Purpose:To study the RNA interference-mediated inhibition of survivin gene on the proliferation of human breast cancer SKBr-3 cells. Methods:SKBr-3 cells were transfected with a pSUPER-S1 vector plasmid that expressed survivin-targeted small interfering RNA, and the mRNA and protein levels of survivin gene were measured with RT-PCR and indirect immunofluorescence, respectively. The proliferation of transfected SKBr-3 cells was investigated through colony forming assay and MTT assay. The cell cycle phases were determined by flow cytometry(FCM). Results:The mRNA and protein levels of survivin declined markedly in pSUPER-S1-transfected SKBr-3 cells. And the colony forming rate of those cells(38?16.70)% was significantly lower than that of the control cells(90.3?4.04)%.The growth of the pSUPER-S1-transfected cells was decelerated and the cell cycle was mainly blocked at G_(1) phase(74.03?8.91)%. Conclusions:survivin gene silencing by RNA interference contributed to a distinctive inhibition of the proliferation of human breast cancer SKBr-3 cells in vitro.

Objective To investigate distribution characteristics of blood culture pathogens,and provid a basis for clinical prevention and control in bloodstream infections.Methods The data of the patients with positive blood culture and the nonduplicate strains were retrospectively analyzed with WHONET 5.6 software in the First People's Hospital of Shangqiu from January 2011 to December 2014.Results The total number of positive strains of blood culture was 1 306 from 25 397 blood cultures,and the positive rates were 5.1％,of which gram positive cocci and gram negative organisms accounted for 64.8％ (846/1 306) and 31.2％ (408/1 306),respectively.Candida accounted for 4.0％ (52/1 306).Gram positive bacteria were mainly Coagulase negative Staphylococcus (CNS) of 564 (43.2％) strains,Staphylococcus aureus of 96 (7.4％) strains,Enterococcus faecium of 48 (3.7％) strains and Streptococcus pneumoniae of 31 (2.4％) strains.The isolated rate of Enterococcus faecium was more than Enterococcus faecalis 20 strains (1.5％).The isolated rate of Escherichia coli,Klebsiella pneumoniae,Acinetobacter Bauman and Pseudomonas aeruginosa were 12.5％ (163 strains),6.2％(81 strains),2.0％(26 strains) and 1.8％(23 strains),respectively.Conclusion The isolated rate of Enterococcus faecium more than Enterococcus faecalis in blood culture,the main pathogens are gram positive cocci in children group.Isolates of gram positive bacteria in the proportion of infants,children and adults with blood culture were 85.2％,87.0％,46.5％.There were 264 strains bacteria isolated in infants and young children's intensive care unit,accounting for 67.3％ of all bacteria isolated from infants and young children.There were 122 strains bacteria isolated in pediatric intensive care unit,accounting for 56.7％ of all bacteria isolated from childen.There were 255 strains bacteria isolated in adult intensive care unit,accounting for 36.5％ of all bacteria isolated from adult.Severe basic diseases were the vulnerable groups of bloodstream infections.

Objective To observe the clinical efficacy of Ziyin Yikang Recipe plus hexadecadrol on anti-HCG antibody positive secondary sterility and the influence on anti-HCG antibody titers. Methods Sixty patients with anti-HCG antibody positive secondary sterility were randomly divided into treatment group (TG) and control group (CG), 30 cases in each. Patients in the TG were treated with Ziyin Yikang Recipe plus hexadecadrol, while patients in the CG were given hexadecadrol only. The Chinese medicine symptom scores and changes of anti-HCG antibody titers were evaluated and compared between the two groups.Results The Chinese medicine symptom scores in the two groups decreased compared with those before treatment (P<0.05), and the TG showed a more significant decrease than the CG (P<0.05). The anti-HCG antibody titers in both groups dropped compared with those before treatment (P<0.05, P<0.01). There was no statistically difference between the two group after treatment (P>0.05). The total effective rate in the TG was higher that that in the CG (P<0.05), with 63.33% (19/30) in the TG and 47.66% (14/30) in the CG.Conclusion The treatment of Ziyin Yikang Recipe plus hexadecadrol on anti-HCG positive secondary sterility can obviously improve the clinical syndrome of yin deficiency and fire hyperactivity, and promote pregnancy success.

