CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES

Sustained challenge to malaria elimination in China: imported malaria

Jun CAO, Yao-bao LIU, Yuan-yuan CAO, Guo-ding ZHU, Shui-sen ZHOU

2018, 36(2): 93-96.

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Malaria is the most important parasitic disease that seriously endangers human health. Remarkable progress has been achieved towards the elimination of malaria in China since the National Malaria Elimination Programme launched in 2010, the number of indigenous malaria cases has decreased continuously. However, with the increasing number of migrant workers, businessmen, tourists and international exchange activities in recent years, the number of imported malaria cases remained at a high level in China. Imported malaria not only threatens people’s health and life, but also becomes most serious challenge for achieving the goal of malaria elimination in China. This paper analyzes the current situation of imported malaria in China and the challenges, and discusses the key points and measures to control it.

Analysis of mutations of Plasmodium falciparum multidrug resistance gene 1 and K13 gene in imported Plasmodium falciparum in Henan Province

Cheng-yun YANG, Su-hua LI, Ya-lan ZHANG, Rui-min ZHOU, Ying LIU, Dan QIAN, Yu-ling ZHAO, Bian-li XU, Hong-wei ZHANG, Yan DENG

2018, 36(2): 97-102.

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Objective To identify mutations of Plasmodium falciparum multidrug resistance gene 1 （Pfmdr1） and K13 gene in imported P. falciparum in Henan Province during 2013-2015. Methods Blood samples and epidemiological information were collected from 386 workers returning from Africa, who were diagnosed as imported falciparum malaria in Henan Province during 2013-2015. Plasmodium DNA was extracted, followed by nested PCR to amplify the Pfmdr1 and K13 gene. The PCR products were sequenced and sequence alignment was carried out to analyze gene mutations. Results The 386 patients returned from 26 African countries and were predominantly males. The top 3 disease-exporting counties were Angola (22.5%, 87/386), Equatorial Guinea (13.7%, 53/386), and Nigeria (13.2%, 51/386). Pfmdr1 and K13 genes were amplified from all the 386 blood samples. Sequencing results indicated mutation rate of 23.6% (91/386) and 3.1% (12/386) at loci 86 and 1246 of Pfmdr1, respectively, and 5.4% (21/386) for K13 gene. The 21 blood samples with K13 gene mutations were from 10 countries, in which no C580Y mutation was found, but the R539T and P574L mutations (0.3%, 1/386 for each), which are associated with artemisinin resistance, were found in samples from Angola and Equatorial Guinea. The mutation rate for Pfmdr1 locus 86 was 28.8% (36/125), 23.4% (32/137) and 18.6% (23/124) in each year during 2013-2015, respectively (χ² = 6.438, P < 0.05), and that for locus 1246 was 4.0% (5/125), 3.7% (5/137), 1.6% (2/124), respectively (χ² = 1.384, P > 0.05). The mutation rate of K13 gene was 3.2% (4/125), 8.8% (12/137) and 4.0% (5/124) in each year during 2013-2015, respectively (χ² = 4.631, P > 0.05). Conclusion The mutation rate at loci 86 and 1246 of Pfmdr1 is 23.6% (91/386) and 3.1% (12/386), respectively, and that of K13 gene was 5.4% (21/386). Besides, two mutations R539T and P574L associated with artemisinin resistance are detected.

Analysis of genes associated with antifolate drug resistance in Plasmodium vivax from different infection sources

Ying DONG, Yan DENG, Meng-ni CHEN, Yan-chun XU, Xiang-hua MAO, Jian WANG, Ai-ming SUN, Jing-bo XUE

2018, 36(2): 103-111.

