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1.
Article in English | WPRIM | ID: wpr-922407

ABSTRACT

OBJECTIVES@#To study the effect of high-fat diet for maternal Sprague-Dawley rats at different stages on glucose and lipid metabolism in offspring and related mechanisms.@*METHODS@#According to the diet before pregnancy and during pregnancy and lactation, maternal rats were randomly divided into four groups (@*RESULTS@#Compared with the control diet groups (CC and CH groups), the groups with high-fat diet before pregnancy (HC and HH groups) had a significant increase in body weight (@*CONCLUSIONS@#High-fat diet for rats at different stages before and after pregnancy has different effects on glucose and lipid metabolism of offspring rats, and high-fat diet before pregnancy and during pregnancy and lactation has the greatest effect. The effect of high-fat diet on glucose and lipid metabolism of offspring rats is considered associated with the changes in the expression of genes involved in glucose and lipid metabolism.


Subject(s)
Animals , Body Weight , Diet, High-Fat/adverse effects , Female , Glucose/metabolism , Insulin , Insulin Resistance , Lipid Metabolism , Liver/metabolism , Male , Pregnancy , Rats , Rats, Sprague-Dawley
2.
Article in English | WPRIM | ID: wpr-829011

ABSTRACT

Objective@#High PM concentration is the main feature of increasing haze in developing states, but information on its microbial composition remains very limited. This study aimed to determine the composition of microbiota in PM in Guangzhou, a city located in the tropics in China.@*Methods@#In Guangzhou, from March 5 to 10 , 2016, PM was collected in middle volume air samplers for 23 h daily. The 16S rDNA V4 region of the PM sample extracted DNA was investigated using high-throughput sequence.@*Results@#Among the Guangzhou samples, , , , , and were the dominant microbiota accounting for more than 90% of the total microbiota, and was the dominant gram-negative bacteria, accounting for 21.30%-23.57%. We examined the difference in bacterial distribution of PM between Beijing and Guangzhou at the genus level; was found in both studies, but was only detected in Guangzhou.@*Conclusion@#In conclusion, the diversity and specificity of microbial components in Guangzhou PM were studied, which may provide a basis for future pathogenicity research in the tropics.


Subject(s)
Air Microbiology , Air Pollutants , Bacteria , Classification , China , Cities , Environmental Monitoring , Microbiota , Particle Size , Particulate Matter , RNA, Bacterial , RNA, Ribosomal, 16S
3.
Article in English | WPRIM | ID: wpr-828995

ABSTRACT

Pseudorabies virus (PRV), a veterinary pathogen that infects domestic animals as well as wild animals such as wild boar and feral swine, was recently reported to infect human and led to endophthalmitis and encephalitis. A retrospective seroepidemiologic survey was conducted using 1,335 serum samples collected from patients with encephalitis and ELISA positive rates were 12.16%, 14.25%, and 6.52% in 2012, 2013, and 2017, respectively. The virus neutralizing antibody titers of positive samples correlated well with ELISA results. The pseudorabies virus antibody positive rate of patients with encephalitis were higher than that of healthy people in 2017. The above results suggest that some undefined human encephalitis cases may be caused by PRV infection.


Subject(s)
Adult , Animals , Antibodies, Viral , Blood , China , Encephalitis , Allergy and Immunology , Virology , Enzyme-Linked Immunosorbent Assay , Female , Herpesvirus 1, Suid , Allergy and Immunology , Humans , Male , Middle Aged , Prevalence , Pseudorabies , Blood , Allergy and Immunology , Virology , Retrospective Studies , Seroepidemiologic Studies , Young Adult
4.
Article in English | WPRIM | ID: wpr-781430

ABSTRACT

OBJECTIVE@#The current outbreak of Zika virus (ZIKV) poses a severe threat to human health. Two ZIKV strains were isolated from mosquitoes collected from the Dejiang prefecture in China in 2016, which was the first isolation of ZIKV in nature in China.@*METHODS@#In this study, serum samples were collected from 366 healthy individuals and 104 animals from Dejiang prefecture in 2017, and the plaque reduction neutralization test (PRNT) was used to evaluate the seroprevalence of ZIKV.@*RESULTS@#None of the 366 residents from whom the samples were collected were seropositive for ZIKV. None of the 11 pigs from whom the samples were collected were seropositive for ZIKV, while 1 of 63 (1.59%) chickens and 2 of 30 (6.67%) sheep were seropositive for ZIKV.@*CONCLUSION@#The extremely low seropositivity rate of ZIKV antibodies in animals in the Dejiang prefecture, Guizhou province in this study indicates that ZIKV can infect animals; however, there is a low risk of ZIKV circulating in the local population.