Objective To study the DNA extraction and its typing in cigarette butts. Method Chelex 100 extraction method was used to extract DNA of 170 cigarette butts, STR loci were typed after PCR amplification by Profiler Plus kit. Results 21 cigarette butts from one volunteer have no results, the others were all obtained typing results. The samples which were added a little tobacco have no results. Sometimes DNA in sponge contacted with mouth was typed. The small fragment loci were typed in cigarette butts which had been reserved for six months. Conclusion Cigarette butt can be typed by DNA analysis, which is useful in forensic identification.

Objective To investigate the effect of Danhong injection combined with edaravone in the treatment of patients with acute cerebral infarction,and its influence on cytokines,cerebral hemodynamics and vascular endothelial function.Methods From March 2018 to March 2019,142 patients with acute cerebral infarction treated in the People's Hospital of Yuhuan were randomly divided into treatment group and control group according to the digital table,with 71 cases in each group.The treatment group was treated with Danhong injection combined with edaravone,while the control group was only treated with edaravone.Both two groups were treated for 2 weeks.The therapeutic effects,changes of cytokines,cerebral hemodynamics,vascular endothelial function,activity of daily living index(Barthel index) and neurological deficit score(NIHSS score) before and after treatment were compared between the two groups.Results The total effective rate of the treatment group was 90.14％ (64/71),which was higher than 74.65％ (53/71) of the control group,the difference was statistically significant(x2 =5.874,P ＜ 0.05).After treatment,the serum levels of CRP [(5.43 ± 1.20) mg/L] and IL-6 [(32.15 ± 7.39) ng/L] in the treatment group were lower than those in the control group [(9.38 ± 1.74) mg/L and (67.43 ± 10.29) ng/L] (t =15.747,23.465,all P ＜0.05).After treatment,the Vp [(69.83 ± 3.24) v ·-1 · s-1] and Vm [(35.24 ± 2.10) v ·-1 · s-1] in the treatment group were higher than those in the control group [(63.81 ± 2.68) v ·-1 · s-1 and (32.18 ± 1.73) v ·-1s-1],while the PI in the treatment group [(0.72 ± 0.04)] was lower than that in the control group [(0.83 ±0.07)],the differences were statistically significant (t =12.064,9.477,11.497,all P ＜ 0.05).After treatment,the serum level of ET-1 [(60.17 ± 5.46) mg/L] in the treatment group was lower than that in the control group[(73.21 ±6.78)mg/L],while the NO level in the treatment group[(72.15 ±7.39) ng/L] was higher than that in the control group [(61.43 ± 10.29) ng/L],the differences were statistically significant (t =12.622,7.130,all P ＜0.05).After treatment,the Barthel index score of the treatment group [(68.93 ± 7.83) points] was higher than that of the control group [(54.57 ± 7.38)points],while the NIHSS score of the treatment group [(9.34 ± 1.97)points] was lower than that of the control group [(14.54 ± 2.89) points],the differences were statistically significant (t =11.246,12.528,all P ＜ 0.05).Conclusion Danhong injection combined with edaravone in the treatment of acute cerebral infarction is effective,which can alleviate inflammation and improve cerebral hemodynamics and vascular endothelial dysfunction.

