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    30 April 2013, Volume 31 Issue 2
    Preliminary Observation on the Effect of Mefolquine against Egg Granuloma Formation in the Liver of Mice Infected with Schistosoma japonicum
    XIAO Shu-hua*,ZHANG Chao-wei
    2013, 31(2):  1-81-86. 
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    Objective  To explore whether mefolquine possesses the effect on granuloma formation induced by Schistosoma japonicum eggs.  Methods  Seventeen out of twenty-eight mice infected with 20 S. japonicum cercariae for 35 days were treated orally with mefloquine at a single dose of 200 mg/kg, and groups of 2-3 mice were sacrificed at various intervals post-treatment. The livers removed from each group of mice were fixed in 10% formaldhyde. While the remained 11 untreated mice divided into 6 groups(1-2 mice per group) were sacrificed at the same time periods as groups of mice treated with mefloquine, and their livers served as untreated corresponding controls. The granulomas with single egg in the center were counted and their diameters were measured using an ocular micrometer. The liver tissue sections were stained with hematoxylin and eosin(HE), Foot’s or Mallory’s methods for observation on histopathological alteration of egg granulomas, and on the appearance of reticular and collagen fibers within the granulomas.  Results  After infected mice were treated with mefloquine for 3, 7, 14, 21, 28 and 35 days, i.e., 38, 42, 49, 56, 63, and 70 days post-infection, the mean diameters of granuloma with single egg measured in the liver tissues section were (161±19), (175±13), (195±9), (171±40), (180±13), and (145±25) ?滋m, respectively, and each of them was significantly lower than that of its corresponding control group of (189±18), (197±11), (211±12), (208±19), (203±16), and (207±36) ?滋m (P<0.01 or P<0.05). Histopatological observation showed that in mice treated with mefloquine, the eosinophil-predominant inflammatory cells around the egg granuloma were sustained to 14-21 d post treatment (49-56 d post infection), which was significantly different from the corresponding control groups that all the eggs were surrounded by fibroblasts at 42 d post infection. Up to 28-35 d post treatment (63-70 d post infection), the boundary of egg granulomas distributed in the liver tissues of mefloquine treated groups was neater in comparison to the corresponding control groups. Further observation on the reticular and collagen fibers within the egg granulomas by using specially staining methods demonstrated that in groups of mice treated with mefloquine for 2 weeks, the emergence and amount of the two kinds of fibers were delayed and less in comparison with corresponding control groups. After infected mice treated with mefloquine for 21 d(56 d post infection), the amount of the two kinds of fibers revealed in some egg granulomas was similar to the corresponding control group, but no further increase in the amount of the fibers, and seldom spread over the boundary of egg granuloma were seen 28 d and 35 d after treatment(63 d and 60 d post infection). While in corresponding control groups, the two kinds of fibers increased continuously with time post infection to become thick, and spread over the boundary of granuloma to further interconnect with the fibers stretched from the adjacent granuloma, and separate the liver tissue to form the grid-like structure.  Conclusion  Preliminary observation demonstrates that mefloquine possesses suppressive effect on granuloma formation induced by S. japonicum eggs.
    Expression of Smad Proteins in the Process of Liver Fibrosis in Mice Infected with Schistosoma japonicum
    ZHANG Bin-bin1 *, CAI Wei-min2, TAO Jun3, ZHENG min2, LIU Rong-hua2
    2013, 31(2):  2-89-94. 
