Prevalence of intestinal protozoan infections among rural children in Henan Province from 2014 to 2015 / 中国血吸虫病防治杂志

Objective To investigate the prevalence and influencing factors of intestinal protozoan infections among rural children in Henan Province. Methods A total of 104 survey sites were sampled from 35 counties (cities) in Henan Province using the stratified cluster sampling method to investigate the prevalence of intestinal protozoan infections among rural children from 2014 to 2015. The trophozoites and cysts of intestinal protozoa were identified using the iodine staining method and the physiological saline direct smear method (one detection for one stool sample). The prevalence of intestinal protozoan infections was compared among rural children with different characteristics, and the factors affecting intestinal protozoan infections among rural children were identified. Results The overall prevalence of intestinal protozoan infections was 0.60% (40/6 771) among rural children in Henan Province from 2014 to 2015. There were 7 species of intestinal protozoa identified, and there was no species-specific prevalence (χ2 = 37.732, P = 0.000). No significant differences were found in prevalence of intestinal protozoan infections among rural children in terms of gender (χ2 = 1.793, P = 0.181), age (χ2 = 1.443, P = 0.486), occupation (χ2 = 0.219, P = 0.896) or ecological region (χ2 = 1.700, P = 0.637). In addition, terrain (χ2 = 2.311, P = 0.510), economic level (χ2 = 4.322, P = 0.229), source of drinking water (χ2 = 0.731, P = 0.393), eating raw vegetables (χ2 = 1.134, P = 0.287) and deworming (χ2 = 1.089, P = 0.297) had no remarkable effects on the prevalence of intestinal protozoan infections among rural children in Henan Province; however, the prevalence of intestinal protozoan infections varied significantly among rural children living in regions with different coverage of non-harmless toilets (χ2 = 10.050, P = 0.018). Conclusion The prevalence of intestinal protozoan infections is low among rural children in Henan Province.

Prevalence and influencing factors of intestinal parasitic diseases among rural children in Henan Province / 中国血吸虫病防治杂志

Objective To understand the epidemic status and influencing factors of intestinal parasitic diseases among rural children in Henan Province. Methods According to the Scheme for The National Survey on Current Status of Major Human Parasitic Diseases in China, the survey counties were selected based on the ecological zones and economic levels in Henan Province between 2014 and 2015. Then, the included counties were stratified according to the topography and economic levels. A township was randomly sampled from each stratum, and a village was randomly sampled from each township as the study site. Finally, a total of 104 study sites from 35 counties were enrolled for the survey of intestinal parasitic diseases in children. At least 250 fresh stool samples were collected from each study site for detection of intestinal helminth eggs with the Kato-Katz technique, for the identification of Necator americanus and Ancylostoma duodenale with the fecal culture method, and for the detection of intestinal protozoa trophozoite and cyst with the physiological saline smear and iodine staining techniques. In addition, the Enterobius vermicularis and tapeworm eggs were detected in children aged 3 to 6 years using the adhesive cellophane-tape perianal swab method. Results The overall prevalence of intestinal parasitic infections was 3.21% (214/6 671) among rural children in Henan Province, and the prevalence of intestinal helminthes (2.62%, 175/6 671) was higher than that of intestinal protozoa (0.60%, 40/6 671). A total of 12 species of intestinal parasites were found, including 4 nematodes species, one trematode species, and 7 protozoa species, and the highest infection was seen in E. vermicularis (2.47%, 161/6 671). Among the four ecological zones in Henan Province, the greatest prevalence of intestinal parasitic infections was detected among children in the Qinba Mountain Ecological Zone (5.85%, 90/1 538). There was no gender-specific difference in the prevalence of intestinal parasitic infections in children (P > 0.05); however, there were age- (χ2 = 32.762, P < 0.05) and education level-specific differences in the prevalence of intestinal parasitic infections in children (χ2 = 67.507, P < 0.05), with the greatest prevalence of E. vermicularis infection seen in all species of intestinal parasites in children at all age groups. Multivariate non-conditional logistic regression analysis showed that high education level, high coverage of harmless toilets, drinking tap water and deworming were protective factors for intestinal parasitic infections in children in Henan Province. The overall prevalence of intestinal parasitic infections appeared a tendency towards a gradual decline among children in Henan Province as compared to the previous two surveys. Conclusions The overall prevalence of intestinal parasitic infections shows a tendency towards a remarkable decline among children in Henan Province. E. vermicularis infection should be given a priority for future parasitic disease control activities among rural children in Henan Province.

