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    30 October 2018, Volume 36 Issue 5
    Orginal Article
    Commemorating Professor MAO Shou-bai, a pioneer of the national program for schistosomiasis control--A commentary for the sixtieth anniversary of the issuance of Chairman MAO Ze-dong’s poem Farewell to the God of Plague
    Sen-hai YU
    2018, 36(5):  429-431. 
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    This commentary recalled the brilliant contribution of Professor MAO Shou-bai in opening up the cause of schistosomiasis control in China, and emphalized that his spirit of dedication, sense of duty and rigorousness must be carried on.

    Emerging challenges: inheritance and innovations of human parasitology education
    Lin CHEN, Jun CAO, Dong-hui ZHANG, Kun YANG, Jing-fan QIU, Zhi-peng XU, Min-jun JI, Guan-ling WU
    2018, 36(5):  432-435. 
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    The most important parasitic diseases have been virtually controlled in China, but are still a major threat to public health in the world, especially in developing countries. With the increased international communications, there have been increases in cases of imported parasitic infections in China. The interest of the global community as raised by the “the Belt and Road Initiatives" also poses a new task for medical education. Human parasitology is a basic course in clinical and preventive medicines, and more importantly, it is imperative to re-think about the development of parasitology discipline. This review summarizes the opportunities and challenges for current parasitology, and proposes that teachings in human parasitology to be strengthened rather than weakened, inherited yet with innovations. Suggestions are also made on reforms to and innovations of parasitology teaching from perspectives of teaching content and form.

    Diagnosis and treatment of human African trypanosomiasis
    Qin LIU, Mu-xin CHEN, Yuan-yuan LI, Yi ZHANG, Shi-zhu LI, Jia-xu CHEN, Xiao-nong ZHOU
    2018, 36(5):  435-442. 
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    Human African trypanosomiasis (HAT) is a neglected tropical disease. The diagnosis and treatment of HAT is quite complicated. With the large-scale migration of populations and the globle economic activity, the imported cases of HAT are increasing these years. Therefore, the training of professionals and technicians on the related knowledge and technology is particularly important. In this paper, the diagnosis and treatment schemes of HAT are systemically summarized for the reference of clinicians and laboratory technicians.

    Evaluation of the molluscicidal effect and cost of three molluscicides combined with a herbicide
    Deng CHEN, Kuang-ji ZHU, Jian-feng ZHANG, Xiao-lan YAN, Hong-xin WANG, Hai-juan DU, Feng-yao SHAO, Xiao ZHANG, Qing-xiang HUANG, Lan-jin ZHANG, Li-yong WEN
    2018, 36(5):  443-449. 
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    Objective To observe the molluscicidal effect and the cost of molluscicides: 25% niclosamide ethanolamine salt (SCN), 26% suspension of metaldehyde and niclosamide ethanolamine salt(MNSC), and 50% niclosamide ethanolamine salt (WPN) used in combination with glyphosate, in order to evaluate their application value. Methods The SCN, MNSC and the glyphosate were prepared into effective doses of 2 μl/L (for laboratory immersion test), 2 ml/m3 (for field immersion test) and 2 mL/m2 (for spraying test), and the wettable powder of 50% WPN was prepared into effective doses of 2 mg/L (for laboratory immersion test), 2 g/m3 (for field immersion test) and 2 g/m2 (for laboratory and field spraying test). There were 5 experimental groups in this study: SCN, SCN in combination with glyphosate (1 : 1), MNSC, MNSC in combination with glyphosate (1 : 1), and WPN in combination with glyphosate (1 : 1). WPN was used as a drug control. Glyphosate and water control groups were also set. In the laboratory immersion experiment, 30 snails were deposited in each group, and in the field immersion test, 9 bags of snails were deposited, each containing 30 snails. At 24 h, 48 h and 72 h after drug administration, 3 bags of snails were taken back to calculate the snail death rate. In the laboratory spraying test, 100 snails were deposited in each group and the snail death rate was calculated on days 1, 3 and 7 after drug administration. In the field spraying test, a 100 m2 area of snail-containing ditch was administered with drugs, and on days 1, 3 and 7 after drug administration 10 boxes of snails were selected to calculate the snail death rate. The cost-benefit was analyzed based on the cost of molluscicides, labor cost, and herbicide labor cost. Results In the laboratory immersion experiment, the snail death rate reached 100% in all the experimental groups at 24 h, 48 h and 72 h, except for the SCN in combination with glyphosate group at 24 h, in which the death rate was 96.7% (29/30). There was no significant difference between experimental groups and the drug control group. In the laboratory spraying test, the snail death rate reached 100% in all the experimental groups on days 3 and 7, except for the SCN in combination with glyphosate group on day 1 (91.0%, 91/100) and on days 3 and 7(≥95%) at 24 h. There was no significant difference between experimental groups and the drug control group. In the laboratory immersion experiment, the snail death rate reached 100% in all the experimental groups and the drug control group at 24 h, 48 h and 72 h, except for the SCN in combination with glyphosate group (>95%). There was no significant difference between the groups (P > 0.05). In the field spraying test, the snail death rate reached 100% in MNSC in combination with glyphosate group on days 1, 3 and 7, higher than those in the drug control group (91.8%, 258/281; 95.4%, 206/216; 94.5%, 147/155) (P < 0.05). There was no significant difference between the other 4 experimental groups and the drug control group (P > 0.05). The costs of SCN, MNSC and WPN without glyphosate per 100 m2 were 101.00 Yuan, 101.80 Yuan and 103.90 Yuan due to the need of more labor input, while the costs of SCN compounding form, MNSC compounding form and WPN compounding form per 100 m2 were 69.50 Yuan, 70.30 Yuan and 72.40 Yuan, all less than the former.Conclusion On consideration of molluscicidal effect and the application cost, 2 g/m2 or 2 ml/m2 compound drugs of the three types of molluscicides are recommended in field snail control by the method of spraying.