5.
Article in English | WPRIM | ID: wpr-781424

ABSTRACT

West Nile virus (WNV) causes West Nile fever and West Nile encephalitis. Because infection by WNV creates serious public health problems, its simple, rapid, and visual detection is very important in clinical practice, especially in resource-limited laboratories. We have developed a rapid, specific, and highly sensitive internally controlled reverse transcription recombinase-aided amplification (RTRAA) assay to detect WNV, using both real-time fluorescence and the lateral flow dipstick (LFD) at 39.0 °C for 30 min. The analytical sensitivity of the RT-RAA assay was 10 plasmid copies and 1.6 pfu per reaction with real-time fluorescence, and 1,000 plasmid copies per reaction with the LFD. No crossreaction with other control viruses was observed. Compared with the RT-qPCR assay, the RT-RAA assay demonstrated 100% sensitivity and 100% specificity for WNV.

6.
Article in Chinese | WPRIM | ID: wpr-699372

ABSTRACT

Objective:To explore influence of preoperative nicorandil application on coronary no-reflow during emer-gency percutaneous coronary intervention(PCI)in patients with acute anterior myocardial infarction(AAMI). Methods:A total of 116 AAMI patients were randomly and equally divided into routine treatment group and nic-orandil group(received nicorandil based on routine treatment).TIMI grade,incidence rate of no-reflow,ECG ST-segment elevation resolution(STR)and incidence rate of major adverse cardiovascular events(MACE)after PCI were compared between two groups.Results:Compared with routine treatment group after PCI,there were signifi-cant rise in percentage of infarct related artery(IRA)TIMI grade 3(77.6% vs.91.4%)and STR on 2h after PCI[(0.15 ± 0.05)mm vs.(0.18 ± 0.05)mm],and significant reductions in incidence rates of no-reflow(13.8% vs. 3.4%)and MACE(17.2% vs.5.2%)in nicorandil group after PCI,P<0.05 or <0.01. Conclusion:Preoperative nicorandil application can reduce incidence rate of no-reflow after PCI,improve myocardial perfusion and reduce in-cidence rate of major adverse cardiovascular events.

7.
Article in English | WPRIM | ID: wpr-690669

ABSTRACT

<p><b>OBJECTIVE</b>To detect Japanese encephalitis virus (JEV) rapidly and distinguish its genotypes, a TaqMan-based reverse transcriptase quantitative polymerase chain reaction (RT-PCR) detection system was developed.</p><p><b>METHODS</b>By aligning the full-length sequences of JEV (G1-G5), six sets of highly specific TaqMan real-time RT-PCR primers and probes were designed based on the highly conserved NS1, NS2, and M genes of JEV, which included one set for non-specific JEV detection and five sets for the detection of specific JEV genotypes. Twenty batches of mosquito samples were used to evaluate our quantitative PCR assay.</p><p><b>RESULTS</b>With the specific assay, no other flavivirus were detected. The lower limits of detection of the system were 1 pfu/mL for JEV titers and 100 RNA copies/µL. The coefficients of variation of this real-time RT-PCR were all < 2.8%. The amplification efficiency of this method was between 90% and 103%.</p><p><b>CONCLUSION</b>A TaqMan real-time RT-PCR detection system was successfully established to detect and differentiate all five JEV genotypes.</p>


Subject(s)
Animals , Culicidae , Virology , Encephalitis Virus, Japanese , Genetics , Polymerase Chain Reaction , Methods , Reproducibility of Results , Sensitivity and Specificity
8.
Article in English | WPRIM | ID: wpr-690667

ABSTRACT

Japanese encephalitis (JE) is a serious public health issue. This study was undertaken to better understand the relationship between JE distribution and environmental factors in China. JE data from 2005 to 2010 were retrieved from National Notifiable Disease Report System. ArcGIS, remote sensing techniques, and R software was used to exhibit and explore the relationship between JE distribution and environmental factors. Our results indicated that JE cases were mostly concentrated in warm-temperate, semitropical and tropical zones with annual precipitation > 400 mm; Broad-leaved evergreen forest, shrubs, paddy field, irrigated land, dryland, evergreen coniferous forest, and shrubland were risk factors for JE occurrence, and the former five were risk factors for counties with high JE incidence. These findings will inform the effective allocation of limited health resources such as intensive vaccination, surveillance and training in areas with high environmental risk factors.