Background and purpose:More and s’more evidence has demonstrated that DNMT1 was expressed at high levels in many different kinds of human tumor tissues or cells,suggesting that high expression of DNMT1 was closely associated with occurrence and development of tumors.In this study,effect of down-regulation of DNMT1 on cell proliferation and migration ability of esophageal squamous cell carcinoma(ESCC) cell line EC9706 cells was studied and its related mechanism was explored.Methods:Cell proliferation assay was investigated using CCK-8 Kit,cell migration ability was analyzed using Boyden chamber and the expressions of DNMT1 and MMP-2 were detected by Real-time PCR and Western blotting methods.Results:The result of cell proliferation experiment showed that down-regulation of DNMT1 could markedly inhibit cell proliferation in EC9706 cells.After transfection with DNMT1 siRNA,invasiveness and metastasis ability of EC9706 cells displayed an obvious decrease(P

Objective To analyze strains of Proteus mirabilis (P .mirabilis )and Proteus vulgaris (P .vulgaris ) isolated in a hospital,detect resistance to commonly used antimicrobial agents,and provide reference for rational ap-plication of antimicrobial agents in clinic.Methods 172 P .mirabilis isolates and 68 P .vulgaris isolates isolated between January 1 ,2011 and June 30,2013 were analyzed,antimicrobial resistance susceptibility testing were per-formed by disk diffusion method,data were analyzed with WHONET 5.4 software.Results P .mirabilis strains were mainly isolated from wound secretion(26.74%),sputum(22.68%)and urine(18.61 %),P .vulgaris were mainly from wound secretion(48.53%),urine(17.65%)and sputum(11 .77%).The resistance rates of P .mirabilis to ampicillin and trimethoprim/sulfamethoxazole were both>45.00%;the resistance rates of P .vulgaris to cefazo-lin and trimethoprim/sulfamethoxazole was 86.76% and 41 .18% respectively;the resistance rates of P .mirabilis and P .vulgaris to piperacillin/tazobactam,cefotaxime,ceftazidime,cefepime,carbapenems (ertapenem and mero-penem)and amikacin were all <20.00%.Conclusion The resistance rates of P .mirabilis and P .vulgaris to pip-eracillin/tazobactam,cefotaxime,ceftazidime,cefepime,ertapenem,meropenem and amikacin are all high,and can be used as the empirical medication for the treatment of related infection.

OBJECTIVETo explore the role of S100A4 in the epithelial-mesenchymal transition (EMT) in esophageal squamous cell carcinoma and its possible molecular mechanism.

METHODSThree chemically synthesized S100A4 siRNA sequences were transiently transfected into esophageal carcinoma EC9706 cells. EC9706 cells transfected with negative siRNA, lipofectamine 2000, and vacant EC9706 cells were used as control. Fluorescence quantitative RT-PCR and Western blot were used to detect the inhibition rate of S100A4 siRNA. S100A4 siRNA2 with the best inhibition rate was chosen to transiently transfect into EC9706 cells under the same conditions. The EC9706 cells transfected with negative siRNA, lipofectamine 2000 and vacant EC9706 cells were also used as control. Fluorescence quantitative RT-PCR and Western blot were used to detect the mRNA and protein expressions of E-cadherin, vimentin and snail. The morphology of EC9706 cells was observed under an inverted microscope. Boyden chamber and scratch test were used to detect the invasion and migration ability of EC9706 cells, and CCK8 assay was used to detect the proliferation ability of EC9706 cells. EC9706 cells transfected with S100A4 siRNA2 were further transfected with snail eukaryotic expression vector. The EC9706 cells transfected with S100A4 siRNA, EC9706 cells transfected with snail eukaryotic expression vector and vacant EC9706 cells were used as control. The above indexes of all the groups were observed, too.