Laboratory diagnosis of an imported case of visceral leishmaniasis in Henan Province / 中国血吸虫病防治杂志

Objective To analyze the epidemiological and clinical data of an imported case of visceral leishmaniasis in Henan Province, and explore the method of laboratory diagnosis of kala-azar. Methods The epidemiological and clinical data of an imported visceral leishmaniasis patient were analyzed. Leishmania donovani bodies in bone marrow smears were observed microscopically. The antibody was detected by using rK39 dipstick test strips. Two pairs of specific primers, K13A-K13B and LITSR-L5.8S, were used to amplify kinetoplast DNA and internal transcribed spacer of rDNA of the parasite, respectively. Results The patient had been in the epidemic area of visceral leishmaniasis, and had symptoms such as irregular fever, splenomegaly, pancytopenia, and inversed ratio of albumin and globulin. The amastigotes of L. donovani were found in the bone marrow smears, and rK39 test strip was positive, and the PCR products of K13A-K13B and LITSR-L5.8S were 87 bp and 285 bp respectively. The similarities of the two fragment sequences to the corresponding sequences of L. donovani were 94% and 100%, respectively. Conclusion The case is diagnosed as visceral leishmaniasis according to the epidemiological data, clinical manifestations and laboratory test results of the patient, and the pathogen is L. donovani.

Investigation on human intestinal parasitic diseases in ecological regions of Qinba Mountains in Henan Province in 2015 / 中国血吸虫病防治杂志

Objective To understand and analyze the epidemic status of human intestinal parasitic diseases in the ecological region of the Qinba Mountains in Henan Province, so as to provide the evidence for the control of the diseases in this region. Methods According to the methods of National Survey Scheme on Major Human Parasitic Diseases, 26 survey spots were selected in 8 counties (cities) in ecological regions of Qinba Mountains in Henan Province in 2015. After collecting the human fecal samples, the eggs of intestinal helminthes were detected with the modified Kato-Katz thick smear method, the species of Ancylostoma of the hookworm egg-positive samples were identified with the tube fecal culture method, the eggs of Enterobius vermicularis were detected by using the adhesive tape method in children aged 3 to 6 years, and the cyst and trophozoite of intestinal protozoa were examined by iodine staining. The infections of intestinal parasites in different populations and the related influencing factors were analyzed. Results Totally 6 706 residents were recruited in this study, 8 kinds of parasites were found including 3 species of helminthes and 5 species of protozoans. The overall infection rate of intestinal parasites was 3.39%, covering 2.92% for helminthes and 0.49% for protozoans. The infection rate of E. vermicularis in children aged 3 to 6 years was 3.93%. The 3 identified helminthes were Ascaris lumbricoides, E. vermicularis, and Dicrocoelium dendriticum, with the highest infection rate of E. vermicularis (2.68%, χ2 = 306.362, P < 0.05). The 5 identified protozoans were Entamoeba hartmani, E. coli, Endolimax nana, Giardia lamblia, and Blastocystis hominis, and the infection rate of E. nana was the highest (χ2 = 23.842, P < 0.05). Among the 8 counties (cities), the highest infection rate of helminthes, mainly with Enterobius infection, existed in Ruyang County, and the infection rate of protozoans in Xin’an County was the highest, mainly with E. nana infection. There was a significant difference in the infection rates of helminthes and protozoans among the 8 counties (cities) (helminthes: χ2 = 357.525, P < 0.05; protozoans: χ2 = 38.795, P < 0.05). There was no significant difference in the helminth infection rate between males and females ( χ2 = 0.034, P > 0.05), but the protozoan infection rate of the males was higher than that of the females ( χ2 = 12.946, P < 0.05). The helminth infection rate of the 0-9 years old group was the highest ( χ2 = 62.600, P < 0.05), up to 6.60%, mainly with E. vermicularis infection. There was no significant difference in the infection rate of intestinal parasites among nationalities ( χ2 = 0.212, P > 0.05). The helminth infection rate of preschool children was the highest and up to 8.43%, mainly with E. vermicularis infection. The infection of A. lumbricoides was mainly happened among farmers, with the infection rate of 0.31%. There was a significant difference in the helminth infection rate among different occupations (χ2 = 84.333, P < 0.05). The intestinal parasites infection rate of the illiterate population was the highest and up to 7.63%, there was a significant difference among the populations with different educational levels (χ2 = 72.013, P < 0.05). The helminth infection rate of plain areas was higher than that of valley and hill ( χ2 = 11.690, P < 0.05). The helminth infection rate of people with low-income was the highest and up to 10.32% (χ2 = 244.999, P < 0.05). The protozoan infection rate of people with middle-income was the highest, who had the annual per capita net income of 6 000 to 7 999 Yuan ( χ2 = 24.749, P < 0.05). The helminth infection rate of people drinking well water was much higher than that of people drinking tap water (χ2 = 62.255, P < 0.05). The protozoan infection rate of people without insect repellent was higher than that of people with insect repellent (χ2 = 5.235, P < 0.05). Conclusions The infection rates of human intestinal parasites in the ecological region of the Qinba Mountains in Henan Province have decreased sharply. E. vermicularis infection in children should be the main point of prevention and treatment of parasitic diseases in the future.

