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1.
Article in Chinese | WPRIM | ID: wpr-777945

ABSTRACT

Objective To investigate rickettsiae infection from host animals and vector arthropods in some areas of Yunnan Province. Methods Rat clip and cage traps were used to capture mice. Chiggers from body surface of mice and ticks from body surface of farm cattle were collected. DNAs were extracted from mice spleens, chiggers and ticks. Rickettsiae groEL segment were amplified by nested-polymerase chain reaction (nPCR), sequenced to analyze the homology with other known sequences. Results A total of 410 samples were collected for rickettsiae groEL segment detection with nPCR and 19 samples (4.63%) showed positive for rickettsiae groEL segment . Among them, 2.68%(11/410)were positive for Orientia tsutsugamsushi (Ot) groEL segment, and 1.22%(5/410)were positive for spotted fever group rickettsia (SFGR) groEL segment, and 0.49%(2/410)were positive for rickettsia mooseri (Rm) groEL segment, and 0.24%(1/410)were positive for rickettsia endosymbiont(Re) groEL segment. When analyzed the homology with other known sequences, 11Ot strains with 93.6%-100% similarities among them in this study shared the highest similarity with other Ot strains from GenBank respectively, reached up to 96.1%-100%; The groEL segments of 5 SFGR strains with 92.1%-99.5% similarities among them in this study shared highest similarity with other SFGR strains from other GenBank respectively, reached up to 98.9%-100%; In this study groEL segments of 2 Rm strains all showed 100% similarity with Wilmington strain (GenBank No:AE017197); One groEL segment of Re showed 98.9% similarity with Re strain (GenBank No:EU435143). Conclusion There were kinds of rickettsiaes infection in host animals and vector arthropods in Yunnan Province, so the monitoring and prevention of the Rickettsiosis should be strengthened.

2.
Military Medical Sciences ; (12): 917-922, 2017.
Article in Chinese | WPRIM | ID: wpr-694281

ABSTRACT

According to the topics of general interest during the Eighth Review Conference of the Biological Weapons Convention,including the implementation of the convention,dual use research,bioterrorism and bio-technology misuse,we can become informed of the current situation of international bio-safety.In line with the reality of China's current bio-safety strategy,corresponding recommendations are made arms control,regarding implementation of the convention,legal systems,and institutional mechanism.

3.
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
4.
Article in Chinese | WPRIM | ID: wpr-324267

ABSTRACT

<p><b>OBJECTIVE</b>To study the sympathetic skin response (SSR) to the effects of N-hexane on autonomic nerves function in patients with chronic N-hexane poisoning.</p><p><b>METHODS</b>The subjects in present study included 30 controls and 37 cases with chronic N-hexane poisoning. Also 37 patients were divided into 3 subgroups (mild, moderate and severe poisoning) according to diagnostic criteria of occupational diseases. All subjects were examined by SSR test and nerve conduction velocity (NCV) test. All patients were reexamined by SSR and NCV every 1 ∼ 2 months. The differences in SSR parameters (latency, amplitude) among groups were observed. In the severe poisoning subgroup, the changes of SSR and NCV parameters (conduction velocity, amplitude) in different poisoning stages were observed.</p><p><b>RESULTS</b>There were significant differences in SSR latency of upper extremity among groups and the significant differences in SSR amplitude of upper and lower extremity among groups (P < 0.05). No significant differences in SSR parameters were found between the adjacent groups (P > 0.05). There were significant differences in SSR latency of upper extremity during different periods and the significant differences in SSR amplitude of upper and lower extremity during different periods among all groups (P < 0.05). The change of SSR parameters consistent with that in NCV. The longest SSR latency of upper extremity and the smallest SSR amplitudes of upper and lower extremity appears 1 - 2 months earlier than that of the smallest action potential amplitude.</p><p><b>CONCLUSION</b>The damage of autonomic nerves induced by N-hexane increased with poisoning progresses. The damage of autonomic nerves corresponded with the damage of myelin sheath of large myelinated nerves, but which appeared 1 - 2 months earlier than the damage of axon of large myelinated nerves. SSR test may serve as a method to detect the damage of autonomic nerves function in patients with chronic N-hexane poisoning.</p>


