This study aims to perform a survey of genetic variation in neuraminidase (NA) gene of influenza A/H3N2 virus, as well as related resistance to NA inhibitors, in Qinghai Province of China, 2010 to 2012. Strains of influenza A/H3N2 isolated during an influenza survey from 2010 to 2012 in Qinghai were enrolled by random sampling. Viral RNA was extracted and amplified by RT-PCR. Purified PCR products were sequenced thereafter. Genetic analysis of nucleic acid and the derived amino acid sequences was performed by MEGA 4.0. Phylogenetic trees were also constructed. Strains isolated during 2010-2011 in this study clustered closely with World Health Organization (WHO) 2010-2012 reference vaccine strain A/Perth/16/2009 and 2008-2010 reference vaccine strain A/Brisbane/10/2007 on the phylogenetic tree, while the 2012 isolates were located on another branch. In analysis of derived amino acid sequences, the 2010 isolates mutated at K81T, the 2011 isolates mutated at I26V and D127N, while the 2012 isolates mutated at E41K, P46A, I58V, T71N, L81P, D93G, D127N, D151N, and I307M. The D151N mutation added a glycosylation site to the activity center of NA. No significant variation was discovered in H3N2 NA gene of 2010-2011 isolates in Qinghai, China. Isolates of 2012 were found with significant mutation, which has the potential of inducing minor resistance to NA inhibitors like zanamivir and oseltamivir.
Amino Acid Sequence ; China ; Humans ; Influenza A Virus, H3N2 Subtype ; classification ; enzymology ; genetics ; isolation & purification ; Influenza, Human ; virology ; Molecular Sequence Data ; Neuraminidase ; chemistry ; genetics ; Phylogeny ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics

OBJECTIVETo study the genetic characterizations of VP1 gene of human enterovirus 71 (HEV71) isolated from clinical specimens of Hand, Foot and Mouth Disease (HFMD) patients in Qinghai Province in 2008.

METHODS335 clinical samples including stools, throat swabs and vesicle fluids were collected from HFMD patients in Qinghai Province. Viral isolation was performed, and molecular typing was performed with the positive isolates. Then 30 identified HEV71 isolates were performed for entire VP1 coding region amplification and sequencing.

RESULTSAmong the 355 clinical samples, 45 human enteroviruses were isolated, and among them, 30 were identified as HEV71. Then 30 HEV71 positive isolates were performed by nucleotide sequencing. It showed that there was some difference in the nucleotide and the amino acid among the 30 HEV71 strains, the homology were 95.2%-100% and 96.6%-100%, respectively. But they all closed to HEV71 strains isolated in China after 1998, and from the phylogenetic tree constructed with 30 Qinghai HEV71 strains and other 35 HEV71 strains represented all known genotype and subgenotype HEV71 strains available from GenBank, it revealed that the 30 Qinghai HEV71 strains clustered within the C4a evolution branch of C4 subgenotype.

CONCLUSIONHEV71 was isolated in HFMD patients in Qinghai province, and the HEV71 strains causing HFMD outbreaks in Qinghai province in 2008 were all belong to C4a evolution branch of C4 subgenotype with several transmission chains.

China ; Enterovirus A, Human ; genetics ; isolation & purification ; Hand, Foot and Mouth Disease ; virology ; Humans ; Molecular Typing ; methods ; Sequence Analysis, DNA ; methods

To study the influence of IFN-alpha on function of CML-DC cultured in vitro and expression of chemokine and its chemokine receptor, bone marrow mononuclear cells from 13 CML patients were cultured in the fetal calf serum culture system supplemented with rhSCF, rhFlt-3L for expansion system, and adding rhGM-CSF, rhTNF-alpha, rhIL-4, with or without rhIFN-alpha to induce DCs. After incubation for two weeks, the phenotypes of CML-DC were analyzed by direct immunofluorescence and flow cytometry. The concentration of MIP-3beta expressed by CML-DC in the supernatant were analyzed by ELISA. The proliferative ability of T cells from healthy volunteers stimulated by CML-DCs were measured by MTT assay. The results showed that expression of CD86, CD83, CD40, MHC-I class molecules, CCR7, the concentration of MIP-3beta expressed by CML-DC, and the proliferative ability of T cells stimulated by CML-DCs in IFN-alpha group were all significantly higher than that in control group (P < 0.01). It is concluded that the immunophenotype of CML-DCs can be partially changed by IFN-alpha to accelerate the maturation of CML-DCs, enhance the capacity of CML-DCs, and stimulate allogeneic T lymphocyte proliferation.
Adult ; Aged ; Antigens, CD ; analysis ; B7-2 Antigen ; analysis ; Bone Marrow Cells ; drug effects ; metabolism ; pathology ; CD40 Antigens ; analysis ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chemokines ; biosynthesis ; Dendritic Cells ; drug effects ; metabolism ; pathology ; Female ; Flow Cytometry ; Fluorescent Antibody Technique, Direct ; Humans ; Immunoglobulins ; analysis ; Interferon-alpha ; pharmacology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; blood ; Leukocytes, Mononuclear ; drug effects ; metabolism ; pathology ; Male ; Membrane Glycoproteins ; analysis ; Middle Aged ; Receptors, Chemokine ; biosynthesis