Investigation on human intestinal parasitic diseases in ecological regions of Qinba Mountains in Henan Province in 2015 / 中国血吸虫病防治杂志

Objective To understand and analyze the epidemic status of human intestinal parasitic diseases in the ecological region of the Qinba Mountains in Henan Province, so as to provide the evidence for the control of the diseases in this region. Methods According to the methods of National Survey Scheme on Major Human Parasitic Diseases, 26 survey spots were selected in 8 counties (cities) in ecological regions of Qinba Mountains in Henan Province in 2015. After collecting the human fecal samples, the eggs of intestinal helminthes were detected with the modified Kato-Katz thick smear method, the species of Ancylostoma of the hookworm egg-positive samples were identified with the tube fecal culture method, the eggs of Enterobius vermicularis were detected by using the adhesive tape method in children aged 3 to 6 years, and the cyst and trophozoite of intestinal protozoa were examined by iodine staining. The infections of intestinal parasites in different populations and the related influencing factors were analyzed. Results Totally 6 706 residents were recruited in this study, 8 kinds of parasites were found including 3 species of helminthes and 5 species of protozoans. The overall infection rate of intestinal parasites was 3.39%, covering 2.92% for helminthes and 0.49% for protozoans. The infection rate of E. vermicularis in children aged 3 to 6 years was 3.93%. The 3 identified helminthes were Ascaris lumbricoides, E. vermicularis, and Dicrocoelium dendriticum, with the highest infection rate of E. vermicularis (2.68%, χ2 = 306.362, P < 0.05). The 5 identified protozoans were Entamoeba hartmani, E. coli, Endolimax nana, Giardia lamblia, and Blastocystis hominis, and the infection rate of E. nana was the highest (χ2 = 23.842, P < 0.05). Among the 8 counties (cities), the highest infection rate of helminthes, mainly with Enterobius infection, existed in Ruyang County, and the infection rate of protozoans in Xin’an County was the highest, mainly with E. nana infection. There was a significant difference in the infection rates of helminthes and protozoans among the 8 counties (cities) (helminthes: χ2 = 357.525, P < 0.05; protozoans: χ2 = 38.795, P < 0.05). There was no significant difference in the helminth infection rate between males and females ( χ2 = 0.034, P > 0.05), but the protozoan infection rate of the males was higher than that of the females ( χ2 = 12.946, P < 0.05). The helminth infection rate of the 0-9 years old group was the highest ( χ2 = 62.600, P < 0.05), up to 6.60%, mainly with E. vermicularis infection. There was no significant difference in the infection rate of intestinal parasites among nationalities ( χ2 = 0.212, P > 0.05). The helminth infection rate of preschool children was the highest and up to 8.43%, mainly with E. vermicularis infection. The infection of A. lumbricoides was mainly happened among farmers, with the infection rate of 0.31%. There was a significant difference in the helminth infection rate among different occupations (χ2 = 84.333, P < 0.05). The intestinal parasites infection rate of the illiterate population was the highest and up to 7.63%, there was a significant difference among the populations with different educational levels (χ2 = 72.013, P < 0.05). The helminth infection rate of plain areas was higher than that of valley and hill ( χ2 = 11.690, P < 0.05). The helminth infection rate of people with low-income was the highest and up to 10.32% (χ2 = 244.999, P < 0.05). The protozoan infection rate of people with middle-income was the highest, who had the annual per capita net income of 6 000 to 7 999 Yuan ( χ2 = 24.749, P < 0.05). The helminth infection rate of people drinking well water was much higher than that of people drinking tap water (χ2 = 62.255, P < 0.05). The protozoan infection rate of people without insect repellent was higher than that of people with insect repellent (χ2 = 5.235, P < 0.05). Conclusions The infection rates of human intestinal parasites in the ecological region of the Qinba Mountains in Henan Province have decreased sharply. E. vermicularis infection in children should be the main point of prevention and treatment of parasitic diseases in the future.