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    Objective  To study the expression of Smads proteins involved in TGF-β1 signal transduction during the process of liver fibrosis in BALB/c mice infected with Schistosoma japoncum.  Methods  Thirty-four BALB/c mice were each infected with (20±1) S. japonicum cercariae. The mice were sacrificed at 8, 12, 16 and 24 weeks post-infection. Ten healthy BALB/c mice served as normal control group. The liver tissues were fixed in 10% formaldehyde for histology and immunohistochemistry assay. The single-egg granuloma area was measured in hematoxylin-eosin stain section. The degree of liver fibrosis was determined by Sirius red staining. Immunohistochemistry was used to observe the expression of Smad protein.  Results  The area of single-egg granuloma peaked at 8th week post-infection [(5.33±1.03) mm2], and with time passing, the area diminished, and the area of granuloma reduced to(2.94±1.69)mm2 at 24 weeks post-infection. The difference was significant among the 4 periods after infection in single-egg granuloma area(P<0.05). Collagen fibers appeared around granulomas at 8 weeks (2.03±0.52) and increased gradually. At 24 weeks post-infection, the degree of liver fibrosis reached a peak(6.90±1.57), and the liver fibrosis degree was significantly different among infection groups(P<0.05). Immunohistochemistry showed low expression level of Smad2/3 and Smad7 and inconspicuous level of Smad4 in livers of the normal mice. The expression of Smad2/3 was found mostly in the cytoplasm and nucleus of cells around granulomas at 8th week post-infection, and the positive area of Smad2/3 was (7.24±1.64)% by semi-quantity. At 12 weeks post-infection, the Smad2/3 protein expression level around granulomas and liver sinus reached the peak[(10.01±1.07) %], and there was significant difference between infection groups and the control [(2.13±0.32)%]. A significant difference in the Smad2/3 protein expression level was found between 12 weeks post-infection group and 8 weeks or 16 weeks post-infection groups. The expression level of Smad4 was(8.81±1.13)% at 8th week post-infection, higher than that in the control[(4.83±1.15)%](P<0.05). There was no difference among the infected mice at different periods in the level of Smad4(P>0.05). After 8 weeks post infection, Smad7 protein sparsely appeared around the granuloma [(4.15±1.26)%] while it disappeared around liver sinus. At 12 weeks post-infection, the level of Smad7 protein was higher[(6.34±1.5)%], but with prolonged infection time, no significant difference was revealed (P>0.05). The level of Smad7 in infected mice was higher than that in the control(P<0.05).  Conclusion  Results show high expression for Smad2/3 and Smad7 and low expression level of Smad4 during the process of liver fibrosis in BALB/c mice infected with Schistosoma japoncum.
    Role of CD8+ T cells in the Tumor Growth Delay Induced by Toxoplasma gondii Excreted-secreted Antigen in B16F10 Mouse Melanoma Model
    JIAO Yu-meng1, FANG Qiang1 *, XIA Hui1, WANG Xue-mei1, TAO Zhi-yong1, CHEN Xing-zhi1, SHEN Ji-long2
    2013, 31(2):  3-95-98. 
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    Objective  To observe the role of CD8+ T cells in the tumor growth delay induced by Toxoplasma gondii excreted-secreted antigens (TgESA) in B16F10 mouse melanoma model in the early stage.  Methods  TgESA were prepared by incubating T. gondii tachyzoites for 12 h in vitro. 15 C57BL/6 mice were randomly assigned to group A, B, and C (5 mice per group). Each mouse in group B and C was subcutaneously injected in right flank with 2×105 B16F10 cells. Mice in group C were intraperitoneally injected with TgESA (100 μl per mouse) at 7 d after B16F10 cells injection. Mice of group A were only injected with PBS. On the 13th day after melanoma cell injection, the mice were sacrified and spleen was removed. The percentage of CD8+ T cells in the spleen was analyzed by flow cytometry. CD8+ T cells were isolated from spleen cells by using immunomagnetic beads. The activity of CD8+ T cells against B16F10 melanoma cells was determined by LDH release assay at different effect-to-target cell ratios (2.5 ∶ 1, 5 ∶ 1, and 10 ∶ 1). Other 30 C57BL/6 mice were randomly divided into group E, F, and G. Each mice were injected with 2×105 B16F10 cells. At the same time, mice in group F and G were simultaneously injected via the tail vein with CD8+ T cells isolated from mice in group B and C. Tumor growth, mortality and survival time of mice were observed and recorded during 35-d observation period.  Results  The percentage of CD3+CD8+ T cells in the spleen cells of group C[(15.74±0.28)%] was significantly higher than that of group B[(14.18±0.27)%] and A [(13.86±0.13)%](P<0.05). At different effect-to-target cell ratios, the activity of CD8+ T cells against B16F10 cells in group C was significantly higher than that of group B (P<0.05). The average time of tumor formation in group G [(14.9±1.2) d] was longer than that in group F [(11.9±0.7) d] and E [(9.4±1.2) d](P<0.05). The tumor size in these groups increased, but there was no obvious difference in the tumor growth rate among the three groups. The tumor size of group G was significantly smaller than the other two groups (P<0.05). In group E, F and G, mice began to die on the 26th day, the 29th day and the 30th day after tumor inoculation, and the number of survival mice was 3, 5 and 7, respectively, at the 35th day after injection.  Conclusions  TgESA may up-regulate the quantity and function of CD8+ T cell in B16F10 melanoma mouse model, which plays a role of delaying tumor growth in early stage.
    Study on Immune Response in BALB/c Mice Induced by ROP2 Protein of Toxoplasma gondii with Cimetidine
    CHEN Xing-zhi1,2 *, YANG Xiao-di1, YANG Wen1, CHEN Yong1, LIU Li-li2, SHEN Ji-long2, SUN Xin1
    2013, 31(2):  4-99-103. 