OBJECTIVETo investigate the analgesic and sedative effects of inhaling a mixture of nitrous oxide and oxygen on burn patient during and after dressing change.

METHODSA total of 240 burn patients hospitalized in the Institute of Burn Research of Changhai Hospital Affiliated to the Second Military Medical University, Department of Burns of the First People's Hospital in Zhengzhou, and Department of Burns and Plastic Surgery of General Hospital of Ningxia Medical University from October 2011 to September 2012 were enrolled in our study, and they were all in accordance with the inclusion criteria. The 240 patients were divided into control group (n = 60, treated with inhalation of oxygen during dressing change) and treatment group (n = 180, treated with inhalation of a mixture of 65% nitrous oxide and oxygen during dressing change) according to the computer-generated list of random number. The other treatments in control group and treatment group were the same. Before, during, and after dressing change, heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP), oxygen saturation (SO2), and adverse effects were observed. The degree of pain and anxiety felt by the patients were respectively evaluated with the visual analogue scale (VAS) and Chinese version of the burn specific pain anxiety scale (C-BSPAS) at the same time points as above. Data were processed with analysis of covariance, chi-square test, analysis of variance, and rank sum test.

RESULTSThere were no significant differences between control group and treatment group in the levels of HR, SBP, DBP, and SO2 before dressing change (with F values respectively 0.76, 0.06, 1.11, 0.70, P values all above 0.05). Compared with those of control group, the levels of HR, SBP, DBP, and SO2 in treatment group were significantly ameliorated during dressing change (with F values respectively 81.78, 146.36, 226.44, 205.62, P values all below 0.01). After dressing change, the levels of DBP in the two groups were close (F = 0.31, P > 0.05), but the levels of HR, SBP, and SO2 showed statistical differences (with F values respectively 7.02, 8.69, 12.23, P < 0.05 or P < 0.01). Before dressing change, the VAS scores were approximate between control group and treatment group (Z = 0.21, P > 0.05). Compared with those in control group (9.4 ± 0.7, 1.7 ± 2.5), the VAS scores were significantly lowered in treatment group during and after dressing change (1.6 ± 1.3, 0.7 ± 1.1, with Z values respectively 11.84, 3.35, P values all below 0.01). There was no significant difference in C-BSPAS score between control group and treatment group before dressing change (Z = 0.62, P > 0.05). Compared with those in control group (75 ± 13, 73 ± 12), the C-BSPAS scores in treatment group were decreased during and after dressing change (9 ± 15, 9 ± 14, with Z values respectively 11.91, 12.28, P values all below 0.01). There were no obvious adverse effects in two groups before, during, and after dressing change.

CONCLUSIONSA mixture of nitrous oxide and oxygen seems to have obvious analgesic and sedative effects on burn patients during dressing change, and it can be widely used.

Administration, Inhalation ; Adolescent ; Adult ; Aged ; Analgesia ; methods ; Bandages ; Burns ; surgery ; Female ; Humans ; Hypnotics and Sedatives ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Nitrous Oxide ; administration & dosage ; therapeutic use ; Oxygen ; administration & dosage ; therapeutic use ; Young Adult

OBJECTIVETo investigate the genetic polymorphism of 15 autosomal short tandem repeat (STR) loci, i.e. D5S818, FGA, D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX and D18S51, in Tibetan population of Changdu area, and to analyze the genetic relationship between this population and other Asian populations.

METHODSThe 15 loci were amplified simultaneously using a multiplex PCR typing system. Amplified STR fragments were analyzed with an ABI PRISM 3100 Avant DNA Analyzer. Arlequin software version 3.1 was used to obtain allele frequencies and polymorphism parameters.