Subject(s)
China , Epidemiology , Encephalitis, Japanese , Epidemiology , Virology , Environment , Epidemiological Monitoring , Humans , Incidence , Risk Factors
9.
Article in English | WPRIM | ID: wpr-296524

ABSTRACT

Fifteen pediatric cases of suspected Japanese encephalitis (JE) were reported in Beijing Children's Hospital during the late summer of 2013. The clinical manifestations in most cases included high fever, seizures, and abnormal magnetic resonance imaging findings. Twelve of 15 cases were laboratory-confirmed as JE cases by pathogen identification. Epidemiological investigations showed that five of the 12 laboratory-confirmed patients had an incomplete JE vaccination history. Follow-up investigations after discharge indicated that seven laboratory-confirmed JE patients without JE vaccinations had relatively poor prognoses, with an average Modified Rankin Scale (MRS) score of 2.6 when compared with the other five laboratory-confirmed, JE-vaccinated patients with an average MRS score of 0.5. The observation of pediatric JE cases among those with a history of JE vaccination warrants further attention.


Subject(s)
Beijing , Epidemiology , Child , Child, Preschool , Encephalitis Virus, Japanese , Physiology , Encephalitis, Japanese , Diagnosis , Epidemiology , Virology , Female , Humans , Japanese Encephalitis Vaccines , Male , Prognosis
10.
Article in English | WPRIM | ID: wpr-264574

ABSTRACT

A real-time RT-PCR (RT-qPCR) assay for the detection of Tahyna virus was developed to monitor Tahyna virus infection in field-collected vector mosquito samples. The targets selected for the assay were S segment sequences encoding the nucleocapsid protein from the Tahyna virus. Primers and probes were selected in conserved regions by aligning genetic sequences from various Tahyna virus strains available from GenBank. The sensitivity of the RT-qPCR approach was compared to that of a standard plaque assay in BHK cells. RT-qPCR assay can detect 4.8 PFU of titrated Tahyna virus. Assay specificities were determined by testing a battery of arboviruses, including representative strains of Tahyna virus and other arthropod-borne viruses from China. Seven strains of Tahyna virus were confirmed as positive; the other seven species of arboviruses could not be detected by RT-qPCR. Additionally, the assay was used to detect Tahyna viral RNA in pooled mosquito samples. The RT-qPCR assay detected Tahyna virus in a sensitive, specific, and rapid manner; these findings support the use of the assay in viral surveillance.


Subject(s)
Animals , Culicidae , Virology , Encephalitis Virus, California , Real-Time Polymerase Chain Reaction , Methods , Reverse Transcriptase Polymerase Chain Reaction , Methods , Sensitivity and Specificity
11.
Article in English | WPRIM | ID: wpr-270609

ABSTRACT

<p><b>OBJECTIVE</b>To determine the molecular characterization of full-length genome of Japanese encephalitis virus (JEV) genotype V.</p><p><b>METHODS</b>The full-length nucleotide sequences of JEV strains isolated from different locations and sources were used in sequence and phylogenetic analysis.</p><p><b>RESULTS</b>The full-length genome of genotypes V JEV, XZ0934, and Muar strain were composed of 10 983 and 10 988 nucleotides respectively and shared a lower level of identity with JEV genotypes I-IV, ranging from 78.4% (G I, KV1899) to 79.7% (G III, JaGAr01), for the nucleotide sequences, and from 90.0% (G I, KV1899) to 91.8% (G III, JaGAr01) for the amino acid sequences. The open reading frame (ORF) of JEV genotype V spanned nucleotides 96 to 10 397 and encoded 3 433 amino acids. Interestingly, a comparison with JEV genotype I-IV revealed that 3 nucleotides (encoded with a serine residue) were inserted in the NS4A gene of JEV genotype V, and the insertion of nucleotides was also found in downstream of the ORF stop codon in 3'-untranslated region. Moreover, numerous amino acid mutations were observed in 3 functional domains of the E gene of JEV genotype V.</p><p><b>CONCLUSION</b>The molecular characterization of JEV genotype V is significantly different from that of the known genotypes I-IV. The mutations located in the coding region and the non-coding region may be molecular markers of JEV genotype V and warrant further studies to determine their effects on biology and immunogenicity of genotype V strains.</p>


