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    30 August 2015, Volume 33 Issue 4
    Genetic Polymophism and Evolution of SRPN14 Gene in Anopheles sinensis(Diptera ∶ Culicidae)
    FENG Xin-yu1,4, MA Ya-jun1 *, XU Jian-nong2, LIANG Jiang-tao3, XIA Ai3
    2015, 33(4):  1-241-246. 
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    Objective  To identify and locate the serine protease inhibitor 14(SRPN14) gene of Anopheles sinensis, and analyze its genetic polymorphism among populations as well as the selective pressure during evolution.  Methods  Primers were designed based on the genomic sequencing data of An. sinensis, and PCR amplification system for the SRPN14 gene was established. The chromosomal location of SRPN14 gene was determined by fluorescence in situ hybridization. The SRPN14 gene of An. sinensis populations collected from 18 sampling sites in 12 provinces(municipality)was sequenced, its genetic variations within and among populations calculated, and the selective pressure during adaptive evolution evaluated.  Results  The amplified part of the SRPN14 gene of An. sinensis was 429 bp in length, and had 77%(nt) and 88%(aa)similarities with An. gambiae. The SRPN14 gene located on 2L: 23C of salivary gland chromosomes of An. sinensis. The sequences of 411 individuals from 13 An. sinensis populations were analyzed. In the 411 individuals, the total number of alleles of the SRPN14 gene was 204, among which 51(25.00%) showed inter-population consistency. The ranges of SRPN14 allel number and genetic polymorphism were from 11 (Liaoning) to 33 (Chongqing), and from 0.008 (Liaoning) to 0.024 (Hainan), respectively. AMOVA result showed that genetic divergence within populations was significantly higher than that among populations; variation within populations was 95.79% of the total variation. The genetic divergence among populations was small, with FST value of 0.042. The number of synonymous substitutions in SRPN14 was significantly higher than that of non-synonymous substitutions sites, and ω was less than 1 in all populations.  Conclusion  Genetic polymorphism occurs in SRPN14 gene of An. sinensis populations, and its evolution is under the negative selective pressure.

    Analysis of MSP1 Allelotypes in Imported Cases of Plasmodium falciparum Malaria from Africa
    ZHOU Shui-mao, YANG Yan, WU Kai, CHE Zhi, Xu Ming-xing, JIA Xi-shuai
    2015, 33(4):  2-247-250. 
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    Objective  To understand the allelotype characteristics of the merozoite surface protein 1(MSP1) in imported Plasmodium falciparum.  Methods  Blood samples were collected from P. falciparum-infected patients returning from African malaria endemic areas. Nested PCR was used to amplify gene fragments of MSP1 coding for block 2 and block 3 motifs of MSP1 of P. falciparum by using the MSP1-specific primers. Then the allelotype of MSP1 was analyzed.  Results  The  MSP1 allelotype was detected in 117 of 135 blood samples, with a detection rate of 86.7%. In the 117 cases with successful PCR amplification, the detection rates for MAD20, K1, RO33, MAD20+K1, MAD20+RO33, K1+RO33 and MAD20+K1+RO33 were 6.0%(7/117), 36.8%(43/117), 20.5%(24/117), 6.8%(8/117), 3.4%(4/117), 17.1%(20/117) and 9.4%(11/117), respectively, wherein the mixed infection accounted for 36.8%(43/135). The mean multiplicity of infection(MOI) of MSP1 allelotype was 1.46. There was no significant difference in the proportion of patients with major severity of illness among the MAD20, K1 and RO33 genotypes. The proportions of patients with major severity of illness were 25.7%(19/74) and 32.6%(14/43) in 74 cases of singular infection and 43 cases of mixed infection, respectively. The two infection types of patients had 241±176 days and 285±216 days of stay abroad, with no significant difference between them.  Conclusion  The three genotypes of MSP1 and their four types of combination exist in imported cases of P. falciparum malaria from Africa. K1 and RO33 are the dominant genotypes.