Subject(s)
Adolescent , Adult , Autonomic Pathways , Case-Control Studies , Female , Galvanic Skin Response , Hexanes , Poisoning , Humans , Male , Neural Conduction , Occupational Diseases , Skin , Sympathetic Nervous System , Young Adult
5.
Article in Chinese | WPRIM | ID: wpr-642263

ABSTRACT

Objective To develop a sensitive,specific, simple and rapid quantitative real-time PCR (Q-PCR) assay for detection of Staphylococcus aureus with SmartCycler.Methods According to the nuc gene sequences specific to S.aureus, a pair of primers and one TaqMan probe were designed. An internal amplification control (IAC) which is a chimeric double-stranded DNA constructed from a fragment of the Listeria monocytogenes hly gene flanked by the nuc-specific target sequences was added to the reaction system. This IAC was detected using a second TaqMan probe labeled with a different fluorophore. The performance of the nuc-IAC Q-PCR was evaluated using artificially contaminated drinking water and commercial UTH whole milk samples spiked with ATCC 6538.Results The nuc-IAC assay could be used reliably for detection with a sensitivity of 5 copies of linear plasmid DNA per reaction, 10 fg of genomic DNA in 62.5% of the reactions or 50 cfu/ml S.aureus cells with 50% probability. The quantification was linear (r~2≥0.998) over a 6-log dynamic range, with a PCR efficiency over 0.967. The 5×10~2 CFU per 25 ml mimic sample of drinking water or milk could be detected by this assay consistently and quantifiably.Conclusion The nuc-IAC Q-PCR assay for S.aureus is developed. It could not only be applied for the quantitative detection of S.aureus, but also prevent the false negatives and underestimations of contamination loads due to PCR failure.

6.
Article in Chinese | WPRIM | ID: wpr-642527

ABSTRACT

Objective:To develop a rapid and quantitative detection method for abrin using colloid gold immunochromatographic assay. Methods:The rapid detection method was established with double-antibody sandwich assay. Quantification was realized by constructing a standard curve. The specificity and sensitivity of the method were tested, and its feasibility was evaluated by various abrin-added food samples. Results and Conclusion:This established method could accomplish qualitative and semi-quantitative detection in 15 minutes; the sensitivity was up to 30 ng/ml with a linear range from 30 ng/ml to 600 ng/ml. The recovery rate was 80%-110%, and the variation coefficient was less than 15%.The colloidal gold immunochromatographic assay is rapid, specific, sensitive,accurate and suitable for field detection.

7.
Chinese Journal of Virology ; (6): 173-177, 2009.
Article in Chinese | WPRIM | ID: wpr-334755

ABSTRACT

5 strains of virus isolated from Culex tritaeniorhynchus, Anopheles sinensis and Armigeres subalbatus, which caused cytopathic effect in C6/36 cells, had been obtained in the survey of arboviruses in Northwestern Yunnan Province. China. The virus particles displayed 70 nanometers diameter (n=7) with no envelope but spikes on the surfaces. RNA-PAGE of the genomes of the isolates showed 6-5-1 profile. A fragment of the 12th segment sequence was amplified by a pair of specific primers for Kadipiro virus strain JKT-7075 in RT-PCR. The full length of the 12th segment was 758 nucleotides, BLAST analysis revealed the highest identity was 90% to JKT-7075. Phylogenetic analysis demonstrated that the isolates appeared to be Kadipiro viruses (Family Reoviridae). It was the first report of kadipiro virus isolation in China.


Subject(s)
Amino Acid Sequence , Animals , Anopheles , Virology , Cell Line , China , Coltivirus , Classification , Genetics , Culex , Virology , Molecular Sequence Data , Phylogeny , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA
8.
Chinese Journal of Virology ; (6): 83-87, 2009.
Article in Chinese | WPRIM | ID: wpr-334742