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Objective To analyze the mutation characteristics of dihydrofolate reductase gene (Pvdhfr) and dihydropterroate synthase gene (Pvdhps), which are associated with antifolate drug resistance, in Plasmodium vivax from different infection sources in Yunnan Province. Methods Dry blood spots on filter papers were collected from vivax malaria patients reported through the Reporting System of Chinese Center for Disease Control and Prevention from August 2012 to December 2016 and the corresponding epidemiology data of the patients were collected as well. DNA was extracted from P. vivax, and the Pvdhfr and Pvdhps fragments were amplified with nest-PCR. The PCR products were sequenced, and all sequences were aligned with the reference sequences (GenBank ID: X98123 and PVX_123230). The haplotype number, expected heterozygsity (He), and genetic differentiation (Fixation index) of the two genes in different Plasmodium populations were analyzed with MEGA 5.04 and Arlequin 3.5.1 softwares. The haplotype media evolution tree was constructed with the Network 4.6.0 software. Results A total of 1 203 blood samples were collected, including 1 060 with an infection source in Myanmar, 70 in Africa and 64 from indigenous patients in Yunnan. The PCR results showed that Pvdhfr and Pvdhps were amplified and sequenced in 272 and 708 samples, respectively. There were 53 haplotyes for Pvdhfr in the 272 DNA sequences, with an He of 0.243, and the Yunnan indigenous isolates had the highest He value (0.667). The case of all 12 loci of Pvdhfr gene being wild-type had a frequency of 8.1% (22/272). All the other 52 haplotypes harbored various types of missense mutations at the 12 loci, including single-locus, double-loci, triple-loci, quadruple-loci, quintuple-loci and sixfold-loci mutations. There were 35 haplotyes in 708 DNA sequences of Pvdhps gene, with an He of 0.153, and the Myanmar-source isolates had the highest He (0.142). The case of all the 5 loci of Pvdhps gene being wild-type had a frequency of 36.2% (256/708). All the other 34 haplotypes harbored 29 types of missense mutations at the 5 loci, including single-locus, double-loci, triple-loci, and quadruple-loci mutations. The media network diagram constructed from all the haplotypes of the two P. vivax genes showed that the haplotypes occurred initially as wild-type, and then evolved from single mutation to multiple mutations. In blood samples with wild-type and mutated Pvdhfr, the percentages of Pvdhps mutations were 18.2% (4/22) and 65.6% (147/224), respectively. The blood samples confirmed with Pvdhps mutations and Pvdhfr mutations were from 14 and 9 prefectures, respectively, and predominated by the Myanmar source samples that constituted 97.3% (442/452) and 95.4% (144/151), respectively. Conclusion Both Pvdhfr and Pvdhps have a high mutation frequency and a high degree of mutation in P. vivax from different infection sources in Yunnan Province.

Composition of anopheline larvae in the China-Myanmar border region in Yingjiang County of Yunnan Province

CHEN Tian-mu, ZHOU Hong-ning, LUO Chun-hai, ZENG Xu-can, GUO Xiang-rui, LIN Zu-rui, TU Hong, WANG Xue-zhong, ZHANG Shao-sen, ZHOU Shui-sen

2018, 36(2): 112-118.

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Objective To understand the species composition of anopheline larvae in the China-Myanmar border region in Yingjiang County, Yunnan Province of China. Methods The Kayahe Village in Nabang Town and Huqueba Village in Pingyuan Town were selected for determining the breeding-place of anophelines. The composition of anopheline larvae was investigated once a month from June to October, 2016, in potential habitats of slack water, puddle, pond, moor, and paddy. The values of pH and area, and the numbers of first-, second-, third-, and fourth-instar larvae as well as pupae of mosquitoes were recorded in each habitat. The species of late third-instar and fourth-instar anopheline mosquito larvae were identified under microscope using commonly accepted guidelines. Pupae were morphologically identified after the emergence. Species composition of anopheline larvae was analyzed using six indices, including density(Ds), Simpson diversity index(D), Shannon diversity index(H), Berger-Parker dominance index(d), and Shannon evenness index (E). The Morisita-Horn similarity index (C) was employed to calculate the similarity of species composition between the two towns. Results A total of 394 larvae samples were collected in 91 specimen collection sites, comprising 313 samples (268 Anopheles and 45 Culex) from Nabang Town and 81 samples(55 Anopheles and 26 Culex) from Pingyuan Town. The Anopheles in Nabang Town were predominated by An. sinensis(183, 68.28%), followed by An. peditaeniatus(33, 13.31%) and An. minimus (25, 9.33%), and those in Pingyuan Town were mainly An. sinensis(27, 49.09%), An. peditaeniatus (24, 43.64%), and An. vagus(2, 3.64%). In Yingjiang County, the Ds of Anopheles was 0.33 per dip, with D, H, d and E being 0.54, 1.16, 0.65 and 0.50, respectively. The values of N, H, and d were relatively higher in Nabang Town, with an E value approaching 0.5, albeit with a slightly lower D value. The values of Ds, N, and H in Nabang Town were higher than those in Pingyuan Town in each month. Puddles were rich in various species of anopheline larvae, with a smaller d and a higher N. D and H were highest in this habitat, and the value of E was most approaching 0.5. Large water habitats (with an area > 100 m²) had more abundant species, with higher D and H values, smaller d, and an E value more approaching 0.5. Habitats with slightly acidic water(pH < 7) had a complicated vector composition. Conclusion There are abundant species of anopheline larvae in habitats with a large area and in puddles with slightly acidic water in the China-Myanmar region in Yunnan Province, with a complicated structure of composition. There is also a high similarity in vectors between the towns.