Subject(s)
Amino Acid Sequence , Animals , Base Sequence , Culex , Virology , Encephalitis Virus, Japanese , Genetics , Genome, Viral , Genotype , Humans , Male , Molecular Sequence Data , Phylogeny , Sequence Analysis, DNA , Sequence Homology, Nucleic Acid , Tibet , Young Adult
12.
Chinese Journal of Epidemiology ; (12): 428-432, 2013.
Article in Chinese | WPRIM | ID: wpr-318382

ABSTRACT

<p><b>OBJECTIVE</b>To understand the epidemiologic characteristics of dengue fever, imported from Myanmar to the border of Yunnan province, China. Viral molecular epidemiologic features were also studied.</p><p><b>METHODS</b>Questionnaires were used on each diagnosed, suspected dengue fever, case or unknown cases with fever when coming from Myanmar entering the port and hospitals in Ruili city of Yunnan province. Serum samples of these patients were collected to detect IgM antibody against dengue virus and RT-PCR assay. Homology and phylogenetic tree based on the whole nucleotide sequence of PrM-C and NS5 gene of dengue virus were further analyzed.</p><p><b>RESULTS</b>A total of 103 sera were collected from patients at acute stage in Ruili city in July to November 2008. Among them, 49 cases were confirmed for dengue fever according to IgM and nucleic acid testings. Except one, other 48 cases were all imported into Ruili, from Myanmar. Of those, 18 patients were residents from Mujie city of Myanmar and hospitalized in Ruili and the rest 30 patients were Chinese citizens who had finished business and returned from Myanmar. Two isolates of serum samples from the imported cases were identified and both homology and phylogenetic analysis were performed, using the nucleotide sequences of PrM and NS5 genes. They were divided into dengue type 1 (RLB61) and dengue type 3 (RLC31) and were closer to the dengue virus strains isolated from Southeast Asia countries.</p><p><b>CONCLUSION</b>It is confirmed that an epidemic of dengue fever which was imported from Myanmar to Ruili city of Yunnan province, China. Evidence also showed that both type I and III epidemic strains of dengue virus did exist in Mujie city of Myanmar in 2008.</p>


Subject(s)
Adolescent , Adult , Child , Child, Preschool , China , Epidemiology , Dengue , Epidemiology , Dengue Virus , Genetics , Female , Genotype , Humans , Infant , Male , Middle Aged , Molecular Epidemiology , Myanmar , Epidemiology , Phylogeny , RNA, Viral , Genetics
13.
Chinese Journal of Virology ; (6): 57-62, 2012.
Article in Chinese | WPRIM | ID: wpr-354771

ABSTRACT

Multiplex reverse transcription-polymerase chain reaction (mRT-PCR) is currently available in virus detection and defined as the simultaneous amplification of two or more DNA/RNA targets in a single reaction vessel. In this study, we attempted to modify the conventional mRT-PCR technique on a basis of GenomeLab Genetic Analysis System (GeXP). Initially, we optimized the analytical validation of the GeXP analyzer and its design of workflow and simultaneously detected eight arboviruses that related to epidemic encephalitis by verifying the specificity of mRT-PCR with Japanese encephalitis virus(JEV) cell cultures and positive strains identified previously and determining the sensitivity with in vitro-transcribed RNA of serial dilutions. The GeXP system after optimization could amplify the specific fragments related to the viruses and exposed specifically a total of 13 target genes out of eight types of arboviruses at the level of 10(2) copies/microL, and the findings suggest that the novel protocol we developed can be high-throughput and highly specific and sensitive as well as quickness in screening of the encephalitis viruses, and is promising in detection of encephalitis-associated viruses for molecular epidemiological studies.


