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    30 April 2014, Volume 32 Issue 2
    Progress and Challenges of the National Schistosomiasis Control Program during the Period of the 12th Five-Year Plan
    LEI Zheng-Long, ZHOU Xiao-Nong
    2014, 32(2):  1-81-85. 
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    The achievements of the national schistosomiasis control program during the period of the 12th Five-Year Plan were reviewed, in particular, the reduction of the prevalence and progress in control activities were evalucated among different regions of China. Moreover, current difficulties of schistosomiasis control and gaps to achieve the transmission interruption of the disease in China were analyzed, which provide more evidences to formulate the future efforts and work-plan to eliminate the disease in the country.
    Prokaryotic Expression and Function Analysis of Schistosoma japonicum Calpain
    DU Xiao-feng1,XU Bin2,LIU Jian1,WANG Ji-peng1,LIU Mu1,LIU Xiu-feng1,MA Xiao-lin1,HU Wei1,2,JU Chuan2 *
    2014, 32(2):  2-86-95. 
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     Objective  To clone and express recombinant calpain of Schistosoma japonicum(Sjcalpain), observe the distribution of Sjcalpain in S. japonicum cercariae and analyze its role in skin invasion.  Methods  The primers were designed according to the full-length sequence of calpain(GenBank accession No. AB016726). The genes encoding catalytic domain and Ca2+ binding domain of Sjcalpain were amplified by PCR, and the target fragments were subcloned into pET-28a. The recombinant proteins were expressed in E. coli BL21(DE3)and purified by Ni-NTA resin. The rabbit polyclonal antibodies were prepared with the two purified recombinant proteins by immunizing New Zealand white rabbits. ELISA was used to detect the titer of rabbit antiserum. Immunolocalization was used to investigate the distribution of Sjcalpain in S. japonicum cercariae. Cercariae were incubated with specific inhibitor before infection of mice and the worm reduction rate was calculated.  Results  The recombinant expression vector Sjcalpain catalytic domain/pET28a and Sjcalpain Ca2+ binding domain/pET28a were constructed and the recombinant proteins were successfully expressed in E. coli BL21(DE3)(about Mr 43 000 and Mr 39 000, respectively). The two target proteins were expressed as inclusion bodies. The purified target proteins were obtained through Ni-NTA affinity purification. ELISA result showed that the titer of prepared rabbit polyclonal antibodies was higher than 1 ∶ 80 000. Immunolocalization study demonstrated that Sjcalpain protein was mainly expressed in the head of cercariae. Inhibition assays suggested that the average number of adult worms in calpain inhibitor-incubation group and control group was 19 and 23, respectively, with a worm reduction rate of 17.4%.  Conclusion  Sjcalpain is mainly expressed in the head of S. japonicum cercariae. Inhibition of Sjcalpain could reduce the number of invading cercariae in infected mice, which suggest that Sjcalpain may play a role in skin invasion by cercariae.
    Cloning, Expression and Transcription Specificity Analysis of Two Tyrosinases from Schistosoma japonicum
    LI Na-na1,WANG Ji-peng2,3 *,DU Xiao-feng2,3,HU Wei2,3
    2014, 32(2):  3-95-100. 
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    Objective  To clone and express the recombinant proteins based on the whole open reading frame of two tyrosinases(tyrosinase 1 and tyrosinase 2) from Schistosoma japonicum, and study the transcription specificity of the two tyrosinases in different sex and developmental stages of S. japonicum.  Methods  The full-length of SjTYR1 and SjTYR2 were amplified with specific primers and subcloned into pSJ2. The recombinant plasmids were transformed into E. coli Rosetta Gami strains and induced with IPTG for expression. The recombinant proteins were purified by Ni-NTA agarose. The recombinant proteins SjTYR1 and SjTYR2 were used to produce the specific antibodies by immunizing the rabbits. The immunogenicity of the recombinant proteins SjTYR1 and SjTYR2 were detected by Western blotting using sera of recombinant proteins-immunized rabbits and S. japonicum-infected rabbit serum as the primary antibody, respectively. The reactivity of sera from recombinant proteins-immunized rabbits was analyzed by Western blotting against the native protein of S. japonicum worm. Total RNA was extracted from 14, 16, 18, 20, 22, 24, 26, and 28-day male and female worms. Transcription levels of the two tyrosinases in different sex and different stage were determined via RT-PCR method.  Results  The expression vector of SjTYR1/pSJ2 and SjTYR2/pSJ2 were constructed and the recombinant proteins SjTYR1 and SjTYR2 were expressed in inclusion body in E. coli(about Mr 55 000 and Mr 56 800). The sera of S. japonicum-infected rabbits reacted positively with the purified recombinant protein SjTYR1, but not with recombinant protein SjTYR2. The native protein of S. japonicum worm could be recognized by sera of rSjTYR1-immunized rabbits (Mr 100 000), but not by sera of rSjTYR2-immunized rabbits. Transcription levels of the two tyrosinases in male worms were nearly zero. In female worms, the transcription levels of the two tyrosinases increased sharply from the 24th day post-infection and reached maximum on the 28th day.  Conclusion  The recombinant proteins of SjTYR1 and SjTYR2 show immunogenicity and immunoreactivity. SjTYR1 and SjTYR2 are both expressed specifically in female worms and the transcription levels increase in 24-28 days after infection.