    Pharmacokinetics of tribendimidine in rat plasma and bile
    Bin JIANG, Yi TAO, Li-li XU, Le-le HUO, Yu-fen WEI, Ni LIU, Meng YIN, Hao-bing ZHANG
    2018, 36(5):  449-454. 
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    Objective To study the pharmacokinetics of tribendimidine in rat plasma and bile. Methods Twenty male Sprague Dawley rats were randomly assigned into control and tribendimidine groups (n = 10 in each). Rats in the tribendimidine group were further divided into two batches after oral gavage of 200 mg/kg tribendimidine: 5 were used for orbital blood collection at 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 8.0, 10.0, 24.0 and 36.0 h after administration, while the other 5 were used for bile collection at 0.5-1.0, 1.0-1.5, 1.5-2.0, 2.0-2.5, 2.5-3.0, 3.0-3.5, 3.5-4.0, 4.0-4.5, 4.5-5.0, 5.0-5.5, 5.5-6.0, 6.0-6.5, 6.5-7.5, 7.5-8.5, 8.5-9.5, and 9.5-10.5 h. Rats in the control group received same operations except that no tribendimidine was administered. After optimizing HPLC detection conditions, concentrations of p-(1-dimethylamino ethylimino) aniline (dADT) in rat plasma and bile were measured. The concentrations were plotted against time, and curves for bile excretion rate and cumulative excretion were drawn. Results The time for maximum dADT (Tmax) and the maximum concentration of dADT (Cmax) in plasma were (0.9 ± 0.2) h and (8.1 ± 2.1) mg/L, respectively, and those in the bile were (1.2 ± 0.4) h and (11.2 ± 2.1) mg/L, respectively. The bile excretion rate was highest during 0.5-3.0 h after administration, and the cumulative excretion quantity of dADT was(40.1 ± 12.0) μg within 11 h, with a cumulative excretion rate of 0.57‰. Conclusion The tribendimidine metabolite dADT shows a higher concentration and a longer stay in the bile than in the plasma, which may be beneficial for the treatment of Clonorchis sinensis that parasitizes in the biliary tract.

    Effect of oxymatrine on Toll-like receptors on intestinal mucosa of mice infected by Cryptosporidium parvum
    Rui JI, Rui-wen LIANG, Zhi-yu GUAN, Rui-fang LI, Yu-rong FU, Hong-yan WANG
    2018, 36(5):  455-460. 
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    Objective To study the effects of oxymatrine (OMT) on Toll-like receptors 2 (TLR2) and Toll-like receptors 4(TLR4) on intestinal mucosa of mice infected with Cryptosporidium parvum and investigate the molecular mechanisms of cryptosporidiosis treatment by OMT. Methods Thirty healthy BALB/c male mice were randomly divided into the infection group, OMT treatment group and non-infection group(n = 10 in each group). Mice in the infection group and OMT treatment group received intragastrical inoculation of C. parvum oocysts (1 × 105/gram) to establish the model of C. parvum intestinal infection, while mice in the non-infection group were fed with normal diet and water. Mice in the OMT treatment group received daily intragastrical inoculation of 50 mg/kg for 2 weeks. The numbers of mean oocysts per gram of mouse feces were counted using the auramine-phenol modified anti-acid staining method. All mice were sacrificed after 2 weeks of treatment. The pathological changes of intestinal mucosa were observed by HE staining, and the height of intestinal villi and the depth of crypt were measured. RNA was extracted from intestinal mucosa, and transcribed into cDNA. The relative expression of TLR2 mRNA and TLR4 mRNA in intestinal mucosa was assessed by real-time fluorescence PCR. The relative expression of TLR2 and TLR4 in intestinal mucosa was assessed by Western blotting. Result The auramine-phenol modified anti-acid staining revealed increased numbers of mean oocysts in mouse feces in the infection group, peaking at day 7 with(179.24 ± 21.12) oocysts/gram, infection intensity was level 4, and reaching a plateau afterwards. In the OMT treatment group, the numbers decreased from day 5, and infection intensity was approaching level 0 after 2 weeks of treatment. HE staining showed atrophy and shedding of intestinal villi in the infection group, with submucosal edema and a significant gap between muscle layers, while the intestinal villi were intact in the non-infection group. The intestinal villi were restored in the OMT treatment group. The villi heights in the non-infection group, infection group and OMT treatment group were (348.1 ± 18.2), (278.0 ± 52.9) and (346.1 ± 19.2) μm, respectively, OMT treatment group was higher than infection group (P < 0.01), and there was no significant difference between OMT treatment group and non-infection group (P > 0.05). The crypt depths in the non-infection group, infection group and OMT treatment group were (149.9 ± 27.4), (173.1 ± 11.1) and (155.4 ± 5.3) μm, OMT treatment group was lower than infection group (P < 0.05), and there was no significant difference between OMT treatment group and non-infection group (P > 0.05). The ratios of villi height/crypt depth in the non-infection group, infection group and OMT treatment group were 2.4 ± 0.6, 1.6 ± 0.3 and 2.2 ± 0.2, OMT treatment group was higher than infection group (P < 0.01), and there was no significant difference between OMT treatment group and non-infection group (P > 0.05). qRT-PCR showed that the relative expression of TLR2 mRNA in intestinal mucosa were 1.0 ± 0.0, 3.0 ± 0.1 and 1.4 ± 0.3 in the non-infection group, infection group and OMT treatment group, and that of TLR4 mRNA were 1.0 ± 0.0, 3.1 ± 0.3 and 1.5 ± 0.1, respectively. That of OMT treatment group was lower than infection group (P < 0.01), and there was no significant difference between OMT treatment group and non-infection group (P > 0.05). Western blotting showed that the relative expression of TLR2 in the intestinal mucosa were 0.2 ± 0.1, 0.6 ± 0.1 and 0.2 ± 0.0, and that of TLR4 were 0.2 ± 0.1, 0.6 ± 0.0 and 0.3 ± 0.1 in the non-infection group, infection group and OMT treatment group respectively. That of OMT treatment group was lower than that of infection group (P < 0.01), and there was no significant difference between OMT treatment group and non-infection group (P > 0.05). Conclusion OMT improves intestinal mucosal repair through down-regulating TLR2 and TLR4 expression in mice infected with C. parvum.