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    Objective  To observe the immune response induced by ROP2 protein of Toxoplasma gonddi with cimetidine in mice.  Methods  Eighty BALB/c mice were randomly divided into 4 groups: PBS(group A), ROP2 protein (group B), ROP2 protein-Freund’s adjuvant (group C) and ROP2 protein-cimetidine(group D). Mice were immunized with PBS, ROP2 protein, ROP2 protein and Freund’s adjuvant, ROP2 protein and cimetidine[200 mg/(kg·d)] by subcutaneous injection three times at an interval of 2 weeks, respectively. Experimental mice were immunized with 100 μg ROP2 proteins, which were diluted to a final volume of 200 μl in PBS. On day 13, 27 and 41 after immunization, mice sera were collected for determination of antibody IgG and cytokine IFN-γ by ELISA. Two weeks after the final immuni-zation, T cells subpopulation was detected by flow cytometry and splenocyte proliferation activity was determined with CCK-8. Another 12 immunized mice in each group were intraperitoneally challenged with 5×104 tachyzoites of T. gondii and the survival time was observed.  Results  Two weeks after final immunization, compared with groups A[(659.750±239.962) pg/ml] and B [(872.750±197.011) pg/ml], the level of IFN-γ significantly increased in groups C [(1 600.750±480.680) pg/ml] and D [(1 494.375±451.655) pg/ml] (P<0.01). Similarly, compared with groups A (0.636±0.108) and B (0.871±0.089), the level of IgG was also higher in groups C(1.068±0.111) and D(1.046±0.147) (P<0.01). The proliferation of splenocytes rose in group C(0.831±0.130) after immunization, and similarly in group D (0.762±0.089), and were both significantly higher than that of groups A (0.504±0.078) and B (0.592±0.160)(P<0.01). Moreover, ratio of CD4+/CD8+ in groups C (0.831±0.130) and D (0.762±0.089) were higher than that of groups A(0.504±0.078) and B(0.592±0.160)(P<0.05). After challenge with violent virulence strain of tachyzoites, the median survival time of mice in groups A, B, C, and D were 96, 108, 132, and 132 h, respectively. The mean survival time of mice in groups C and D were longer than that of groups A and B(P<0.05). There was no significant difference in 5 parameters between C and D: the level of IFN-γ and IgG, CD4+/CD8+ ratio, splenocyte proliferation, and survival time of mice(P>0.05).  Conclusion  Cimetidine can enhance the humoral and cellular immune response induced by ROP2 protein.
    Effect of Toxoplasma gondii Prugniaud Strain Infection in Pregnant Mice on the Learning Ability of the F1 Generation
    WANG Zheng-rong1,BAO Huai-en2 *
    2013, 31(2):  5-104-109. 
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     Objective  To study the effect of Toxoplasma gondii prugniaud strain infection on female reproductive toxicity in mice and learning ability of their F1 generation.  Methods  Thirteen ICR mice were each infected intragastrically with 10 T. gondii cysts on the 15th day of pregnancy(late stage pregnancy). 12 mice were treated with physiological saline as control. The time from conception to birth and the number of offspring were recorded. Three mice from each group were sacrificed when pregnant 20 d, placentas from the sacrificed and output stillbirth mice were examined by using histopathology and immunohistochemistry. DNA extraction was performed from placenta tissue, and then T. gondii B1 gene was amplified by PCR. The F1 generation mice from experiment group and control group were tested by Morris water maze test. Statistical analysis on learning and memory ability was made by SPSS 13.0 software.  Results  The time from conception to birth in experiment group[(19.2±1.751)d]was shorter than that in control group [(21.0±1.732)d](P<0.05). No significant difference was found in the number of offspring between experiment group(70) and control group(85)(P>0.05). Microscopic examination with HE staining showed multiple T. gondii among placental villi, the increase of the number of Hofbauer cells, blood sinus expansion and hyperemia, and visible nucleated erythrocytes. Immunohistochemically, T. gondii antigen was detected in placenta tissue. T. gondii B1 gene was detected in placenta tissue(194 bp). On the third and fourth day of the Morris water maze test, the latency of experiment group [(29.92±4.28) s, (27.69±6.23) s] was longer than that of the control[(24.07±5.32) s, (22.25±7.94) s](P<0.05). In the spatial probe test, the distance across the platform quadrant of experiment group[(384.66±41.33) cm] was shorter than that of the control[(426.12±46.48) cm](P<0.05).  Conclusion  T. gondii Prugniaud strain infection in late stage pregnancy of mice may induce reproductive toxicity and affect the learning and memory capability of the F1 generation.