    Expression of Toll-like Receptors 2 and 6 in Mice Liver during Schistosoma japonicum Infection
    TANG Juan-juan, JI Xiao-fang , ZHU Xun-min, HUANG Hua-yi, LI Zi*
    2014, 32(2):  4-101-105. 
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    Objective  To investigate the expression of hepatic Toll-like receptor 1(TLR1), TLR2 and TLR6 on mice with Schistosoma japonicum infection.  Methods  Fifty BALB/c mice were infected with 20±3 S. japonicum cercariae through abdominal skin. At 6 weeks post-infection, the mice (n=10) in treatment group were administered intragastrically with praziquantel [250 μg/(g·d)] for 3 d. The livers of mice (n=10) were collected at pre-infection and 5, 6, 8 and 12 weeks post-infection, and then the mRNA expression levels of hepatic TLR1, TLR2, TLR6 gene were detected with reverse transfer PCR. Hepatic TLR2, TLR6 protein levels were detected by immunohistochemical staining.  Results  The mRNA levels of TLR1, TLR2, and TLR6 on 5, 6, 8 and 12 weeks post infection were significantly higher than that of uninfected mice. After praziquantel treatment, the mRNA level of TLR2 and TLR6 in murine liver of treatment group was lower than that of infection group, but the level of TLR1 mRNA had no obvious change. Furthermore, immunohistochemistry results revealed that the expression of TLR2 and TLR6 proteins in murine liver was up-regulated at 5, 6, 8 and 12 weeks post-infection. After praziquantel treatment, the percentage of TLR2 positive area in liver of infected mice without and with praziquantel treatment were (44.2±4.3)%, (8.8±3.1)%, respectively, and TLR2 protein level was considerably down-regulated(P<0.01). The percentage of TLR6 positive area in liver of infected mice without and with praziquantel treatment was (48.4±5.4)%, (37.4±3.5)%, respectively, and TLR6 level decreased slightly(P<0.05).  Conclusion  The expression level of TRL2 and TLR6 in murine liver increases after Schistosoma japonicum infection. While compared with TLR2, the role of TLR6 in this progress is a weaker one.
    Changes of IFN-γ, IL-4 and T Cells in Schistosoma japonicum- infected Mice after Praziquantel Treatment
    LIU Xiang-qin1,2,TIAN Xi-feng1,ZHANG Jin2,XIN Xiao-fang2,ZHANG Ying2 *
    2014, 32(2):  5-106-109. 