Prevalence and influencing factors of intestinal parasitic diseases among rural children in Henan Province / 中国血吸虫病防治杂志

Objective To understand the epidemic status and influencing factors of intestinal parasitic diseases among rural children in Henan Province. Methods According to the Scheme for The National Survey on Current Status of Major Human Parasitic Diseases in China, the survey counties were selected based on the ecological zones and economic levels in Henan Province between 2014 and 2015. Then, the included counties were stratified according to the topography and economic levels. A township was randomly sampled from each stratum, and a village was randomly sampled from each township as the study site. Finally, a total of 104 study sites from 35 counties were enrolled for the survey of intestinal parasitic diseases in children. At least 250 fresh stool samples were collected from each study site for detection of intestinal helminth eggs with the Kato-Katz technique, for the identification of Necator americanus and Ancylostoma duodenale with the fecal culture method, and for the detection of intestinal protozoa trophozoite and cyst with the physiological saline smear and iodine staining techniques. In addition, the Enterobius vermicularis and tapeworm eggs were detected in children aged 3 to 6 years using the adhesive cellophane-tape perianal swab method. Results The overall prevalence of intestinal parasitic infections was 3.21% (214/6 671) among rural children in Henan Province, and the prevalence of intestinal helminthes (2.62%, 175/6 671) was higher than that of intestinal protozoa (0.60%, 40/6 671). A total of 12 species of intestinal parasites were found, including 4 nematodes species, one trematode species, and 7 protozoa species, and the highest infection was seen in E. vermicularis (2.47%, 161/6 671). Among the four ecological zones in Henan Province, the greatest prevalence of intestinal parasitic infections was detected among children in the Qinba Mountain Ecological Zone (5.85%, 90/1 538). There was no gender-specific difference in the prevalence of intestinal parasitic infections in children (P > 0.05); however, there were age- (χ2 = 32.762, P < 0.05) and education level-specific differences in the prevalence of intestinal parasitic infections in children (χ2 = 67.507, P < 0.05), with the greatest prevalence of E. vermicularis infection seen in all species of intestinal parasites in children at all age groups. Multivariate non-conditional logistic regression analysis showed that high education level, high coverage of harmless toilets, drinking tap water and deworming were protective factors for intestinal parasitic infections in children in Henan Province. The overall prevalence of intestinal parasitic infections appeared a tendency towards a gradual decline among children in Henan Province as compared to the previous two surveys. Conclusions The overall prevalence of intestinal parasitic infections shows a tendency towards a remarkable decline among children in Henan Province. E. vermicularis infection should be given a priority for future parasitic disease control activities among rural children in Henan Province.

Laboratory diagnosis of an imported case of visceral leishmaniasis in Henan Province / 中国血吸虫病防治杂志

Objective To analyze the epidemiological and clinical data of an imported case of visceral leishmaniasis in Henan Province, and explore the method of laboratory diagnosis of kala-azar. Methods The epidemiological and clinical data of an imported visceral leishmaniasis patient were analyzed. Leishmania donovani bodies in bone marrow smears were observed microscopically. The antibody was detected by using rK39 dipstick test strips. Two pairs of specific primers, K13A-K13B and LITSR-L5.8S, were used to amplify kinetoplast DNA and internal transcribed spacer of rDNA of the parasite, respectively. Results The patient had been in the epidemic area of visceral leishmaniasis, and had symptoms such as irregular fever, splenomegaly, pancytopenia, and inversed ratio of albumin and globulin. The amastigotes of L. donovani were found in the bone marrow smears, and rK39 test strip was positive, and the PCR products of K13A-K13B and LITSR-L5.8S were 87 bp and 285 bp respectively. The similarities of the two fragment sequences to the corresponding sequences of L. donovani were 94% and 100%, respectively. Conclusion The case is diagnosed as visceral leishmaniasis according to the epidemiological data, clinical manifestations and laboratory test results of the patient, and the pathogen is L. donovani.

Outbreak of dengue Fever in central China, 2013 / 生物医学与环境科学（英文）

In 2013, the first dengue fever (DF) outbreak in central China was reported in the central of Henan province, northern temperate regions, although they have been sequentially recorded in Southern China. 106 suspected DF cases were reported and 73 patients were confirmed dengue virus type 3 (DEN-3) infections. 62/392 (15.8%) local health persons showed DEN antibodies positive. To this day Henan is the northernmost province in China which has been reported about outbreak of DF and what is important is that it warns us the endemic range of DF has been expanded geographically in China.

Genome sequences of coxsackievirus B5 isolated from viral encephalitis patients in Henan province, 2011 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To analyze the complete genome sequences of two coxsackievirus B5 (CVB5) isolated in Henan province, 2011.</p><p><b>METHODS</b>Specimens were collected from viral encephalitis patients and followed by viral isolation on them. RNA were extracted from positive isolates and the amplified products were sequenced. The full-length genomes of them were acquired by assembling the fragments, using DNAStar 5.01 software while phylogenetic analysis were performed with Mega 5.05 and other software.</p><p><b>RESULTS</b>The genomes RNA of 03001N and 17Y showed 7408 bp and 7404 bp long, and the 5'- and 3'-untranslated regions were 747 bp, 743 bp and 103 bp, 103 bp, respectively. BLAST analysis of these two isolates, based on the complete genome, showed 97% identity, with both of them having the highest similarity(98%, 99%)to the CVB5 strain isolated from Henan in 2010 rather than other CVB5 strains. Coding regions of both isolates were 6558 bp, code for a polyprotein of 2185 amino acids (aa) and both of them showed 99% amino acid identity. Phylogenetic tree in VP1 region showed that the two isolates belonged to the same clade with other strains isolated from all over the country in the past years, except for some CVB5 strains isolated from Henan and Shandong province in 2009 that formed the other cluster.</p><p><b>CONCLUSION</b>It seemed that more than one group of CVB5 were circulating in Henan province and these two isolates appeared the main epidemic strains circulating in the past years.</p>