    Preliminary Genetic and Functional Analyses of Schistosoma japonicum Chymotrypsin-like Protease
    CHAI Ri-yi1, WANG Ji-peng1, LI Jian1, ZHANG Rui-xiang1, LI Qing1,
    2015, 33(4):  3-251-257. 
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     Objective  To investigate the gene expression, localization and potential functions of Schistosoma japonicum chymotrypsin-like protease(SjCTRL) in the host, and evaluate its potential immune-protection efficacy against S. japonicum infection in mice.  Methods  The physicochemical properties of SjCTRL and its phylogenetic relationship with homologous genes from other species were analyzed with bioinformatic software. The distribution of SjCTRL transcripts in 26-day-old worms was investigated using whole-mount in situ hybridization. The transcriptional levels of SjCTRL in male and female worms at four development stages(14, 18, 22, and 26 days after infection) were measured with quantitative real-time PCR. The SjCTRL-dsRNA was prepared and used to induce RNA interference(RNAi) in 26-day-old worms via soaking in vitro, and confocal microscopy was used to observe the morphological changes of worms after RNAi. Primers were designed to amplify the encoding sequence(excluding the transmembrane region) from the S. japonicum cDNA. The truncated gene was subcloned into the pET-28a plasmid, transformed into E. coli BL21(DE3) for expression. Mice were immunized with the purified recombinant protein and challenged with cercariae. The worms and mouse liver were collected on day 35 after the challenge, and the worm-reduction rate and egg-reduction rate were calculated.  Results  The in situ hybridization results showed that SjCTRL mRNA was located in the posterior segment of intestinal tract of female worms, and having abundence only in 26-day-old female worms. After RNAi with SjCTRL-dsRNA, the mRNA expression was reduced to 25.7% (P<0.05), without significant morphological changes. Using the recombinant plasmid pET-28a/SjCTRL, expression of insoluble SjCTRL protein was induced. Mice immunized with this protein gained a worm-reduction rate of 25.4% and an egg-reduction rate of 80.5% in liver after being challenged with cercariae.  Conclusion  This study proves a high transcriptional level of SjCTRL in the posterior segment of intestinal tract in 26-day-old female worms, which can be reduced by RNAi treatment in vitro. Immunization with the SjCTRL protein can reduce adult worms and liver eggs.

    Clinical Analysis of 36 Elderly Patients with Schistosome-Induced Liver Disease Complicated by Hepatitis E
    YU Xiao,DENG Min,ZHANG Kan,XIE Xin-sheng
    2015, 33(4):  4-258-263. 
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    Objective  To explore the clinical characteristics of schistosome-induced liver disease complicated by hepatitis E in elderly patients.  Methods  Thirty-six senile patients with schistosome-induced liver disease complicated by hepatitis E(the observation group) and 38 senile patients with simple hepatitis E(the control group) were from the First Hospital of Jiaxing City. The blood routine measurements, indicators of liver function and blood coagulation function, serum markers of liver fibrosis, and B-ultrasonic measurements of hepatic portal vein diameter and spleen hilus inside diameter of splenic vein, were analyzed in combination with clinical manifestations, disease course and outcomes.  Results  All the 74 cases had acute onset of hepatitis E. The patients in the observation group had severer conditions, which were prone for aggravation. In this group, the incidences of nausea(91.7%, 33/36), vomit(88.9%, 32/36) and pruritus(33.3%, 12/36) were all higher than those in the control group(P<0.05). The levels of average total bilirubin and alanine aminotransferase at admission were (153.22±29.71) mol/L and (705.14±356.55) U/L respectively, in the observation group, significantly higher than those in the control group[(109.89±26.28) mol/L and (485.74±297.49) U/L](P<0.01). In contrast, the levels of albumin, prothrombin activity, white blood cell, hemoglobin and platelet at admission were significantly lower in the observation group than in the control(P<0.01). In addition, the levels of four serum markers of liver fibrosis, i.e, the transparent hyaluronic acid, laminin, type III serum procollagen peptide and type IV collagen, were all significantly higher in the observation group than in the control(P<0.01). Further, B-ultrasonic measurements of hepatic portal vein diameter and spleen hilus inside diameter of splenic vein in the observation group were (16.56±3.38) mm and (10.28±3.31) mm, significantly higher than those in the control[(14.82±3.48) mm and (8.26±3.27) mm]. The recovery of liver function in the observation group was largely prolonged compared with the control[hospitalization duration, (43.7±7.1) days versus(29.5±5.1) days, P<0.01].  Conclusion  The elderly patients with schistosome-induced liver disease complicated by hepatitis E show severer liver injury than those with pure hepatitis E, thus needing intensive care and treatment.