ABSTRACT

By RT-PCR and TAIL-PCR, the full coding region of Batai virus isolated in China (YN92-4 strain)was sequenced for the first time. According to the results, the genome of the virus contained three segments S, M and L of 947, 4,371 and 6,860 nucleotides, respectively. The S segment coded a nucleoprotein of 234 amino acids and a nonstructural protein of 102 amino acids, the M and L segments coded a precursor protein of 1 ,435 amino acids and RNA polymerase of 2,239 amino acids, respectively. Compared with the full coding sequence of Batai viruses isolated out of China, the S and M segments of YN92-4 and ON-7/B/01 showed the highest homology in nucleotide and amino acid sequenes with similarity of 97.7% (100%) and 95.7% (98%), respectively. Since there was no full coding sequence information on the L segment in GenBank for the reference, the L segment of YN92-4 was compared with that of Bunyamwera virus and the homology of nucleotide and amino acid was 73.5%and 81.6%, respectively. Phylogenetic analysis showed YN92-4 strain was clustered into one group with the prototype of Batai virus (MM2222). The results suggested that the YN92-4 strain had no occurrance of genetic reassortment (like Ngari virus) and was close to the Batai virus (ON-7/B/01 strain) isolated from cattle serum in Japan.


Subject(s)
Amino Acid Sequence , Animals , Base Sequence , Bunyamwera virus , Genetics , Cattle , China , Clinical Laboratory Techniques , Cloning, Molecular , Genome, Viral , Genetics , Hemorrhagic Fever with Renal Syndrome , Molecular Sequence Data , Open Reading Frames , Polymerase Chain Reaction , Reassortant Viruses , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Sequence Alignment , Sequence Analysis, DNA
9.
Chinese Journal of Epidemiology ; (12): 239-242, 2009.
Article in Chinese | WPRIM | ID: wpr-329485

ABSTRACT

Objective To determine the hosts of hantavirus (HV) and its molecular epidemiological characteristics, to provide evidence for prevention and control on hemorrhagic fever with renal syndrome (HFRS). Methods Rodents were captured by a special trap within the residential area. The antigens of HV in lung tissues were detected by direct immuno-fluorescence assay (DFA). Nucleotide sequences of HV were amplified by RT-PCR with HV genotype-specific primer. The amplified genes were then sequenced. Phylogenetic tree were built on nucleotide sequence with Clusta1X 1.83 software. Results 1421 rodents were captured and classified into 8 species of 4 Genera in the epidemic area within 10 counties of Chuxiong prefecture, Yunnan province, between 2005 and 2006. Out of the 1421 rodents, 1056 (74.31%) of them were Rattus norvegicas and 280 (19.70%) belonged to Rattus flavipectus. The antigens of HV were detected by DFA in lung tissues and the total positive rate of HV was 5.15% (53/ 1029). After applying the sequencing nucleotide method to the 53 positive specimens, data showed that 21 specimens were positive and all of them belonged to Seoul type ( 15 samples were from Rattus norvegicus, 4 samples Rattasflavipectas, 2 samples Rattus nitidas). The partial S segments from 12 specimens were sequenced which appeared homologic with R22, L99 and HLD65 from GenBank in relatively high level (87.1%-99.7%). When compared to 76-118 strain of Hantaan type, their homologic degree was only 64.4%-69.1%. Results from Phylogenetic analysis showed that 12 specimens belonged to Seoul type. As for their homology, they were significantly similar to Seoul type and could be tentatively divided into two subtypes S1 and S3. Conclusion It was confirmed that the Seoul type virus, as HFRS' s pathogenetic agent mainly carried by rats, prevailed widely in Chuxiong prefecture. Owing to the local ecological environment, we also noticed the characteristics of different HV subtypes among Seoul type.

10.
Article in Chinese | WPRIM | ID: wpr-316055

ABSTRACT

<p><b>OBJECTIVE</b>To investigate arboviruses in Wenshan and Hekou county which are the Sino-Vietnam frontier regions of Wenshan, Yunnan province, China.</p><p><b>METHODS</b>In September 2007, 6091 Culicines, 1334 Anophelines, 848 Aedes vexans and 53 Armigeres obturbans were collected from 5 field sites. Mosquitoes were collected and stored in liquid nitrogen after classification. The mosquito pools were homogenized, and centrifuged, then the supematant was inoculated onto C6/36 and BHK-21 cells, and the viral isolates were identified by serological and molecular biological methods.Sequence alignment and phylogenetic analysis on the viral isolates were carried out using Clustal X 1.85, GENEDOC and MEGA4 software.</p><p><b>RESULTS</b>A total of 4 pairs of virus isolated with C6/36 cells cytopathic effect were observed, and other mosquito species have not cytopathic effect.Strain WS0704-2 was Banna virus which identified by antibody response and PCR. Strain WS0704-1, WS0708-1, WS0708-2 were Culex pipens pallens densovirus (CppDNV) which identified by PCR. The phylogenetic analysis the 12th segment showed significant difference between the new Banna virus and other strains isolated in China.</p><p><b>CONCLUSION</b>There are many mosquito vectors in frontier regions (China and Vietnam) of Wenshan in Yunnan province of China, and mosquito-borne arbovirus, such as BAV were isolated here.</p>


