Table of Content

30 December 2011, Volume 29 Issue 6

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Malaria Situation in the People’s Republic of China in 2010

ZHOU Shui-Sen, WANG Yi, LI Yu

2011, 29(6):  1-401-403. 

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Reactive Epitope Analysis of AgB Subunit Antigens of  Echinococcus granulosus by Using Synthetic Peptides

JIANG Li, LI Xiong, ZHANG Yao-Guang, MA Xiao-Jiang, NIU Xin-Ling, HE Yan-Yan, FENG Zheng

2011, 29(6):  2-404-409. 

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Objective   To analyze reactive epitope of three subunit antigens AgB1，AgB2 and AgB4 of Echinococcus granulosus by using synthetic peptides.  Methods   Five synthetic peptides，KK36，RK30，B4-1，B4-2，and B4-3，derived from the sequences of AgB1，AgB2，and AgB4 subunit of E. granulosus，and the three recombinant subunits were used for the detection of serum antibodies by ELISA. A panel of 209 serum samples from patients with cystic echinococcosis （115），alveolar echinococcosis （54），cysticercosis （22），and healthy persons （18） was used in the study. The diagnostic efficiency of the recombinant subunits and peptides for serum detection was estimated and compared using receiver-operating characteristics （ROC） curve.  Results   The sensitivity and specificity of peptides KK36 and RK30 in patients with cystic echinococcosis were 89.2% and 62.5%，85.0% and 59.4%，respectively. Their diagnostic efficiency （84.8% and 80.4%） was similar to AgB1 and AgB2 antigen （84.5% and 81.2%）. The ROC curves of peptides KK36 and RK30 were well fitted by that of recombinant subunit AgB1 and AgB2. For the three peptides derived from AgB4 subunit，serum detection indicated that the diagnostic efficiency of B4-1，B4-2 and B4-3 were 49.4%，57.9%，and 77.4%，respectively. Peptides B4-3 showed best reactivity and B4-2 also showed certain reactivity to the sera from patients with cystic echinococcosis.   Conclusion   Peptides KK36 and RK30 contain the reactive epitope region of AgB1 and AgB2 subunits. B4-2 and B4-3 contain partial region of the reactive epitope of AgB4. The epitope region of AgB4 may be in the central and back part.

Inhibition of Invasive Growth and Metastasis of Hepatic Alveolar Echinococcosis by Anti-Osteopontin Antibody

ZHANG Shi-Jie, ZHANG Long, ZAO Jiang-Qiao, ZHANG Yong-Guo, WU Xiang-Wei, PENG Xin-Yu, CAO Yu-Wen, YANG Hong-Jiang, LV Hai-Long, SUN Gong, CHEN Xiao-Beng

2011, 29(6):  3-410-414. 

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Objective   To observe the inhibitive effect on invasive growth and metastasis of Echinococcus multolocularis by exogenous anti-osteopontin（OPN）antibody.  Methods   180 gerbils were infected with 20% E. multolocularis suspension（approximately 400 protoscoleces in 0.1 ml per gerbil）through abdominal opening injection in liver，and then divided into model group，experiment group and control group. Experiment group and control group each with 60 gerbils were injected via the tail vein with 0.15 ml of anti-OPN antibody（1 ∶ 32）and rabbit serum，respectively. All gerbils in the two groups received injections，with 2-day interval for the first seven injections，and then at 7-day interval for the remaining injections. Model group were without any treatment. The three groups were subdivided into six groups each with 10 gerbils. The gerbils from each group were sacrificed on day 1，20，40，60，80，and 100 after infection，respectively. Hepatic echinococcus cyst and metastasis tissue were observed. The expression of OPN was measured by immunohistochemistry staining（SP method）. Serum samples were collected at 100 d post-infection, and the content of OPN in  sera was measured by ELISA.  Results  There were no significant difference in cyst weight and metastatic rate of thoracic lymph node among the three groups at 1，20，40，60，and 80 d post-infection（P＞0.05），while at 100 d post-infection，cyst weight and metastatic rate of thoracic lymph node in experiment group［（7.28±0.38） g，20%］were lower than that of control group［（9.70±0.61） g，70%］and model group［（9.32±0.73） g，70%］（P＜0.05）. Expression of OPN was found at different levels in echinococcus cysts. OPN was located in the cytoplasm, and mainly distributed in the germinal layer. The OPN positive expression levels were not significantly different between experiment group and other groups on day 1，20，40，60，and 80 afer infection（P＞0.05）. At 100 d post-infection，OPN positive expression rate and serum OPN content in experimental group［20% and（30.90±2.25） ng/μl， respectively］was lower than that of control group［80% and （41.03±2.76） ng/μl］and model group［80% and （42.39±2.85） ng/μl］（P＜0.05）.   Conclusion   Anti-osteopontin antibody can reduce OPN concentration in hepatic echinococcus cyst and serum，and inhibit the invasive growth and metastasis of E. multolocularis.