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    Objective  To investigate the serum levels of IFN-γ and IL-4, and the dynamic changes of IFN-γ-specific and IL-4-specific lymphocytes in mice with Schistosoma japonicum infection after treatment by praziquantel.  Methods  Ninety BALB/c mice were randomly divided into three groups(n=30) named as infection group, treatment group and control group. The mice in treatment group and infection group were infected with (25±2) S. japonicum cercariae through the abdominal skin. At 6 weeks post-infection, the mice in treatment group were administered orally with praziquantel[300 mg/(kg·d)] for 3 d. At 4, 6, 8 and 12 weeks post-treatment, the mice were weighed, and serum samples were collected. Serum levels of IFN-γ and IL-4 were measured by ELISA. At the same time, the spleens were aseptically removed to prepare cell suspension, and the counts of IFN-γ and IL-4 specific lymphocytes were examined by ELISPOT after stimulation of Schistosoma japonicum soluble egg antigen(SEA).  Results  From 4 to 12 weeks after praziquantel treatment, the body weight of mice in treatment group were significantly heavier than that of infection group (P<0.05), but no significant difference was found between treatment group and control group(P<0.05). At 4 weeks post-treatment, there was no significant difference in serum levels of IFN-γ and IL-4 between treatment group and infection group(P>0.05). At 6, 8, and 12 weeks after treatment, the serum levels of IFN-γ(0.038±0.013, 0.028±0.001, and 0.027±0.007) and IL-4(0.051±0.020, 0.045±0.019, and 0.043±0.016) in treatment group were significantly lower than that of infection group(IFN-γ: 0.057±0.004, 0.060±0.023, and 0.052±0.017; IL-4: 0.150±0.014, 0.148±0.014, and 0.123±0.017)(P<0.05). Serum IFN-γ and IL-4 levels in treatment group and infection group were significantly higher than that of control group(P<0.05). ELISPOT results showed that at 4, 6 weeks post-treatment, there was no significant difference in the number of IFN-γ-specific lymphocytes between treatment group and infection group(P>0.05). While at 8 and 12 weeks after treatment, the IFN-γ-specific lymphocytes in treatment group (39.9±22.8 and 38.5±6.2) were significantly less than that of infection group (141.9±39.3 and 106.8±28.6)(P<0.05). At 4-week post-treatment, the IL-4-specific lymphocytes in treatment group were much more than that of infection group(175.6±62.3)(P<0.05), and then began to decline. At 8 and 12 weeks after treatment, the IL-4-specific lymphocytes (111.3±14.3 and 113.0±44.2) in treatment group were sig-nificantly less than that of infection group(220.3±107.1 and 208.1±17.2)(P<0.05). The IFN-γ-specific and IL-4-specific lymphocytes in treatment group and infection group were significantly more than that of control group(P<0.05).  Conclusion  After praziquantel treatment, the serum levels of IFN-γ and IL-4 in mice with S. japonicum infection decrease, and the number of IFN-γ and IL-4 specific lymphocytes reduces.
    Application of Space-time Scan Statistics in the Analysis of Spatial and Temporal Distribution of Oncomelania hupensis Snails in Gaoyou County,Jiangsu Province
    WANG Qiang1,GAO Jin-bin2,XU Jing1,HUANG Ya-min2,HE Yong2,GAO Yang3,YANG Kun3,QIAN Ying-jun1,FU Qing1,LI Shi-zhu1,ZHOU Xiao-nong1 *
    2014, 32(2):  6-110-115. 
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    Objective  To investigate the distribution features of Oncomelania hupensis infested areas in Gaoyou County so as to formulate surveillance and intervention strategies.  Methods  A database was established through collecting data of the snail infested areas during 1970-2009 in the County. The data were input into SaTScan 9.2 software for spatial-temporal cluster analysis to determine the spatial and temporal cluster of the snail habitats. The results were displayed by ArcGIS 10.1 software.  Results  There were historically 720 snail habitats in the County in 1970-2009 including 521 in plain region with water networks and 199 in lake & marshland region. Those in water networks covered an area of 456.62 ha distributing mainly in the northern towns/townships of the County, and the latters distributed in the Xinmin Beach between Gaoyou Lake and Shaobo Lake, and Qiaojian Beach close to Tianchang County of Anhui Province with an area of 4 495.75 ha. The spatial-temporal cluster analysis revealed that among all the historical snail habitats, there were two prominent spatial-temporal clusters with a relative risk of >3. One cluster appeared in Xinmin Beach in 1983-2002 and another one located in the north of Gaoyou in 1970-1973. Separate analysis was performed by the regions of water network or lake & marshland, indicating 2 clusters in each of the regions. During 1970-2009, 244 snail habitats were newly found in the County with 130 in water network region and 114 in lake & marshland region. Again, the spatial-temporal cluster analysis displayed 2 prominent clusters. By separate analysis, 2 clusters existed in each of the regions.  Conclusion  The space-time scan statistics can be applied in detecting the cluster of snail infested areas in two dimensions, which will provide information for guiding specific measures of surveillance and control.
    Diagnostic Potential of Five Natural Antigens from Echinococcus granulosus in the Patients of Cystic Echinococcosis
    JIAO Wei1 *,FU Cheng1,LIU Wan-li1,WANG Ying1,GAO Chun-hua2,CHAI Jun-jie1
    2014, 32(2):  7-116-122. 