Establishment and evaluation of colloid gold-labeled immunochromatographic test strip for rapid detection of antibody against Necator americanus

Feng SHI, Yi YANG, Jun-yun WANG, Yue-tao YANG, Chun-hua GAO

2018, 36(2): 119-123.

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Objective To establish and evaluate colloid gold-labeled immunochromatographic test strip for rapid and convenient detection of antibody against Necator americanus. Methods Adult worms of N. americanus was ground in liquid nitrogen, immersed in membrane protein surfactant, followed by repeated freezing and thawing in liquid nitrogen and ammonium sulfate precipitation, to extract soluble antigen. The colloid gold-labeled immunochromatographic test strip was established using the soluble antigen as a coating antigen and the colloidal gold-labeled G protein as the detective probe. This test strip was used to examine sera from patients infected with N. americanus, Ascaris lumbricoides, Trichuris trichiura, Schistosoma japonicum and Toxoplasma gondii and sera from healthy participants, to evaluate the sensitivity and specificity of the assay, accompanied by ELISA as a parallel control to evaluate the detection efficacy. Results The colloid gold-labeled immunochromatographic test strip for detecting antibody against N. americanus was established. This test was performed to examine sera from patients infected with N. americanus (95 samples), A. lumbricoides (10 samples), T. trichiura (11 samples), S. japonicum (10 samples) and T. gondii (10 samples) and 74 serum samples from healthy participants, accompanied by ELISA as a parallel. The sensitivity of the colloid gold-labeled immunochromatographic strip test and ELISA in detecting N. americanus was 88.4% （84/95) and 90.5% （86/95), respectively, with false positive rates of 4.1% （3/74） and 6.8% （5/74） in 74 healthy serum samples, with rates of cross reaction with ascariasis being 2/10 and 4/10, rates of cross reaction with trichuriasis being 1/11 and 4/11. The strip test had no cross reaction with schistosomiasis or toxoplasmosis, while ELISA had a cross reaction rate of 1/10 with schistosomiasis but no cross reaction with toxoplasmosis. The specificity of the strip test and ELISA was 94.9% (109/115) and 88.7% (102/115), and the diagnostic efficacy was 91.9% and 89.5%, respectively. There was no significant difference in sensitivity （P > 0.05） or specificity （P > 0.05） between the strip test and ELISA. Conclusion The colloid gold-labeled immunochromatographic test strip shows relatively high sensitivity and specificity in diagnosis of ancylostomiasis.

The change and role of interleukin-33 in mouse allergic asthma

Qiang CHAI, Hong-yu SONG, Chao-pin LI

2018, 36(2): 124-129.

