Objective To analyze malaria situation and evaluate the effect of control program in Henan Province during 1990-2005. Methods Data were collected and analyzed on the measures and effects of malaria control, vector surveillance, blood examination for cases with fever and serological surveillance in the province during 1990-2005. Results In the 16 years, a total of 802 700 people were given pre-transmission season treatment with chloroquine and primaquine for a radical cure of vivax malaria, chemoprophylaxis was given to 764 300 people at high risk during the transmission season, treatment or presumptive treatment was given to 43 891 cases. 11 216 100 cases with fever were tested and 11 213 (0.10%) were found positive accounting for 29.01% (11 213/338 654) of all malaria cases. A total of 1 332 800 bed nets were treated with insecticide and 1 999 300 people were protected in 1990-1992 and 1996-1999. 34 846 residents including pupils were tested with IFAT in 1990-2000 and 1149 (3.30%) were positive. The parasite rate amongst 71 234 local residents including pupils was 0.40% (286/71 234). The principal transmitting vectors were Anopheles sinensis and An.anthropophagus. The man-biting habit for An. sinensis and An.anthropophagus was 0.060 8 and 0.314 3 respectively, and the vectorial capacity of An.anthropophagus was 22.4 times higher than that of An.sinensis. In this period, 38 654 malaria cases were reported in the province and the annual malaria incidence was 2.62 per hundred thousand, the lowest annual incidence was in 1992 (0.37 per hundred thousand). 70.05% (27 076/38 654) of these malaria cases were from areas where An. anthropophagus was present. Conclusions In general, the malaria control activities have been effective and the epidemiological situation kept stable in Henan Province, although in some areas the situation is unstable and outbreak spots or focal epidemics occur.

A highly sensitive immuno-PCR assay based on sandwich ELISA and PCR was developed to detect the circulating antigen in trichinellosis. Antigens were purified from the muscle larvae of Trichinella spiralis, and the myeloma cells were fused with spenocytes immunized with T. spiralis antigens to product the specific monoclonal antibodies. Indirect ELISA was used to select the antibody-secreting hybrodoma cells. By this method of procedure, monoclonal antibody F4C6 against the T. spiralis ES antigen was obtained, which was used as the indicator antibody, while the rabbit polyclonal antibodies against T. spiralis were to be used as capturing antibodies. The plasmid Bluecript Ⅱ KS was amplified by PCR with biotin-labeled primer M13-20, and thus the biotin-labeled DNA was obtained. Both the second antibody and DNA labeled with biotin were to be linked with 100 ng/ml avidin. The whole procedures of assay consisted of two steps, in which the circulating antigens were captured by monoclonal antibody through sandwich ELISA in the first step, and the DNA linked by monoclonal antibody was amplified by PCR in the second step. The sensitivity of this method was compared with that of the ELISA assay. It was found that the measuring ranges to detect the circulating antigens in trichinellosis were 50 pg/L to 0.005 pg/L for the immuno-PCR assay, and 5 μg/L to 0.05 μg/L for ELISA assay, the former was quite higher than that of the latter. It is evident that this method is highly sensitive for the detection of circulating antigens in trichinellosis.

Objective To evaluate different detection methods in the diagnosis of severe fever with thrombocytopenia syndrome (SFTS), and find the most quick and accurate one for the identification of new bunyavirus infection. Methods Real-time PCR and ELISA-IgM were used to detect serum samples of 158 patients with acute phase of SFTS, which were collected from the special monitoring system of SFTS in Henan Province in 2014. IgM and IgG antibodies were detected by ELISA in 109 acute and convalescent paired serum specimens. The differences of the positive rates were compared between the three methods, and the influence of the collected interval time on the detection results was analyzed. Results For 158 acute phase serum samples of SFTS patients, the positive rate detected by real-time PCR (76.58%) was higher than that of ELISA-IgM (47.47%), and the difference was statistically significant (χ2=34.13, P < 0.05). For 109 cases with acute and convalescent paired serum samples, there was no significant difference in the positive rates between ELISA-IgG ( 75.23%) and real-time PCR (72.48%) detections (χ2=0.18, P>0.05). In both the acute phase and convalescent phase, the positive rate of IgM was higher than that of IgG, and the difference was statistically significant (χ2=41.68 and 6.25, P<0.05). With the extension of collected interral time, the positive rates of IgM and IgG antibodies were both increased ( Z=6.42 and 10.08, P < 0.05). Conclusion Real-time PCR is the most sensitive method for the early diagnosis of the SFTS. ELISA-IgG is suitable for the detection of SFTS at recovery period. ELISA-IgM can be used as an assistant method to guide clinical diagnosis.

