinant protein with relative molecular weight of 96 000 was analyzed by SDS-PAGE followed by Coomassie blue staining. The GST tag in GST-ROP17 and the antigenicity of ROP17 were detected efficiently by Western blotting with the GST primary antibody and with the prepared antiserum against T. gondii, respectively.   Conclusion   The recombinant GST-ROP17 protein has been produced in E. coli and shows specific antigenicity.

Dynamic Change of IL-10 and TGF-β1 in the Liver of Echinococcus multilocularis-Infected Mice

ZHAO Hui1，BAI Xue2，NIE Xiao-han2，WANG Jiang-tao2，WANG Xi-xian2，PANG Nan-nan1，LI Yu-jiao1，MA Xiu-min3，WEN Hao1，DING Jian-bing 3*

2012, 30(1):  6-32-35. 

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  Objective   To observe the dynamic expression and function of IL-10 and TGF-β1 in liver of BALB/c mice infected with Echinococcus multilocularis (Em).  Methods   Sixty female BALB/c mice were randomly divided into experiment group and control group. Mice in the experiment group were each injected with 0.2 ml Em protoscolex suspension (containing about 400 protoscoleces), while those in control group received same volume of normal saline. At 2, 8, 30, 90, 180, and 360 d after infection, 5 mice from each group were sacrificed and liver specimens were collected for pathological examination and immunohistochemical detection for IL-10 and TGF-β1.  Results   In mice of the experiment group, Em cysts in different sizes were found in the abdominal cavity and the liver tissue, which gradually enlarged with the time. HE staining showed infiltration of lymphocytes in liver tissue, pathological change between the cyst wall and hepatic cells. In the control, the liver lobules showed integrity and inflammatory cells were seen occasionally. The level of IL-10 expression in liver tissue of the infected mice increased with the time, and reached a peak  [(16.39±1.73)%]  at 90 d post-infection and maintained a high level thereafter. The expression of TGF-β1 also reached the highest level [(23.69±2.29)%]. Both were significantly higher than the control (P<0.01), though a low level expression was found in the control at 90 d post-injection.   Conclusion   The expressions of IL-10 and TGF-β1 both increase in the middle and late stages of the infection. Besides, their inhibited functions do not be helpful for clearing and controlling Echinococcus multilocularis infection in livers.

Genotype Analysis and Isoenzyme Patterns of Ten Isolates of Blastocystis hominis from Guangxi

ZHAN Ting-zheng，SHI Huan-huan*，HE Shan-shan，LIU Teng

2012, 30(1):  7-36-40. 

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Objective   To analyze genotypes and lactate dehydrogenase（LDH) and esterase(EST) patterns in 10 isolates of Blastocystis hominis collected from Guangxi.  Methods   Ten B. hominis isolates（BhGX1~BhGX10） were obtained from the fecal specimens of patients and cultivated in vitro, and then the genomic DNA was extracted. The isolates were genotyped by PCR using seven pairs of known sequenced-tagged site （STS） primers. Isoenzyme patterns of LDH and EST were investigated by SDS-PAGE.  Results   Out of the 10 isolates, 8 were identified as genotype Ⅰ and the genotypes of the other two （BhGX4 and BhGX7） were unknown which were negative to all the STS primers. Among the ten isolates, 10 LDH bands were found, more with Rm37, Rm49, Rm57, Rm68 and Rm92. 12 bands showed in EST patterns: Rm14, Rm18, Rm23, Rm27, Rm35, Rm41, Rm45, Rm50, Rm55, Rm68, Rm77 and Rm82. Difference existed with the LDH and EST patterns among the isolates.   Conclusion   Genotype Ⅰ is the major one in the 10 B. hominis isolates from Guangxi, and the isolates show different isoenzyme patterns for LDH and EST.

In Vitro Sensitivity of Plasmodium falciparum Isolates from China-Myanmar Border Region to Chloroquine, Piperaquine and Pyronaridine

ZHANG Cang-lin1，2， ZHOU Hong-ning2，WANG Jian2， LIU Hui2

2012, 30(1):  8-41-44. 