    Orginal Article论著
    Regulations of IL-9 by the TGF-β/Smads signaling pathway in human Echinococcus granulosus infection
    Nan-nan PANG, Meng-ting AN, Feng-bo ZHANG, Xiao ZHAO, Fang YAN, Jian-bing DING
    2018, 36(5):  460-464. 
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    Objective To examine mRNA expression of TGF-β, Smad3, Smad7, PU.1 and Interleukin-9 (IL-9) in livers of patients with cystic echinococcosis, in order to explore the regulations of Th9 cells by the TGF-β/Smad signaling in Echinococcus granulosus infection. Methods The liver mononuclear cells were isolated from liver tissues obtained during surgery from 30 patients with cystic hydatid disease and 20 patients with hepatic hemangioma (control group) in the First Affiliated Hospital of Xinjiang Medical University using the Percoll method. Real-time fluorescent quantitative PCR was performed to detect the mRNA expression of TGF-β, Smad3, Smad7, PU.1 and IL-9 in the mononuclear cells. The relationship between Smad3 mRNA expression and PU.1 mRNA expression was assessed by Spearman correlation analysis. The Smad3 inhibitor SIS3 or PBS was added to the mononuclear cells of the infected group, then PU.1 and IL-9 mRNA expression were determined by real-time fluorescent quantitative PCR, and IL-9 level in the supernatant of cell culture was assessed by ELISA. Results The relative mRNA expression of TGF-β, Smad3, PU.1 and IL-9 in CE patients were 2.55 ± 1.32, 2.26 ± 1.35, 1.71 ± 0.66 and 1.67 ± 0.75, respectively, all significantly higher than those in the control group (1.54 ± 0.76, 1.34 ± 0.51, 1.27 ± 0.62 and 1.01 ± 0.50, P < 0.05 or 0.01). The relative mRNA expression of Smad7 in CE patients (0.49 ± 0.37) was significantly lower than that in the control group (0.88 ± 0.50; P < 0.01). The Smad3 mRNA expression and PU.1 mRNA expression were positively correlated (r = 0.397, P < 0.05). After blocking the TGF-β/Smad signaling pathway by SIS3, the expression of PU.1 and IL-9 mRNA in liver mononuclear cells (1.00 ± 0.57, 0.85 ± 0.50) both decreased compared with the PBS treatment (1.46 ± 0.64, 1.32 ± 0.72) (P < 0.01). ELISA results showed that the IL-9 level in the supernatant of cell culture [(128.45 ± 41.97) pg/ml] also reduced compared with the PBS treatment [(166.64 ± 57.90) pg/ml; P < 0.05]. Conclusion The expression of TGF-β, Smad2, Smad3, PU.1 and IL-9 up-regulated in patients with cystic hydatid disease and the TGF-β/Smad pathway may regulate the expression of IL-9 after Echinococcus granulosus infection.

    Orginal Article
    Effects of recombinant autophagy related 5 protein on the maturation of dendritic cells stimulated by Toxoplasma gondii
    Shang-hua WU, E-yan GENG, Jing ZHANG, Heng ZHANG, Zhi-qiang SHI, Shan WANG, Wei LU, Yi-zhen WU, Gui-jun WANG, Yong WANG, Qian-ming XU
    2018, 36(5):  464-468. 
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    Objective To investigate the effect of autophagy related 5 (ATG5) protein on the maturation of dendritic cells (DCs) stimulated by Toxoplasma gondii. Methods Total RNA was extracted from Kunming mice using TRIzol, reverse transcribed into cDNA, and amplified by PCR to receive ATG5 gene. The recombinant plasmid pET-32a-ATG5 was constructed and transformed into Escherichia coli Rosetta strain. The expressed recombinant ATG5 protein was purified by urea and identified by Western blotting with anti-His tag mAb. Mouse DCs were isolated, and then infected with T. gondii, which were treated by recombinant ATG5 protein in vitro. There were three groups: control group, T. gondii group (T. gondii + DCs), and ATG5 protein group (ATG5 + T. gondii + DCs), each with 3 replicates. The T. gondii tachyzoites (1 × 104) were added to the DCs and incubated at 37 ℃ under 5% CO2 for 2 h. The T. gondii in the ATG5 protein group were pre-treated with ATG5 protein (50 μg) in vitro for 2 h. The percentage of CD11cCD83 mature DCs among all DCs was assessed by flow cytometry. The effect of ATG5 protein on the maturation of DCs stimulated by T. gondii was evaluated. Result The amplified product of ATG5 was 729 bp, and the relative molecular mass (Mr) of the recombinant ATG5 protein was 46 000. Western blotting showed that the expressed recombinant ATG5 protein had good reactivity. Results of flow cytometry showed that the percentages of CD11cCD83 mature DCs in the control group, T. gondii group, and ATG5 protein group were (13.3 ± 0.5)%, (52.5 ± 0.3) % and (47.8 ± 0.6) %, respectively, with a significant difference between the latter two groups (P < 0.01), and between the control group and the other two (P < 0.01). Conclusion The ATG5 protein can reduce the ability of T. gondii to stimulate the maturation of DCs in vitro.