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Objective To investigate the change and the role of interleukin-33 (IL-33) in mice with allergic asthma induced by Dermatophagoides farinae group 1 allergen (Der f 1). Methods Forty female BALB/c mice were randomized into four groups using a random number table method: asthma group (sensitized by Der f 1 allergen), SIT group (specific immunotherapy with Der f 1), inhibitor administration group and negative control group (PBS group) (n = 10 for each). All mice except the PBS group were intraperitoneally injected with Dermatophagoides farinae allergen extract containing 100 μg/ml Der f 1 on days 0, 7 and 14, and those in the inhibition group also received an intraperitoneal injection of inhibitor (soluble ST2) at the same time. From day 21, mice in the three groups were sensitized through aerosol inhalation of the extract (0.5 μg/ml for 30 min , once per day for consecutive days). Mice in the immunotherapy group began to receive an immunotherapy through intraperitoneal injection of 200 μl of Der f 1 protein solution (100 μg/ml) 0.5 h prior to aerosol inhalation, from day 25 on, while in the PBS group, PBS was used instead of Der f 1. Twenty-four hours after the final aerosol inhalation on day 27, all the mice were sacrificed to collect the bronchoalveolar lavage fluid (BALF) and eyeball blood. The total eosinophils in BALF were counted and pulmonary histopathological sections were prepared. ELISA was performed to measure the levels of cytokines IL-5, IL-13, IFN-γ and IL-33 in the BALF, as well as serum levels of IgE and IgG2a. Results All groups except for the PBS group displayed different degrees of asthma symptoms since day 21. From day 25, the asthma symptoms in the immunotherapy group began to show remission. The total numbers of BALF eosinophils in the immunotherapy group and the inhibitor group were (1.43 ± 0.14） × 10⁵/ml and (2.73 ± 0.33） × 10⁵/ml, respectively(t = 24.50, P < 0.01 between the two groups). The numbers of eosinophils in SIT group（1.43 ± 0.14） × 10⁵/ml and the inhibitor administration group (2.73 ± 0.33）× 10⁵/ml were both significantly lower than that in the asthma group（F = 129.72,P < 0.01）. ELISA results showed that the levels of IFN-γ in BALF in the asthma group, the SIT group, the inhibitor administration group and the negative control group were （83.06 ± 11.38）,（277.97 ± 22.46）,（175.13 ± 13.41）and（224.77 ± 19.97）pg/ml, respectively. The BALF IFN-γ levels in the SIT group and the inhibitor administration group were both significantly higher than that in the asthma group (t = 17.31, t = 11.71, P < 0.01). The levels of IL-5 in BALF in the asthma group, the SIT group, the inhibitor administration group and the negative control group were （208.64 ± 11.55）,（106.87 ± 11.39）,（140.71 ± 14.58）and（90.15 ± 9.49）pg/ml, respectively. The BALF IL-5 level in the asthma group was significantly higher than that in the other groups (F = 97.19, P < 0.01). There was a significant difference in the BALF IL-13 level between the asthma group [(308.37 ± 13.67) pg/ml] and the SIT group [(175.66 ± 11.79) pg/ml] (P < 0.01), and that in the inhibition group was significantly lower than that in the asthma group (t = 16.44, P < 0.01). The BALF IL-13 level in the PBS group was (97.57 ± 18.38) pg/ml. Compared with the asthma group, mice in the SIT group showed improved inflammation in the lungs. Although the inhibition group also showed eosinophilia, epithelial cell shedding and bronchial epithelial cells around the bronchus, the symptoms were much attenuated compared with the asthma group. Serum IgE levels in the asthma group, the SIT group, the inhibitor administration group and the PBS group were （31.97 ± 3.48）,（1 2.86 ± 2.22）,（1 8.43 ± 2.30）and（9.68 ± 1.27）IU/ml, respectively. The serum IgE levels in the SIT group and the inhibitor administration group were both significantly lower than that in the asthma group (t = -7.77, P < 0.01). In contrast, the IgG2a level in the SIT group [(35.06 ± 2.57) μg/ml] was significantly higher than that in the asthma group [(26.94 ± 2.96) μg/ml] (t = 6.55, P < 0.01). The IgG2a level in the PBS group was(10.31 ± 1.48) μg/ml. Conclusion The IL-33/ST2 signaling pathway may play a critical role in allergic asthma by ELISA.

The epidemiological characteristics and influencing factors for Blastocystis hominis infection among human immunodeficiency virus seropositive individuals in Tengchong of Yunnan Province

Xue-jiao TENG, Yan-hong CHU, Cheng-cheng ZHAI, Ying-fang YU, Yu-chun CAI, Shao-hong CHEN, Lin AI, Li-guang TIAN, Jia-xu CHEN

2018, 36(2): 129-134.

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Objective To investigate the epidemiological characteristics, genotype distribution and influencing factors for Blastocystis hominis infection among human immunodeficiency virus （HIV） seropositive individuals in Tengchong of Yunnan Province, China. Methods In the cross-sectional survey, stool specimens from HIV-seropositive individuals were collected in a hospital in Tengchong City, and the small-subunit ribosomal RNA (SSU-RNA) of B. hominis was examined by PCR. The positive results were further verified by sequencing. The evolutionary tree was constructed to analyze genotype distribution. A structured questionnaire was applied to collect basic information. The influencing factors for co-infection with HIV and Blastocystis hominis were analyzed with the single-factor analysis and logistic regression analysis. Results A total of 324 stool specimens were collected. PCR results showed that 12 samples had a specific band of SSU-RNA for B. hominis at 1 100 bp, with a positive rate of 3.7% (12/324). The evolutionary tree showed that the B. hominis subtypes ST1 (n = 3, 25%), ST3 (n = 2, 16.7%), ST4 (n = 3, 25%), ST7 (n = 3, 25%), and ST12 (n = 1, 8.3%) were the main subtypes in the HIV-infected individuals. Single-factor analysis revealed that the type of tap water (χ² = 4.398, P < 0.05), livestock-raising (χ² = 7.448, P < 0.05), frequent contact with animals (χ² = 6.276, P < 0.05), CD4⁺ cell count (χ² = 4.414, P < 0.05), and HIV-RNA viral load (χ² = 11.829, P < 0.05) were influencing factors for B. hominis infection among HIV-seropositive individuals. Logistic regression model showed that drinking unboiled tap water (χ² = 6.595, P < 0.05), livestock-raising (χ² = 6.740, P < 0.059), CD4⁺ cell count ≤ 500 cells/μl (χ² = 3.864, P < 0.05), and an HIV-RNA viral load > 50 copies/ml(χ² = 9.561, P < 0.05) were the influencing factors for B. hominis infection among HIV-seropositive individuals. Conclusion The prevalence of B. hominis infection among HIV-seropositive individuals is 3.7% in Tengchong of Yunnan Province. The B. hominis mainly has five genotypes. The main influencing factors for B. hominis infection among HIV-seropositive individuals include drinking unboiled tap water, raising of livestock and low immune function.

