ObjectiveTo identify the first cases of cholera in Huainan,0139 epidemic causes and biological properties. MethodsConventional methods of serological and bacteriological methods of the strains were identified by polymerase chain reaction (PCR) to detect cholera virulence genes, susceptibility testing modified KB.Results1 ,was isolated and serum agglutination results,2 patients were all positive stool samples,6 were in close contact with all the negative rectal swab;turtle pond-like 18,5 were positive;turtle egg in battle were like 9,2 were positive ; turtle eggs 7, the positive three copies; turtle waste 11, were negative. 2, or more positive culture drug sensitivity test of the pioneer B, doxycycline resistant; to ampicillin, tetracycline in sensitive;on norfloxacin, gentamicin, ciprofloxacin sensitive. 3,or more positive cultures gene DNA PCR test results,cholera toxin(CT) ,toxin coregulated pilus (TCP) all were positive. ConclusionFrom the outbreak of cholera 0139 is water pollution caused by turtle ponds,The trip types of 0139 Vibrio cholera were highly homologous with that isolated from green turtle;the bacteria of the pioneer B, doxycycline resistance and carrying cta, tcpa virulence genes.

To study on the phylogenetic characterization of the VP1 genes of coxsackievirus A16 (CVA16) causing hand-food-mouth disease (HFMD) isolated from Anhui province in 2014. A total of 413 throat swab specimens from HFMD patients were collected during January to November, 2014 for the isolation and identification of enteroviruses using real-time RT-PCR assays. The VP1 regions of CVA16 isolates were amplified using RT-PCR and sequenced. And the phylogenetic tree was constructed among the VP1 regions of those isolates, the different genotypes and sub-genotypes of CVA16 strains. A total of 97 enteroviruses were isolated from 413 samples, the positive rate was 23.49% (97/413), including seventeen CVA16, seventy six HEV71 and four other enteroviruses. The results of the phylogenetic tree showed that 17.CVA16 strains isolated from Anhui in 2014 clustered within B1b evolution branch of B1 genotype. The nucleotide and amino acid sequence identities were 95.30%-100% and 98.70%-100% among the isolates, respectively, but within B1b branch of 17 strains formed several small transmission chains. The nucleotide acid of 17 CVA16 isolates in Anhui province were closed to the strains isolated from Yunnan, Hunan, Guangdong, Tibet and Jiangsu, especially from Hunan in 2013 and from Shenzhen of Guangdong in 2014, the identity were 96.40%-99.70%. The CVA16 strains isolated from Anhui in 2014 were all belong to genetic subtype B1b of B1 genotype was dominant, and among those isolates, several small virus transmission chains had formed with co-circulating and evolution.
China ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; virology ; Genotype ; Humans ; Molecular Sequence Data ; Phylogeny ; Viral Proteins ; genetics ; metabolism

Objective To investigate the effect of hypoxia-inducible factor-1? gene on the proliferation and differentiation of neural stem cells after focal cerebral ischemia in rats,and to explore the mechanism of the effect. Methods Middle cerebral artery occlusion(MCAO) and reperfusion models were established and divided into sham group,NS group,AD group and Ad-HIF-l? group.NS,AD and Ad-HIF-l? were injected into the ischemic ventricle respectively.The mNSS was evaluated the expression of EPO was observed,and the number of BrdU positive cells in subventricular zone and that of BrdU/NF200,BrdU/GFAP double labeled cells in cortex were calculated by immumofluorescence method.Results The mNSS were statistically different between Ad-HIF-l? group and the other three groups;the expression of EPO was higher in Ad-HIF-l? group;the number of BrdU positive cells increased obviously in Ad-HIF-l? group;the cellular rebirth and differentiation demonstrated that there existed a significant difference(P

Objective To display space occupying lesions of spine,mediastina and lungs behind heart and small lesions in bilateral pulmonary fields in the same chest film without changing the exposure doses which were now used for routine chest images.Methods The distribution of current intensifying screens' intensifying materials were changed and reasonable technical processes were given.High-velocity and middle-velocity intensifying materials were placed in center and in two sides of the screen respectively.Results Tests in seven volunteers using the new kind intensifying screens demonstrated that diseases could be better shown than normal intensifying screens in chest images.Conclusion The renovation can provide more and clearer and richer stratifications of image information to help chest X-ray diagnoses.