Subject(s)
Arboviruses , Genetics , Encephalitis Virus, Japanese , Genetics , Encephalitis Viruses , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Methods , Sensitivity and Specificity
14.
Article in Chinese | WPRIM | ID: wpr-305070

ABSTRACT

<p><b>OBJECTIVE</b>To understand molecular characteristics of Japanese encephalitis virus (JEV) isolated from the major Japanese encephalitis epidemic areas in Sichuan Province, and to provide the foundation for JEV prevention.</p><p><b>METHODS</b>13 JEV strains were isolated from mosquitoes in Sichuan during 2007-2010, E genes and preM genes were sequenced and phylogenetic analyses were performed using MEGA5 molecular software.</p><p><b>RESULTS</b>Phylogenetic analysis indicated that all 13 JEV strains from Sichuan belonged to genotype I, homologies at nucleotide level and deduced amino acid level in PreM gene were 97%-100% and 98.7%-100%, and 97.8%-99.9% and 99.6%-100% in E gene, respectively. Homologies at nucleotide level and deduced amino acid level in PreM gene between 13 JEV strains and JEV isolated in 2004 in Sichuan were 96.2%-99.1% and 97.5%-98.7%, and were 97.7%-99.6% and 98. 6%-100% in E gene, respectively. By comparison with vaacine strains P3 and SA14-14-2, homologies at nucleotide level and deduced amino acid level were 84.1%-85.8% and 93.7%-96.2% in PreM gene, and were 87.6%-88.3% and 97%-97.8% in E gene, respectively. The neurovirulence-related 8 amino acid sites encode by E gene remained unchanged in 13 JEV strains.</p><p><b>CONCLUSION</b>JEV with genotype I predominated in Sichuan, nucleotide sequences and deduced amino acid sequences in PreM gene and E gene were highly conserved, key neurovirulence-rerlated sites remained unchanged. It suggested currently used vaccine is still capable of preventing JEV infection.</p>


Subject(s)
Amino Acid Sequence , Animals , China , Epidemiology , Culicidae , Virology , Encephalitis Virus, Japanese , Chemistry , Classification , Genetics , Encephalitis, Japanese , Epidemiology , Virology , Genotype , Humans , Molecular Sequence Data , Phylogeny , Sequence Alignment , Viral Proteins , Chemistry , Genetics
15.
Article in Chinese | WPRIM | ID: wpr-246175

ABSTRACT

<p><b>OBJECTIVE</b>To grasp the infection rate and genotypes of Japanese encephalitis virus (JEV) in mosquito in Fujian province.</p><p><b>METHODS</b>Mosquito specimens in Sanming city, Jianyang city and Fuzhou city in Fujian province were collected in 2010. RT-PCR was used to detect the JEV sequence from the mosquitoes by specific primers. The sequence splicing and the differentiation analysis for nucleotides, deduced amino acid sequence and phylogenetic tree were performed by the software of ATGC, Clustal X (1.83), MegAlign, GeneDoc 3.2 and Mega (4.0).</p><p><b>RESULTS</b>Totally 6987 mosquitoes were collected and main species was Culex tritaeniorhynchus and Anopheles sinensis. The infection rate of JEV in mosquitoes in Sanming, Jianyang and Fuzhou were 1.25%, 1.76% and 0.65%, respectively. One full genome in the positive specimens was sequenced. And further study showed that the positive JEV sequences belonged to genotype I.</p><p><b>CONCLUSION</b>Genotype I Japanese encephalitis virus is the main genotype in mosquitos in Fujian province.</p>


Subject(s)
Animals , Culicidae , Virology , Encephalitis Virus, Japanese , Classification , Genetics , Genotype , Phylogeny , Time Factors
16.
Article in Chinese | WPRIM | ID: wpr-246174