    Construction and Expression of the Echinococcus granulosus Recombinant BCG-EgG1Y162
    Zulipiye·TUERXUN1,Delixiati·YIMITI2,CAO Chun-bao2,MA Hai-mei2,LI Yu-jiao1,ZHOU Xiao-tao2,ZHU Ming2,MA Xiu-min1,WEN Hao1,DING Jian-bing1,2 *
    2013, 31(2):  6-110-113. 
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    Objective  To construct and express Echinococcus granulosus recombinant bacille Calmette-Guerin (BCG) strain rBCG-EgG1Y162.  Methods  The encoding gene of the antigen EgG1Y162 of E. granulosus was recombined with E. coli-Mycobacterium shuttle expression plasmid vector pMV361 by genetic engineering technique, and transformed into E. coli for amplification. The recombinant plasmid rpMV-EgG1Y162 was identified by PCR, double digestion with restriction enzymes, and sequence analysis. The confirmed rpMV-EgG1Y162 was transformed into BCG strain via electroporation technique to construct the recombinant rBCG-EgG1Y162. After identification by PCR and double digestion with restriction enzymes, the recombinant strain was cultured for about 2 weeks. In order to induce the expression of target protein, the rBCG was placed in 45 ℃ for 30 min. SDS-PAGE and Western blotting were used to analyze the expressive protein.  Results  The product of recombinant plasmid rpMV-EgG1Y162 was approximately 360 bp by PCR amplification and double digestion with restriction enzymes, consistent with the expected fragment length. Sequencing results showed that the inserted sequence was correct. The rBCG-EgG1Y162 grew well and the identification of PCR and enzyme digestion revealed accuracy. The results of SDS-PAGE and Western blotting showed that the relative molecular weight(Mr) of the protein was about 71 000.  Conclusion  The E. granulosus rBCG-EgG1Y162 strain is constructed and expressed.
    Taxonomic Composition of Metagenomic Community in the Larval Gut of Mosquito Anopheles sinensis(Diptera ∶ Culicidae)
    NAN Chun-yan1,2,MA Ya-jun2 *,XU Jian-nong3,LIANG Jian1
    2013, 31(2):  7-114-119. 
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    Objective  To investigate the bacteria diversity in larval gut of field-collected Anopheles sinensis.  Methods  The 16S rDNA V4 region of An. sinensis larvae collected from paddy on Jiading District of Shanghai (L1/L2)and small seeping water on Wenchang City of Hainan(AS) was sequenced by high-throughput pyrosequencing. Using Qiime and Mothur softwares, the number of sequences and operational taxonomic units(OTUs) for each sample was sorted and calculated, the species abundance and distribution, Alpha diversity index and difference times of species abundance among samples were analyzed.  Results  The number of sequences and OTUs for each sample were 253 724/3 930(L1), 225 203/4 312 (L2) and 73 990/2 380 (AS). The rarefaction curves showed that adequate sampling was achieved. The number of OTUs was close to actual situation. The value of richness index was 5 942.61/6 534.88(L1), 6 328.17/7 235.89(L2) and 4 228.66/5 651.20 (AS); diversity index was 4.63/0.03 (L1), 5.10/0.02 (L2) and 0.14/3.94 (AS). The dominant species of An. sinensis larvae gut microbiota all belonged to the phylum Proteobacteria, with a percentage of 87%(AS) and 90% (L). In addition, the dominant phyla among them were Firmicutes, Bacteroidetes and Actinobacteria. The comparison of bacterial abundance between L and AS showed that there were 18 phyla with significant difference, except the Proteobacteria and Deinococcus-Thermus; only 9 phyla were different significantly between L1 and L2.  Conclusion  Evenness and richness of bacteria flora in the An. sinensis larvae gut collected from paddy and small seeping waters were obtained.
    Cloning and Expression of the Mucin-Related Protein1(Aamucin1)from Salivary Gland of Aedes albopictus
    LIU Jian1,CHENG Jin-zhi1,SUN Yu2,ZHU Ru-fang3,ZHANG Liang4,WU Jia-hong1 *
    2013, 31(2):  8-120-123. 
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    Objective  To clone the mucin-related protein(Aamucin1)gene from salivary gland of Aedes albopictus Guangzhou isolate, and analyze the expression difference due to blood-feeding.  Methods  Total RNA was extracted from the salivary gland. The coding region of Aamucin1 was amplified with a pair of specific primers by RT-PCR. The product was sequenced and analyzed by bioinformatics. Expression analysis was conducted by real-time RT-PCR.  Results  The product of RT-PCR was 849 bp with encoding 283 amino acids. To compare with that from Ae. albopictus Rome strain, 13 amino acids were deleted at the C end, and Aamucin1 in Guangzhou isolate shared 58% identity in amino acids with that of Rome isolate. In addition, an alternative splicing was found in Aamucin1 and located in a proline enrich area by Protscan. To compare with that of non-blood-feeding(group SG), Aamucin1 was significantly down-regulated with 0.39 fold expression at zero time after engorged (group BSG_0, mosquitoes with abdominal distention from the first 2 hours after blood-feeding, P<0.01) and 0.61 fold expression at the 24th hour after engorged(group BSG_24,mosquitoes from the 24th hours after blood-feeding, P>005).  Conclusion  The full length of Aamucin1 gene of Ae. albopictus is cloned and it can be modulated by blood-feeding.