    Cloning and Prokaryotic Expression of Fusion Gene of Group Ⅱ Allergen Der p2 T Cell Epitope from Dermatophagoides pteronyssinus
    DUAN Bin-bin, SONG Hong-yu, LI Chao-pin*
    2015, 33(4):  5-264-268. 
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    Objective  To express and purify the T cell epitope fusion peptide of the major allergen Der p2 from Dermatophagoides pteronyssinus.  Methods  Nucleotide sequences reported to encode four T-cell epitopes(T1-T4) of Der p2 of D. pteronyssinus were linked in the rank of T1-T2-T3-T4. In this way, the chimeric gene was synthesized, named as Der p2 T. The gene of Der p2 T was amplified by PCR, purified, and cloned into the pET-28a(+) vector, forming the prokaryotic recombinant expression vector pET-28a(+)-Der p2 T. This formation was verified by double digestion. The pET-28a(+) -Der p2 T vector was transfected into E. coli strain BL-21, and its expression was induced by addition of IPTG. The recombinant protein was purified and collected by Ni-NTA affinity chromatography, and prepared for SDS-PAGE and Western blotting analysis. ELISA was used to evaluate the binding ability of Der p2 T cell epitope fusion peptide to serum IgE from patients with house dust mite allergy.  Results  Double digestion results confirmed the construction of the pET-28a(+)-Der p2 T vector. SDS-PAGE revealed the expression of recombinant Der p2 T cell epitope fusion peptide with Mr of 10 000. Western blotting confirmed the purification of Der p2 T cell epitope fusion peptide. The binding ability of Der p2 T cell epitope fusion peptide to serum IgE from patients with house dust mite allergy [(37.70±9.89) μg/ml] decreased significantly in comparison to that of Der p2 [(85.89±9.63) μg/ml](P<0.01).  Conclusion  The Der p2 T cell epitope fusion peptide is prepared, and its binding ability to serum IgE from patients with house dust mite allergy significantly decreases than that of Der p2.

    Survey on Echinococcosis in Maqing County of Qinghai Province
    MA Xiao1 *,WANG Hu1,HAN Xiu-min1,ZHANG Jing-xiao1,LIU Yu-fang1,ZHAO Yan-mei1,WANG Yong-shun1,MA Jun-ying1,LIU Hai-qing1,GANG Jian2
    2015, 33(4):  6-269-272. 
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    Objective  To understand the status of echinococcosis in Maqing County of Qinghai Province in order to facilitate echinococcosis control in this region.  Methods  Ultrasonic scanning and indirect hemagglutination assay were used to detect echinococcosis infection in residents >1 year old, according to the People’s Republic of China Health Industry Standard--Diagnostic Criteria for Hydatid Disease(WS257-2006). Meanwhile, ELISA was used to detect the Echinococcus antigen in dog’s feces collected in Youyun, Dangluo and Xiadawu townships.  Results  Ultrasonic scanning showed that the prevalence of hydatid disease in the residents was 7.4% (116/1 561), cystic hydatid disease 5.3% (82/1 561), alveolar hydatid disease 2.2% (34/1 561). The serum positive rate in human population was 23.8%(307/1 288). Of the 82 cases of cystic hydatid disease, 23 cases (28.1%) had the hydatid cyst with a diameter of >10 cm. The prevalence in males and females in the county was 5.3% (40/753) and 9.4% (76/808), respectively (Ρ<0.05). Among populations with different occupations, the highest prevalence of hydatid disease fell into houseworkers (11/61, 18.0%), monks(5/41, 12.2%) and herdsmen(84/758, 11.1%). Among the age groups, the groups of >60 years(24/132, 18.2%) and 30-40 years(31/302, 10.3%) had higher prevalence of hydatid disease. The three townships with the higher prevalence were Youyun(29/247, 11.7%), Changmahe (6/63, 9.5%) and Dangluo(54/645, 8.4%). Of the 199 samples of dog’s feces, 54 were positive for Echinococcus antigens(27.1%), with a positive rate of 40.4% (23/57) in Youyun towship, being significantly higher than in the other two(Ρ<0.05).  Conclusion  Maqin county is a co-endemic area of cystic echinococcosis and alveolar echinococcosis. The prevalence is higher in females and those over 60 years-old.