RESULTSThe S100A4 mRNA and protein expression levels of the S100A4 siRNA2 group were 0.417 ± 0.041 and 0.337 ± 0.039, the transmembrane cell number was 61.608 ± 8.937, the scratch healing distance was (0.216 ± 0.064) mm, the A value was 0.623 ± 0.084, the E-cadherin mRNA and protein levels were 0.619 ± 0.032 and 0.495 ± 0.034, the vimentin mRNA and protein levels were 0.514 ± 0.032 and 0.427 ± 0.028, the snail mRNA and protein levels were 0.573 ± 0.029 and 0.429 ± 0.041. These data were significantly different with the liposome group, the negative control group and the blank group (P < 0.05 for all). After the S100A4 siRNA2 treatment for 24 h, the appearance of EC9706 cells changed to epithelial cell morphology. The transmembrane cell number and the scratch healing distance of the S100A4 siRNA2+snail eukaryotic expression vector group were (69.382 ± 9.666) cells and (0.274 ± 0.029) mm, the A value was 0.823 ± 0.101, the snail mRNA and protein levels were 0.704 ± 0.037 and 0.625 ± 0.031, the vimentin mRNA and protein levels were 0.712 ± 0.046 and 0.609 ± 0.038, and these data were significantly higher than those of the Sl00A4 siRNA2 group (P < 0.05 for all). The E-cadherin mRNA and protein levels of the S100A4 siRNA2+eukaryotic expression vector group were 0.437 ± 0.038 and 0.381 ± 0.031, significantly lower than those of the S100A4 siRNA2 group (P < 0.05 for all). However, snail had no effect on the morphology of EC9706 cells.

CONCLUSIONSS100A4 may be involved in the EMT process of esophageal squamous-cell carcinoma by regulating the expression of snail and then plays a role in the invasion and metastasis of esophageal carcinoma.

Cadherins ; analysis ; Carcinoma, Squamous Cell ; metabolism ; pathology ; physiopathology ; Cell Line, Tumor ; Epithelial Cells ; Epithelial-Mesenchymal Transition ; Esophageal Neoplasms ; metabolism ; pathology ; physiopathology ; Humans ; Indicators and Reagents ; Lipids ; RNA, Messenger ; analysis ; RNA, Small Interfering ; analysis ; physiology ; S100 Calcium-Binding Protein A4 ; S100 Proteins ; antagonists & inhibitors ; genetics ; physiology ; Snail Family Transcription Factors ; Transcription Factors ; analysis ; genetics ; Transfection ; Vimentin ; analysis ; genetics

Objective To understand the distribution and antimicrobial resistance of pathogenic bacteria isolated from patients in the department of infectious diseases in Xiangya Hospital.Methods The distribution and antimicrobial susceptibility testing results of pathogenic bacteria isolated from patients in this department in 2011 -2015 were analyzed retrospectively.Results A total of 560 strains were isolated during 5 years,of which gram-posi-tive bacteria and gram-negative bacteria accounted for 44.1 % (n =247)and 55.9%(n =313)respectively.69.8%(81/116)of coagulase-negative staphylococcus and 24.3%(9/37)of Staphylococcus aureus were methicillin-resistant (MRCNS,MRSA)respectively.Enterococcus was highly susceptible to vancomycin,linezolid,and phosphonomy-cin (>81 %).Enterobacteriaceae remained highly susceptible to carbapenems (88.9%-100.0%),and was suscep-tible to amikacin,cefoperazone/sulbactam,and piperacillin/tazobactam (>84%).Acinetobacter baumannii was the major isolated multidrug-resistant organism (MDRO),isolation rate of imipenem-resistant Acinetobacter baumannii increased from 50.0% in 2011 to 77.8% in 2015,its resistance rate to imipenem was 64.9%.Conclusion The majority of clinically isolated pathogenic bacteria from this department is gram-negative bacilli,and detection rate of MDROs showed an upward trend;antimicrobial agents should be chosen according to distribution and antimicrobial resistance of pathogenic bacteria.

Objective To study the relationship between the changing law of cytokine gene expression profile and tumor-related fatigue and to verify the preliminary theory conclusion that the pathogenesis of tumor-related fatigue may be due to the up-regulation of the TNF,TGF-?,etc.Methods The changing characteristics of gene expression profile of cytokines were studied in peripheral blood mononuclear cells samples with gene chip.Results The gene expression level of TNF-? and TGF-?1 was up regulated.Conclusions The experimental results conform to the theory research conclusion and show that the pathogenesis of tumor-related fatigue due to the up-regulation of the gene expression level of TNF and TGF-?.