Therapeutic Effect of Hepatic Artery Infusion with Albendazole Microspheres on Hepatic Alveolar Echinococcosis in Rats

FAN Yu-Xiang, LIN Wei-Xin, DI Li-Mu-La-Chi-·Ba-Wu-Dong, GU Dun-Feng, HU Xiao-Dong, ZHANG Hai-Xiao, JI Wei-Zheng, JIANG Chao, WEN Hao

2011, 29(6):  4-415-418. 

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Objective   To observe therapeutic effect of hepatic artery infusion of albendazole solid dispersionchitosan microspheres on hepatic alveolar echinococcosis （HAE） in rats.  Methods   After the establishment of hepatic alveolar echinococcosis model，30 rats were randomly divided into control group（A），blank microspheres group（B），and albendazole microspheres group（C） with 10 rats in each group. 0.3 ml normal saline，2.7 mg/kg blank microspheres and 2.7 mg/kg albendazole solid dispersionchitosan microspheres with 0.3 ml normal saline were injected through hepatic artery of rats in the groups of A，B and C，respectively. At 1 d，3 d，7 d，14 d，and 42 d after injection，venous blood were collected，and white blood cells （WBC），apartate aminotransferase （AST），and alanine aminotransferase（ALT）were evaluated. All the rats were sacrificed on 42 d after injection，and HAE pathological changes were observed.  Results   Transient elevation of white blood cells was observed in all groups at 1 d after infusion ［Group A （86.11±19.14）×109/L，B （117.11±21.76）×109/L，C （118.11±24.52）×109/L］，at 7 d WBC fell to normal level ［A （7.85±6.57）×109/L，B （11.73±4.85）×109/L，C （8.49±1.36）×109/L］. In groups B and C，AST，ALT reached their peaks on day 3 after infusion ［B：AST （193.15±21.57） U/L，ALT （78.39±9.78） U/L；C：AST （189.91±14.06） U/L，ALT （88.43±9.23） U/L］， and decreased to normal level at 14 d after infusion ［B：AST （109.31±15.48） U/L，ALT（47.855±9.49） U/L；C：AST（105.37±8.16） U/L，ALT（49.53±6.75）U/L］. Histopathological examination at 42 d after infusion showed that in groups A and B structure of the cysts was virtually normal，but in group C most cysts showed necrosis in germinal layer.   Conclusion   Hepatic artery infusion with albendazole solid dispersionchitosan microspheres shows certain therapeutic effect on hepatic alveolar echinococcosis in rats.

Effect of Different Drugs on Echinococcus multilocularis Metacestodes in vitro

MAI Hepiretihan·Ai Erken，WANG Yun-hai，ZHAO Jin-ming，BAI Lei

2011, 29(6):  5-419-424. 