Investigation on human intestinal helminth infection in urban areas of Henan Province in 2015

Ya-lan ZHANG, Yan-kun ZHU, Wei-qi CHEN, Yan DENG, Xi-meng LIN, Peng LI, Hong-wei ZHANG, Bian-li XU

2018, 36(2): 135-139.

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Objective To understand the endemic status of intestinal helminth infection in urban residents of Henan Province. Methods According to the protocol of The Third National Survey on Major Human Parasitic Diseases, the random cluster sampling method was used to select 62 residents committees as the survey spots from 37 counties in 14 prefectures, including Anyang, Hebi, Jiaozuo, etc., from January to May, 2015. Fecal samples were collected from residents aged > 1 year and detected by the Kato-Katz thick smear method. Cluster analysis was performed on families with Enterobius vermicularis infection. Results A total of 15 893 residents were examined, the total infection rate was 1.09% (174/15 893), and the standardized rate was 1.08%, 6 species of intestinal helminths were found, all showing light infection. No co-infection cases were found. E. vermicularis had the highest infection rate (0.92%, 147/15 893) （χ² = 117.648, P < 0.01), followed by Ascaris lumbricoides （0.07%, 11/15 893）, Ancylostoma sp. （0.06%, 10/15 893）, Trichuris trichiura（0.03%, 4/15 893）, and Strongyloides stercoralis as well as Clonorchis sinensis （0.01%, 1/15 893）. Among the 14 prefectures, the infection rate was highest in Luoyang （3.40%, 17/500） and lowest in Sanmenxia (0.20%, 1/501). There was no significant difference in the infection rate between males (1.03%, 77/7 477) and females (1.15%, 97/8 416). The population of 60- years had the highest infection rate (1.64%, 1/1 895) (χ² = 17.698, P < 0.05); the population of 50- years were detected with 6 species, which was the highest among age groups. The population of high school or above had the highest infection rate (1.48%, 58/3 912), significantly differed from preschool children （χ² = 8.145, P < 0.01). There was a significant difference in infection rate among different educational groups （χ² = 12.135, P < 0.05）. The medical personnel had the highest infection rate （4.76%, 10/210）, inclusive of E. vermicularis. C. sinensis and S. stercoralis were only found in farmers. There was a significant difference in infection rate among different occupations （χ² = 34.534, P < 0.05). A total of 147 cases of E. vermicularis infection were distributed in 124 households, and the results of the binomial distribution shows that the infection of E. vermicularis presented a characteristic of households clustering. Conclusion The infection of E. vermicularis is relatively high in urban areas of Henan Province.

A survey on Enterobius vermicularis infection among pre-school children in Guoyang County of Anhui Province

Xiao-li WANG, Qi ZHOU, Ao SHI, Liang LI, Yuan-yuan WANG, Shou-feng HU, Jie CUI

2018, 36(2): 139-143.

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Objective To understand the status of Enterobius vermicularis infection among pre-school children in Guoyang County of Anhui Province and its risk factors Methods E. vermicularis infection was surveyed using a stratified random sampling method from June to October, 2016. The whole county was divided geographically into east, south, west and north parts, and in each part 2-3 kindergartens were selected to reach a total of 10. In each kindergarten, age- and sex-matched classes of junior level, middle level, and senior level were randomly selected (n = 1 class for each level). E. vermicularis infection was examined by using adhesive cellophane anal swab in the morning with permission from parents. Information on family and school sanitary conditions and personal health habit was collected through a questionnaire, in order to analyze potential risk factors. Results A total of 1 035 children were surveyed, and 1 011 (97.6%) valid questionnaires were received. The overall infection rate of E. vermicularis was 6.8% (69/1 011). The infection rate in females and males was 5.6% (28/498) and 8.0% (41/513), respectively (χ² = 2.231, P > 0.05). Children in the senior class had highest infection rate of 10.1% (36/355), followed by the middle class 6.4% (22/345) and the junior class 3.5% (11/311) (χ² = 11.534, P < 0.05). There was no significant difference in the infection rate between public (6.4%, 32/497) and private kindergartens (7.2%, 37/514) (χ² = 0.229, P > 0.05). The infection rate in town- and county-level kindergartens was 8.0% (53/661) and 4.6% (16/350), respectively (χ² = 4.275, P < 0.05). The infection rate in rural areas (8.7%, 51/585) was significantly higher than that in urban areas (4.2%, 18/426) (χ² = 7.824, P < 0.05). The infection rate among those with a maternal education level of senior high school and above had the lowest infection rate (5.6%, 48/854). Unlike the significant difference among those with different maternal education levels (χ² = 17.154, P < 0.01), no significant difference was found among those with different paternal education levels (χ² = 3.813, P > 0.05). There was no significant difference in the infection rate between children with and without habit of washing hands before dinner (5.3%, 26/487; 8.2%, 43/524), sucking finger (8.3%, 35/423; 5.8%, 34/588), or biting toys (9.0%, 25/279; 6.0%, 44/732) (χ² = 3.263, 2.402, 2.764, all P > 0.05). There was a significant difference in the infection rate between children in kindergartens with a high sterilization frequency (4.7%, 20/422) and those with a low sterilization frequency (8.3%, 49/589) (χ² = 4.955, P < 0.05 ), and between children having and having not a habit of washing hands after pooping (4.8%, 32/670; 10.9%, 37/341) (χ² = 13.112, P < 0.01). Multivariate logistic regression analysis revealed risk factors of grade level, place of residence, maternal educational level, kindergarten sterilization frequency, and a habit of washing hands after pooping for E. vermicularis infection (P < 0.05). Conclusion There is a high E. vermicularis infection rate among pre-school children in Guoyang County, and grade level, place of kindergarten, place of residence, maternal educational level, kindergarten sterilization frequency, and a habit of washing hands after pooping are risk factors for the infection.