    Cloning, expression and identification of cysteine protease genes AcCBL1 and AcCBL2 of Angiostrongylus cantonensis
    Hui-fang BAI, Qi-wen KE, Yun-qiu CHEN, Zhi-qing LIU, Ling-min ZHANG, Chun-yun WU, Gui-xiu YUAN, Xi-mei ZHAN, Mei CHENG
    2018, 36(5):  469-473. 
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    Objective To clone and express the cysteine protease genes AcCBL1 and AcCBL2 of Angiostrongylus cantonensis, and analyze the immunoreactivity of the purified recombinant proteins. Methods Two cysteine protease genes AcCBL1 and AcCBL2 were identified from Angiostrongylus cantonensis fourth-stage larvae cDNA library, and cloned into pET-28a vector to generate recombinant plasmids pET-28a-AcCBL1 and pET-28a-AcCBL2, respectively. The constructed recombinant plasmids were transformed into(E. coli) BL21(DE3) respectively for expression under the induction of isopropyl β-D-1-thiogalactopyranoside(IPTG). The recombinant proteins rAcCBL1 and rAcCBL2 were purified by Ni-NTA affinity chromatography and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The immunoreactivity of rAcCBL1 and rAcCBL2 were analyzed by Western blotting with 6 × His tag antibody, as well as anti-sera from mice infected with A. cantonensis and patients. Results PCR generated a specific band at 1 000 bp and 1 100 bp for AcCBL1 and AcCBL2, respectively, consistent with the theoretical values. The constructed vector pET-28a-AcCBL1 and pET-28a-AcCBL2 were confirmed by gene sequencing. SDS-PAGE revealed that rAcCBL1 was soluble with relative molecular mass of 37 800, and rAcCBL2 was expressed as inclusion body in E. coli with relative molecular mass of 40 800. Western blotting showed that the two purified recombinant proteins can be recognized by 6 × His tag antibody, as well as anti-sera from mice infected with A. cantonensis and patients. Conclusion AcCBL1 and AcCBL2 genes have been expressed in prokaryotic expression system. The two recombinant proteins both show adequate immunoreactivity.

    Morphological observation and phenotypic detection of dendritic cells from peripheral blood of patients with alveolar echinococcosis
    Yong FU, Ru MENG, Hai-fang XUE, Hai-ning FAN, Hai-feng NIU, Zi-jia ZHOU, Hong-bin WANG
    2018, 36(5):  474-477. 
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    Objective To understand the morphological and phenotypic characteristics of dendritic cells (DCs) from peripheral blood of patients with alveolar echinococcosis. Methods Peripheral blood was collected from 10 patients with alveolar echinococcosis (AE), 10 healthy Han participants (HH) (n = 10) and 10 healthy Tibetan participants (TH), respectively. Monocytes were isolated, and cultured by adherent culture. The monocytes were induced into DCs with recombinant human granulocyte macrophage colony stimulating factor and recombinant human interleukin 4. DCs were collected on days 1, 3, 5 and 7 of culture and cell morphology was observed by inverted microscopy and scanning electron microscopy. The positive rate of cell surface markers was detected by flow cytometry on day 7. Data were analyzed with SPSS 22.0 software. Results On day 1 of in vitro induction, the majority of DCs in the 3 groups had a single round shape with clear boundary, bright cytoplasm, and a small volume, floating in the culture medium. On day 3 of induction, the three groups of DCs aggregated into semi-suspension and the cells enlarged with an irregular shape. On day 5 of induction, the three groups of DCs formed colonies, and most DCs had a rough surface. On day 7 of induction, the three groups of DCs grew into semi-suspension with a filamentous cell edges. Compared to the HH and TH groups, the number of DC colonies in group AE was smaller, and the irregular protuberances of DC were not obvious, displaying an atypical DC morphology. The positive expression rates of CD1a, CD80 and CD86 on the surface of DCs induced differentiation in AE group (12.73% ± 1.73%, 12.41% ± 2.83% and 16.34% ± 3.59%, respectively) were significantly lower than those in the HH and TH groups (P < 0.05), while those of HH group (18.40% ± 1.20%, 20.77% ± 3.40%, 31.78% ± 5.02%) and TH group(17.50% ± 1.44%, 23.75% ± 5.33%, 33.20% ± 2.47%) were not significantly different (P > 0.05). Conclusion The induced DCs in the AE group have an atypical morphology and a decreased level of cell surface markers, indicating the occurrence of maturation disorder.