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Objective   To test the effect of 6 drugs or combination on the in vitro cultured Echinococcus multilocularis（Em）metacestodes.  Methods   The drugs used included albendazole（ABZ），itraconazole（ITZ），ivermectin，miltefosine，nitazoxanide（NTZ），and rifampin. The Em metacestodes were cultured for 5 weeks and divided into 17 groups each with 120-140 cysts：ABZ groups（1 μg/ml and 10 μg/ml），ITZ group（0.7 mg/ml），ivermectin group（1.75 mg/ml），miltefosine groups（0.5 μg/ml，2.5 μg/ml and 7.5 μg/ml），NTZ groups （0.1 μg/ml，1 μg/ml and 10 μg/ml） and rifampin group（10 μg/ml）；groups of NTZ combined with ABZ at respective concentration （10 μg/ml +10 μg/ml，10 μg/ml+1 μg/ml，1 μg/ml+10 μg/ml，1 μg/ml+1 μg/ml）；and 2 groups of control： ① experiment culture containing 40 μl of dimethyl sulfoxide（DMSO） per 20 ml and ② blank culture medium. In the 6 weeks culture period，the drug effect on the Em cysts was observed. Drug combination groups were continually observed for 3 weeks，3 months and 6 months after discontinuation of drugs. In order to determine the viability of metacestodes，the cysts were injected intraperitoneally into female BALB/c mice，and 8 weeks later，the mice were sacrificed for collecting the parasites.   Results   Ivermectin，miltefosine and rifampin showed no effect on metacestodes. Albendazole，itraconazole，nitazoxanide and albendazole combined with nitazoxanide all inhibited or killed the parasites in the in vitro culture system（P＜0.05）. High dose of nitazoxanide（10 μg/ml） was more effective in the destroying parasites than others. No re-growth of cysts was observed in the 4 groups of combination treatment with nitazoxanide and albendazole continually cultured for 3 weeks，3 months，or 6 months after drug discontinuation. In the in vivo viability test，no metacestodes were found in all mice in the nitazoxanide group（10 μg/ml） and 2 mice in albendazole group，but the parasites developed in all mice at 8 weeks after injection in other single drug groups. Also，no metacestodes from all the combination treatment groups were found re-developed in mice.   Conclusion   This investigation indicates certain inhibitory effect of itraconazole and nitazoxanide on Em cysts，and a combination of albendazole with nitazoxanide may enhance the parasitocidal effect.

Inhibition of Culture Supernatant of Mesenchymal Stem Cells on Macrophages RAW264.7 Activated by Soluble Egg Antigen of Schistosoma japonicum

XU Hui-Juan, QIAN Hui, ZHU Wei, ZHANG Xu, YAN Yong-Min, ZHANG Lei-Lei, MAO Fei, HU Wen-Rong

2011, 29(6):  6-425-430. 