To investigate the animals infection situation of novel bunyavirus in Xinyang City ,Henan Province ,China , animal serum samples such as cattle ,dog ,swine ,mice were collected in Shangcheng County and Guangshan County in Xinyang City .All the serum samples were detected by novel bunyavirus ELISA and real time RT-PCR method .A total of 292 animal serum samples were collected including 5 kinds of animals .The result of all the animal serum samples were negative by using real time RT-PCR ,and the positive rate was 45 .19% (141/312) by ELISA method .Of the 5 animal serum samples including mice ,cattle ,goats ,swine and dogs ,the positive rate were detected to be 1 .06% ,100 .00% ,76 .27% ,3 .57% ,and 75 .00%respectively .There was significant difference in results among 5 kind of animal serum antibodies .Animals such as cattle and dog may be the host of novel bunyavirus which were detected novel bunyavirus antibodies in cattle and dog in Xinyang City , Henan Province ,China .

Objective To investigate the training needs of health emergency professionals in centers for disease control and prevention (CDC) ,and to provide evidence for making training plan .Methods Totally 66 health emergency professionals who par‐ticipated in health emergency training class of CDC were surveyed with questionnaires in July ,2014 .The items included training content ,mode ,time ,teachers ,assessment forms and graduation way .Results Forty one persons (62 .12% of all subjects) selected health emergency disposal of all kinds of emergencies as the training contents ,and case analysis as the training mode .There were no statistical significances for the differences of the proportions of the professional staffs between different genders ,education levels ,ti‐tles ,categories and agencies (P>0 .05) .52 persons (78 .79% ) considered that training frequency of 1-2 times per year was appro‐priate ,and 53 persons (80 .30% ) considered that the most appropriate duration for each training was 2 -3 days .Domestic experts as a training teacher had the highest proportion (56 .06% ) ,followed by health emergency management cadres (34 .85% ) ,and foreign experts (6 .06% ) .The proportion of selecting university professor as a training teacher was lowest (3 .03% ) .71 .21% (47 persons) selected analog dealing with practical problems as assessment form ,and 71 .21% (47 persons) selected granting credits as graduation way .Gender and agencies were two important influencing factors for selecting different graduation ways (P< 0 .05) . Conclusion Training program of health emergency should be made according to the training needs .Appropriate training content and form should be selected in order to improve the quality and effectiveness of training ,and to improve the ability of the health e‐mergency professionals .

To explore the relationship between meteorological factors and incidences of fever, thrombocytopenia and leukopenia syndrome (FTLS), incidence data of FTLS in Xinyang City were collected and described. The relationship between FTLS and monthly meteorological factors such as average atmospheric pressure, average temperature, average relative humidity, average wind speed and precipitation was analyzed with simple correlation and multi-stepwise linear regression. The results showed that there was a significant negative correlation between incidence of FTLS and monthly average atmospheric pressure (P<0.01), and there were significant positive correlations between monthly average temperature, monthly average relative humidity, monthly precipitation and incidence of FTLS (P<0.05). The multi-stepwise regression showed that the regression equation was Y (monthly cases) =-12.70 + 0.28X2 (monthly average temperature), and the coefficient of determination was 0.76. The incidences of FTLS are correlated with temperature.