The epidemiology,clinical feature, radiology and outcome of 8 cases with cerebral sparganosis mansoni

Shan-ke YE, Lie XU, Qin HUANG, Yun LING

2018, 36(2): 144-147.

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Objective To analyze the epidemiological characteristics, clinical features, laboratory testing data, imaging features and prognosis of cerebral sparganosis mansoni. Methods Clinical information of 8 patients with cerebral sparganosis mansoni (as determined through detection of antibody for sparganosis mansoni in serum by the National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention) admitted in Shanghai Public Health Clinical Center (SPHCC), China, from January 2013 to December 2016, was obtained. The epidemiological characteristics, clinical features, laboratory testing data, imaging features and prognosis were analyzed. Results A total of 8 patients were enrolled in this study, with the antibody for sparganosis mansoni in serum and cerebrospinal fluid. All the patients were from outside of Shanghai, and 6 of them may be infected by inadequately cooked food. The patients were found with headache (5 cases), epilepsy (3 cases), weakness (2 cases), character change (1 cases), and of decline intellecteual (1 cases). The number of eosinophils in cases 1, 4, 5, 6 and 7 was higher than normal before treatment and normal after treatment. The number of eosinophils in cases 2, 3 and 8 was normal before treatment and increased after treatment. All the cases were showed a mass lesion by brain magnetic resonance imaging (MRI). With the praziquantel [15~20 mg/(kg·d),2 times/d] plus albendazole [15~20 mg/(kg·d),2 times/d] treatment, 5 cases were found with the local foci disappeared, indicating fully recovered, 3 cases were recurrent through repeated treatment. Conclusion Praziquantel plus albendazole treatment is effective for cerebral sparganosis mansoni.

Follow-up of a patient with Leishmania and human immunodeficiency virus co-infection who received prolonged sodium stibogluconate treatment

SONG Shi-hui，GUI Xi-en*

2018, 36(2): 148-152.