Subject(s)
Animals , Arboviruses , Classification , Genetics , China , Culicidae , Virology , Densovirus , Genetics , Phylogeny , Sequence Alignment , Sequence Analysis, DNA
11.
Chinese Journal of Epidemiology ; (12): 899-904, 2008.
Article in Chinese | WPRIM | ID: wpr-298356

ABSTRACT

Objective To understand the epidemiological features of two rabies cases in Baoshan city year 2006 and 2007 and to analyze its source of infection.Methods Questionnaires were used to do the epidemiologieal survey on each of the rabies cases.Brain timue samples of rabies patients were collet to detect the rabies virus by direct immunofluoreseence assay(DFA)and RT-PCR assay.Homology and phylogenetic tree were analyzed.based on the whole nucleotide and deduced amino acid sequence of P,M and N gene of rabies virus followed by molecular epidemiological analysis.Results In July 2006,one human rabies case was identified in Longyang district,and another one in Tengchong county in Baoshan city in 2007.The degrees of exposure of these two patients was all at degreeⅢ.Two brain tissue samples among the dead patients(No.CYN0601H and CYN0701H)were confirmed positive by both DFA and RTPCR assay.The homology analysis of P,M and N gene sequences among CYN0601H,CYN0701H and other rabias strains isolated from other provinces and other counties.showed that the samples in Baoshan city shared the highest homology with the strains in Thailand.Phylogenetic analysis indicated that the two samples were very dose and all belonged to genetype 1 Lyssavirus,with the closest relationship between samples in Baoshan city and strains in Thailand.Conclusion It Was confirmed on the virus molecular level that the two patients in Baoshan city were both suffered from rabies.The prevalent strains in Baoshan city WaS probably imported from foreign country,suggesting that prevention and control measures on rabies virus in the boarder areas of Yunnan should be strengthened.

12.
Article in Chinese | WPRIM | ID: wpr-254135

ABSTRACT

<p><b>OBJECTIVE</b>Explore the genotype of the epidemic Japanese encephalitis virus (JEV) in Yunnan Province from molecular level and to understand the molecular differences of the virus isolated from Yunnan at different time, locality and host.</p><p><b>METHODS</b>Three-day suckling mice were inoculated with viruses continuously and when the disease developed and the mice were dying, the brain was taken and DNA was extracted from the supernatants of the brain after grinding; then the gene fragments of Prm-C region were amplified by RT-PCR. The viral gene sequences were compared with those of other 72 strains of JEV originated from both China and abroad at different times. Finally the genotypes were analyzed with the method which was established by Woan-Ru Chen.</p><p><b>RESULTS</b>All the 3-day suckling mice which were inoculated with the virus died within 78 h. The results of the nucleic acid-sequence analysis showed that 17 strains of the experimental virus belonged to genotype 1 and 2 strains belonged to genotype 3. The difference between genotype 1 and type 3 were more than 15%. While the difference between 17 strains of genotype 1 which were separated at different time, location and hosts were only 3.8%-5.2%.</p><p><b>CONCLUSION</b>The above results suggest that the genotype of the epidemic JEV in Yunnan Province are type 1 and 3 and the latter is the main type.</p>


Subject(s)
Animals , Animals, Newborn , Brain , Pathology , Virology , China , Encephalitis Virus, Japanese , Classification , Genetics , Encephalitis, Japanese , Virology , Genotype , Mice , Phylogeny , RNA, Viral , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Viral Envelope Proteins , Genetics
13.
Article in Chinese | WPRIM | ID: wpr-282314