Objective To investigate the effeets of insulin on the apoptosis of vascular endothelial cells cuinduced by burn$eruln in order to explore its possible mechamsm.Method Cultured human ECV304 cells were randomly divided into 33x)ups:control group,the ECV304 cells hured by 15%(V/V)rat normal,qertlm(t=6);bum semm group,the ECV3()4 cells simulated by 15%(V/V)self-made burn semm collected from rats with 30%TBSA full-thickness burns on the,back(n=6);and burn Serum+insulin group.the ECV304 cells cultared by insulin(10-7mol/L)and 15%(V/V) seIf-made rat bum serum(n=6).The transferase mediated nick end labeling(TUNEL) method was employed to measure the apoptosis of endothelial cells at 6 hours after stimulation.Meanwhile.immanohistochemical technique and Western blotting were used to determine the protein expressions of bcl-2 and eNOS.Data are expressed ills mean ±SEM.Statistical comparison was made using oneway analysis of vtriance.Significance was accepted at P<0.05.Results Compswith the control group,bum$erunl induced the apoptusis(18.5±3.1%)and down-regalated bcl·2(O.36±0.12)and p-eNOS(O.55±0.28)protein expressions of HUVECs(P<0.01).Burn 9AJ'unl+insulin significantly decreased the apoptosis(9.6 4-2.8%)and up-regulated bcl-2(0.944-0.25)and p-eNOS(0.89±0.16)protein expressions ofHU-VECs in comparison with the bum serllm group(P<0.01).eNOS showed no significant differences in three groups.Conclusions Insulin could markedly inhibit the apoptosis and up-regalate bcl-2 protein expression of HUVECs induced by bum serum,and its mec,harfism might involve the protein expression ofphosphorylated eNOS.

BACKGROUND: Mesenchymal stem cells (MSCs), with low immunogenicity, can regulate cellular immunity and mitigate graft rejection, which has a good prospect in tissue engineering. However, it is rarely present in bone marrow. OBJECTIVE: To explore an isolation and culture method of the rabbit bone marrow-derived MSCs, to observe the biological characteristics and differentiation potential of bone marrow-derived MSCs.METHODS: MSCs were isolated from rabbit tibia bone marrow by combination of gradient centrifugation and different adherent method, then proliferation in vitro. Morphology was examined by phase contrast microscopy, and the growth curve of cultured MSCs was drawn via MTT results. MSCs were treated with osteogenetic inductor (L-DMEM/F12, 10% fetal bovine serum, 0.1 μmol/L dexamethasone, 200 μmol/L vitamin C, 10 mmol/L β-phosphoglycerol), adipose inductor (L-DMEM/F12, 10% FBS, 1 μmol/L dexamethasone, 200 μmol/L antifani, 0.5 mmol/L IBMX, 10 μg/mL insulin), and chondrocytes inductor (L-DMEM/F12, 10% FBS, 10 μg/L TGF-β1, 0.1 μmol/L dexamethasone, 50 μmol/L vitamin C, 6.25 mg/L insulin) to differentiated into osteoblast, dipocytes and chondrocytes. And the differentiated cells were identified by alkaline phosphatase staining, oil red O staining, and toluidine blue staining, respectively.RESULTS AND CONCLUSION: Bone marrow-derived MSCs can be isolated and cultured by the combination of gradient centrifugation and different adherent method in vitro, which have the better potentiality of proliferation and multi-directional differentiation. Mostly of the primary and passaged cells were spindle-shaped. After osteogenetic induction, cells were positive to alkaline phosphatase staining. Oil red O staining showed that red lipid droplet existed in adipose cells, and toluidine blue staining showed that toluidine blue was positive after chondrocytes induction.

China ; Humans ; Streptococcus pneumoniae

Objective: To analyze the genotype diversity and phylogenetic characterization of norovirus(NoV) in patients with diarrhea from Anhui province. Methods: NoV positive fecal specimens from sentinel hospitals were collected from January, 2016 to December, 2017. The samples were detected by Real-time fluorescent quantitative PCR. Positive samples were of randomly selected and amplified by RT-PCR and the products were sequenced. Phylogenetic tree was constructed by the Neighbor-Joining method based on partial VP1 gene regions of NoV to perform phylogenetic analysis. Results: A total of 263 NoV positive samples were genotyped, of which 239 belonged to genogroup II, 24 belonged to genogroup I. Fifty-five positive samples were successfully sequenced. There were 6 NoV GII genotypes, which included GII.2, 3, 4/Sydney_2012, 13, 17 and 21, while NV GII.17 and GII.4 were the dominant genotypes from 2016 to 2017. The predominant genotype was GII.4/Sydney 2012 (47.27%, 26/55), followed by GII.17 (23.64%, 13/55) and GII.2 (14.55%, 8/55). Phylogenetic tree showed that 26 strains belonged to genotype GII.4/Sydney 2012, NoV. The nucleotide homology among the 26 VP1 genes was 97.8% to 100%. Analysis of the partial VP1 genes of 26 strains showed that it shared the highest homology of 98.9% with the strain of GII.4Sydney2012 (GenBank ID: KU720515). However, the prevailing genotype in the Anhui province has shifted on two separate occasions, the GII.17 strain was dominant in 2016, and the GII.4/Sydney 2012 strain was dominant in 2017. Conclusions NoV GII was the major pathogen causing sporadic diarrhea in Anhui province during from 2016 to 2017, the genotypes are widely distributed, and shifted into the two predominant strains.