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the molecular basis of pathogenicity of Japanese encephalitis virus (JEV) by sequencing of complete nucleotide sequence and analyze the characteristics of full-length genome of genotype I Japanese encephalitis virus strains (GZ56) which was isolated from the first cerebrospinal fluid (CSF) of Japanese encephalitis patients.</p><p><b>METHODS</b>The complete nucleotide sequence was obtained by RT-PCR and sequencing was performed directly. Bioinformatics was used to analyze the nucleic acid data, deduced amino acid sequence and phylogenetic trees.</p><p><b>RESULTS</b>The result of sequence analysis showed that the genome of GZ56 strains had 10 965 nucleotides, which coded for a 3432-amino acid polyprotein. Phyolngenetic analysis based on full-length genome showed that GZ56 strains and M-28 strains which were the first isolated from mosquitoes in Yunnan in 1977 were in the same evolutionary branch. GZ56 strains belongs to genotype I of Japanese encephalitis virus, the homology of genome ranged from 96.2% to 98.6% in nucleotide and from 98.2% to 99.7% in amino acid sequences respectively when compared with selected genotype I of JEV strains in GenBank. There were 11 amino acid divergences in E protein when compared with the JEV inactivated P3 strain but they are not the key virulence sites. However, there were 14 amino acid divergences in E protein when compared with the JEV live attenuated vaccine SA14-14-2 strain and 8 amino acid divergences were the key virulence sites.</p><p><b>CONCLUSION</b>This study indicated that the full length of genome GZ56 strains had no ignificant change. It can be hypothesized from genomic level that the currently available JEV vaccines(inactivated and live attenuated) can protect against GZ56 strains infection, meanwhile, the JEV live attenuated vaccine (SA14-14-2) formulation conferred higher levels of protection.</p>


Subject(s)
Computational Biology , Encephalitis Virus, Japanese , Classification , Genetics , Encephalitis, Japanese , Virology , Enzyme-Linked Immunosorbent Assay , Genome, Viral , Genotype , Japanese Encephalitis Vaccines , Allergy and Immunology , Phylogeny , Sequence Analysis, DNA
17.
Article in Chinese | WPRIM | ID: wpr-246229

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the prevalence characteristics and influence factors of Japanese encephalitis (JE) neutralizing antibody in healthy people.</p><p><b>METHODS</b>Xinyang and Luoyang is the two cities in Henan Province. In 2010, healthy people of these two cities were selected by random sampling method to eight age groups: less than one year old, 1 -2 years old, 3 -4, 5 -6, 7 -14, 15 -19, 20 -59,and above 60 years old, their blood specimens were collected in May before JE infection and in November after JE infection, then followed with epidemiological investigation for JE neutralizing antibody by MCPENT.</p><p><b>RESULTS</b>519 healthy people were surveyed, 1008 effective blood specimens were collected and tested. The JE neutralizing antibody positive rate was 59.52% in men, and 67.39% in male, these two rates had no statistical significance (chi2 = 3.41, P > 0.05). The JE neutralizing antibody was 58.66% in May, and 61.20% in November, these two rates had no statistical significance (chi2 = 0.68, P > 0.05). The JE neutralizing antibody positive rate of 0 - 14 years old age group was 55.19% in Xinyang, and 45.03% in Luoyang,these two rates had no statistical significance (chi2 = 3.53, P > 0. 05). The JE neutralizing antibody positive rate of above 15 years old age group is 97.78% in Xinyang,and 48.94% in Luoyang, these two rates had statistical significance (chi2 = 55.42, P < 0.05). The JE neutralizing antibody positive rate of JE vaccination was 56.85%, and 38.35% in no JE vaccination, these two rates had statistical significance (chi2 = 10.88, P < 0.05).</p><p><b>CONCLUSION</b>The JE neutralizing antibody positive rate was showing significant differences in people above 15 years old between Xinyang and Luoyang. The JE neutralizing antibody positive rate was showing significant differences between JE vaccination and no vaccination. Xinyang and Luoyang City, recommended strengthening the 0 - 14 year-olds immunized, and at the same time, exploring and paying attention to JE immunization strategy of people above 15 years old in Luoyang.</p>


Subject(s)
Adolescent , Adult , Aged , Antibodies, Neutralizing , Blood , Antibodies, Viral , Blood , Child , Child, Preschool , China , Encephalitis Virus, Japanese , Allergy and Immunology , Female , Humans , Infant , Male , Middle Aged
18.
Article in Chinese | WPRIM | ID: wpr-316904