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Objective To observe the curative effect and prognosis of prolonged sodium stibogluconate treatment in a case of Leishmania and human immunodeficiency virus(HIV) co-infection. Methods Clinical information of the patient was collected. The curative effect and prognosis were followed-up. Results The patient was working in Jiuzhaigou County of Sichuan Province （an endemic area of visceral leishmaniasis） in 1990. At the end of 1992 he presented with fever, hepatomegaly and splenomegaly, and was diagnosed with liver cirrhosis by local hospitals. The symptoms were not improved after symptomatic treatment, with continued hapastosplenomegaly, accompanied by severe anemia and weight loss. He was later diagnosed with visceral leishmaniasis via bone marrow examination in 1994, and given intravenous infusions of sodium stibogluconate (pentavalent antimony 600 mg/d for 6 days). After 6 days of treatment, his condition got improved. In August 2010, an abdominal mass was found in the left upper quadrant, accompanied by abdominal distension. Leishmania amastigotes were found in the bone marrow smear. Again conventional treatment of pentavalent antimony was given for 6 days and his condition was improved. During hospitalization, the patient was screened for HIV antibody and found positive, which was further confirmed by Western blotting analysis. The CD4+ T lymphocytes was as low as 42 cells/ml. He was diagnosed with acquired immunodeficiency syndrome (AIDS). From July 2011, he began to receive conventional treatment including lamivudine (3TC, 300 mg/d) + zidovudine（AZT, 600 mg/d） + efavirenz (EFV, 600 mg/d). Two months later, AZT was replaced by tenofovir (TDF, 300 mg/d) due to severe anemia. On 25 July 2012, he was admitted to our hospital for “abdominal distension for two years”. Physical examination showed severe pallor and hepatosplenomegaly (the liver was 2 cm below the costal edge and the spleen was palpable 7 cm below the umbilical level). Leishmania was found in the bone marrow aspirate. Therefore, anti-HIV treatment with 3TC + TDF + EFV was continued, combined with sodium stibogluconate (pentavalent antimony 600 mg/day for 4 days, and 1 200 mg/day for an additional 4 days). After the liver and spleen began to shrink, the usage of stibogluconate therapy (pentavalent antimony 600 mg per time for 22 times) was changed every other day. Another two phase treatment with stibogluconate (pentavalent antimony 600 mg for 18 times) was given at one month and three months after the end of first phase treatment, respectively. The patient was given pentavalent antimony therapy with a total dose of up to 42 g during the three phase treatment, while also receiving conventional treatment on AIDS. Fifty months after pentavalent antimony withdrawal in April 2017, Leishmania was not found in the bone marrow, the CD4+ T lymphocytes was 225 cells/ml, and HIV-RNA was below detection limit. Conclusion Prolonged stibogluconate and conventional treatment can cure visceral leishmaniasis co-infected with HIV.

Comment on the essentials of writing English medical research articles

Xiao-nong ZHOU, Jin CHEN, Hui-feng SHENG, Pin YANG

2018, 36(2): 153-156.

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With the enhancement of our overall national strength and the rapid development of global health disciplines, the increasing number of medical experts and scholars write English scientific papers and publish them in English academic journals. This paper gives a comparative analysis of the differences between English and Chinese medical journals in terms of access type, review comment and writing methodology; discusses writing patterns and skills of writing English medical scientific paper, namely, background, method, result, discussion and conclusion with specific examples, so as to assist researches and experts to write and publish English research articles.

Meta-analysis on severity of hydatidosis

Meng-yuan ZHANG, Li-ying WANG, Ya-yi GUAN, Wei-ping WU

2018, 36(2): 156-160.

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By searching China Hospital Knowledge Database, WANGFANG DATA, and Pubmed, literature on clinical manifestations (signs/symptoms) of hydatidosis published from the foundation date of the databases to August, 2016, was obtained. The publications were included in the meta-analysis according to the inclusion criteria. Meta-analysis was performed using Microsoft Office Excel 2007 and R 3.2.3 software to calculate the echinococcosis types, infection rate of organs, severity of the disease and mortality. A total of 165 papers were included in this study, adding up to 18 613 patients. The patients with cystic echinococcosis (CE) and alveolar echinococcosis (AE) constitute 93.74% (17 447/18 613) and 5.78% (1 076/18 613), respectively. The lesions occurred more frequently in the liver (63.70%, 11 857/18 613), followed by the lungs (24.13%, 4 492/18 613). The disease severity was classified as asymptomatic (6.17%, 1 139/18 458), mild and moderate (66.12%, 12 204/18 458), and severe (27.71%, 5 115/18 458). The severity of AE was higher than that of CE (χ² = 164.560, P < 0.05). The one-year case fatality rate after surgery was 0.83% (155/18 613). More specifically, the one-year case fatality rate of AE (2.27%, 19/842) after surgery was higher than that of CE (0.70%, 102/14 587) (χ² = 24.811, P < 0.05). In conclusion, liver is the major involved organ in hydatidosis. The severity and mortality of AE are higher than those of CE.

Research progress on proteins associated with Plasmodium vivax invasion of erythrocytes

Xin-xin ZHANG, Rui-lin CHU, Ying-hua XUAN, Yang CHENG

2018, 36(2): 161-165.

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Different Plasmodium species utilize different invasion pathways and specific sets of ligand-receptor interactions to exert distinct invasion behaviors of merozoites. Most studies on host cell invasion by Plasmodium have concentrated on invasion of erythrocytes. With completion of the whole-genome sequencing of P. vivax, proteins involved in erythrocyte invasion, such as the duffy binding protein, the reticulocyte binding protein, and tryptophan-rich antigens, have been identified. This review summarizes the role of these proteins in erythrocyte invasion.

Taxonomy and molecular epidemiology of Echinococcus granulosus complex causing cystic echinococcosis

Jing-ye SHANG, Guang-jia ZHANG, Wei HE, Wen-jie YU, Sha LIAO, Qi WANG, Fan CHEN, Rui-rui LI, Qian WANG, Bo ZHONG, Yan HUANG

2018, 36(2): 166-174.