Objective To understand genomic characteristics of 2 strains of influenza A (H9N2) virus isolated from human infection cases in Anhui province in 2015.Methods Two human infection with H9N2 virus were confirmed by national influenza surveillance laboratory network in Anhui through viral isolation in April and September,2015,respectively.The full genomic sequences of the two viral isolates were analyzed in this study by using molecular bioinformatics software Mega 6.0.Results Human infection with H9N2 virus was first reported in Anhui province.The analysis of genomic sequence showed that the HA and NA genes of the two H9N2 isolates belonged to A/Chicken/ Shanghai/F/98(H9N2)-like lineage,and shared high identity with H9N2 virus circulating in poultry in 2013.The PB2 and MP genes belonged to the A/quail/Hong Kong/G 1/97-like lineage,and shared high homology with H7N9,H10N8 or H6N2 viruses.The amino acid sequence alignment results showed that several mutations for human infection tropism presented in the two virus strains,including Q226L,H183N and E190T in HA;S31N in M2;63-65 deletion in NA.In addition,the H9N2 influenza virus strains possessed the PSRSSR\GL motif in HA.Meanwhile several human-like signatures,including PA-100A,PA-356R and PA-409N were also found in the two virus strains.Conclusion The H9N2 viruses isolated from human infection cases in Anhui province belonged to a reassortant virus originated from different lineage H9N2 avian influenza virus.The virus has possessed several human susceptibility locus.

OBJECTIVE：To investigate inhibitory effects of Qing fei baoyu an capsule on airway inflammation in rats with chronic obstructive pulmonary disease （COPD），and its effects on NLRP 3 signaling pathway. METHODS ：Totally 60 SD male rats were randomly divided into blank control group ，model group ，dexamethasone group （positive control ，0.2 mg/kg），Qingfei baoyuan capsule high-dose ，medium-dose and low-dose groups （1 232.0，616.0，308.0 mg/kg），with 10 rats in each group. Except for blank control group ，other groups were fumigated for 28 days and given intratracheal dripping of lipopolysaccharide twice to induce COPD model. Since the 29th day after modeling ，blank control group and model group were given constant volume of normal saline intragastrically ，and administration groups were given related medicine intragastrically. The administration volume was 10 mL/kg，once a day ，for consecutive 28 days. After last administration ，the lung function was detected. The pathological changes of lung tissue were observed by HE staining. The content of interleukin- 1β（IL-1β）in bronchoalveolar lavage fluid （BALF）were detected by ELISA ，and the number of leukocytes was counted ；the expression of NLRP 3 and Cleaved caspase- 1 in lung tissue of rats were detected by Western blotting assay. RESULTS：Three，two，one，one and two rats died in model group， dexamethasone group ， Qingfei baoyuan capsule high-dose，medium-dose and low-dose groups ，respectively. Compared with blank control group ，FEV0.3/FVC of rats in # model group was significantly decreased （P＜0.01）. A large number of inflammatory cells infiltration we re found in the lung tissue ，and lung tissue lesion was obvious. The content of IL- 1β and white blood cell count in BALF，relative expression of NLRP3 and Cleaved caspase- 1 protein in lung tissue were increased significantly （P＜0.05 or P＜0.01）. Compared with model group，FEV0.3/FVC of administration groups were increased significantly （P＜0.05 or P＜0.01）；lung tissue lesion of them were improved to different extents. The content of IL- 1β and white cell count in BALF，relative expression of NLRP 3 protein（except for Qingfei baoyuan capsule low-dose group ）and Cleaved caspase- 1 protein（except for Qingfei baoyuan capsule medium-dose and low-dose groups ）in lung tissue were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：Qingfei baoyuan capsule can relieve lung tissue lesion and improve lung function in COPD model rats ，the effects of which may be associated with inhibiting inflammation reaction by inhibiting NLRP 3 signaling pathway.