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Objective  To observe the inhibitive effect of rat mesenchymal stem cells (MSC) culture supernatant on macrophages activated by soluble egg antigen （SEA） of Schistosoma japonicum.  Methods   To select optimal SEA effecting concentration and time，macrophages RAW264.7 were induced by 5，10，20 or 40 μg/ml SEA for 12 h，or by 20 μg/ml SEA for 4，8，12 or 24 h before examination of TNF-α mRNA by RT-PCR. Macrophages were divided into five groups，i.e. negative control group，SEA group，SEA+MSC supernatant group （MSC group），SEA+NRK-52E supernatant group （NRK-52E group） and SEA+DMEM group （DMEM group）. Except negative control group, macrophages in other four groups were induced by 20 μg/ml SEA for 12 h. SEA was then removed from MSC group，NRK-52E group and DMEM group and replaced with MSC supernatant，NRK-52E supernatant and DMEM，respectively. Morphology of macrophages in each group was observed by microscope after cultured with supernatant for 12 h. TNF-α mRNA in macrophages was detected by real-time quantitative PCR after cultured with supernatant for 12 h and 24 h. TGF-β1 in macrophages was observed by Western blotting analysis after cultured with supernatant for 12 h. Macrophage proliferation was tested by MTT method after cultured with supernatant for 24 h and 48 h.  Results  The optimal SEA concentration and time for macrophage activation was 20 μg/ml and 12 h，respectively. Compared with SEA group，NRK-52E group，and DMEM group，macrophages in MSC group were round and small with less pseudopodia after cultured with supernatant for 12 h. TNF-α mRNA after cultured with MSC supernatant for 12 h and 24 h was （1.0±0.4） and （1.0±0.5） fold of negative control group，respectively，significantly less than NRK-52E group ［（10.4±3.9） and （16.5±5.0） fold］ （12 h: P<0.05；24 h：P<0.01） and DMEM group ［（6.0±2.1） and （2.4±0.7） fold］（P<0.05）. The grey density image analysis of TGF-β1/GAPDH was 0.31±0.10 in MSC group，much lower than 0.88±0.10 in NRK-52E group （P<0.01） and 0.58±0.06 in DMEM group （P<0.05） after cultured with supernatant for 12 h. After 48 h culture,  A490 of macrophages in MSC group was 0.22±0.05，much lower than 0.53±0.02 in NRK-52E group and 0.31±0.03 in DMEM group （P<0.05）.   Conclusion   MSC supernatant can inhibit activation and proliferation of macrophages which were induced by SEA of S. japonicum.

Construction of Eukaryotic Recombinant Plasmid of Periodic Brugia  malayi CPI Gene and its Immunity

ZHANG Sai-Nan, FANG Zheng, LIU Shi-Juan, WANG Hui, XU Bang-Sheng

2011, 29(6):  7-434-438. 

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Objective   To observe the immune responses elicited in BALB/c mice by DNA vaccine encoding cysteine protease inhibitor （CPI） of periodic Brugia malayi cloned in vector pcDNA3.1.  Methods   Specific primers were designed on the basis of known sequences of CPI gene from periodic B. malayi. The desired gene fragment was amplified by PCR from cDNA, inserted into cloning vector，pGEM-T，and sub-cloned into pcDNA3.1 to construct pcDNA3.1-BmCPI. Forty-eight mice were randomly divided into 4 groups, i.e. normal control group， pcDNA3.1(+) group， pcDNA3.1-BmCPI group， and pcDNA3.1-BmCPI/CpG group injected with PBS 100 μl， pcDNA3.1 100 μg， pcDNA3.1-BmCPI 100 μg and pcDNA3.1-BmCPI 100 μg+CpG 30 μg， respectively on left hind leg of each mouse. All mice received three immunizations with 2-week interval. At the 4th week after the last immunization the muscle around injection spot was collected， in which the level of BmCPI mRNA was detected by RT-PCR. The stimulation index (SI) of spleen lymphocytes was measured by MTT method and the levels of secreted IL-4 and IFN-γ in serum were detected by ELISA.   Results   The recombinant plasmid pcDNA3.1-BmCPI was constructed and the length of the gene fragment was 621 bp. The results showed that BmCPI gene in the muscle of the immunized mice was detected by PCR. At the 4th and 6th weeks after immunization， the SI of the two immunized groups was significantly higher than normal control group and pcDNA3.1(+) group (53.789±1.937, 59.735±4.139, and 61.975±1.029) (P<0.05). No significant difference existed between pcDNA3.1-BmCPI group and pcDNA3.1-BmCPI/CpG group (P>0.05). Serum IFN-γ in pcDNA3.1-BmCPI group and pcDNA3.1-BmCPI/CpG group increased from the 2nd to the 6th week after the last immunization with the value of 69.544±3.145 and 106.069±7.518，120.019±5.968 and 136.229±7.198，149.109±2.700 and 178.429±1.126，respectively. The levels of IFN-γ in serum from the immunized mice were significantly higher than those of normal control group and pcDNA3.1(+) group (28.264±1.129, 35.179±1.029, and 40.110±1.176， respectively) （P<0.05）. There was a significant difference between the two immunized groups at the 2nd and the 6th weeks after the last immunization （P<0.05）. The level of IL-4 in serum from the immunized mice was significantly higher than those of normal control group and pcDNA3.1（+） group at the 4th and the 6th weeks after the last immunization （P<0.05）. No significant difference was noted in IL-4 level between pcDNA3.1-BmCPI group and pcDNA3.1-BmCPI/CpG group （P>0.05）.   Conclusion   The recombinant eukaryotic plasmid pcDNA3.1-BmCPI was transcripted in vivo and elicited immune responses in mice.