    Effect of Feeding on Different Tissues on Larva Development of Lucilia sericata
    WANG Yao,WAN Li-hua*,LI Xue-bo
    2013, 31(2):  9-124-126,130. 
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    Objective  To observe the effect of feeding on different pig tissues on the development of Lucilia sericata larvae.  Methods  Under a constant temperature of 25 ℃, about 200 larvae each were reared on four different substrates, i.e. pig’s brain, liver, muscle and a mixture of minced pork muscle and fat (6 : 4). Length and weight of larvae and pupae were measured at 12 h interval 16 h after eclosion. The time of development, mortality, sex ratio of adults were recorded.  Results  Compared to the other groups, the larvae of liver and mixture groups grew slower, time of reaching maximum length and weight was delayed for 12-24 h. The duration of larva development of liver group [(284.0±12.6) h] was longer than that of brain group[(257.0±11.9) h], muscle group [(258.0±10.2) h] and mixture group [(260.0±9.8) h] (P<0.05). The mean maximum larva length and weight in mixture group[(11.85±0.36) mm, (40.4±0.2) mg] and liver group[(12.01±0.43) mm, (42.8±0.4) mg] was statistically less than that of brain group and muscle group(P<0.05). The pupal length and weight in mixture group[(7.81±0.60) mm, (38.4±2.4) mg] was less than that of other three groups (P<0.05). The larval and pupal mortality of mixture group[(9.8±2.4)% and (10.3±1.8)%] was statistically higher than that of other three groups (P<0.05). There was no significant difference in the sex ratio among the four groups(P>0.05).  Conclusion  The development duration of the larvae fed on liver tissue is longer than other groups, and the larvae body length and weight of liver group are less than other groups. The body length and weight of larvae and pupae fed on mixture diet are less than other groups with higher mortality.
    Laboratory Detection on Two Cases with Imported Plasmodium ovale Infection
    ZHOU Rui-min, ZHANG Hong-wei*, DENG Yan, QIAN Dan, LIU Ying, CHEN Wei-qi, YAN Qiu-ye, SU Yun-pu, ZHAO Xu-dong, XU Bian-li
    2013, 31(2):  10-127-130. 
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    Objective  To compare the laboratory tests of the imported Plasmodium ovale infection and analyse the genetic character.  Methods  After Giemsa staining and microscopy,  CareStartTM rapid detection and nested PCR were used to detect two cases with P. ovale infection returning from Congo(Brazzaville) in Henan Province. Sequencing was performed after PCR amplification using the 18S rRNA genus-specific primers. Their genetic characteristics were analyzed and the sequence homology analysis was performed in the NCBI.  Results  The two cases were confirmed as P. ovale infection by morphological examination microscopically. Amplified bands were produced by 18S rRNA nested PCR, which was the same with P. ovale in size, whereas the results of CareStartTM rapid detection test were all negative. A sequence of 906 bp in length was obtained by sequencing their 18S rRNA genes in which GC accounted for 35.4%, and the sequence showed 99% homology to the corresponding part of the known P. ovale 18S rRNA gene(GenBank accession No. AB182492). Conclusion  Both the nested PCR and microscopy confirm the infection of P. ovale. A negative result of CareStartTM rapid detection can not ruled out the Plasmodium infection.
    Analysis Report of the National Technique Competition for Diagnosis of Parasitic Diseases in 2012: Ⅰ. Capability Analysis of Plasmodium Detection
    ZHANG Shao-sen,XIA Zhi-gui *,YIN Jian-hai,YAN He,ZHOU Shui-sen,LI Shi-zhu,ZHENG Xiang,HUANG Fang,LI Mei,CHEN Hai-tang,WANG Qiang,ZHANG Li,LIU Wei,XIAO Ning,ZHOU Xiao-nong
    2013, 31(2):  11-131-134. 