    Preliminary Analysis of the Genetic Loci of SAG1 and SAG3, the Surface Antigens of Toxoplasma gondii, in HIV-positive People in Dali of Yunnan Province
    LENG Li1,2, CHEN Ling-juan3,4, LI Wei3, HE Yan-hong3,LUO Mi5, GAO Ju6, MA Ning1, SHEN Li-jie1 *
    2015, 33(4):  7-273-276. 
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    Objective  To identify the genotypes of Toxoplasma gondii that infects HIV-positive people in Dali of Yunnan Province through analyzing the genetic loci of the surface antigens SAG1 and SAG3.  Methods  A total of 291 blood samples from HIV-positive cases were collected from the HIV/AIDS Prevention and Control Institution in Yunnan. Nested PCR was used to amplify SAG1 and SAG3 genes in the blood samples. The products were digested with restriction enzymes Sau96Ⅰ, HaeⅡ and NciⅠ, and sequenced.  Results  Of the 291 HIV-positive blood samples, 64 showed successful amplification of SAG1 gene, and 42 of SAG3 gene, with product sizes of 390 bp and 225 bp, respectively. Enzymetic digestion of the PCR products resulted in fragments of 350 bp and 50 bp for SAG1, and -200 bp band for SAG3, consistent with RH, a particular type Ⅰ strain of T. gondii. Sequencing of the SAG1 and SAG3 PCR products showed that their sequence identities with SAG1 (Accession No. GQ253073) and SAG3(Accession No. JX218225.1) of the type Ⅰ strain of T. gondii were 99.98%-100% and 99.96%-99.98% respectively.  Conclusion  The Toxoplasma gondii in HIV-positive cases in Dali of Yunnan Province is the type I strain of T. gondii.

    Identification of Trypanosoma Species in Rattus flavipectus and R. losea using Microscopy and PCR Method
    ZHANG Ling-ling1,NI Qing-xiang2,YAO Li-nong1,ZHANG Xiao-he2,CHEN Hua-liang1,FENG Yan1,ZHANG Xuan1,YU Ke-gen1,RUAN Wei1 *
    2015, 33(4):  8-277-280. 
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    Objective  To identify two suspected Trypanosoma species in Rattus found in Zhejiang Province using microscopy and PCR method.  Methods  Blood samples were collected from Rattus losea and R. flavipectus. Blood smears were prepared, and observed under microscope. The morphological indices of trypanosomes were measured and calculated. The genomic DNA was extracted from the trypanosomes, and the specific fragment of Trypanosoma 18S rRNA gene was amplified using PCR. The products were further sequenced and submitted to GenBank. Blast analysis was performed on line in NCBI.  Results  Blood samples from Rattus flavipectus and R. losea were collected from Lucheng District and Wencheng County of Wenzhou, respectively. The parasites from R. losea and R. flavipectus were found to possess the characteristic features of Trypanosoma species, such as nucleus, free flagellum, and kinetoplast, etc. The body length was 27.50 μm and 23.80 μm, and the free flagellum length was 9.60 μm and 9.20 μm, respectively. The nucleus index was 0.74 and 1.05, the kinetoplast index was 1.40 and 1.57, respectively. Based on the morphological characteristics and host specificity, the parasites from R. losea and R. flavipectus were identified as Herpetosoma species, mainly found in rodents. The amplified products were about 700 bp by 18S rRNA gene PCR with the DNA isolated from the trypanosomes. The products were further sequenced, and the resulting sequences were submitted to GenBank with assession numbers of KP098535(from R. losea) and KP098536(from R. flavipectus). Blast analysis showed that KP098535 was completely homologous with the sequences from Herpetosoma subgenus (AY491765.1, AY491764.1, and AJ223568.1), and KP098536 was completely homologous with Trypanosoma lewisi (AB242273.1, AJ009156.1).  Conclusion  The Trypanosoma species found from Rattus flavipectus is Trypanosoma lewisi, and the other one belongs to Herpetosoma subgenus, which may be named as Trypanosoma lewisi-like trypanosome.