Clinical trial on the prevention of diarrhea by oral BIFICO for infants aged 1-6 years / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the prevention of diarrhea by oral BIFICO for infants aged 1-6 years.</p><p><b>METHODS</b>490 cases of infants were randomly divided after age stratification: the experimental group (n = 247) and the control group (n = 243). Based on principles of randomized double-blind and placebo-controlled, the infants were given BIFICO (dedicated clinical research)therapy for 4 consecutive days, then observed for 21 days. 25 days composed a cycle. They were observed total 5 cycles. During the study period, principles for "the diarrhoea patients must be detected", follow-up visited the participant infants and conducted etiology detection by way of sampling for diarrhea infants. Evaluate the prevention efficacy of diarrhea by oral BIFICO for infants aged 1-6 years.</p><p><b>RESULTS</b>A total of 480 completed all study. 120 and 95 infants in the control group and experimental group were detected with diarrhea. The incidence of diarrhea was 50.85% and 38.93% in these two guoup, respectively. The difference has statistical significance (chi2 = 4.175, P = 0.041). In the third observation period, the infants in the control group had a higer incidence of diarrhea compared with which in the experimental group (chi2 = 4.415, P = 0.036). 14 strains of rotavirus, 3 strains of norovirus, 3 strains of sappovirus, 2 strains of adenovirus, 5 strains of salmonella and 4 strains of Shigella were check out in 128 samples.</p><p><b>CONCLUSION</b>Oral BIFICO can paly certain preventive role on diarrhea, and decrease the incidence of diarrhea in infants aged 1-6 years.</p>

Comparison of different molecular assays for the rapid detection of enterovirus 71 (EV71) / 病毒学报

Molecular detection of enterovirus (EV)71 RNA based on PCR methods is a quick and sensitive approach. At present, different PCR-based methods for EV71 RNA detection are available, but comparisons of results obtained using different approaches are limited. This study is to compare the analytical sensitivity and specificity of different real-time reverse transcription-polymerase chain reaction (rRT-PCR) and conventional reverse transcription-polymerase chain reaction (cRT-PCR) assays for enterovirus and EV71 detection, Altogether, three rRT-PCR assays and one cRT-PCR assay targeting the 5'UTR gene for universal detection of enterovirus; two rRT-PCR assays andone cRT-PCR assay targeting the VP1 gene for specific detection of EV 71 were examined. All assays showed good specificity. The detection sensitivity ranged from 8.19 x 10 to 8.19 x 10(5) copy equivalents. In general, rRT-PCR assays were more sensitive than cRT-PCR assays. All rRT-PCR assays showed 100% sensitivity for clinical specimens.

Survey on ticks and detection of new bunyavirus in some vect in the endemic areas of fever, thrombocytopenia and leukopenia syndrome (FTLS) in Henan province / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the distribution, species, seasonal fluctuation of ticks and detect new bunyavirus in some hematophagus in the endemic areas of fever thrombocytopenia and leukopenia syndrome (FTLS) in Henan province.</p><p><b>METHODS</b>From March to December 2011, the free ticks were collected manually with white cloth from the grassland and the parasitic ticks were collected from the host skin by hand searching in Xinyang and Jiyuan. The density and seasonal fluctuation of ticks were analyzed after classification of the specimen. The hematophagus were collected including gadfly (38 in 16 groups), cattle lice (224 in 16 groups), mosquitoes (238 in 17 groups) and ticks (825 in 77 groups), then RNA of new bunyavirus were detected by RT-PCR.</p><p><b>RESULTS</b>A total of 12 388 ticks were collected in Xinyang and Jiyuan, consisting of 2 families, 5 geniuses and 6 species. In Xinyang city, 622 ticks were identified, consisting of 2 families, 3 geniuses and 3 species, including 2 (0.32%) Ornithodoros lahorensis, 451 (72.51%) Haemaphysalis longicornis and 117 (18.81%) Boophilus microplus. In Jiyuan city, 11 766 ticks were identified, consisting of 1 family, 4 geniuses and 5 species, including 7718 (65.60%) Haemaphysalis longicornis, 164 (1.39%) H.anatolicum anatolicum and 710 (6.03%) other ticks such as H. detritum, Boophilus microplus and Rhipicephalus sanguineus. Haemaphysalis longicornis were found in both districts as the predominant species in Henan province. Ticks were active from March to October. The average density was 160 per person hour and the peak was from May to July with density 278, 209 and 542 per person hour respectively. The results was positive in RNA detection of new bunyavirus in 11 groups of tick and 3 groups of gadfly by RT-PCR. The results were negative in all other hematophagus.</p><p><b>CONCLUSION</b>Ornithodoros lahorensis, Haemaphysalis longicornis, Boophilus microplus, H.anatolicum anatolicum, Rhipicephalus sanguineus and H. detritum were found in Henan province. Haemaphysalis longicornis was the predominant species. The density of ticks varied with the seasons. The detection of new bunyavirus by PCR was positive in some ticks and gadflies.</p>