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    Objective  To analyze the result of the national technique competition for diagnosis of parasitic diseases in 2012, so as to understand the capability of detection on Plasmodium parasites among professionals from institutes for disease control and prevention at different levels.  Methods  Four professionals from institution were selected as contestants (age≤45 and at least two contestants from county-level institution). The content of the competition included making thick and thin blood slides of Plasmodium(3 slides in 30 min, 10 scores as full marks and 6 as passing score) and identification of species and number with microscopy(5 slides, 8 min per slide, 30 scores as full marks and 18 as passing score). All contestants were grouped by gender, age, professional title, level of institution, classification according to malaria endemicity, geographical location and economic developement of the province. Their scores were statistically analyzed by SPSS 16.0 software.  Results  The average score of blood smear making test in 120 contestants from 30 provinces was 8.7, the highest was 10 and the lowest was 5.8, 118(98.3%) contestants passed the test. The average score of blood smear reading was 16.0, the highest was 29 and the lowest was 0, 52(43.3%) contestants passed the test. There were no significant differences for the scores among genders, ages(≤30, 31-40, >40), professional titles (junior, intermediate and senior), institution levels (provincial, municipal or county level)(P>0.05). However, there was a significant difference among provinces with different malaria endemicity, geographical location and development status (P<0.05). For the blood slide-making and film-reading, scores of contestants from malaria endemic provinces including Class Ⅰ(9.29±0.41,18.17±6.42), Class Ⅱ (8.92±0.79, 18.31±6.94) and Class Ⅲ (8.61±0.89, 15.63±7.52) were higher than those from non-endemic provinces (7.95±1.00, 10.19±7.01)(P<0.01). Scores of contestants from southern provinces(9.16±0.61, 18.82±6.78) were significantly higher than that from northern ones(8.30±0.99, 13.23±7.45)(P<0.01). The film-reading scores were significantly higher in those from eastern provinces(18.20±6.88) than those from western(13.39±7.60) (P<0.05), while no significant difference was found in blood slide-making (P>0.05).  Conclusion  The capability of malaria parasite detection is imbalanced.
    Clinical Analysis of 137 Patients with Visceral Leishmaniasis
    GAO qin,LIU Yan-bin,ZHONG Ce-jun,LV Xiao-ju*
    2013, 31(2):  12-135-139,142. 
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    Objective  To analyze the clinical and epidemiological characteristics of visceral leishmaniasis cases in Sichuan.  Methods  The medical records of 137 patients with visceral leishmaniasis were reviewed between January 2000 and April 2012 in West China Hospital. The epidemiological data, clinical manifestations, laboratory features, diagnosis, therapeutic procedures and outcome of the patients were retrospectively analyzed.  Results  Eighty-eight(64.2%) out of 137 cases were the residents in the endemic area of Sichuan Province and adjacent areas, and 49(35.8%) were non-endemic area residents with a history of visiting endemic area. Patients living in rural areas accounted for 84.7%(116/137), in town for 15.3%(21/137). Visceral leishmaniasis should be strongly suspected in a patient with prolonged fever, marked hepatosplenomegaly, lymphadenectasis, cytopenia and hypergammaglobulinemia. All patients showed positive in rk39 dipstick test, and were treated with antimony sodium gluconate. Among these patients, 86.1% (118/137) were cured by drug, 2.9% (4/137) received splenectomy, and 6.6%(9/137) relapsed. The misdiagnosis rate was 23.4%(32/137).  Conclusion  Bone marrow smear staining and biopsy, combined with rk39 antibody detection and epidemiological history are crucial for early diagnosis and treatment of visceral leishmaniasis. Antimonials is still an effective therapeutic choice.
    Report on 55 Cases of Small Intestine Hookworm Disease Diagnosed by Capsule Endoscopy
    WANG Pu1,LI Rong-zhi2,HUANG Zhi-yin1,TANG Cheng-wei1 *
    2013, 31(2):  13-140-142. 
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    Objective  To demonstrate the diagnostic value of capsule endoscopy for small intestine hookworm disease.  Methods  A retrospective study was carried out to analyze the clinical data and capsule endoscope image of 55 patients with small intestine hookworm disease in the hospital from June 2006 to June 2012.  Results  Among these patients, 40 cases manifested as gastrointestinal bleeding, 7 had iron deficiency anemia, 6 had chronic abdominal pain, and 2 showed abdominal distension or discomfort. Hookworm eggs were found in stool specimens of 2 cases, 6 cases showed peripheral eosinophilia, 46 cases were found to be fecal occult blood positive. Out of the 55 cases investigated, 44 showed anemia (11 severe, 26 moderate, and 7 mild). All patients were definitely diagnosed by capsule endoscopy. The hookworms were translucent and about 5-10 mm in length. Hookworms in most cases were diffusely distributed, but 12 patients suffered massive and severe hookworm infection. In most cases, hookworms were found in the proximal small intestine, and 6 in the distal intestine. Erosion and injury in intestinal mucosa around the hookworm were observed in several cases.  Conclusion  Capsule endoscopy is an effective and safe diagnostic technique for hookworm disease in small intestine.