RESULTSThere were 135 STR alleles in the 15 STRs of Changdu Tibetan, with allele frequencies ranging from 0.0065 to 0.5455. The average heterozygosity was 0.7340, the discrimination power was higher than 0.8 except for TPOX and TH01. The accumulative discrimination power was more than 0.9999998, and the accumulative non-parental exclusion probability was 0.99999997.

CONCLUSIONThe 15 STR loci of Changdu Tibetan possess characteristics of high genetic diversity. They can be employed in minority genetics investigation, and individual and paternity test in forensic medicine.

Asian Continental Ancestry Group ; ethnology ; genetics ; Female ; Humans ; Male ; Microsatellite Repeats ; Pedigree ; Polymorphism, Genetic ; Tibet ; ethnology

OBJECTIVETo investigate the influences of Shensu Yin to RAW 264.7 on the expression of TLR3, TLR4 and the factors of the downstream in RAW 264. 7 cells.

METHODRAW 264.7 cell line was stimulated with Lipopolysaccharide and POLY I: C, respectively, and treated with the drug serum of Shensuyin simultaneously. 24 hours later, collected the supernatant and measured the inflammatory factors TNF-alpha and IFN-beta, extracted mRNA and measured the expression of TLR3, TLR4 and other correlated indexes of the downstream, analyzed and evaluated Shensu Yin's substance basis of pharmacodynamic actions.

RESULTShensu Yin drug serum depressed the expression of TLR4, MyD88, TRAF-6, TRAM and TRIF mRNA, as a result, it decreased the amount of TNF-alpha and IFN-beta.

CONCLUSIONDepressing the expression of TLR3, MyD88, TRAM and TRIF mRNA may be the elementary basis of Shensu Yin to play heat-clearing and detoxicating effect.

Adaptor Proteins, Vesicular Transport ; genetics ; Animals ; Cell Line ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Interferon-beta ; secretion ; Lipopolysaccharides ; pharmacology ; Macrophages ; cytology ; drug effects ; metabolism ; Male ; Mice ; Myeloid Differentiation Factor 88 ; genetics ; Plants, Medicinal ; chemistry ; Poly I-C ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Interleukin ; genetics ; Signal Transduction ; drug effects ; Toll-Like Receptor 3 ; genetics ; Toll-Like Receptor 4 ; genetics ; Tumor Necrosis Factor-alpha ; secretion

The study was aimed to investigate the influence of interferon alpha (IFN-alpha) on the expressions of Fas and Fas ligand (FasL) in dendritic cells (DCs) from patients with chronic myeloid leukemia (CML). In addition to adding stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor alpha (TNF-alpha) and interleukin 4 (IL-4), the IFN-alpha was added to the serum-free medium for DCs. After culturing for 10 - 14 days, cell phenotype and percentage of Ph(1) chromosome were detected by different methods. The expression of Fas or FasL on CML-DCs and cell cycle of DCs labeled with propidium iodine (PI) were measured by flow cytometry. The concentration of sFas in supernatants was analyzed by enzyme-linked immunosorbent assay (ELISA). The results indicated that the expression of co-stimulatory molecules were improved significantly while the percentages of Ph(1) positive cells decreased. The level of Fas on cells was up-regulated and the concentration of sFas decreased. However, the expression of FasL was negative. The ratio of apoptosis rose gradually while the concentration of IFN-alpha increased. It is concluded that IFN-alpha can accelerate the apoptosis of Ph(1) positive cells through Fas/FasL pathway, so the number of Ph(1) negative cells increases relatively.
Adolescent ; Adult ; Apoptosis ; drug effects ; Cells, Cultured ; Child ; Culture Media ; pharmacology ; Dendritic Cells ; cytology ; metabolism ; Fas Ligand Protein ; genetics ; metabolism ; Female ; Humans ; Interferon-alpha ; pharmacology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; Male ; Middle Aged ; Philadelphia Chromosome ; Young Adult ; fas Receptor ; genetics ; metabolism

