1.Experimental analysis the first cases of cholera epidemic detection of 0139 of Huainan
Shoujie YU ; Xiaomei LIU ; Min CAO ; Shoukui HU ; Wanfu HU
Chinese Journal of Primary Medicine and Pharmacy 2011;18(16):2166-2167
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.
2.Phylogenetic Analysis of the VP1 Region of Coxsackievirus A16 Strains Isolated in Anhui Province, 2014.
Yonglin SHI ; Xian WANG ; Guoping CHEN ; Jin ZHANG ; Wanfu HU
Chinese Journal of Virology 2015;31(6):660-664
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
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Enterovirus A, Human
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classification
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genetics
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isolation & purification
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Enterovirus Infections
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virology
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Genotype
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Humans
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Molecular Sequence Data
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Phylogeny
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Viral Proteins
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genetics
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metabolism
3.EFFCT OF HYPOXIA-INDUCIBLE FACTOR-1? GENE ON THE PROLIFERATION AND DIFFERENTIATION OF NSCs AFTER FOCAL CEREBRAL ISCHEMIA IN RATS
Wanfu WU ; Xiu CHEN ; Changlin HU ; Wenqin CAI
Acta Anatomica Sinica 1955;0(03):-
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
4.Develop of a New Wide-range X-ray Intensifying Screen
Dingzheng LIU ; Ming GU ; Qinghua WANG ; Zhaofen HU ; Lin HUANG ; Wanfu LUO
Journal of Practical Radiology 1991;0(03):-
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.
5.Isolation, culture and multiple differentiations of rabbit bone marrow-derived mesenchymal stem cells in vitro
Chengfeng XU ; Dahai HU ; Zhouting ZHAO ; Wanfu ZHANG ; Xiaozhi BAI ; Weixia CAI
Chinese Journal of Tissue Engineering Research 2010;14(6):1002-1005
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.
6.Effect of insulin on the apoptosis of vascular endothelial cells induced by burn serum
Wanfu ZHANG ; Dahai HU ; Genfa LV ; Yunchuan WANG ; Xiongxiang ZHU ; Feng GAO
Chinese Journal of Emergency Medicine 2009;18(1):56-59
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.
7.Molecular etiological characteristics of the gastroenteritis outbreak associated to norovirus infection in Anhui province
Yonglin SHI ; Xiangyu KONG ; Miao JIN ; Jincai FANG ; Xudong SHI ; Wanfu HU
Chinese Journal of Experimental and Clinical Virology 2015;29(4):310-312
Objective To study the genotype and phylogenetic characterization of the pathogens associated to the epidemic outbreak of acute gastroenteritis.Methods A total of 19 anal swab and feces samples from acute gastroenteritis outbreak in a junior middle school were collected in November 2014,Anhui province.Norovirus (NoV) nucleic acid was detected by Real-time PCR method,and the partial capsid gene of the all positive specimens were amplified by conventional RT-PCR and sequenced.Phylogenetic tree was constructed by the Neighbor-Joining method based on partial capsid gene sequences of norovirus to perform phylogenetic analysis.Results Of the 19 specimens,12 (12/19) were positive for NoV,and the positive samples of those were sequenced.Phylogenetic analysis showed that 11 strains were belong to genotype GⅡ.2,and one strain was belong to genotype GⅡ.6 norovirus.The nucleotide identity of the partial capsid gene sequences were 95.60% between 11 genotype GⅡ.2 strains identified and Melksham strain of norovirus.Conclusion The epidemic outbreak of acute gastroenteritis from Anhui province was caused by genotype GⅡ.2 and GⅡ.6 norovirus co-infection,and genotype GⅡ.2 norovirus was predominant strains in this outbreak.
8.Study of molecular of 80 clinical streptococcus pneumoniae strains in Maanshan area.