Stable Green Fluorescent Protein Expression in Toxoplasma gondii Mutant

WU Liang, YUAN Yi-Wei, JIANG Xu-Gan, FU Hang-Li, DAI Zhao-Chi, CHU Guo-Hua, LI Li, CHEN Cheng-Xia, ZHOU Gong

2011, 29(6):  8-439-442. 

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Objective   To construct a Toxoplasma gondii mutant for stably expressing green fluorescent protein （GFP），and establish method to determine the rate of mutant-infected HeLa cells.  Methods   Freshly lysed-out tachyzoites of T. gondii RH strain were transfected with plasmid ptubP30-GFP/sag-CAT. Stable transformants were selected with chloramphenicol and limited dilution. The expression of GFP in mutant tachyzoite was determined by RT-PCR and fluorescence microscopy. When infected with 1×10⁴-1×10⁷ mutant tachyzoites respectively for 24 h，the total number of HeLa cells with green fluorescence was determined by fluorescent microscope in 10 high-power fields，and the rate of HeLa cells with parasitophorous vacuole was determined by flow cytometry.  Results   Untransfected tachyzoites were killed by chloramphenicol，while the stable transformants showed resistance to chloramphenicol. The expression of GFP gene was detected by RT-PCR. The P30-GFP transfectants displayed fluorescence outside the parasite. The rate of mutant-infected HeLa cells increased with the rise of the number of mutant for infection. When infected with 1×104-1×107 tachyzoites，the numbers of HeLa cells with fluorescence were （14±6），（133±45），（332±93）  and （443±90），and the rates of infected cells were （0.49±0.09）%，（8.76±0.50）%，（21.02±1.49）% ，and （39.00±3.47）% by flow cytometry，respectively.   Conclusion   T. gondii mutant with GFP tag is constructed，which provides a new method to determine the proliferation when cultured in host cells.

Taxonomic Status of Anopheles yatsushiroensis in Rongcheng City of Shandong Province

ZHOU Xiao-Jun, SHI Wen-Qi, ZHANG Yi, YUE Ai-Ping

2011, 29(6):  9-443-446. 

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Objective  To clarify the taxonomic status of Anopheles yatsushiroensis in Rongcheng City of Shandong Province.  Methods   By the end of August 2009，Anopheles yatsushiroensis were collected in the western suburbs of Wangjia Village of Rongcheng City，and raised for the next generation of various types of adult mosquitoes. Through morphological identification of filial generation various types, mosquito feet of 1-2 mosquitoes  of various types were taken for complete DNA extraction，and complete sequence analysis of rDNA-ITS2 alignment was made by PCR. Multiple sequnence comparison was carried out with those previously documented by DNAstar7.1，including An. yatsushiroensis in Shandong Province（YSD，AY306128），An. yatsushiroensis in Sichuan Province（YSC，AY170925），An. yatsushiroensis in Liaoning Province（YLN，AY170923） and An. yatsushiroensis from South Korea （YK，AF146749）.  Results    56 An. yatsushiroensis with blood meal were collected in Rongcheng City，of which 7 successfully laid eggs, and received 354 adult mosquitoes of filiar generation，including 240 An. yatsushiroensis （Y type），8 An. pullus （P type） and 106 hybrid type（H）. The rDNA-ITS2 sequence alignment homology between single-parent female offspring of adult mosquitoes and various types of An. yatsushiroensis segments （JN865249，JN865246，JN865247，JN865248）was 98.7-100%，and the rDNA-ITS2 sequence alignment homology to those from Sichuan（YSC），Liaoning （YLN） and Korea （YK） was 98.5%-99.3%，98.7%-99.6% and 98.7 to 100%，respectively, while to the original An. yatsushiroensis from Rongcheng（YSD）, it was only 67.1%-68.5%.   Conclusion   There are Y，P and H types of An. yatsushiroensis in Rongcheng City，Shandong Province，but the rDNA-ITS2 sequence is close to An. pullus. The local distribution of An. pullus is deduced.