    Investigation on species and sporozoite rate of Anopheles spp. in Champasak Province of Laos
    Wei-li WANG, Rui YANG, Chun-hai LUO, Xiao-fang GUO, Jin-yong JIANG, Hong-ning ZHOU, SORCHAMPA Somphad, VANNAPASEUTH Phoxay, Cang-lin ZHANG
    2018, 36(5):  478-482. 
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    Objective To investigate the species of Anopheles and rate of Plasmodium sporozoites in anopheline mosquitoes in Champasak Province of Laos, so as to provide essential data for malaria control. Methods Mosquitoes were captured by overnight trapping with light traps and by outdoor human landing catches in Batong County of Champasak Province in July 2017. After morphological identification, genomic DNA was extracted from some female adult mosquitoes, and 18S rRNA-based nested-PCR was performed to detect soprozoites of Plasmodium falciparum and P. vivax. The sporozoite positive rate of Anepheles was calculated. Results A total of 34 687 mosquitoes were captured, belonging to 29 species of 8 genera, dominated by the genus Culex (92.6%, 32 110/34 687), followed by genus Anopheles (5.6%, 1 947/34 687). The genus Anopheles was dominated by An. vagus (65.5%, 1 275/1 947), followed by An. kochi (12.1%, 235/1 947), An. philippinensis (11.9%, 232/1 947), An. sinensis (5.1%, 100/1 947). A total of 336 anopheline female mosquitoes (comprising 234 An. kochi, 100 An. sinensis and 2 An. dirus) were examined by nested-PCR, of which 8 were detected to be positive for P. vivax, including 6 An. kochi (positive rate 2.6%, 6/234) and 2 An. sinensis (2.0%, 2/100). No soprozoites of P. falciparum were found. Nested-PCR amplification showed a band of ~120 bp, sequence of which had 99%-100% homology to those of P. vivax (GenBank Accession No: KT991325, KT991317, MG708221, MF540772, LT635613, KU569498, AF145335, KT991314 and KX007932). Conclusion Both An. kochi and An. sinensis mosquitoes carry P. vivax sporozoites with a positive rate of 2.6% and 2.0% respectively.

    Epidemiological analysis of visceral leishmaniasis in Hancheng City of Shaanxi Province in 2017
    Feng-ping WANG, Zheng ZHANG, Wen LV, Dong-li LIU, Tian-hai WANG, Yi-qun WANG, An-li WANG, Sheng-zhen LI, Lei SHI, Lin MA, Lei CAO, Feng WU
    2018, 36(5):  483-488. 
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    Objective To understand the current endemic status of visceral leishmaniasis in Hancheng City and provide basis for the establishment of endemic surveillance and control measures. Methods The endemic information and survey reports of visceral leishmaniasis in Hancheng City in 2017 were collected from the Disease Surveillance Reporting System of the Chinese Center for Disease Control and Prevention. Blood was collected from the patients. Six villages that have reported cases since 2012 were selected as the survey spots, including the Chengjiadong Village, Yangwan Village, and Shuicaota Village in Sangshuping Town, Caojiashan Village in Xizhuang Town, Dongjia Village in Zancun Town, and Xiyuan Village in Longmen Town. The former four villages located in the hilly regions while the latter two in the Loess plateau area. Based on the family information on disease incidence, anticoagulation and normal blood samples (3 ml each) were collected randomly from residents and domestic dogs. Bone marrow was collected from suspected dogs. Dogs with positive serological results were sacrificed to collect bone marrow swab. The sandfly density was monitored manually from middle June to late September. The rK39 immunochromatographic test strip and ELISA kit were used to detect the antibody for Leishmania in human serum and dog serum, respectively. The positive samples were further confirmed by rK39 immunochromatographic test strip. The dog bone marrow underwent pathological testing. DNA was extracted from blood, bone marrow and sandfly specimens. PCR was performed for kinetoplast target gene of Leishmania spp. The positive samples were further used for amplification of internal transcribed spacer 1(ITS-1) sequence, small subunit ribosomal RNA(SSU rRNA) and DNA repeats, and cluster analysis was made to identify the species. Result A total of 10 cases of visceral leishmaniasis in Hancheng City were reported in 2017, of whom 8 were workers from other cities and 2 were indigenous kids. The positive rate of visceral leishmaniasis among residents was 3.1% (7/224). No positive results of PCR were found in the asymptomatic residents. The positive rate of visceral leishmaniasis in dogs was 34.3% (115/335), which was higher in hilly mountain area (51.5%, 105/204) than that in plain area(7.6%, 10/131)(χ2 = 68.000, P < 0.05). The average sandfly density from June to September was 16.7/man hour, with a peak of 122/man hour. The average sandfly density in the hilly area was 23.6/man hour, significantly higher than 0.3/man hour in plateau area (t = 2.19, P < 0.05). A total of 366 sandflies were captured, including 168 males and 168 females, and 228 were identified, of which 29.4% (67/228) were Phlebotomus chinensis. The positive rate of Leishmania in sandflies was 1.5% as revealed by PCR. Leishmania was isolated from the bone marrow samples from 1 dog and 2 patients, and identified as Leishmania infantum by ITS-PCR, SSU rRNA-PCR and reiterated DNA sequence. Conclusion Leishmania infection is present among residents, dogs and sandflies in Hancheng City. The infection rate in hilly areas is higher than that in the plain areas.