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    Objective  To analyze the characteristics of serum antibody reactivity of cystic echinococcosis (CE) patients with different clinical status towards five native antigens obtained from Echinococcus granulosus (Eg).  Methods  The protoscolex somatic soluble antigen (EgPS), crude hydatid cyst fluid antigen (EgHF), partially purified hydatid fluid antigen (Burstein′s antigen, EgBu), adult somatic soluble antigen (EgAs) and the native antigen B (EgAgB) were prepared. 369 serum samples from CE patients and 281 sera samples from healthy individuals were examined for the antibodies against 5 native antigens with indirect ELISA. The serologic results were classified according to clinical status, and the statistical analyses were carried out to understand the relationship between the results of different antigen-ELISA and the clinical status of patients.  Results  The results of EgBu, EgAS and EgAgB-ELISA showed that the antibody positive rate in hepatic CE patients [74.1% (212/286), 73.4% (210/286), 63.6% (182/286)] was significantly higher than that of other groups (including renal CE and pelvic CE, 1/8, 2/8, 1/8)(P<0.05). Except EgAS, the S/N value of other groups examined by the rest four antigen-ELISA (EgPS: 3.10, EgHF: 2.40, EgBu: 1.60, EgAgB: 2.38) was also significantly lower than that of hepatic CE patients (3.73, 3.65, 4.40, and 3.61) (P<0.05). EgBu, EgAS and EgAgB-ELISA results showed that the antibody positive rate in sera of recurrent CE patients [82.4% (150/182), 86.3% (157/182), 70.9% (129/182)] and the S/N value (5.54, 3.23, 3.75) were significantly higher than that of primary patients [positive rate: 67.4% (126/187); 63.6% (119/187); 57.2% (107/187); S/N value: 4.20, 2.70, 3.75] (P<0.05). The S/N value detected by EgPS-ELISA and the positive rate examined by EgAgB-ELISA significantly increased with the increasing of the number of operations (P<0.05), reached 4.23 and 91.7% (11/12), respectively, in the patients with ≥4 times of operations. The positive rate and S/N value of EgAS-ELISA and EgAgB-ELISA increased with the number of hydatid cysts in patients (P<0.05), reached 90.5% (19/21), 76.2% (16/21), and 3.97, 4.42, respectively, in patients with at least 4 cysts. Among the five antigen-ELISA, the positive rate increased with the cyst diameter (P>0.05). The S/N value of EgHF-ELISA and EgAS-ELISA increased significantly with the cyst diameter (P<0.05), reached 3.66 and 3.69, respectively, when the cyst diameter was ≥15.1 cm. ROC analysis result showed that among the 5 native antigen-ELISA, the AUCROC was highest in patients with cysts at CE2 stage (EgPS: 0.988±0.009, EgHF: 0.957±0.013, EgBu: 0.969±0.011, EgAs: 0.910±0.024, EgAgB: 0.894±0.021), EgAgB-ELISA presented the lowest AUCROC of 0.267±0.031 in patients with cysts at CE5 stage. Except EgAgB, the positive rate of another 4 antigen-ELISA in detection of patients with cysts at CE 2 stage [EgPS: 97.2% (69/71), EgHF: 93.0% (66/71), EgBu: 88.7% (63/71), EgAs: 85.9% (61/71)] was slightly higher than that of the patients with cysts at CE1 stage, and then promptly reduced in patients with cysts at CE5 stage (EgPS: 56.3%, EgHF: 43.8%, EgBu: 12.5%, EgAs: 12.5%). In the patients with cysts at CE5 stage, the S/N value of the five antigen-ELISA was lowest (EgPS: 2.29, EgHF: 1.50, EgBu: 1.11, EgAs: 0.78, and EgAgB: 1.11).  Conclusion  Compared with the other three antigens, the EgPS and EgAgB antigens have higher antigenicity, sensitivity, and specificity. The sera of hepatic CE patients are more reactive to the five native antigens than the other clinical types.
    Effect of Culture Supernatant of Toxoplasma gondii on the Proliferation and Apoptosis of BGC-823 Cells
    LUO Qiang1,SUN Li1 *,TIAN Qing-qing1,REN Hong2,LIU Hua1,YANG Cui-jun1,LI Xin1
    2014, 32(2):  8-123-127. 