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 Objective   To assess the in vitro sensitivity of Plasmodium falciparum to chloroquine, piperaquine and pyronaridine in China-Myanmar border area.  Methods   Fifty-one blood specimens of P. falciparum isolates were collected from Laza City of Myanmar during September to December in 2009, and the sensitivity of the parasites to the drugs was detected by Rieckmann′s in vitro microtest.  Results   Among the 42 blood samples with valid results of sensitivity test,  the resistance rate to chloroquine, piperaquine and pyronaridine was 95.2%, 7.1%, and 54.8%, with a corresponding 50% inhibition dose(ID50 )of 320.5, 128.2, and 96.0 nmol/L, respectively. Pyronaridine-resistant P. falciparum exhibited some degree of cross-resistance to chloroquine [91.3%(21/23)] and piperaquine [13.0%(3/23)], and chloroquine-resistant P. falciparum showed cross-resistance to piperaquine [7.5%(3/40)] and pyronaridine [52.5%(21/40)]. High level of cross-resistance was present to chloroquine （100%） and pyronaridine （100%） in piperaquine-resistant P. falciparum.   Conclusion   In Laza City, P. falciparum shows high resistance to chloroquine, half isolates are resistant to pyronaridine, and most isolates are still sensitive to piperaquine.

Analysis of Surgery Effect on 198 Cases of Pulmonary Echinococcosis

YU Shan-Hai, LI De-Sheng, YI Li-YA-Er-·Xia-He-Ding, WU Ming-Bai, ZHANG Li-Wei-*

2012, 30(1):  9-45-48. 

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 Objective   To summarize the experience in the surgical treatment of pulmonary echinococcosis，and evaluate the operative approach to avoid recurrence.  Methods   A retrospective study was carried out to analyze the clinical data of 198 cases with pulmonary echinococcosis from January 2000 to October 2010 in the First Affiliated Hospital of Xinjiang Medical University，involving the general conditions，clinical course，epidemiology，clinical characteristics，therapeutic approaches and outcomes.  Results   There were 114 male and 84 female patients aged from 2 to 68 years with a mean of 25.5 years. Herdsmen accounted for 74.7%（148/198），and  the juveniles occupied 21.7%（43/198）. The major operative methods were complete removal of endocyst and cystectomy with needle aspiration. All the 198 cases were improved or cured. Postoperative pulmonary infection occured in 9 case（4.5%）， 7 cases（3.5%）suffered from wound dehiscence and 4 cases（2.0%）had wound infection，but cured by dressing. 165 patients（83.3％）were followed up for l-10 years. The recurrence rate was 5.6%（11/198）. Patients with recurrence were cured after operation. 198 patients received 226 operations.   Conclusion   Complete removal of endocyst is the main method for pulmonary echinococcosis with better effect and less recurrence.

Magnetic Resonance Imaging Expression of Central Nervous System due to Angiostrongyliasis cantonensis

WANG Xiao-yan*，LIN Lan，LIU Jiang

2012, 30(1):  10-49-51. 

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Objective   To analyze the MRI characteristics of the central nervous system （CNS） due to infection of Angiostrongylus cantonensis.  Methods   A retrospective study was carried out to analyze MRI expression of CNS in 25 cases of angiostrongyliasis cantonensis in the First People′s Hospital of Yunnan Province from May 1997 to July 2011.  Results  Among the 25 patients，8 cases were normal on MRI, 7 manifested meningitis，8 showed parenchymal pathological change, and 2 patients showed spinal meningitis and ventriculomegaly，respectively. These lesions diffused or scattered in CNS and appeared as iso- or slightly low signal intensity on T1WI，high signal intensity on T2WI and fluid-attenuated inversion recovery（FLAIR） images. After administration of Gd-DTPA，multiple enhanced round- or oval-shaped nodules appeared. Meningeal involvement revealed linear or nodular enhancement in leptomeninges or ependyma.  Conclusion  MRI image of angiostrongyliasis cantonensis was diverse. Multiple linear and nodular enhancement in the brain and spinal cord and enhancement in the leptomeninges were the main findings，but the MRI findings are mostly nonspecific.

Surveillance on the Prevalence of Soil-transmitted Nematode  Infection in Fujian in 2006-2010

CHEN Bao-jian，LI Li-sha，ZHANG Rong-yan，LI Yan-rong，ZHANG Zhi-fang，ZHENG Guo-bin，FANG Yan-yan，LIN Chen-xin，JIANG Dian-wei

2012, 30(1):  11-52-55. 