    Prevalence of Neospora caninum Infection in the Intestine of Pet Dogs in Some Areas of Anhui and Zhejiang
    LI Wen-chao, LIU De-yi, ZHOU Li, CAO Jia-tong, MEI Nan, ZHANG Heng, CHEN Cheng, CHEN Tao, HAN Min-min, ZHANG Wei, FAN Zi-lai, GU You-fang*
    2015, 33(4):  9-283-286. 
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    Objective  To investigate the prevalence of Neospora caninum infection in the intestine of pet dogs in areas of Anhui and Zhejiang.  Methods  A total of 315 fecal samples from pet dogs were collected in pet clinics from April to December 2013 in Baohe District in Hefei city, Xuanzhou District in Xuancheng city, Fengyang County in Chuzhou city, Longzihu District in Bengbu city, and Si County in Suzhou City in Anhui Province, as well as in Yuhang District in Hangzhou city of Zhejiang province. All samples underwent nested-PCR targeting Neospora-specific gene NCLI-004830. The results were further confirmed by PCR amplification of N. caninum ITS1 followed by sequence analysis.  Results  The rate of N. caninum infection in the 315 samples was 1.59%(5/315). The infection rate in Chuzhou and Bengbu was 3.37% and 6.45%, respectively, and no N. caninum infection was found in the remaining areas. There was no association between the infection rate and the sex or age of the dogs.  Conclusion  N. caninum infection is prevalent in pet dogs in Chuzhou and Bengbu of Anhui.

    The mosquito name of Anopheles ramsayi in China should be corrected to Anopheles pseudojamesi Strickland and Chowdhury, 1927(Diptera ∶ Culicidae)
    Qu Feng-yi
    2015, 33(4):  10-288-289. 
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    The present review deals with the nominate priority of mosquito names between An. ramsayi Covell, 1927 (published in Ind J Med Res, April 1927 number) and An. pseudojamesi Strickland and Chowdhury, 1927 (published in Ind Med Gaz, May 1927 number) and its synonymy reported in India. They adopted the priority by order of monthly number of the periodicals without exact dates. The name ramsayi for the priority used as the valid name long time in literature, and forms a wrong nominate case. The editor of Ind. Med. Gaz.(1936) entered an interesting footnote regarding the exact dates of publication for two names: ramsayi issued in May 15th 1927, while pseudojamesi was May 10th 1927. The fact showed that the name pseudojamesi has its priority. Nurul Huda and Harrison(1985) informed clearly for wrong nominate case, and clarified that the name pseudojamesi has priority over ramsayi. Therefore An. pseudojamesi is elevated to species rank (corrected to valid name), whereas An. ramsayi is regarded as its junior synonym (invalid name). That the rejected name of An. ramsayi now still being used in Chinese lliterature should be corrected to An. pseudojamesi quickly.

    The Role of Adoptive Transfer of Immune Cells in Helminth-induced Regulation of Allergy and Autoimmune Diseases
    DING Yi-han1,ZHOU rui2,YANG Xiao-di1*,ZHANG Li-li1
    2015, 33(4):  11-290-294. 
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    Parasitic worms(helminth) or their derivates can inhibit allergy and autoimmune diseases by inducing the activation of immune cells and thus the release of regulatory factors. A large number of animal experiments have shown that adoptive transfer of lymphocytes can protect against immune deregulation and have potential clinical applications. In this review, we discuss the research progress on the role of adoptive transfer of immune cells in worm-induced regulation of allergy and autoimmune diseases.

    Research Advances on Toxoplasma gondii Virulence Mediating Factors
    XIA Jing,PENG Hong-juan*
    2015, 33(4):  12-297-300. 
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    Toxoplasma gondii strains are grouped into three clonal lineages(TypesⅠ,Ⅱ, and Ⅲ)with different virulence. T. gondii rhoptry proteins ROP18, ROP16, and ROP5 are the major virulence factors, which are secreted into the host cell by T. gondii during invasion and mediate the strain virulence. This paper reviews the research progress of T. gondii virulence mediating factors.