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    Objective  To investigate the effect of culture supernatant of Toxoplasma gondii on the proliferation and apoptosis of human gastric cancer BGC-823 cells.  Methods  Toxoplasma gondii tachyzoites with seed counts of 2×107/ml, 4×107/ml, and 8×107/ml harvested from infected mice were cultured for 24 h, and then the culture supernatant was collected. BGC-823 cells (5×104/ml) at mid-exponential phase were incubated with different concentrations of culture supernatants of Toxoplasma gondii tachyzoites. While in control group, the same volume of DMEM was given. At 24, 48 and 72 h after incubation, the measurement of tumor cell growth inhibition rate was performed by using CCK-8 kit. Cell apoptosis was observed under a fluorescence microscope after 24 h incubation. The DNA ladder zone of apoptosis cells was analyzed with the method of agarose gel electrophoresis. Flow cytometric analysis was used to analyze cell cycle for cell proliferation index and the expression of p53 and Bcl-2.  Results  The culture supernatant of Toxoplasma tachyzoites inhibited the proliferation of BGC-823 cells and the growth inhibition rates increased with the time and the concentration. The highest rate reached at the 72nd hour when the concentration of Toxoplasma tachyzoites was 8×107/ml. Apoptotic bodies were found in experimental group. The amount of apoptotic body was positively associated with the tachyzoite concentration. DNA fragment in the treated cells after 24 h incubation was revealed by agarose electrophoresis. When the proportion of BGC-823 cells in G0/G1 phase increased, the proportion of cells in S phase decreased. Cell proliferation index decreased with the increase of the concentration of tachyzoites. The proliferation index(0.36) of the cells cultured with culture supernatant of 8×107/ml tachyzoites was significantly lower than that of control group(0.6, P<0.05). p53 protein expression was higher in experiment group than that of the control, whereas Bcl-2 protein expression in experiment group was lower than that of the control (P<0.05).  Conclusion  The culture supernatant of Toxoplasma gondii can inhibit the proliferation of BGC-823 cells and cause apoptosis of BGC-823 cells, which may be related with up-regulating p53 expression and down-regulating Bcl-2 expression.
    Preparation and Application of the Polyclonal Antibody of Toxoplasma gondii Autophagy Protein 8(TgAtg8)
    TAN Feng, HUA Qian-qian, LI Xing-pan, LI Xiang-zhi, LIANG Shao-hui*
    2014, 32(2):  9-130-134. 
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    Objective  To prepare and evaluate specific-TgAtg8 polyclonal antibody.  Methods  The known Saccharomyces cerevisiae Atg protein sequences were used to identify Toxoplasma gondii homologous protein through bioinformatics analysis. TgAtg8 cDNA was amplified and cloned into prokaryotic expression vector pGEX-6p-1. The constructed pGEX-6p-1-TgAtg8 was transformed into E. coli BL21 cells and induced with IPTG for expression. The expression product was analyzed through SDS-PAGE and Western blotting. The recombinant TgAtg8 protein with an N-terminal glutathione-S transferase tag was used to immunize rabbits and raise specific polyclonal antibody against TgAtg8. Subsequently, the antibody was applied for Western blotting and IFA assay.  Results  Recombinant expression plasmid of pGEX-6p-1-TgAtg8 was confirmed correct by restriction enzyme digestion and sequencing. SDS-PAGE and Western blotting analysis showed that the recombinant TgAtg8 protein with the predicted molecular weight (Mr 40 000) was expressed highly in E. coli BL21. After immunization, the specific antibodies against TgAtg8 protein were produced. The anti-TgAtg8 polyclonal antibody reacted specifically with TgAtg8 fusion protein or endogenous TgAtg8. Importantly, IFA assay determined that the TgAtg8 signal was generally distributed throughout the cytoplasm of the tachyzoites. However, the green fluorescence signal gathered into one or more green spots after induction of autophagy.  Conclusion  The specific polyclonal antibody against TgAtg8 could be used to observe the dynamics of autophagosome formation in T. gondii, which is useful tool to investigate the autophagic machinery in this parasite.
    Effect of Diets with Different Fat Levels on the Body Size and Development of Lucilia sericata
    LI Xue-bo1,ZHENG Shi-li2,WANG Qing-shan3,DING Chun-li1,LI Hong-wei3 *,YANG Yong-qiang4,WAN Li-hua4
    2014, 32(2):  10-135-138. 