    Laboratory detection of parasitic infections in clinical samples from hospitals in Shanghai during 2014-2017
    Peng SONG, Hao LI, Jian GUO, Lin AI, Yan LU, Yu-chun CAI, Mu-xin CHEN, Hai-mo SHEN, Shao-hong CHEN
    2018, 36(5):  489-494. 
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    Objective To analyze the results of laboratory detection of parasitic infections in clinical samples from hospitals in Shanghai during 2014-2017. Methods Samples included in this study were from hospitals in China which cooperated with National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, consisting of 7 secondary hospitals, 28 tertiary hospitals, and 6 foreign-related hospitals. The samples included whole blood, serum, feces, sputum, urine, body fluids (hydrothorax and ascites), histopathological sections, and worms. Information of the patient’s clinical data was also recorded. Parasitic infection in the samples was examined using direct smear method, modified acid-fast staining method, ELISA, and molecular biological methods. Results A total of 25 549 clinical samples were sent to the institute for examination, including 8 877 samples for pathogen detection, of which 633 (7.1%) were found to be infected with 33 species of parasites. Specifically, 331 samples were with Blastocystis homini (3.7%) and 120 with Entamoeba histolytica (1.4%). In addition, 16 672 samples were examined by the serological method, resulting in positive results for 1 881(10.9%) samples, consisting of 424 samples seropositive for Cysticercus cellulosae (23.4%), 388 for Sparganum mansoni (21.4%), 289 for Schistosoma japonicum (16.0%), 267 for Paragonimus westermani (14.7%), and 260 for Toxoplasma gondii (14.4%). Protozoan infection was detected in 371 samples, most of which were from Shanghai (125 samples), and helminth infection was revealed by ELISA in 87 samples, mainly from Shanghai (29 samples). Conclusion B. hominis and E. histolytica were the major intestinal protozoans in pathogen detection in the samples, and the seropositive cases mainly show possible infection with C. cellulosae and S. mansoni.

    Evaluation on the measures by a grid-based design for wildlife control in hyper-endemic areas for echinococcosis
    Shuai HAN, Qing YU, Shi-jie YANG, Ning XIAO, Ping YE, Jing-bo XUE, Tian TIAN, Wei-ping WU, Xiao-nong ZHOU
    2018, 36(5):  495-498. 
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    Objective To evaluate the preliminary effect of a grid-based design of anthelmintic bait delivery on wild canines control in echinococcosis hyper-endemic areas. Methods Gemeng and Gayi Townships with frequent wild canine activities, of Shiqu County in Garze Tibetan Autonomous Prefecture in Sichuan Province were selected as experimental and control groups, respectively. During April and October in 2016, areas of 20 m × 100 m and 30 m × 100 m were in Gemeng Township to deliver anthelmintic baits based on the grid-based design. The depositing locations were recorded by a handheld GPS system. Eight to 10 pellets of anthelmintic baits (each containing 50 mg praziquantel) were deposited every two months (i.e. in April, June, August and October), meanwhile wild feces within a radius of 1 m from the depositing location were collected. In Gayi Township, an area of 200 m × 1 000 m was selected to collect no less than 50 samples of wild feces simultaneously. No baits were deposited there. The collected feces were identified preliminarily by morphological discrimination, and the fecal antigen positive rate was tested by ELISA. In addition, the density of small rodent intermediate hosts was measured in the two townships in April and August 2016. The density of plateau pika (Ochotona curzoniae) in Gemeng Township was estimated based on the presence of pika holes, while that in Gayi Township was based on the number of pikas captured within a 50 m × 50 m area. The plateau pikas were sacrificed to examine hydateid cyst infection in organs. Results During April and October 2016 in Gemeng Township, 240 spots were recorded with delivery of more than 15 000 baits (including loss). The occurrence rate of feces in Gemeng Township was 35.4%-56.3%, comprising 51.9%, 31.5% and 16.6% from foxes, dogs and wolves, respectively, while the constitution ratio in Gayi Township was 5.5%, 94.5% and 0.0%, respectively. A total of 464 and 418 wild feces were collected in Gemeng and Gayi Townships, respectively, with an antigen-positive rate of 1.08% and 1.20% respectively (P > 0.05). The densities of plateau pika in Gemeng Township were 273/hm2 (1 hm2 = 10 000 m2) in winter and 498/hm2 in summer, with a hydateid cyst infection rate of 6.8% (3/45). The densities of plateau pika in Gayi Township were 784/hm2 in winter and 632/hm2 in summer, with a hydateid cyst infection rate of 3.7% (13/354). ConclusionThe grid-based design can be used to survey the distributions of intermediate and terminal hosts of echinococcosis. However, more time is needed to examine the real effect of baits delivery in recorded spots.

    World Health Organization-external competency assessment of malaria microscopists in China
    Mei LI, He YAN, He-jun ZHOU, Xin-yu FENG, Zhi-gui XIA, Shui-sen ZHOU
    2018, 36(5):  499-504. 
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    In order to understand the weakness of, and develop strategies for malaria microscopy in China, 12 participants of the 2017 World Health Organization-external competency assessment of malaria microscopists (WHO-ECAMM) were included in this study, consisting of 11 experts in provincial institutions, and 1 expert in a national-level institution. Their personal information and judgement results on blood slides were collected. Judgement on each slide was scored for each participant, and the associations of parasite density with the correctness of participants in species identification and parasite counting were analyzed. The associations of the score levels of participants with their work experience in microscopy and the average number of malaria cases in recent 3 years in the relevant province where they worked were analyzed. A total of 42 slides were used for species identification. Results showed that the participants had highest correctness rate in identifying slides with Plasmodium ovale and P. malariae [both (93.8 ± 4.0)%], followed by negative slides [(92.5 ± 2.2)%], P. faciparum [(78.3 ± 22.3)%] and P. vivax [(70.8 ± 14.2)%]. There was no correlation between parasite density and correct rate of participants in identifying P. faciparum slides (r = -0.13). Results of parasite counting showed that the average correctness rate of the participants was (42.2 ± 5.9)%. When the parasite density was less than 2 000/μl, the correctness rate of parasite counting was positively correlated with the density(r = 0.79). There were 2, 4, 3 and 3 participants scored levels 1, 2, 3, and 4, respectively. The level was not correlated with work experience (r = -0.16), but was negatively correlated with the average number of malaria cases in the province where they worked (r = -0.55). These results suggested that it is necessary to strengthen the training and practice in slide reading and species identification among microscopists, to maintain or improve their ability to diagnose malaria with microscopy.