Objective To identify the pathogen that caused an outbreak of viral encephalitis in Henan area in 2011.Phylogenic analysis was carried out on Coxsackie-virus B5 (CVB5) which was isolated during this outbreak.Methods Five throat swab,21 stool and 14 cerebrospinal fluid (CSF) specimens were collected from 29 inpatients during this outbreak.Viral isolation and real time RT-PCR were then performed for all specimens.Viral nucleic acid of enterovirus 71 (EV71),coxsackievirus A 16 (CA16) and pan-enterovirus (PE)were detected by real time RT-PCR.Phylogenetic tree based on entire VP1 sequences was constructed among CVB5 isolates from 2 stool and 3 CSF specimens of 5 inpatients and others published data retrieved from GenBank.Results The real time RT-PCR results showed that the PE nucleic acid positive rates of throat swab,stool and CSF specimens were 60.0％ (3/5),61.9％ (13/21) and 85.7％ (12/14) respectively.All of these specimens were negative for EV71 and CA16.The isolation rates of throat swab,stool and CSF specimens were 20.0％ (1/5),25.0％ (5/21) and 29.0％ (4/14),respectively.BLAST with both VP1 and 5′-UTR sequences and molecular typing indicated that CVB5 was the main pathogen.Analysis among the 5 positve isolates based on the complete VP1 sequences showed 97.9％-99.5％ homology.Data from homologous comparisons indicated that these isolates had the highest nucleotide acid identity with the Changchun CVB5 CC10/10/Changchun strain (97.1％-98.1％) which caused hand,foot,and mouth disease (HFMD) outbreak in Changchun in 2010,and lower identity (89.0％-89.6％ and 91.8％-92.5％) with the COXB5/Henan/2010 and 03001N strain isolated from Pingdingshan,Henan in 2010 and 2012,respectively.Phylogenetic tree in VP1 region showed that isolates of this outbreak belonged to genotype D,the same clade with Changchun strain.Conclusion CVB5 was the major etiological agent correlated with this outbreak.The shift of predominant genotype might serve as one of the causes that associated with this outbreaks.

Objective To analyze the genotyping of hantavirus and investigate the pathogenic features of local rats in Henan province. Methods A total of 600 rats captured in Queshan county, Zhumadian city from 2014-2016 were chosen to find out the major species and density. Rat lung specimens were detected by RT-PCR using partial M and S segment primers, then sequencing and phylogenetic analysis based on M segment (2003-2302 nt) were performed to analyze gene subtype and evolution. Results In the field of Queshan county, major species were sewer rats and apodemus agrarius, and the average density of rats was 1.33%-1.83%. Sewer rats, mus musculus and apodemus agrarius were major species in the residential area, and the average density of rats was 1.36%-1.97%. Hantaviruses were detected by RT-PCR in three captured rats in 2014, and the species were mus musculus, cricetulus triton and sewer rats. Nucleotide homology similarity based M and S segment of three positive products was 100%. Phylogenetic analysis indicated the virus was belonged to S4 subgenotype of Seoul virus, which was similar with the strains in Korea and Hubei province, China. Conclusion The virus from rats in Queshan county, Henan province is seoul virus, S4 subgenotype. It is necessary to take the relevant prevention and control measures to prevent hemorrhagic fever of renal syndrome because of wide host range.

China ; Electrophoresis, Gel, Pulsed-Field ; Escherichia coli O157 ; classification ; genetics ; Genes, Bacterial ; Humans ; Molecular Typing ; Virulence

Objective To develop and validate a method for detecting factor 8 gene (F8) mutations in hemophilia A patients by Ion Torrent semiconductor sequencing .Methods Intron 22 and intron 1 inversions of F8 gene were identified by long distance PCR (LD-PCR), other mutations in the F8 gene were identified by Ion Torrent sequencing.Candidate variants were validated by Sanger sequencing .Sanger sequencing was applied to screen HA carriers from 11 female family members in the 8 pedigrees.One pregnant woman was offered prenatal diagnosis via analyzing the fetal DNA obtained through amniocentesis . Results Four missense mutations ( c.1331A >C, 1648C >T, c.6506G >A, c.6544C >T), two frameshift mutations ( c.2393 _2394insT, c.6320delG), one splicing mutation ( IVS5 +5G >A), one nonsense mutation (c.43C >T) and one Inv22 mutation were identified in all nine probands respectively . Among 11 female family members, 10 females were identified to be HA carriers, and one didn′t carry the maternal pathogenic mutation.Prenatal diagnosis result showed that the fetus inherited the wild -type maternal allele and was predicted to be unaffected by HA .Conclusion The targeted Ion Torrent sequencing is a reliable and efficient method to detect F8 mutations in patients with Hemophilia A disease .