Daoli CHEN ; Machao LI ; Haijian ZHOU ; Guojun LIU ; Yan WANG ; Baiqi YU ; Mingmei SHI ; Xianfeng CHENG ; Ying HONG ; Jin CHEN ; Wanfu HU ; Jun REN ; Shengwei ZHAN
Chinese Journal of Preventive Medicine 2015;49(1):56-59
9. Genetic analysis of norovirus strains detected from sporadic cases in Anhui province, 2016-2017
Yuan YUAN ; Yonglin SHI ; Yong SUN ; Wanfu HU ; Weiwei LI ; Yinglu GE
Chinese Journal of Experimental and Clinical Virology 2019;33(2):136-141
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.
10.Clinical effects of autologous split-thickness skin grafting for prefabricating urethra combined with scrotal flap in repairing middle urethral defect with penile defect
Lin TONG ; Wanfu ZHANG ; Fei HAN ; Zhigang XU ; Dahai HU ; Hao GUAN
Chinese Journal of Burns 2023;39(3):215-220
Objective:To investigate the clinical effects of autologous split-thickness skin grafting for prefabricating urethra combined with scrotal flap in repairing middle urethral defect with penile defect.Methods:The retrospective observational study was conducted. Eight male patients (aged 14 to 58 years) with middle urethral defect and penile defect caused by various injuries who met the inclusion criteria were admitted to the First Affiliated Hospital of Air Force Medical University from January 2015 to January 2022. The length of urethral defect was 3 to 5 cm, and the wound area of penile defect after debridement was 5.0 cm×2.5 cm to 7.0 cm×5.5 cm. All the patients underwent autologous split-thickness skin grafting for prefabricating defect urethra in stage Ⅰ, and urethral anastomosis was performed and unilateral scrotal flap was transferred to reconstruct urethra and penis in stage Ⅱ. The area of scrotal flap was 6.0 cm×3.0 cm to 8.0 cm×6.0 cm. The wound in the donor area of skin graft was covered by oil gauze, and the wound of flap donor area was sutured directly. On the 7 th day after the operation of stage Ⅱ, the survival of the flap was observed. In 3 weeks after the operation of stage Ⅱ, the urinary flow rate was measured by the urinary flow rate detector (urinary flow rate >15 mL/s was regarded as unobstructed urination), the urinary fistula and erectile function were observed, and the self-made therapeutic satisfaction questionnaire was used to investigate the therapeutic satisfaction degree of patients. During follow-up, the appearance of the flap recipient area was observed, the Vancouver scar scale (VSS) was used to evaluate the scar situation in the donor areas of skin graft and flap, the urinary flow rate was detected as before, the urethral stricture, urinary fistula, and erectile function were observed, and the therapeutic satisfaction degree of patients was investigated. Results:On the 7 th day after the operation of stage Ⅱ, the flaps survived completely in 8 patients. In 3 weeks after the operation of stage Ⅱ, the urinary flow rate was 25.3 (18.0, 38.5) mL/s, with unobstructed urination, without urinary fistula and with erectile function, and the score of therapeutic satisfaction degree was 14.3 (14.0, 15.0). During follow-up of 1 to 7 years, the flap recipient area of 8 patients was full in appearance and not swollen, with similar color to the surrounding tissue; the VSS scores of the donor areas of skin graft and flap were 11.5 (10.0, 13.0) and 10.5 (9.3, 12.0), respectively, the urinary flow rate was 24.6 (17.7, 34.1) mL/s, with no urethral stricture, urinary fistula, and erectile dysfunction, and the score of therapeutic satisfaction degree was 13.5 (13.3, 14.8). Conclusions:Autologous split-thickness skin grafting for prefabricating urethra combined with scrotal flap in repairing the urethral and penile defects not only reconstructs the structure of urethra and the shape of penis, but also restores the sensation and erectile function of penis, with few postoperative complications, no obvious scar hyperplasia, and high satisfaction degree of patients, which is worthy of clinical promotion.