    Progress on the Epidemiology of Echinococcosis
    QI Yan-feng,WU Wei-ping*
    2013, 31(2):  14-143-150. 
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    Echinococcosis is a zoonotic parasitic disease caused by the larval stages (metacestodes) of cestodes belonging to the genus Echinococcus. Echinococcosis is a major public health problem in many countries and regions. The epidemiological study of echinococcosis would contribute to the control and elimination of this disease. This paper summarizes the research status and progress on epidemiology of echinococcosis.
    Research Progress on the Mechanism of Host Immune Response Regulated by Trichinella spiralis
    ZHAO Ge, YANG Wen-tao, WANG Chun-feng, YANG Gui-lian*
    2013, 31(2):  15-151-154. 
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    Trichinosis caused by Trichinella spiralis is a parasitic zoonosis with world-wide distribution, which impacts on the development of animal husbandry and food safety, and thus threatens human health. T. spiralis has the ability to evade the host immune response, which results in forming a long-term infection in the host. The previous studies indicated that a changed host immune state due to T. spiralis was an important reason for the evasion. Among the factors, cytokines, dendritic cells and regulatory T cells played an important role in the regulation of the host immune process.
    Research Priorities for the Control and Elimination of Major Helminthiases
    QIAN Men-bao,CHEN Ying-dan,ZHOU Xiao-nong*
    2013, 31(2):  16-155-159. 
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    In 2009, the Disease Reference Group on Helminth Infections(DRG4) was established by the Special Programme for Research and Training in Tropical Diseases(TDR) to comprehensively review recent advances, identify gaps and rank priorities in helminthiases research towards control and elimination. Six major human helminthiases are targeted, namely onchocerciasis, lymphatic filariasis, soil-transmitted helminthiases, schistosomiasis, food-borne trematodiasis and cysticercosis/taeniasis. Systematic reports made by the DRG4 from such aspects as the impact of helminthiases, control and elimination, interventions, diagnostics, social-ecology and health systems, modeling, basic research and capacity building in research was published in PLoS Neglected Tropical Diseases in 2012. Generalized introduction is presented here and further analysis of its influence on the research of the major helminthiases in China is done.
    IL-10 Level in Allergic Rats Infected by Echinococcus granulosus
    AN Ran,HUANG Mou,ZHANG Qin,ZHENG Hong*
    2013, 31(2):  17-86-88. 
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    Forty Wistar rats were randomly divided into 4 groups with 10 in each group. Group A served as normal control. The other 3 groups were injected intraperitoneally with Echinococcus granulosus protoscoleces 5×104 each. Six months later, group D was injected intraperitoneally with antibody against IL-10 2.0 μg/time, twice a day for 2 d. Two days later, rats in groups C and D were injected intraperitoneally with cyst fluid 5 ml each to induce allergic reaction. 30 min later, all the rats were sacrificed to observe the infection status and obtain peripheral blood. The level of IL-10, IgE and histamine in the sera was detected by ELISA. The results showed that 5, 6 and 5 rats in groups B, C and D were infected successfully, respectively, with 2 deaths in group A. The rats in groups C and D appeared nasal itching, sneezing and declined activity. 30 min later, the symptoms in group C got improved, but not for group D. The levels of IL-10 and IgE in groups C and D increased significantly compared to group B(P<0.05). The levels of IL-10 and IgE in group D were (142.61±43.58) pg/ml and (20.67±1.58) μg/ml, respectively, lower than those in group C(P<0.05). The level of histamine was(17.69±3.90) ng/ml, higher than that in groups B and C(P<0.05). There was no significant difference in histamine level between B and C(P>0.05).
    Investigation on the Current Situation of Human Soil-borne Nematode Infection in Shapingba District of Chongqing
    LIU Hong-hong1,YANG Lian-jian1,CHEN Min1,LI Ting-rong1,CHEN Yin-zhi2
    2013, 31(2):  18-107-109. 