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    Objective  To observe the effect of diets with different fat levels on the body size and development of Lucilia sericata.  Methods  Under the constant temperature of 28 ℃, the larvae were reared on the diets containing 0%(G0), 10%(G1), 30%(G3), 50%(G5) and 80%(G8) fat tissues(fat/muscle ratio), respectively. Length and weight of larvae and pupae were measured at 12 h interval since 16 h after eclosion. Length of inter-medial cross vein(m-m) of adult left wing was measured. 10 samples were collected in each group. The developmental duration time, mortality and sex ratios of adults were recorded.  Results  The mean maximal larval length[(13.3±1.2), (12.0±1.1), (10.2±0.9) and (8.8±0.8) mm, respectively] and mean maximal larval weight[(72.8±6.1), (62.2±5.7), (47.2±4.3), and (34.9±5.7) mg] in G1, G3, G5 and G8 groups were significantly less than that of the G0 group[(14.8±1.3) mm and(80.4±8.1) mg](P<0.01). The body size of pupae and adults was also significantly less than that of G0 group(P<0.01). The total duration time of G5 and G8 groups [(293.3±22.2) and (285.2±24.6) h] were significantly shorter than that of G0 group [(312.8±20.1) h](P<0.01). The mortality of larvae [(32.6±5.6)% and (44.3±7.7)%] and pupae[(28.6±5.5)% and (43.5±6.2)%] of G5 and G8 group were also significantly higher than that of G0 group[(5.7±3.3)% and (4.5±1.9)%](P<0.01). There was no significant difference in sex ratio among the 5 groups(P>0.05).  Conclusion  The body size of larvae, pupae and adults of Lucilia sericata is smaller, the development time is shorter and mortality is higher when the food substrate contains more fat tissues.
    Preliminary Survey on the Host of Angiostrongylus cantonensis in Three Plateau Lakes of Yunnan Province
    ZHANG Xiao-xiao1,CUI Li-yun1, CAO Shu-zhen1,WANG Fei1,LI Kun-hua2,WANG Jin-yong3,YANG Yi-mei1 *
    2014, 32(2):  11-139-142. 
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    Objective  To investigated the intermediate hosts of Angiostrongylus cantonensis in three plateau lakes of Yunnan Province, and analyze the effect of temperature on A. cantonensis during 1991-2010.  Methods  An epidemiological investigation of angiostrongyliasis cantonensis in Erhai Lake, Fuxian Lake and Xingyun Lake was conducted from April to September in 2012. Snails were examined for the third stage larvae by enzyme digestion or lung examination. Rodents were captured in the fields, and their hearts and lungs were dissected for adult worms. The potential distribution of A. cantonensis and its main intermediate host Pomacea canaliculata were predicted based on degree-day models using GIS technique.  Results  A total of 4 950 snails were collected, belonging to 4 species, P. canaliculata, Cipangopaludina chinensis, Bellamya aeruginosa, and B. quadrata. 174 rodents were captured, belonging to 5 species. No positive samples were found. The potential distribution map showed that the distribution of A. cantonensis and P. canaliculata in Yunnan would expand with the rise of temperature, and with the passage of time they could complete one generation in the region which couldn’t finish one generation in one year along with time passing.  Conclusion  A. cantonensis are not found in the hosts. The natural environment and ecological system of the three lakes match the condition of A. cantonensis transmission.
    Progress of CD8+ T Cell-Mediated Immune Response to Toxoplasma gondii Infection
    WU Bin, LYU Fang-li *
    2014, 32(2):  12-143-147. 
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    Toxoplasma gondii is an obligate intracellular protozoan parasite, which infects warm-blooded animals including humans. T. gondii infection in the immune-competent individual is largely asymptomatic, but it can cause severe toxoplasmosis in immune-deficiency patients. T cell-mediated immunity plays an important role in resistance against T. gondii. Therefore, this review focuses on the progress of CD8+ T cell-mediated immune response to T. gondii infection.
    The Role of CD4+CD25+ Regulatory T Cells in Helminth Infection Immunity
    XU Yun-fei, YANG Wen-tao, WANG Chun-feng, YANG Gui-lian*
    2014, 32(2):  13-148-151. 
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    Regulatory T cells(Tregs), a class of CD4+ T lymphocytes which are different from the Th1 and Th2 lymphocytes, have immunosuppressive function and play an important role in maintaining immune homeostasis and immune tolerance. Previous studies showed that the generation of Tregs could be induced, and Tregs reduced the immune pathological damage to the host by inhibiting T-cell function in helminth infection.