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Objective   To analyze the prevalent trend of soil-transmitted nematode infection in Fujian Province during the past 5 year surveillance and evaluate the control effect.  Methods   From 2006 to 2010, fecal samples of the inhabitants of 3 years old and above were collected every November and examined for intestinal helminth eggs by the modified Kato's thick smear technique at the 2 surveillance sites: Punan village of Zhangzhou and Gushan village of Shaowu. Cellophane tapes were used to detect pinworm eggs for children aged 3-12. Soil samples were also collected from vegetable field, lavatory, courtyard and kitchen of 20 randomly selected families (in 2 villages) each with stool egg-positive findings and examined for ascaris eggs by a modified saturated sodium nitrate floatation method.  Results   The prevalence of soil-transmitted nematode infection at the surveillance sites decreased from 45.3% (946/2 087) in 2006 to 15.1%(226/1 494)in 2010, with a reduction of 66.6%. Among the infected subjects, hookworm infection occupied 75%-85%, while ascaris or trichuris infections each accounted for less than 10%. In terms of infection intensity, 65.2%-85.5% of the hookworm infection was light, and majority of the infected subjects were farmers. The pinworm prevalence in children were still high although it had dropped down from 46.1% (140/304) in 2006 to 29.8%(36/121)in 2010, declined by 35.4%. In the 5 years, totally 400 soil samples from 100 families were examined and 21 samples were found ascaris egg positive with viable eggs in only one sample.   Conclusion   The 5 year surveillance reveals a decreasing trend of the soil-transmitted nematode prevalence but shows a relatively high hookworm infection rate in the population and pinworm infection in children.

Functional Roles of Macrophage Migration Inhibitory Factor in Anti-parasitic Diseases

LIANG Le1，LIU Hai-peng2，CAO Jian-ping3 *

2012, 30(1):  12-56-60. 

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Macrophage migration inhibitory factor（MIF）is an important proinflammatory and immuno-regulatory mediator，which locates at the apex of the cascade of innate immune response. It also plays an important role in regulating adaptive immune responses and counter-regulating the immunosuppressive effect of glucocorticoid. MIF is actively involved in a series of cell processes such as cell proliferation and differentiation as well as cell apoptosis. MIF is reported to regulate the host immune response to parasitic infection. Interestingly，the ortholog of mammalian MIF was identified in multiple parasites such as Plasmodium spp.，Leishmania major，Brugia malayi and so on， which is actively involved in the regulation of host-pathogen interaction and therefore provides a novel immune evasion mechanism. Therefore，development of small molecular inhibitors and vaccines against MIF will pave a new avenue for the prevention and treatment of related parasitic diseases.

PCR-derived Technology in Gene Identification and  Typing of Toxoplasma gondii

NIE Da-ping1，YOU Ying-xia1，SHEN Li-jie1, 2 *，LI Wei1

2012, 30(1):  13-61-64. 

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Different genotypes of Toxoplasma gondii show a great diversity in pathogenicity and drug sensitivity. Application of the PCR-derived technologies in gene identification and typing of T. gondii provides an important basis to clinical diagnosis and treatment. This article reviews the relevant technologies in gene identification and typing of T. gondii.

Lymphatic Filariasis in Hubei Province：from Prevailing to Elimination

ZHAN Fa-xian*，ZHANG Shao-qing，WANG Li-li，YUAN Fang-yu，ZHANG Hua-xun，DONG Xiao-rong，LI Kai-jie，YU Pin-hong

2012, 30(1):  14-65-70. 

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 Lymphatic filariasis used to highly prevalent in 69 counties （cities） with 29 million people at the risk of infection in Hubei Province. There were 2 million people infected either by B. malayi or W. bancrofti that 1.6 million microfilaremia cases and 0.4 million chronic patients. The average rate of microfilaremia among population was 5.94%. Anopheles sinensis and Culex quinquefasciatus were the principal transmitting vectors. Since 1970s，with the strategy of taking elimination of infection source as a major focus，the average rate of the microfilaremia reduced to 0.048% with a village as the unit in 1988，and reached the standard of transmission interruption. With continuous surveillence for over a decade，the province reached the goal of filariasis elimination in 2001. This paper reviews the prevalence，control and elimination process of filariasis in Hubei Province since 1950s.

Research Progress on the Sugar-feeding Behavior of Mosquitoes and  its Application in Mosquito Control

JIANG Jin-yong1, 2，MA Ya-jun1*，ZHOU Hong-ning2

2012, 30(1):  15-17-19. 

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The energy of the physiological activities for both male and female mosquitoes is provided by feeding sugar outdoors. The attractive toxic sugar bait（ATSB）was developed according to the behavior, and showed a good result in reducing mosquito population size. This paper introduces the progress on the sugar-feeding behavior of mosquitoes and application in vector-borne disease control.

Efficacy of Ablendazole for Treatment of Mice Infected  with Sparganum mansoni

CUI Jing，WANG Ming-ming，ZHAO Yu-wei，GAN Guan-hua，HU Bo-wen，JIANG Peng，QI Xin，LIU Li-na，WANG Zhong-quan*

2012, 30(1):  16-71-72. 