    Associations of natural killer cells and their receptors with liver fibrosis in schistosomiasis
    Xiao-ling WANG, Yuan HU, Jian-ping CAO
    2018, 36(5):  504-509. 
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    Schistosomiasis japonica is a widespread zoonotic parasitic disease. The parasite eggs can deposit in the host liver after infection, causing hepatic fibrosis, which further progresses into cirrhosis. Schistosomiasis severely impairs the life quality of patients and has become a major public health problem. Natural killer (NK) cells are important innate immune cells, which can respond to the target without prior sensitization. Hepatic stellate cells (HSCs) are the major source of myofibroblasts that cause liver fibrosis in schistosomiasis. Recent studies showed that the NK cells can alleviate hepatic fibrosis by killing HSCs. The development of hepatic fibrosis is closely related to the status of NK cell activation, while the latter depends on the expression of their inhibitory or activating surface receptors. In this paper, recent research advances on NK cells and their surface receptors are reviewed, which provides a perspective of studies on the roles of NK cells against schistosomiasis liver fibrosis.

    Research progress on disability weight for major helminthic diseases
    Xin-zhong ZANG, Huan-zhang LI, Men-bao QIAN, Hui-hui ZHU, Chang-hai ZHOU, Ying-dan CHEN, Zhi-qiang QIN, Shi-zhu LI
    2018, 36(5):  510-515. 
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    The helminthic diseases are caused by helminths that parasitize in mammals, which are receiving inadequate attention due to their prevalence in remote, low-income and resource-constrained countries and regions. In 2010, the World Health Organization listed 17 neglected tropical diseases, of which 8 were helminthic diseases. The impact of these helminthiases is less studied. This paper reviews the prevalence, harmful effects and disability weight of common helminthic diseases, and comprehensively analyzes research methods and applications of the disability weight as well as the problems that exist in these processes.

    Research advances on TGF-β/Smad and MAPK signal pathways in Echinococcus
    Ling-ling WANG, Hai-mei MA, Jian-bing DING
    2018, 36(5):  516-519. 
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    Due to the close contact between Echinococcus and the host, both cells of Echinococcus and the host cells secrete various cytokines during infection, which transmit the stimulation signal into the host cells through the cell surface receptors. This process interferes with the immune system of the host and produces immune tolerance, enabling long-term parasitism of Echinococcus within the host. This review summarizes recent findings on the immunoregulatory role of TGF-β/Smad and MAPK signal pathways during Echinococcus infection.

    Research progress on functional genomics of important tapeworms
    Xiang LI, Bo LUO, Bi-ying ZHOU
    2018, 36(5):  520-525. 
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    Taeniasis is a group of parasitic diseases that seriously impairs human health and livestock husbandry. The control measures for the disease still need improvement. In recent years, molecular biology technology has emerged as an effective means to fight against taeniasis. Therefore, it is of significance to study the functions of tapeworm genes. This article introduces several techniques commonly used in functional genomics research, and reviews the progress of some important functional genomics of tapeworms in recent years.

    Research progress on calcium-binding protein in Toxoplasma gondii
    Xu ZHAO, Ting ZHANG, Yi-wei ZHANG, Da-wei WANG, Ying FENG, Na YANG, Ran CHEN, Xiao-yu SANG, Qi-jun CHEN, Ning JIANG
    2018, 36(5):  525-528. 
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    Toxoplasma gondii, a type of apicomplexa protozoan parasite, can infect nucleated cells. Studies have shown that Ca2+ is associated with physiological activity of Toxoplasma gondii such as infection and movement. Ca2+ is abundant in endoplasmic reticulum. There are many calcium-binding proteins important for physiological activity of Toxoplasma gondii. The calcium-binding proteins contain various EF-hand (EF) motifs, and currently there are 55 such proteins according to the records in Toxo DB, and 69 in Plasmo DB. In this review, the roles of Ca2+-binding proteins are discussed.

    Laboratory testing and traceability analysis of a case of transfusion-transmitted falciparum malaria in Guangdong Province
    Qiang MAO, Fu-quan PEI, Yong-zhen CEN, Meng-ran LIU, Hao ZHANG, Zhuo-hui DENG
    2018, 36(5):  529-533. 
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    A clinical case of falciparum malaria with no history of travel but a recent history of blood transfusion, was reported by Guangdong Provincial Center for Disease Control and Prevention in October 2017. We collected clinical data of the patient and carried out an epidemiological traceability survey. Blood samples were collected from the patient and 20 blood donors for malaria rapid diagnostic tests(RDT), microscopic observation of blood smears, and nested PCR. The positive specimens were traced using molecular methods and genotyped. Results of RDT and PCR tests revealed the occurrence of falciparum malaria in the patient and a donor. Their blood samples showed a 100% homology in SSU rRNA sequence, and both were infected with K1 genotype of Plasmodium falciparum. This patient was thus confirmed with Plasmodium falciparum infection via blood transfusion from a donor.