    Preliminary Evaluation of Potential Transmission Risk of Malaria after Earthquake in Cangyuan County, Yunnan Province
    FENG Jun*, XIA Zhi-gui, ZHOU Shui-sen, WANG Qiang, XIAO Ning, ZHOU Xiao-nong
    2015, 33(4):  13-301-305. 
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    Objective  To evaluate the transmission risk and the prevalence potential of malaria in Cangyuan County after earthquake.  Methods  The malaria epidemiological data were collected through the web-based reporting system and the malaria statistics annual reporting system from 2005 to April 2015, and the epidemic factors of malaria after earthquake in the recent three years in Cangyuan County of Yunnan Province were analyzed.  Results  According to the web-based reporting system, 799 malaria cases were reported in Cangyuan County from 2005 to April 2015, of which 652 were vivax malaria, 127 falciparum malaria, and 20 unclassified. The highest number of cases occurred in 2006 (n=326), and one indigenous falciparum malaria case was reported after earchquake. In addition, the cases were mainly distributed in Mangka (n=320, 40.1%), Banlao (n=191, 23.9%), Banhong (n=98, 12.3%) and Mengdong (n=92, 11.5%). According to the annual reporting system, there were a total of 519 indigenous cases (58.4%) and 370 imported cases (41.6%) from 2005 to April 2015. The proportion of indigenous case was above 65% in each year except for 2006, when it was less than 15%. The risk indicator in western townships of Canyuan County was higher, especially in Mangka and Mengjiao.  Conclusions  There is a potential risk of malaria transmission in Cangyuan County of Yunnan province, due to the huge natural, social and biological alterations after earthquake and the yet presence of malaria transmission vehicle.

    Survey on Malaria Epidemics in China-Myanmar Border Area
    LI Ben-fu1,LIN Yin-xue2,GUO Xiang-rui2,CHEN Li-fei2,ZHOU Dai-li2,YU Guo-cui2,ZOU Jun2,SUN Xiao-dong1*
    2015, 33(4):  14-261-263. 
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    The malaria epidemics in Laza city of Myanmar and Yingjiang county in Yunnan province of China in 2012 and 2013 was reviewed retrospectively, and a survey on malaria infection was conducted in residents in the border areas. A total of 179 malaria cases were reported in Yingjiang county from 2012 to 2013, with an average annual incidence of 2.9 per 10 000. Of the 179 cases, 77.7% were imported cases and 22.3% were local cases; 79.3% were infected with Plasmodium vivax, 20.1% with P. falciparum, and 0.6% unidentified. In Laza city of Myanmar, 2 069 malaria cases were reported, with an average annual incidence of 322.5 per 10 000. Of them, 73.4% cases were infected with P. vivax, 20.1% with P. falciparum and 6.5% unidentified. In addition, the microscopic results revealed that the malaria parasite rate in the residents in Yingjiang county was 0%, while that in Laza city was 1.5%.

    Clustering Analysis of Hydatid Disease in Gansu Province
    YU Da-wei1,2, DING Guo-wu1*, HOU Yan-dong2, FENG yu2, LI Fan2
    2015, 33(4):  15-280-282. 
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    The prevalence of hydatid disease in human population and livestock, and the positive rate of echinococcal antigen in canine feces were analyzed with sample clustering method, according to the survey on hydatid disease in 72 counties in Gansu province in the database of the National Survey on Prevalence of Echinococcosis in 2012. The prevalence of hydatid disease in huma and livestock, and the positive rate of echinococcal antigen in canine feces were 0-1.59%, 0-15.22%, and 0-16.87% respectively. Clustering analysis revealed four types of prevalence in the 72 counties. The first type existed only in Dunhuang city, with the three indicators being 0.27%, 15.22% and 16.87%; the second in four counties, with the three indicators being 0.43%, 6.57% and 1.83%; the third in 22 counties, with the three indicators being 0.22%, 1.15% and 10.35%; and the fourth in 45 counties, with the three indicators being 0.16%, 0.58% and 1.69%.

    Cloning and Bioinformatics Analysis of Toxoplasma gondii ROP21 Gene
    SHI Shi-jun,CUI Yong,LI Jin,WANG Hong-fa,YIN Kun,WEI Qing-kuan,HUANG Bing-cheng,SUN Hui,LIU Gong-zhen*
    2015, 33(4):  16-294-296. 
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    The full-length gene sequence of Toxoplasma gondii ROP21(TgROP21) gene was amplified with PCR. The signaling peptide and transmembrane domain of TgROP21 protein were predicted by SignaIP and TMHMM online predictive sites, and the hydrophilicity and antigenic index of this protein were ananlyzed with DNAStar software. Meanwhile, the functional domains and tertiary structure were modeled by combined use of ExPASY and PRODATA online sites. As expected, the PCR results revealed one band at 2 022 bp. The signaling peptide, transmembrane domain, hydrophilicity, antigen index, functional domain and 3D structure of TgROP21 were successfully predicted. This work may provide a theoretical foundation for further verification of TgROP21 function. 