Isolation and Identification of Cow-origin Cryptosporidium Isolates in Hefei

SUN Tao, LIU Wei, WANG Ju-Hua, XUE Xiu-Heng, DIAO Chang-Cheng, LI Pei-Yang

2011, 29(6):  10-447-452. 

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Objective   To isolate cow-origin Cryptosporidium in Hefei，and identify its species.  Methods   285 dairy cattle fecal samples collected from a farm in Hefei were examined by using floating saturated solution of sucrose and modified acid-fast staining. Cryptosporidium oocysts were isolated and purified from positive fecal samples. Genetic DNA was extracted to be the template. According to the sequence of 18S rRNA gene and HSP70 gene from Cryptosporidium sp.，the primers were designed and synthesized. The PCR products were amplified by PCR and nested-PCR. The nested PCR products were cloned and sequenced. Homology searches and phylogenic tree construction were done by DNAStar software.  Results   Five fecal samples were positive by morphological methods with an infection rate of 1.8%（5/285）. Oocysts from the 5 positive fecal samples were elliptical or ovoid detected by using floating saturated solution of sucrose and modified acid-fast staining with the size of 7.37 μm×6.13 μm and 7.58 μm×6.20 μm，and a shape index of 1.20 and 1.22，respectively. Nested-PCR resulted in a 18S rRNA and HSP70 gene fragments with approximately 250 bp and 325 bp，respectively. The five isolates showed a high level of nucleic acid identity with sequence data of the 18S rRNA gene of Cryptosporidium andersoni （DQ989573），and they were clustered in the same clade. The highest HSP70 gene sequence identity was found among the five isolates and other reported C. andersoni isolates （AY954892 and DQ989576），and they were placed into the same clade.   Conclusion   The cow-origin Cryptosporidium isolates derived from Hefei is Cryptosporidium andersoni.

Isolation and Purification of Oncomelania hupensis Agglutinin and Determination of its Molecular Weight

ZHOU Shu-Lin, PENG Huai-ming, LI Chao-Pin, LIU Hui, ZHAO Jin-song, CAO Zhi-Guo

2011, 29(6):  11-453-457. 

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Objective   To isolate and purify agglutinin from Oncomelania hupensis snail and determine its molecular weight.  Methods   Agglutinin was preliminarily isolated from snail tissue homogenate by 0%-40% saturated ammonium sulfate，and then successively purified with Sephadex G-75 gel filtration and Sepharose 4B affinity chromatography. Bradford assay was used to determine the protein content. The agglutination activity was determined by rabbit erythrocytes. The purity of agglutinin preparations was assessed by SDS-PAGE. The molecular weight of agglutinin subunit was determined by Sephadex G-75 gel filtration. Results   The specific activity of snail tissue homogenate was 21.74 titer/mg. After ammonium sulfate precipitation， Sephadex G-75 gel filtration and Sepharose 4B affinity chromatography， the specific activity of snail agglutinin from the homogenate solution increased to 61.93 titer/mg， 75.89 titer/mg and 963.86 titer/mg， respectively. SDS-PAGE analysis indicated that snail agglutinin （M_r 53 000） was purified by Sephadex G-75 gel filtration and Sepharose 4B chromatography. The molecular weight of the snail agglutinin produced by Sephadex G-75 gel filtration was M_r 78 000.   Conclusion   Combined use of salt fractionation，gel filtration and affinity chromatography can be efficient for extraction and purification of agglutinin from Oncomelania hupensis species. The snail agglutinin is characterized as monosubunit protein with a molecular weight of M_r 78 000.