    Epidemiological analysis of malaria in Yantai City of Shandong Province during 2010-2017
    Jing FENG, Hong-jie ZHANG, Ji-ping XIANG, Hai-yun LIU, Shao-yi YU, Shu-na QU, Ying XU, Qian-qian WANG
    2018, 36(5):  533-535. 
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    The reported data concerning malaria during 2010-2017 in Yantai City were collected and analyzed with regard to prevalence, sources of infection, species of Plasmodium and distribution of the disease. A total of 129 malaria cases were reported in Yantai City during 2010-2017, consisting of 103 cases(79.8%) of falciparum malaria, 12 (9.3%) vivax malaria, and 14 (10.9%) ovale malaria. All of them were imported from other countries, with a majority from Africa (93.8%, 121/129) and a minority from other Asian nations (6.2%, 8/129). Blood test was conducted in the City for 116 034 febrile patients, with a positive detection rate of 0.1%. The disease onset showed no significant seasonal and regional variations. The reported cases comprised 127 male cases, concentrated in the 39-49 age group. They were mainly workers, peasants, and unemployed persons. The median interval from symptom onset to diagnosis was 3 days, and 15 (11.6%) patients received definite diagnosis within 24 h. After medication, among the 129 malaria cases, 1 patient died and others recovered after regular treatment. The cases were mainly reported by clinical medical institutions (90.7%, 117/129), while 5.4% (7/129) were reported by disease control institutions.

    Malaria epidemiological characteristic in Baoshan City in 2017
    Jia-zhi WANG, Qiang MA, He-xian YANG, Jia-quan LI
    2018, 36(5):  536-538. 
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    This paper analzyed the characteristics of malaria cases in Baoshan in 2017. Data of malaria situation in Baoshan in 2017 were collected, and analyzed by regions, sex, age, occupation, months, countries of infection source, and hospital visit time using SPSS 20.0 software. Results showed that there were 54 cases of malaria reported in 2017 in Baoshan, and all imported from other countries, consisting of 53 cases of tertian malaria and 1 falciparum malaria. Among regions, Tengchong ranked top in the number of imported cases (35, 64.8%), followed by Longyang County (9, 16.6%), Longling County (8, 14.8%), Shidian County (1, 1.8%) and Changning County (1, 1.8%). Most of the cases were males (74.1%, 40/54). The ages ranged 4-52 years, mostly concentrated between 40-44 (13, 24.1%), then between 35-39 (10, 18.5%). Most of the cases were farmers (77.8%, 42/54), then labor workers (14.8%,8/54), students (3.7%, 2/54), children (1.9%, 1/54) and drivers (1.9%, 1/54). The number of cases peaked in February (12, 22.2%), with other months having distributions of cases except December. The country sources of imported cases were Myanmar (46, 85.2%), Laos (7, 13.0%), and Republic of Guinea (1, 1.9%). The shortest interval from onset to hospital visit was 0 day (4, 7.4%), and the longest was 22 days (1, 1.8%). More cases visited hospitals on days 3 and 4 (10, 18.5% for each day). These results suggested that the monitoring and management of imported malaria should be strengthened on males, 40-44 age population, and farmers in Tengchong city.

    Epidemiological analysis of malaria in Xuchang City of Henan Province during 2005-2016
    Jin-hua PENG, Yan WANG, Liu-ye NIU, Ning WU, Xiao-li LI, Xiao-lu ZHANG
    2018, 36(5):  539-541. 
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    To understand the epidemiological characteristics and trends of malaria in Xuchang City of Henan Province during 2005-2016, a database was established based on data of malaria epidemic situation from China Information System for Disease Prevention and Control, in combination with work summary and case investigation reports on malaria in the period. Results showed that 94 malaria cases were reported in Xuchang City, consisting of 77(81.9%) imported cases and 17(18.1%) indigenous cases, One case died from falciparum malaria. No cases of local infection have been reported since 2012. There was incidence of malaria throughout the months, with no significant seasonal variations. Most of the patients were males of 22 to 49 years (85.1%, 80/94). The top 3 occupations with regard to malaria incidence were farmers (36.2%), workers (19.1%), and labor workers (8.5%).

    Status of Enterobius vermicularis infection in preschool children in rural areas of Ningxia during 2015-2016
    Xiang-lin WU, Rong-ting QI, Hong-ju DUAN, Xue-ping MA, Yi-ren FU
    2018, 36(5):  541-543. 
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    To investigate the status of Enterobius vermicularis infection in preschool children in rural areas in Ningxia, 48 villages were selected in 13 counties (cities) as spots using the stratified cluster sampling method. A transparent adhesive tape test was performed in 3-6 years children to examine the eggs of Enterobius vermicularis. Results showed an overall prevalence of 1.3%(12/911). The infections were detected only in Jingyuan, Longde and Pingluo counties, with prevalence of 14.3% (5/35), 4.6% (5/110) and 3.0% (2/67), respectively (P < 0.05). There was no significant difference in prevalence between boys (1.7%, 8/472) and girls (0.9%, 4/439) (P > 0.05), or between Han (1.4%, 7/499) and Hui (1.2%, 5/412) ethnicities (P > 0.05). The prevalence showed a trend of decrease with age (P < 0.05), being highest among 3-year-old children (2.9%, 4/173). The results suggest a clear regional bias of prevalence among preschool children in rural areas of Ningxia. Efforts should be made to take measures specifically in these areas.