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In order to observe the efficacy of albendazole for the treatment of mice infected with Sparganum mansoni，a total of 72 mice were divided into 8 groups （9 mice each），each mouse was orally infected with 5 plerocercoid. At one week after infection，groups A-C were treated with a 7-day course of albendazole （1 700，2 500，and 3 300 mg/kg，twice daily），and sacrificed at 1 week post-treatment；groups E-G were treated with the second course of albendazole with the same dosage at 1 week interval after the first course，and sacrificed at 1 week after the second course；the groups D and H were used as control for A-C and E-G，respectively. After the infected mice were sacrificed，the mean number of worms recovered was observed and worm reduction rate was determined. When treating with one course of albendazole at 1 week post in-fection，the worm reduction rate in groups A-C was 20.0%，20.0% and 24.9%，respectively （χ²=0.351，P>0.05）. After treatment with two courses of albendazole，the worm reduction rate in groups E-G was 22.3%，36.4% and 31.9% （χ²=1.812，P>0.05）. The difference of the worm reduction rate in the infected mice treated with 1 and 2 courses of albendazole showed no statistical significance （P>0.05）. The results indicated albendazole has no obvious efficacy for treating sparganosis in mice.

Dynamic Pathologic Changes in Pigs Infected with Taenia solium

FANG Wen1，XIAO Liang-liang2，BAO Huai-en2 *，MU Rong2

2012, 30(1):  17-73-75. 

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Four three-way crossed hybrid pigs were infected each with 80 000 Taenia solium eggs. One pig was sacrificed respectively at 40，80，120，and 150 d post-infection，and tissue samples containing cysticerci were collected for pathological sections. The results showed that in 40-80 days，cysticerci in the liver were surrounded by membrane formed with the hyperplastic fibrous connective tissue and inflammatory cells. On 120th day，a large number of the cys-ticerci were dead or calcified. On 150th day，all cysticerci died or calcified. On 40th day，the inflammatory cells infil-trated around the cysticerci in the tongue and skeletal muscle，and the inflammatory responses near the scolex were more serious than that near cyst wall. In the 80-150 days，the inflammatory cells showed bandlike distribution. The pathological response in cardiac muscle was similar to that of the skeletal muscle and the tongue，but with more inflammatory cells. On 40th day，there were a few inflammatory cells infiltrating around cysticerci in the brain，in the 80-150 days，a large number of inflammatory cells aggregated around the cysticerci. The inflammatory response in the tissues progressed with the duration of infection and the inflammation in the liver was significantly stronger than that in other tissues.

Cloning and Bioinformatics Analysis of PFC0460w in Plasmodium falciparum

ZHOU Hong-chang*，DAI Jia-li，FAN Cheng-cheng，ZHONG Hui-ya，SHEN Yu-qing

2012, 30(1):  18-76-77. 

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The gene fragment of PFC0460w was amplified from RNA of Plasmodium falciparum 3D7 strain with RT-PCR，and cloned into pGEM-T easy vector. The recombinant plasmid was transformed into E. coli DH5α and the positive clones were selected，which were identified by PCR and sequencing. The results showed that there were three sequences of PFC0460w fragment, respectively with length of  618，597 and 543 bp. The 618 bp fragment was completely consistent with the sequence published in PlasmoDB（GenBank No. XM_001351147），and the 597 bp and 543 bp fragments were submitted to GenBank with Accession No. of JF799872 and JF799873，respectively. 205 amino acids were encoded by the 618 bp fragment，and five kinds of protein structure were predicted by Robetta.

Bioinformatics Prediction of EgA31 Recombinant Antigen Epitopes of Echinococcus granulosus

LI Yu-jiao1，YANG Jing2，ZHAO Hui1，JIA Hai-ying3，ZHANG Li-na4，LIU Xiao-xia4，MA Xiu-min4，WEN Hao1，DING Jian-bing1，4 *

2012, 30(1):  19-78-80. 

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Specific primers were designed and synthesized based on the reported EgA31 gene of Echinococcus granulosus （GenBank Accession No. AF067807）. Total RNA was extracted from E. granulosus and its EgA31 gene was amplified by reverse transcription-polymerase chain raction （RT-PCR）. The PCR product was purified and cloned into plasmid pUCM-T，then transformed into Escherichia coli DH5α. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. The positive recombinant plasmid pUCM-T/EgA31 was confirmed by sequencing and homology comparison. Five parameters and methods were used to predict B-cell epitopes in amino acid sequence of EgA31. The amplified DNA fragment （636 bp） had an identity of 100% with the EgA31 gene sequence of E. granulosus. B-cell and T-cell epitopes of EgA31 were probably at or adjacent to 32-79，79-95，105-124 and 141-154 in its amino acid sequence.

Two cases of misdiagnosed toxoplasmosis

LIU Yang，NING Wei-juan，ZHANG Xu，ZHENG Li-juan

2012, 30(1):  20-5,11. 

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