This study was aimed to investigate the influences of interferonalpha (IFN-alpha) on expressions of CCR7, interleukin10 (IL-10) and IL-12p70 in dendritic cells (DCs) from patients with chronic myeloid leukemia (CML). In addition to stem cell factor (SCF), granulocyte-macrophage colony stimulating factor (GM-CSF), tumor necrosis factor-alpha (TNF-alpha) and IL-4, IFN-alpha was added to the serum-free medium of DCs. After culture for 10-14 days, phenotypes and function of CML-DCs were evaluated respectively by flow cytometry and methyl thiazolyl tetrazolium (MTT) assay. Chromosome of DCs was analyzed by displaying G banding assay. The concentrations of IL-10 and IL-12P70 in supernatants were evaluated by enzyme-linked immunosorbent assay (ELISA). The results showed that the expressions of CD40, CD83, CD86 and CCR7 and the OD value in allogeneic mixed-lymphocyte reaction (MLR) in group with IFN-alpha (300 U/ml) were twice as high as those in group without IFN-alpha. The percentage of Ph1 positive cells and concentrations of IL-10 and IL-12 P70 were reduced in group with IFN-alpha. It is concluded that the defective phenotypes and functions of CML-DCs can be recruited partly by IFN-alpha. The mechanism may lie in the facts that expression of CCR7 and co-stimulatory molecules is promoted and the inhibitory effect of IL-10 on CML-DCs is relieved partly through the regulation of IFN-alpha.
Cells, Cultured ; Dendritic Cells ; cytology ; Humans ; Interferon-alpha ; pharmacology ; Interleukin-10 ; genetics ; metabolism ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; immunology ; Philadelphia Chromosome ; drug effects ; Receptors, CCR7 ; genetics ; metabolism

OBJECTIVE: To investigate the frequencies of allele and genotype of HLA-DRB1 gene in Luoba ethnic group. METHODS: HLA-DRB1 genes from 92 individuals of Luoba ethnic group in Tibet Autonomous region were investigated, using PCR-SSO (polymerase chain reaction -sequence specific oligo-nucleotide) genotyping methods. We compared allele frequencies of Luoba population in HLA-DRB1 locus with that of various ethnic groups distributed worldwide, constructed the phylogenetic tree by UPGMA (unweighted pair group method with arithmetic mean), and counted genetic distance by Nei measure. RESULTS: Of the 11 DRB1 alleles detected, the two most common genes were DRBI * 04( gene frequency: 27.20% ) and DRB * 12 (25.50%) , which covered 52.70% of the total alleles detected from Luoba ethnic group. The frequencies of HLA-DRB1 * 14(15.20% ), DRB1 * 15(9.80% ) and DRB1 * 08(8.20% ) were more than 5%. Luoba was unique in the distribution of HLA alleles. The genetic distance between Luoba and Tibetan was closer than that with other population. A dendrogram based on the DRB1 genes by cluster analysis suggested that Luoba ethnic group might cluster with Tibetan firstly, then cluster with other population living in China (except Uygr) , and cluster with Caucasian and Black finally. CONCLUSION The HLA-DRBI gene frequency of Luoba individuals in Tibet Linzhi had some differences compared with that of other Chinese population. The kindred relation between Luoba and Tibetan was closer than that with other population, which was coincident with the results of ethnology, history and sociology.
China ; ethnology ; Female ; Gene Frequency ; Genotype ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Male ; Phylogeny ; Polymorphism, Genetic ; Tibet ; ethnology

Objective To understand the epidemic situation and the source of infection of the reemerge human rabies in Qinghai. Methods We collected the data on human rabies and the data on the cases of multi- victims bitten by the identical dog, and also the laboratory data of the nucleoprotein ( N) gene of rabies virus from the samples which were detected by reverse transcription-polymerase chain reaction (RT-PCR) and direct immunofluorescence assay (DFA) from 2012 to 2017, to describe the epidemiological characteristics of human rabies and the prevalence of rabies virus in host animals, and to explore the source of infection of reemerge human rabies. Results A total of 7 human cases were reported in 2012-2017 in Qinghai province, among which 1 was bitted by wolf, 2 were bitted by stray dogs, 3 were bitted by domestic dogs which injured by stray dogs or wolfs. A total of 892 canine brain tissue samples were collected, from which 46 positive samples were detected with the positive rate of 5.16% (95% CI：3.70%-6.61%). The positive samples were collected from the nomadic region, which were consistent had the location of the human rabies. The samples collected from the cases of multi-victims bitten by the identical dog/animal had the positive rate of 73.08%, and 4 out of 7 human rabies were exposed to the cases of multi-victims bitten by the identical dog/animal. Genetic sequencing of the rabies virus detected from canine brain tissue samples were belong to China IV lineage, which was closely related to the Arctic clade. Conclusions The reemerging rabies happened in nomadic region of Qinghai province could be a consequence of spillover from wildlife especially from wolfs. The better surveillance system covering the human, livestock and wildlife should be set up to mitigate the rabies virus spread from the wildlife.