ABSTRACT

<p><b>OBJECTIVE</b>To study the viremia formation in guinea-pigs infected with wild type and attenuated Japanese encephalitis virus (JEV).</p><p><b>METHODS</b>Guniea pigs were inoculated intraperitoneally with different wild JEV strains and the attenuated vaccine strain and its parent virulent strain. Viremia was detected on different days following virus inoculation.</p><p><b>RESULTS</b>All the guinea-pigs inoculated with the wild JEV strains induced different levels of viremia (1.00-3.40 Lg pfu) on the 1st and 3rd day post inoculation. Using a virus titer of 10(4) pfu for inoculation, the animals inoculated with the SA14 parent strain induced relatively high viremia (10(2.4)-10(3.4) pfu), however no viremia coulds be detected on any tested days.</p><p><b>CONCLUSION</b>The degree of viremia in guinea pigs can be used as a new method to evaluate the attenuation of JEV.</p>


Subject(s)
Animals , Disease Models, Animal , Encephalitis Virus, Japanese , Virulence , Physiology , Encephalitis, Japanese , Virology , Guinea Pigs , Humans , Japanese Encephalitis Vaccines , Vaccines, Attenuated , Viremia , Virology , Virulence , Virus Replication
19.
Article in Chinese | WPRIM | ID: wpr-291518

ABSTRACT

<p><b>OBJECTIVE</b>To sequence and analyze the complete genome of two new Japanese encephalitis virus (JEV) strains isolated from mosquitoes collected in Hubei province in 2008, and to understand the molecular biological characteristics of JEV in this area.</p><p><b>METHODS</b>RT-PCR was used to amplify the fragments of HBZG08-09 strain and HBZG08-55 strain with 16 pairs overlapping primers after they had been recovered and identified, then the full-length genome was obtained by sequencing and splicing. Biological sequence alignment, nucleotide and amino acid sequence analysis, phylogenetic analysis and analysis of amino acid differences were performed by the software of Clustal X (1.83), MegAlign, Mega (4.0) and Genedoc (3.2).</p><p><b>RESULTS</b>The genome of two new strains were both 10 965 nucleotides in length with a single open reading frame from 96 to 10 392 coding for a 3432 amino acid poly-protein, the homology of nucleotide and amino acid sequence between two isolates were 98.2% and 99.7% respectively. Further study showed that the new strains were both belonging to genotype I. Two new strains were most closely related to isolates obtained from Henan and Zhejiang province in recent years. Compared with the live attenuated vaccine strain SA-14-14-2 in China, HBZG08-09 strain had 82 amino acid divergence; HBZG08-55 had 84 amino acid divergences. But the amino acid difference occurred in sites were not the key ones affecting the toxicity or antigenic of JEV.</p><p><b>CONCLUSION</b>Two new JEV isolates were both belonging to genotype I, and the key sites of amino acid were not changed.</p>


Subject(s)
Animals , China , Culicidae , Virology , Encephalitis Virus, Japanese , Classification , Genetics , Genome, Viral , Genotype , Molecular Sequence Data , Phylogeny , RNA, Viral , Sequence Alignment
20.
Article in Chinese | WPRIM | ID: wpr-329537

ABSTRACT

Objective To isolate and identify arboviruses from mosquito pools in some regions of Liaoning province.Methods Mosquitoes were collected from Shenyang,Yingkou,Panjin,Jinzhou and Dandong cities of Liaoning province in 2006.Viruses were isolated by inoculating the specimens onto C6/ 36 and BHK-21cells.The new isolates were identified using serological and molecular biological methods.Results 5410 mosquitoes were collected from the five cities in total.Three isolates produced CPE in C6/ 36 cell and five isolates produced CPE in both C6/36 and BI-IK-21 cell.Three isolates (LN0684,LN0688 and LN0689) were identified as Banna virus and one isolate (LN0636) was identified as Getah virus.Phylogenetic analysis showed that the three Banna virus strains were clustered into the same evolution branch as the other Chinese isolates.The identity of nucleotide sequence was between 91.2% and 94.7%,compared with other Banna virus strains.The new isolated Getah virus was clustered into the same branch with the strain of South Korea (swine).The identity of nucleotide sequence was 99.2%,when comparing with the strain of South Korea and was 95% to 99% with the strains fi'om Russia,mainland of China and Taiwan region.Conehmion Eight virus isolates,including three Banna virus,one Getah virus and four unknown virus strains were isolated from mosquitoes in Liaoning province.Banna virus and Getah virus were reported for the first time in Liaoning province,while Getah virus showed the highest nucleotide homology with the South Korea strains.

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