Detection and Species Identification of Two Imported Cases of Cutaneous Leishmaniasis

YANG Yue-Tao, ZHANG Min, GAO Chun-Hua, SHI Feng, GUAN Li-Ren, WANG Jun-Yun

2011, 29(6):  12-461-464. 

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Objective   To diagnose and identify pathogen of two suspected cases of cutaneous leishmaniasis.  Methods   Two cases of dermatosis with several major ulcers on the skin were examined， who worked and returned from Algeria （case 1） and Saudi Arabia （case 2）， respectively. The stained smears of skin tissue from lesions were observed by microscope. Extravasate from lesions was cultured in NNN medium to search protozoan parasites， which were obtained by centrifugation. Two pairs of species-specific primers， ITS1-ITS2 and K13A-K13B， were used to amplify internal transcribed spacer of rDNA and kinetoplast DNA， respectively. The products were sequenced and analyzed by Blast.  Results  There were Leishmania amastigotes in the tissue smear of case 2， while none in that of case 1. Promastigotes were found in culture medium of both cases. The PCR products of ITS1-ITS2 and K13A-K13B from 2 cases were about 330 bp and 120 bp with respective homology of 100% and 96% to corresponding sequences of Leishmania major. The accession numbers of 4 sequences were JF831924-JF831927.   Conclusion   Two cases of dermatosis are diagnosed as imported cutaneous leishmaniasis and the pathogen is L. major.

Transcriptional Regulation by Silence Information Regulator 2

ZHANG Xia, JU Hong-Mei, LI Ya-Jie

2011, 29(6):  13-465-468. 

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Silence information regulator 2（Sir2）family is a group of conserved deacetylases，widely existed in organisms from archaea to mammals，and characterized by NAD+-dependent deacetylase and ADP-ribosyltransferase activities. Sir2 plays an important role in various life progresses，such as chromatin silence，gene regulation，metabolic regulation，life span of cells and so on. Through its deacetylase activity and/or interaction with other proteins，Sir2 can regulate chromatin structure or modify transcription related factors，thus regulating the transcriptional process. This article emphasizes on the progress of Sir2-dependent regulation of gene transcription.

Progress in Transgenics on Toxoplasma gondii

ZHANG Yan-Lei, ZHANG Hou-Shuang, ZHOU Jin-Lin

2011, 29(6):  14-469-472. 

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The application of transgenetics in Toxoplasma gondii research facilitates its genetic analysis. This article reviews the progress in construction of transgenetic vector, the protocol of electroporation and application of transgenics in the research of T. gondii.

Research Progress on the Role of Schistosomiasis in Regulating  Autoimmune and Allergic Diseases

DU Jiu-Wei, WANG Xue-Feng

2011, 29(6):  15-473-476,479. 

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Schistosome infection down-regulates the Th1 cell-mediated autoimmune diseases and Th2 cell-mediated allergic diseases. It was revealed recently that a novel pathogenic T cell subset （Th17） was also involved in the pathogenicity of autoimmune and inflammatory diseases, and schistosome infection was reported to suppress Th17 response in autoimmune diseases. Here we summarize research advances on the effect of schistosome infection on Th1-，Th2-，Th17-mediated autoimmune or allergic diseases，and discuss the possible mechanisms of schistosome-induced suppression.