Analysis of the epidemic characteristics of fever and thrombocytopenia syndrome in Henan province, 2007 - 2011 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze the epidemiological characteristics of fever thrombocytopenia and leukopenia syndrome (FTLS) in Henan province, China in 2007 - 2011.</p><p><b>METHODS</b>Data from specific surveillance system for FTLS in Henan and Information Management System of Chinese Center for Disease Control and Prevention were used to collect the information of the cases.Descriptive epidemiological methods were used to analyze the surveillance data during 2007 - 2011. Patients' sera were collected to detect new bunyavirus using fluorescent RT-PCR and virus isolation.</p><p><b>RESULTS</b>During 2007 - 2011, 1021 FTLS cases were reported in Henan province. The fatality rate was 2.25%with 23 deaths. The cases reported in Xinyang city were 1007, accounting for 98.75%.Cases were mainly occurred between April and October, accounting for 96.47% (985/1021). Epidemic peak was May to July, accounting for 59.16% (604/1021). The second peak occurred in September, accounting for 12.05% (123/1021). The age of the cases ranged from 1 to 88 years old with the median age of 59. Sex ratio (male:female) was 1:1.50 (408:613). In all cases, 93.73% (957/1021) were farmers. In 465 patients' sera, the positive rate of new bunyavirus was 69.25% (322/465) using fluorescent RT-PCR. In 164 patients' sera, 67 strains of new bunyavirus were isolated with isolation rate of 40.85% (67/164).</p><p><b>CONCLUSION</b>FTLS in Henan province is caused mainly by the new bunyavirus and has certain regional and seasonal characteristics. Most cases are female older farmers.</p>

Analysis on the diagnosis and treatment of a cluster of cases infected by new bunyavirus / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze and summarize the clinical characteristics, experience of diagnosis and treatment of cases infected by new bunyavirus, which occurred in Henan province in 2010.</p><p><b>METHODS</b>The clinical characteristics and effect of diagnosis and treatment of 5 cases were analyzed using descriptive epidemiological method. Blood specimens were detected by RT-PCR and pathogen separation.</p><p><b>RESULTS</b>PCR testing was positive for all 5 cases. New bunyavirus were isolated from 2 cases. In 5 cases, fever (5/5), the whole body aches (5/5), fatigue (5/5), anorexia (5/5), nausea (5/5), the chills (4/5), cough (4/5), expectoration (4/5), vomiting (3/5), conjunctival hyperemia (3/5); Leukocyte reduction (5/5), thrombocytopenia (5/5), elevated alanine aminotransferase (4/5), elevated aspartate aminotransferase (4/5), elevated lactate dehydrogenase (5/5), creatine kinase elevations (4/5), urinary protein (3/5). By symptomatic and supportive treatment and prophylactic antibiotics, the first case died and the other 4 cases were cured. The average course of disease was 15.4 days.</p><p><b>CONCLUSION</b>Cases infected by new bunyavirus have complicated clinical feature and multiple organ damage. If symptomatic treatment is in time, prognosis will be good.</p>

Establishment of indirect immunofluorescence assay (IFA) for detection of IgG antibody against new bunyavirus / 中华预防医学杂志