ABSTRACT

<p><b>OBJECTIVE</b>To identify the effects of NMDA receptor subunits NR2A and NR2B expression in rat's hippocampus after exposure to 1800 MHz radiofrequency radiation.</p><p><b>METHODS</b>Four-week old female Wistar rats were randomly divided into four groups, with 12 animals for each. The subjects in two experimental groups had been continuously exposed to 1800 MHz microwave radiation (CW) with respective power density of 0.5 mW/cm(2) and 1.0 mW/cm(2) 12 hours each day for 21 days. Meanwhile, sham-controls were carried out. The brain tissue sections were performed by immunohistochemistry to demonstrate both expressions of NR2A, NR2B immune-activity in the hippocampal CA1, CA3 and DG by using computer-assisted image analysis system.</p><p><b>RESULTS</b>In NR2A: the expression of 0.5 mW/cm(2) power density group was significantly lower than 0 mW/cm(2) power density group in CA3 [(8.5 +/- 1.5) vs (11.1 +/- 1.8), P < 0.01] and had not been significantly changed in CA1 and DG. The expression of 1.0 mW/cm(2) power density group was significantly lower than 0 mW/cm(2) power density group in CA1 and CA3 [(7.9 +/- 1.6) vs (9.7 +/- 1.5); (8.4 +/- 1.7) vs (11.1 +/- 1.8), respective P < 0.05, P < 0.01] and had not been significantly changed in DG. In NR2B: the expression of 0.5 mW/cm(2) power density group was significantly lower than 0 mW/cm(2) power density group in CA1 and CA3 [(16.4 +/- 1.0) vs (17.8 +/- 1.6); (9.6 +/- 1.9) vs (11.2 +/- 2.1), respective P < 0.05]. The expression of 1.0 mW/cm(2) power density group was significantly lower than 0 mW/cm(2) power density group in CA1, CA3 and DG [(13.1 +/- 2.4) vs (17.8 +/- 1.6); (9.3 +/- 1.4) vs (11.2 +/- 2.1); (7.3 +/- 0.1) vs (8.5 +/- 1.0), respective P < 0.01, P < 0.05, P < 0.05].</p><p><b>CONCLUSION</b>There were findings of the effects on NMDA receptor subunits in different hippocampus sections after exposure to 1800 MHz radiofrequency radiation.</p>


Subject(s)
Animals , Female , Hippocampus , Metabolism , Radiation Effects , Immunohistochemistry , Radio Waves , Rats , Rats, Wistar , Receptors, N-Methyl-D-Aspartate
14.
Virologica Sinica ; (4): 74-77, 2001.
Article in Chinese | WPRIM | ID: wpr-635226

ABSTRACT

In July 1997, a strain (GB30) of virus was isolated from 60 samples of brain tissues of Murina aurata (Chiroptera: Vespertilionidae) co llec ted in Gengma county, Yunnan province, China. Isolation of virus was negative fr om 4 samples of brain tissues of Rousettus leschenaulti (Chiroptera: Pteropo did ae) collected in Gengma. GB30 virus strain could regularly cause illness and dea th in suckling mice, produced evident CPE in BHK21 cells. It agglutinated red b lood cells of dove at pH5.75~7.4. This virus has been identified serologically by hemagglutination inhibition and immunofluorescent tests using Japanese enceph alitis (JE), dengue (DEN) type 1,2,3,4, and chikungunya (CHIK) viruses monoclona l antibodies, and JE and sindbis (SIN) viruses immune sera. It showed specific r eaction to JE virus only and no reaction with DEN 1~4, CHIK and SIN viruses. Th erefore it can be identified as JE virus. This is the first report on the isolat ion of JE virus from Murina aurata. The results showed that bats are conside red as the reservoir and amplifier host of JE virus transmission in nature.

15.
Virologica Sinica ; (4): 97-100, 2001.
Article in Chinese | WPRIM | ID: wpr-635207

ABSTRACT

The biological characters of Sindbis virus strain of Yunnan(YN87448 strain) were studied by the test of the filtration, acid-resistant, ether-resistant, CPE, susceptibility of animal, HA, plague, determination of virus titres, and the cross-HI, cross-IFAT and PRNT as well.The results indicated that YN87448 strain belongs to Sindbis virus, Alphavirus, Togaviridae. YN87448 strain virus was plaque purified(PYN87448). The biological character of PYN87448 strain virus was studied too. PYN87448 strain virus will be used in the molecule biological test.

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