    Immunity to Parasitic Infection:The Role of Dendritic Cells
    QU Kai-ge1,ZHAO Quan1,2,JIANG Jing1,2,YANG Gui-lian1 *
    2014, 32(2):  14-152-156. 
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    Dendritic cells are a major class of professional antigen-presenting cells which play an important role in the immune response of parasite-infected host. This article summarizes the role of dendritic cells in the immunity to parasitic infection by analyzing the studies of protozoan, nematode, and trematode infections.
    Retrospective Analysis of Granted Projects of the National Institute of Parasitic Disease, China CDC during 2002-2012
    ZHOU Xiao-jun, GUAN Ya-yi*, ZHANG Min-qi, XIONG Yan-hong, ZHENG Bin
    2014, 32(2):  15-159-160. 
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    In this paper, the granted projects of the National Institute of Parasitic Diseases(NIPD), China CDC, was analyzed. The results showed that from 2002 to 2012, 126 projects were granted to NIPD. 28.6%(36/126) of the projects were at the national level; 27%(34/126) were at provincial and ministrerial level. International cooperation projects and those supported by state key laboratory and enterprises accounted for 28.6%(36/126) and 15.8%(20/126), respectively. 94 projects belonged to applied researches and 32 belonged to basic researches. Most project leaders were young and middle-aged researchers with senior professional titles.
    Hemocyte Morphology and Classification in Oncomelania hupensis
    ZHENG Sheng-bang,ZHOU Yi-biao*,LI Lin-han,WU Jin-yi,SONG Xiu-xia,JIANG QING-wu
    2014, 32(2):  16-91-94. 
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    Hemocytes were collected from Oncomelania hupensis in Junshan, Hunan Province and Puge, Sichuan Province, respectively, and stained with Giemsa solution for light microscopic examination. The cells were classified morphologically. Five types of hemocytes were identified, viz., large acidophilic hyalinocytes, small acidophilic hyalinocytes, basophilic hyalinocytes, basophilic small granulocytes and basophilic large granulocytes. The proportion of small acidophilic hyalinocytes was the most abundant hemocyte [36.7%(229/624) in snails from Junshan and 31.7%(257/810)in snails from Puge], followed by basophilic hyalinocyte[23.1%(144/624)in Junshan and 24.4%(198/810)in Puge]. Basophilic large granulocyte was about 9.3% (58/624) in Junshan and 11.6%(94/810)in Puge. The length of large acidophilic hyalinocytes was the maximum and its nucleocytoplasmic ratio was minimum, followed by small acidophilic hyalinocytes. The length of basophilic cells was shorter and its nucleocytoplasmic ratio was smaller than those of acidophilic cells. There was no significant difference in cellular constituents of hemocytes and the morphological features of hyalinocytes between the snails from Junshan and Puge, while the length and nucleocytoplasmic ratio of granulocytes in Junshan snails were smaller than those of Puge ones.
    Resistance of Anopheles sinensis to Three Common Insecticides in Hainan Province
    SUN Ding-wei, WANG Shan-qing*, ZHUO Kai-Ren, ZENG Lin-hai, LI Shan-gan
    2014, 32(2):  17-127-129. 
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    Anopheles sinensis adults were collected by cow-baited trap in Haikou City, Sanya City, Lingshui County, Changjiang County and Qiongzhong County of Hainan Province during 2011-2013. The mosquitoes were exposed to insecticide impregnated papers with discriminating concentrations of DDT(4%), deltamethrin(0.05%), and malathion(5%) using WHO standard assays. Knockdown rate was recorded at 10, 15, 20, 30, 40, 50, and 60 min, and KT50 values were calculated. Mortality was recorded after 24 hours of exposure. The resistance level was graded as sensitive group(S) with a mortality rate of 98%-100%, preliminary resistance group (M) with mortality rate of 80%-97%, and resistance group (R) with mortality rate of lower than 80%. The results showed that the mortality rate of An. sinensis in Qiongzhong County in 24 h-post-exposure to 0.05% deltamethrin was 95.0% with a resistance degree of M. That to 0.05% deltamethrin in the other 4 sites was 17.0%-63.0%, all with a resistance degree of R. That to 4% DDT in Haikou, Sanya, Lingshui, Qiongzhong and Changjiang was 36.0%, 27.0%, 24.0%, 59.1%, and 82.0%, with a resistance level of R, R, R, R, and M, respectively. That to 5% malathion in Haikou, Sanya, and Lingshui was 16.0%-41.0%, all with a resistance degree of R, while that to malathion in Qiongzhong and Changjiang was 100% and 98.0%, respectively, with a resistance level of S.