    Comparison of Malaria Diagnosis between County and Provincial Laboratories in Guizhou Province
    HUANG Yu-ting*,LU Li-dan,SHE Dan-ya,ZHANG Nian-heng,HUANG Tian-yi
    2015, 33(4):  17-307-308. 
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    Malaria cases reported by county laboratories were further tested in the provincial laboratory in Guizhou province by using PCR test and microscopy. The consistency between PCR and microscopic results in the provincial laboratory was set as the basis for evaluation of microscopic results in county laboratories. In 89 samples, 24 were identified by PCR to be positive for malaria, among which 15 were infected with P. falciparum, 7 with P. vivax, and 2 with P. ovale; all were imported cases. And 21 samples had consistent identifications by PCR test and microscopic examination in the provincial laboratory. The total coincidence rate between county and provincial laboratories was 79.8%(67/84), and the undetected and error rates in county laboratories were 9.5%(2/21) and 23.8%(15/63), respectively. The Kappa value between county and provicial diagnosis was 0.6, being at the medium-to-high level of consistentcy.

    Analysis of Time Distribution of Malaria Incidence in Baoshan City During 1990-2014
    LI Jia-quan, YANG He-xian
    2015, 33(4):  18-309-311. 
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    The malaria epidemic data in Baoshan city from 1990 to 2014 were analyzed with concentration ratio and circular distribution. Results showed that the incidence of malaria in Baoshan city displayed a trend of decrease from 1990 to 2014. The total concentration ration M was 0.36, the circular distribution r was 0.30, and the average angle α was 148.78°. The malaria incidence peaked on May 31, with the epidemic period being from March 2 to August 29, which is inconsistent with the seasonal ebb and flow of local malaria vector.

    Infection of Mice with Normal Immune Function by Taenia asiatica
    LIU Xiao-yan,GUO Guang-wu,CHEN Li-hong,MO Xing-ze,YU Yue-sheng
    2015, 33(4):  19-312-314. 
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    The Taenia asiatica eggs pre-incubated with sodium hypochlorite solution for 4 min, 6 min and 8 mins were subcutaneously injected into mice with normal immune function(groups A1-A3 respectively, n=20 in each) and mice with immunosuppression(groups B1-B3, n=20 in each). All groups of mice began to show body discomfort on day 5 after infection and develop lumps on the back about on day 15. In groups A1-A3, animal death occurred during days 7-15, with a same survival rate of 95.0%(19/20) and infection rate of 89.4%(17/19), 73.6%(14/19) and 47.3%(9/19) respectively. In groups B1-B3, animal death occurred during days 7-50, with survival rate of 60%(13/20), 55%(11/20)and 55%(11/20) and infection rate of 76.9%(10/13), 54.5%(6/11) and 45.4%(5/11) respectively. After the scolex of cysticercus was evaginated with 15% pig bile, four suckers, an apparent rostellum and two distinct hook-like puncta structures were seen. These results indicate that mice with normal immune function can be used as a replacement of immunosuppressive mice to establish a T. asiatica oncosphere infection model. In addition, the T. asiatica eggs pre-incubated with sodium hypochlorite solution for 4 min have the strongest infection ability.

    Inhibitory Effect of the Excretory/Scretory Proteins of Trichinella spiralis on Proliferation of Human Hepatocellular Carcinoma HepG2 Cell line
    LIU Ying-jie1,XU Jing2,HUANG Hong-ying1,XU Guo-qiang1
    2015, 33(4):  20-315-317. 
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    Human hepatocellular carcinoma HepG2 Cell line were cultured with different concentrations of excretory/secretory proteins from Trichinella spiralis, and MTT assay was used to evaluate the cell inhibition rate. After co-cultured with 300 μg/ml excretory/secretory proteins for 24 h, the HepG2 cells were observed under a fluorescence microscope with AO and EB staining. When co-cultured with 75 μg/ml excretory/secretory proteins for 24 h, the HepG2 cells were quantified by flow cytometry using Annexin V-FITC/PI stain, and the expression of cleaved-caspase 9 was detected by immunofluorescence assay. The proliferation of HepG2 cells was inhibited significantly by excretory/secretory proteins in a dosage dependant manner. Under fluorescence microscope, some HepG2 cells presented typical apoptotic morphologic changes and the cleaved-caspase 9 protein expression was higher than that of the control. The early and late apoptotic cells and necrotic ones occupied 17.9%, 7.3%, and 6.6%, respectively.