<p><b>OBJECTIVE</b>To develop an indirect immunofluorescence assay (IFA) for detection of IgG antibodies against new bunyavirus.</p><p><b>METHODS</b>The antigen slides were prepared with 5 new bunyavirus strains isolated using Africa green monkey kidney (Vero) cells. Specificity and sensitivity evaluation of IFA were carried out by optimizing working conditions of IFA. Using established IFA, serum samples from both acute and recovery phases were tested for 126 cases with fever thrombocytopenia and leukopenia syndrome in Xinyang, Henan province in 2007 - 2011. The results were compared with detections by RT-PCR.</p><p><b>RESULTS</b>The new bunyavirus stable immunofluorescence specific WZ69 strain was selected to prepare antigen slides of IFA. The optimum conditions of IFA were: optimum dilution for primary antibody (serum) and secondary antibody (isosulfocyanic acid fluorescence marked goat anti-human IgG antibody) was 1:40 and 1:150 respectively. The optimum dilution for Evans blue in secondary antibody was 1:20 000. Among the 126 patients, 96 paired serum specimens were tested positive to the new bunyavirus and 30 patients were tested negative to the virus. The positive rate of antibodies was 76.19%. There was no significant difference in results between IFA and RT-PCR (72.22% (91/126)) (P > 0.05).</p><p><b>CONCLUSION</b>The IFA has high sensitivity and specificity with easy operation. It can be used in detecting the new bunyavirus infection in patients with fever, thrombocytopenia and leukopenia syndrome.</p>

Culture, isolation and identification of new bunyavirus in African green monkey kidney(Vero) cells / 中华预防医学杂志

<p><b>OBJECTIVE</b>To culture, isolate and identify new bunyavirus in Vero cell line.</p><p><b>METHODS</b>Samples of 164 new bunyavirus positive by real time RT-PCR detection and well preserved serum specimens were selected from cases of fever, thrombocytopenia and leukopenia syndrome (FTLS) in Xinyang, Henan province in 2009 - 2011. These sera were cultured in Vero cell line and new bunyavirus were detected by observing cytopathic effect (CPE), Real-time RT-PCR, indirect immunofluorescence assay (IFA) and thin-section electron microscopy observation. A total of 10 positive PCR products were selected randomly for sequencing and the results were compared with sequence in Genbank.</p><p><b>RESULTS</b>Among 164 FTLS serum specimens cultured in Vero cell line, no special CPE were observed and 67 strains (40.85%) were positive detected by Real-time RT-PCR. Nucleic acid similarity of 10 specimens were 97.8% - 100% and there's also a high similarity (> 99%) between specimens and new bunyavirus isolates (Accession No. HQ141600.1). Among 67 positive strains, 58 of them showed specific fluorescence particles by IFA. The viral particles were observed to be spheres with a diameter of 80 - 100 nm by electron microscopy.</p><p><b>CONCLUSION</b>Vero cell line is suitable for culture, isolation and identification of new bunyavirus.</p>

Etiology surveillance of hand-foot-mouth disease in Henan province between 2008 and 2011 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To understand etiological types and distribution features of hand-foot-mouth disease (HFMD) in Henan province between 2008 and 2011.</p><p><b>METHODS</b>A total of 30 486 specimens of feces, rectal swabs or throat swabs from HFMD patients were collected by each Municipal CDC in Henan from 2008 to 2011. The enterovirus 71 (EV71), coxsackie virus A16 (CA16) and other enterovirus (EV) were detected by RT-PCR or real time RT-PCR. The VP1 gene of EV71 was amplified and the sequences were analyzed by bioinformatics software. A genetic evolution tree of the sequence was constructed as well.</p><p><b>RESULTS</b>The positive rates of EV71, CA16 and other EV were 62.70% (11 209/17 876), 12.03% (2150/17 876), 25.27% (4517/17 876) in 17 876 laboratory diagnosed cases, respectively. The differences were statistically significant (χ(2) = 157.17, P < 0.05). The positive rates of EV71, CA16 and other EV were 63.40% (7370/11 624), 11.58% (1346/11 624) and 25.02% (2908/11 624) in male patients and 61.40% (3839/6252), 12.86% (804/6252) and 25.74% (1609/6252) in female patients, respectively. The differences were statistically significant (χ(2) = 4.06, P < 0.05). The children under 5 years old were high-risk population of HFMD, accounting to 97.67% (17 459/17 876) of the laboratory-diagnosed patients.86.92% (15 537/17 876) cases were children between 1 to 3 years old. Constituent ratio of EV71 changed seasonally during a year, there was a high infection ratio of EV71 between April and June, especially in May, the infection ratio reached 69.34% (2384/3438). The positive rates of EV71, CA16 and other EV were 82.48% (5715/6929), 1.76% (122/6929) and 15.76% (1092/6929) among the 6929 laboratory-diagnosed severe cases, respectively. The positive rates of EV71 was higher than CA16 and other EV (χ(2) = 9259.17, 6170.81, P < 0.05, respectively). There were 117 deaths because of severe HFMD, 55 (47.01%) of which were laboratory confirmed. 50 death cases were infected by EV71, and according to the genetic evolution analysis, the VP1 gene of EV71 strain was belonged to subtype C4 of gene C.</p><p><b>CONCLUSION</b>The EV71 and CA16 were the main pathogens which caused HFMD in Henan province, and EV71 virus was the dominant strain, belonging to C4 subtype of gene C.</p>