    Diagnosis of the First Imported Case of Plasmodium ovale Infection at Guangdong Port
    SHI Yong-xia, HUANG Ji-cheng*, SU Jin-kun, LI Xiao-bo, ZHENG Kui, DING Guo-yun
    2014, 32(2):  18-156-158. 
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    The first imported case of Plasmodium ovale infection in Guangdong Province was identified. The patient worked in Myanmar for one week and had a fever when he arrived at Guangzhou Baiyun International Airport. Epidemiological information and blood sample were collected. The detection was conducted by microscopy, right VIEW rapid malaria test (RDTs) and real-time PCR with Plasmodium genus-specific and species-specific primers and probes. The case showed weak positive RDT result, and was confirmed as P. ovale infection by microscopy and real-time PCR. After treatment with artemether, his symptoms improved.
    Investigation on Lophomonas blattarum Infection in Periplaneta americana in Wuhan City
    YANG Jie-xi,TANG You-yuan,FANG Zheng-ming,TONG Zhen-zhen,LI Yong-long,WANG Ting*
    2014, 32(2):  19-161-162. 
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    An investigation of Lophomonas blattarum infection in Periplaneta americana in Wuhan City were conducted. A total of 110 P. americana were dissected and the intestines were separated. The intestines were washed with 0.6% saline and the washing solutions were smeared on slides. The slides were stained with Giemsa stain and observed under a microscope(×1000). Out of 110 intestine washing solution samples, 44 were suspected of L. blattarum infection. The parasite was oval or pyriform in shape and 20-40 μm in size. A tuft of flagella extended down the central axis of the parasite and a trumpet-shaped calyx enveloped the flagellar area and the nucleus. An axostyle was slender and pointed posterior ends. Based on the above morphological characteristics, the parasite was identified as L. blattarum. The results showed that the infection rate of L. blattarum in P. amerivana in Wuhan City was 40.0% (44/110).
    Survey on Intestinal Nematode Infections among School Students in Tengchong County
    YANG Yuan-he1,SONG Jian-cheng1,LI Xin-he1,LI Rong-xing1,ZHOU He-jun2 *
    2014, 32(2):  20-163-164. 
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    One primary school and one middle school were selected from Gudong Town, Tengyue Town and Puchuan Township of Tengchong County, respectively, by using the lamination stochastic group sampling method. The intestinal parasite infections were investigated with the iodine-stained direct smear method and modified Kato-Katz thick smears method. A total of 1 134 students were investigated and the total infection rate of intestinal nematodes was 12.4%(141/1 134). The infection rate of Ascaris lumbricoides, Trichuris trichura, and hookworm was 9.4%(106/1 134), 2.8% (32/1 134), and 0.3%(3/1 134), respectively. The prevalence of intestinal nematodes among the students of urban(2.2%, 8/363) was lower than those of rural(17.3%, 133/771)(P<0.01). The infection rate in students from Gudong Town was higher than those of Tengyue (2.2%, 8/363) and Puchuan County(2.3%, 8/35)(P<0.01), whereas the economy level of Gudong Town(29.9%, 123/412)was the best in the three towns. After all, the infection rate of the middle school students(13.7%, 59/432) was higher than that of pupils(11.7%, 82/702)(P<0.01). Compared with 2003, the prevalence of nematode infection among the school students in Tengchong County decreased significantly in 2013.
    Composition and Seasonal Fluctuation of Acaroid Mite Communities in Warehousing Environment of Northern Anhui Province
    WANG Hui-yong*,LI Bei-li,ZHANG Yan-xing,ZHA Dao-de,TU Long-xia
    2014, 32(2):  21-165-166,封三. 
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    Samples of stored products were collected every month in 2012 in northern Anhui Province. Acaroid mites were isolated, identified and counted. Among 1 440 samples, 692(48.1%) had mite infestation and 34 species were identified, and the density was 32.1 mites/g. The species richness index(Rmargalef) ranged from 0.48 to 3.30, which was highest in August. The species diversity index (H′) was 1.29-3.32, highest in July. The species evenness index (J) ranged from 0.91 to 0.97, highest in March. The number of species, breeding density, species richness index and species diversity index were stable in the year, while the species evenness index showed irregular change. There were many species of acaroid mites in the storage circumstance. The composition of acaroid mites were diverse, and changed with the seasons.