Pattern Recognition Receptors for Recognizing Hemozoin

QIAN Feng, XU Hu-Ji

2011, 29(6):  16-477-479. 

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Hemozoin（malaria pigment）is a byproduct of malaria parasites due to hemoglobin digestion，which can stimulate host′s innate inflammatory responses. However，data from different studies are controversial about how hemozoin is recognized by the host′s pattern recognition receptors. This article reviews the recent progress in the area.  

Reform and Practice on the Experiment Teaching of Medical Parasitology

ZHAO Jin-Gong, TANG Xiao-Niu, GAO Ti-Yin, WANG Shao-Ku, LI Chao-Pin

2011, 29(6):  17-430-433. 

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A new model of education is investigated to meet the new idea of experiment teaching in university. Therefore the establishment of experiment teaching model of medical parasitology needs to be correspondently reformed. A variety of new management measures are taken to raise the efficiency of experiment teaching in training the students in the College.

Effect of N-acetylcysteine on Malondialdehyde and Superoxide Dismutase in Hepatic Tissue of Mice with Schistosomiasis japonica

FAN Zhi-Gang, LI Kai-Jie, ZHANG Ling-Min

2011, 29(6):  18-457-460. 

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90 mice were randomly divided into six groups: nomal control，infected control，long-term drug use group 1(L1)，long-term drug use group 2(L2)，short-term drug use group 1(S1)and short-term drug use group 2(S2). Mice in all groups except those in the nomal control group were infected with 30 cercariae of Schistosoma japonicum through abdominal skin. N-acetylcysteine（NAC）solution was orally given to mice in L1 and L2 groups，200 mg/kg and 400 mg/kg，respectively，2 times/d from the day of infection，while for S1 and S2 groups，200 mg/kg and 400 mg/kg，respectively，2 times/d from the 42th day after L2 infection. Mice in the groups of nomal control，infected control，L1 and L2 were sacrificed either on day 42 or day 56 after infection，while those in S1 and S2 were sacrificed on day 56 after infection. Number and area of the single egg granuloma were measured with computer image analysis software. The concentration of malondialdehyde（MDA）and the activity of superoxide dismutase（SOD）in serum and hepatic tissue were detected. The number of“+”single egg granulomas in the liver of mice in L1 was the fewest by 3.04，followed by those in S1，by 4.87. The results indicated that the level of MAD in hepatic tissue of L2（9.2-9.3 nmol/mg）was markedly lower than that of L1（P＜0.05），and the level of SOD in hepatic tissue of L1 was 170.00-190.00 U/(g·pro)，similar to those of S1 and L2 at the 42th day（P＞0.05），but the level in L2 at the 56th day was close to that of S2（P＞0.05）. Hence，NAC may retard the formation of single egg granulomas in the liver of infected mice，and may regulate the concentration of MDA and the activity of SOD in the liver.

Investigation of Pinworm Infection among Kindergarten Children in Ninghai County

GU Min-Xia, XU Zhi-gang

2011, 29(6):  19-480-481. 

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6 257 children in 39 kindergartens were examined by adhesive tape wiping in the morning during April，2009 to November，2010， and 460 children were found pinworm infected with a prevalence of 7.4%（460/6 257）， 7.7%（260/3 359）in girls and 6.9%（200/2 898）in boys. Children in junior class had lowest infection rate of 4.6%（77/1 667），while children in senior class had highest rate of 9.7%（252/2 594）（χ2=36.8，P<0.01）. Furthermore, the rate in public kindergartens（6.5%，255/3 927） was lower than that in private kindergartens（8.8%，255/2 330）（χ2=11.4，P＜0.01）, and the rate in urban kindergartens（12.4%，199/1 611）was much higher than that in rural kindergartens（5.6%，261/4 646）（χ2=79.7，P＜0.01）. Evidently，pinworm prevalence is high in children in Ninghai, and it is higher in private and urban kindergartens than in public and rural ones respectively.