Epidemic characteristics and viral antibody level among healthy people of Japanese B encephalitis in Henan province in 2010 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To understand the epidemic characteristics and viral antibody level among healthy people of Japanese B encephalitis (JE) in Henan province in 2010.</p><p><b>METHODS</b>A total of 433 selected JE cases in Henan province in 2010 were analyzed by descriptive epidemiological method. Xinyang and Luoyang were selected as survey sites in 2010.12 administrative villages were randomly selected from both cities. As the investigation objects, 519 healthy people from the two cities were randomly selected by eight age groups:less than one year old, 1 - 2 years old, 3 - 4 years old, 5 - 6 years old, 7 - 14 years old, 15 - 19 years old, 20 - 59 years old, and above 60 years old. A total of 1008 samples of blood specimens were collected both in May and November, and JE viral antibody was detected by micro-cytopathic effect neutralization test.</p><p><b>RESULTS</b>The incidence rate of JE was 0.46/100 000 (433/94 130 434) in Henan province in 2010.97.69% (423/433) of the patients were found between July and September, and 81.06% (351/433) were distributed in Nanyang, Xinyang, Luoyang, Zhumadian and Zhoukou city. Children aged 0 to 14 years were the primarily affected group (82.22%, 356/433), the people above 15 years old accounted for 17.78% (77/433) of whole cases in Henan province, but the same group accounted for 65.79% (50/76) of whole cases in Luoyang city, which obviously higher than the percentage in Henan province (χ(2) = 79.57, P < 0.05). Most patients were scattered children in Henan province, accounting for 58.89% (255/433). In Luoyang city, most patients were peasants, accounting for 44.74% (34/76). The antibody positive rate of JE among health people above 15 years old in Luoyang city was 48.94% (46/94), which was lower than it in Xinyang city at 97.78% (88/90). The difference showed statistical significance (χ(2) = 55.42, P < 0.05). The antibody positive rate among healthy people under vaccination was 50.41% (61/121), which was obviously higher than that among people without vaccination, at 16.67% (6/36) in Luoyang city. The difference showed statistical significance (χ(2) = 12.92, P < 0.05). The antibody positive rate among healthy people under vaccination was 67.11% (51/76) in Xinyang city, which was obviously higher than that among people without vaccination, at 46.39% (45/97). The difference showed statistical significance (χ(2) = 7.40, P < 0.05).</p><p><b>CONCLUSION</b>The incidence of JE showed seasonal and regional characteristics, there were differences among ages and occupations. The difference was consistent with the difference in viral antibody level among health people in Henan province and Luoyang city.</p>

Molecular epidemiological analysis of species B enteroviruses isolated from Henan Province of China during the six months in 2010 / 病毒学报

This report presents an overview of human enterovirus B species in Henan Province. A total of 14 isolates of HEV-B species isolated under HFMD surveillance network during the six months in 2010 were examined. Based on molecular typing results targeting VP1 region, 14 isolates were classified into 6 serotypes of HEV-B. Furthermore, comparison of these 14 isolates with reference strains and strains in mainland China was conducted. The phylogenetic analysis revealed that E25, E11 and E6 showed homology with those from Shandong Province which adjoins Henan Province. E1 and E13 showed homology with those from Yunnan Province, and E30 showed homology with Henan strain isolated in 2008. Cocirculation of two lineages of echovirus 6 was observed.

Genomic characteristics of coxsakievirus A16 isolated in Henan Province / 病毒学报

To reveal the genomic sequence characteristics of coxsackievirus A16 (CoxA16) strain isolated from patients with hand-foot-mouth disease (HFMD) in Henan province. A total of 406 samples were detected by reverse-transcription polymerase chain reaction (RT-PCR) and cell-culture-based isolation of coxsackievirus A16. The whole genome of CoxA16 isolate was amplified using 10 pairs of primers, the sequences were analyzed and phylogenetic tree was generated by bioinformatics software. The full length of HN1162/HN/CHN/2010 genome was 7411bp. Compared with the other CoxA16 strains released in GenBank, the nucleotide similarities were 87.0-97.9%, 77.0%-95.4%, 80.3%-96.9%, 77.9% 96.2%, 80.5-100% in 5'UTR, P1, P2, P3, 3'UTR region, respectively; The similarities of nucleotide and amino acid sequences in VP1 region were 91.4%-96.4% and 99.3%-99.7%, respectively. Phylogenetic tree analysis showed that CoxA16 strains isolated from Henan, Shenzhen, Guangzhou and Fujian belonged to the same cluster. The newly isolated CoxA16 from Henan province belonged to subgenotype C2/B-2. These results will have great significance in monitoring CoxA16 and for prevention and control of hand-foot-mouth disease.