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Cystic echinococcosis is a common parasitic zoonosis caused by the larva of multiple species of Echinococcus. The disease significantly impairs the health of humans and animals, and incurs a severe socioeconomic loss. In this review we summarize the taxonomic status, transmission patterns, host specificity and geographical distribution of the species, in order to provide references for further studies on the etiology, molecular genetics, epidemiology, and control strategies of cystic echinococcosis.

Research advances on the Toxoplasma gondii toxofilin protein

Gong-zhen LIU, Yuan-jing LIU, Xiang-yun PENG, Jing-xuan KOU

2018, 36(2): 174-177.

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Toxofilin is a member of the Toxoplasma gondii rhoptry protein family with a unique property of lack of a typical serine/threonine kinase domain for rhoptrys. Toxofilin secreted into host cytoplasm can interact with phosphopeptide 2C (PP2C) and actin to form a toxofilin-actin-PP2C trimer complex. Toxofilin is phosphorylated upon activation by casein kinase Ⅱ (CKⅡ) and PP2C, which plays an important role during its invasion. This review summarizes the discovery and research advances on function and immune-protection of toxofilin.

Research development on 14-3-3 protein in parasites

Bo LUO, Xiang LI, Bi-ying ZHOU

2018, 36(2): 178-183.

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The 14-3-3 protein plays an important role in parasite development and environmental adaptation. Recent studies have shown that the 14-3-3 protein can bind to a plethora of interacting signaling proteins including phosphatases, kinases and some transmembrane receptors, thus participating in a wide range of important regulatory processes, such as signal transduction, metabolism, cell growth, cell proliferation and differentiation, apoptotic cell death, and cell cycle control. Systematic studies on the 14-3-3 protein can advance our understanding of the biological functions and regulatory mechanisms of parasite growth and development. This review summarizes the gene identification, structure, biological functions and application potentials of the 14-3-3 protein in parasites.

Scanning electron microscopic and transmission electron microscopic observations of the tegument structure of adult Clonorchis sinensis

Yun-liang SHI, Xiao-ling WAN, Zhi-hua JIANG, Xiao-jing CHENG, Yi-chao YANG

2018, 36(2): 184-186.

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To study the ultrastructural characteristics of Clonorchis sinensis, scanning electron microscopy (SEM) was performed to observe the body surface of adult C. sinensis obtained from an infected rat, and transmission electron microscopy (TEM) was used to observe the tegument structure. By SEM, we found a number of nodules connected by filaments on the body surface of the parasite, with unevenly-distributed different sizes of sense papillae protruding from the body surface. The TEM showed that the epidermis of the C. sinensis was composed of tegument, the muscle layer and the cell layer. The surface of the tegument was covered with numerous protrusions containing mitochondria, secretory granules and secretory vesicles. The outermost-layer protrusions showed a tendency to detach from the tegument. The matrix contained spherical and rod-shaped secretory granules, secretory vesicles and mitochondria. The muscular layer was distributed with circular and longitudinal muscles. In the cell body layer, cell nuclei were clearly seen and the cytoplasm contained two types of secretory bodies and secretory vesicles. The cell body layer and the tegument were connected by cytoplasmic bridges.

Development of cytochrome c oxidase subunit 1 gene-based duplex PCR for identifying Clonorchis sinensis and Haplorchis taichui

Zhi-hua JIANG, Qing-li YANG, Yi-chao YANG

2018, 36(2): 187-189.

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Genomic DNA was extracted from Clonorchis sinensis and Haplorchis taichui, primers for cytochrome c oxidase subunit 1(COX1) gene were designed, respectively, and single and duplex PCR were performed. Single PCR for C. sinensis using three pairs of primers, FC1/RC1 and FC3/RC3 primer pairs generated specific bands of 328 bp and 356 bp, respectively, while no band was produced when using the FC2/RC2 primer pair. Similarly, single PCR using FH1/RH1 and FH2/RH2 primer pairs for H. taichui produced specific bands of 200 bp and 190 bp, respectively. Furthermore, duplex PCR combining the FC1/RC1 primer pair with FH1/RH1 or FH2/RH2 primer pair resulted in specific bands for both C. sinensis and H. taichui, with no cross-species reaction.

Cloning and expression of the nucleoside diphosphate kinase gene of Giardia lamblia

YUAN Yi-xue1, ZHENG Guo-xia2, WANG Dan-dan1, ZHUAN Hang1, DU Meng1, WANG Yun-hua1*

2018, 36(2): 190-192.

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