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    By stratified cluster sampling method, 2 urban and 2 rural fields were selected from Shapingba district of Chongqing for survey in December 2009 to February 2010. According to the Administrating Regulations of National Investigation on Important Human Parasitic Diseases, Kato-Katz method was used to examine human intestinal soil?鄄borne nematode eggs, and adhesive cellophane anal swab method was applied to examine Enterobius infection for children under 12 years old. 203 cases were found positive in 2 121 subjects, with an infection rate of 9.6%(203/2 121), and the infection rate of hookworms, Ascaris lumbricoides and Trichuris trichiura with mild infection mostly was 9.3%(197/2 121), 0.4% (8/2 121) and 0.1%(2/2 121), respectively. The rate among people over 50 years old was 15.5%(160/1 030), and the farmers was with 22.3%(113/506). The higher the education level, the lower the infection rate(P<0.01), and there was a significant difference in the prevalence between ruban(2.1%)and rural people(17.3%)(χ2=140.443 5, P<0.01). The infection rate of soil-borne nematodes in Shapingba of Chongqing was much lower than the standard of Ⅱ regions and most infected subjects were with hookworm infection.
    Relationship between Morphology and Pathogenicity of  Blastocystis hominis Trophozoites
    SHEN Ji-qing1,TIAN Chun-lin1,LU Zuo-chao1,WAN Xiao-ling2,LIU Deng-yu1,LIU Xiao-quan1,WANG Jing1,LI Xue-ming2 *
    2013, 31(2):  19-137-139,142. 
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     Six hundred and eighty-six fresh fecal specimens were collected from outpatients(663 well-formed feces and 23 watery feces) during March 2011 to March 2012. All specimens were examined microscopically by direct smear and iodine stained method. B. hominis obtained from the human positive fecal specimens were cultured in LES medium, and inoculated into the abdominal cavity of 10 female mice of 6-8-week old. The abdominal fluid was examined with same methods. 103 of 686 patients were positive (80 well-formed feces and 23 watery feces). Micro-scopically, the granular form and vacuolated form of B. hominis trophozoites could be easily identified by direct smear and iodine staining in well-formed fecal specimens, showing ovoid in shape and about(13.2±0.2) μm in size. The tro-phozoites cultured in LES medium showed similar feature. But in the watery fecal specimens and mice ascites specimen, they were amorphous containing more granules. And their average size was(28.0±0.3) μm which was larger than the former. Moreover, the amebal form of B. hominis trophozoites was also detected in the 23 watery fecal specimen and mice ascites specimen. The trophozoites of B. hominis were varying in shape and size depending on their living environment.
    Sequence Homology Analysis on Lactate Dehydrogenase(LDH)of Plasmodium vivax Anhui Isolates
    HU Ming-jie1,FANG Qiang2 *,WU Shou-wei1,TANG Bi-kui1,ZHANG Jing1,JIAO Yu-meng2,XIA Hui2,SHEN Ji-long3
    2013, 31(2):  20-148-150. 
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     DNA in dried blood spots of 39 vivax malaria patients (2009-2010) from Anhui(Bengbu urban district and counties of Wuhe, Huaiyuan, Mengcheng and Lixin) was extracted. The Plasmodium vivax LDH(PvLDH) gene was amplified, cloned and sequenced. The sequences were subjected to NCBI Blast program. The results showed that the targeted DNA fragment size was 951 bp without difference among the 39 samples(accession No. GU078391), and was more than 99% homologous to the PvLDH sequences in other strains from GenBank. There was only one different amino acid in the protein sequences between the isolates from Anhui and EJEU60134 or MIA061251 strains.
    Field Evaluation of SDBIOLINE Malaria Antigen Plasmodium falciparum/Plasmodium vivax Rapid Test Kit
    LIU Hui1 *, LI Xi-rong2, LI Chun-fu1, LI Xing-liang1, WANG Heng-ye1, NIE Ren-hua1
    2013, 31(2):  21-160-封三. 
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    Four hundred and seventy-five patients with fever within 48 h were detected for Plasmodium using double blind field trials in China-Myanmar border from June to December 2011. The result showed that 202 of 475 were positive by SDBIOLINE kits, with 98 positive of Plasmodium falciparum and 104 positive of Plasmodium vivax. By microscope examination, 206 were positive. Taking the result of microscope examination as the reference standard, the general sensitivity and specificity were 98.1%(202/206) and 97.8%(263/269) respectively, and the general coincidence rate of SDBIOLINE kits with microscopy was 97.9%(465/475). The sensitivity and specificity of P. falciparum were 99.0%(98/99) and 97.8%(263/269) respectively, and the coincidence rate of SDBIOLINE with microscopy was 98.1%(361/368). The sensitivity and specificity of P. vivax were 97.2%(104/107) and 100%(269/269), and the coincidence rate of SDBIOLINE with microscopy was 99.2%(373/376). Therefore, the test results of SDBIOLINE are stable with a high specificity and sensitivity.
    Case report:Fasciola gigantica infection treated by triclabendazole
    FANG Wen, CHEN Feng, YANG Qiong, LIU Yu-Hua, LIU Hong-Kun
    2013, 31(2):  22-封二. 
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