The temperature pressure relationship of mixed gases in the autoclave was analyzed with Clausius Clapeyron equation, Boyle's law and Dalton's law of partial pressure Antoine equations discribing the P T relationship inside the autoclave with different residual air were built up according to data published and a method calculating the temperature from values of manometer was introduced The necessity to efflux the air from the autoclave was explained theoretically

ObjectiveTo determine the effect of multi-criteria decision analysis (MCDA) on the effect of bundle treatment for severe pneumonia.Methods A prospective historical control observation was conducted. Seventy-five patients with severe pneumonia having received MCDA (from January 2013 to August 2014) were assigned as intervention group. MCDA group was set up by the medical staff. Bundled treatment plan was composed of the MCDA evaluation results, anti-infection, phlegm and other conventional treatment measures which was adjust on time until the patient was transferred out of the respiratory intensive care unit (RICU) or died. Seventy patients with severe pneumonia before receiving MCDA (from August 2010 to December 2012) were set as historical control group. Comparison of general condition before treatment and the incidence of hospital infection, average hospitalization cost, duration of RICU stay and mortality between these two groups were performed.Results There were no statistically significant differences in gender, age, past history, and acute physiology and chronic health evaluationⅡ (APACHEⅡ) score at admission between two groups. Compared with control group, the incidence of hospital infection [1.33% (1/75) vs. 11.43% (8/70),χ2 = 4.723,P = 0.030], mean hospitalization cost in RICU (10 thousand Yuan: 3.44±0.79 vs. 3.76±0.91,t = 2.265, P = 0.025), length of RICU stay (days: 15.01±4.22 vs. 16.92±4.79,t = 2.552,P = 0.012) and mortality in RICU [8.0% (6/75) vs. 21.4% (15/70),χ2 = 5.272,P = 0.032] in intervention group was significantly decreased. Conclusions Application of MCDA in the bundle treatment of severe pneumonia could elevate the scientificalness of decision, and reduce the medical cost. Additionally, MCDA is worth to be generalized because the implementation of guidelines can improve the clinical outcome and prognosis of the patients.

Objective To investigate the mechanism related to reduced antibiotic sensitivity of Acinetobacter baumannii inducted by meropenem in vitro.Methods Three strains of clinically isolated carbapenems-sensitive Acinetobacter baumannii were induced by meropenem in vitro, and the mutant strains (MS1, MS2 and MS3) were obtained.Minimal inhibitory concentrations (MICs) of antimicrobial agents to strains before and after induction were determined by automatic drug sensitivity analyzer .The homology of strains was analyzed by Enterobacterial repetitive intergenic consensus -polymerase chain reaction ( ERIC-PCR).Modified Hodge test and EDTA-Na2-double disk synergy test were used to detect carbapenemase and metallo-β-lactamase (MBL), respectively.Main carbapenemase genes were detected by PCR and followed by DNA sequencing.Expressions of adeB and outer membrane proteins in strains before and after induction were detected with fluorescence quantitative PCR and SDS -polyacrylamide gel electrophoresis , respectively.t test was used for data analysis .Results The sensitivity of mutant Acinetobacter baumannii strains to meropenem and most antibiotics was reduced , except to imipenem, amikacin and polymyxin; and the reduced sensitivity to meropenem in MS2 and MS3 was of genetic stability.ERIC-PCR showed 100%homology between the mutant strains and parental strains .Both carbapenemase and metallo -β-lactamase were negative in mutant strains and parental strains , and only OXA-51 gene was found.The expressions of adeB gene in mutant strains were 24.26 ±0.91, while those in parental strains were 22.81 ±0.38, and the difference was not significant (t =2.534, P >0.05).Outer membrane protein with molecular weight 54 000 was missing in MS1, while that with molecular weight 47 000 was missing in MS2 and MS3.Conclusion Reduced antibiotics sensitivity in meropenem -induced Acinetobacter baumannii may be correlated with the deficiency of outer membrane protein with molecular weight 47 000.

Objective To identify the drug resistance-related genes in a clinically isolated strain of Enterobacter cloacae.Methods A strain of Enterobacter cloacae was isolated from sputum of a patient with chronic obstructive pulmonary disease from the Affiliated Hospital of Xuzhou Medical University in March 2013.Modified Hodge test and metal enzyme inhibition test were performed for drug-resistant phenotype screening.Carbapenemase genes blaMUS-1, blaVIM-1, blaVIM-2, blaIMP, blaKPC-2, blaNDM-1, blaOXA-48 and blaGESwere amplified by polymerase chain reaction (PCR), and the positive products were sequenced and analyzed.Plasmid conjugation and transformation experiments were used to confirm that the resistance gene mediated by plasmids.Agar dilution method was used for antibiotic susceptibility test.Results Both modified Hodge test and metal enzyme inhibition test were positive in this strain of Enterobacter cloacae.blaNDM-1 gene and blaKPC-2 gene were detected by PCR, and further confirmed by sequencing.blaNDM-1 gene was carried by IncX plasmid with 54×103 bp, KPC-2 gene was carried by untyping plasmid with 42×103 bp.The strain was only sensitive to tetracycline (MIC=2 μg/mL) and tigecycline (MIC=1 μg/mL).The symptoms were improved after the patient was treated by tigecycline combined with Piperacillin/Tazobactam.Conclusion blaNDM-1 and blaKPC-2 genes in Enterobacter cloacae can be mediated by plasmids, and appropriate therapy for its infection should be based on the result of antibiotic susceptibility test.

Objective To explore molecular mechanism of expression of vascular endothelial growth factor (VEGF) mRNA and secretion of VEGF protein in HL-60 cells induced by all-trans refinoic acid (ATRA). Methods MTr method was used to detect the proliferation of HL-60 cells induced by ATRA,cell cycle and CD11b expression in HL-60 cells were detected by flow cytometry. Expression of VEGF, c-myc, by-poxia-inducible factor(HIF)-lα, matrix metalloproteinase (MMP)-9 and MMP-2 mRNA were detected by semi-quantitative RT-PCR. VEGF protein in HL-60 cells culture supernatant was measured by ELISA before and after being induced by ATRA. Results After treatment with ATRA,the proliferation of HL-60 cells was obviously inhibited, CD11b expression increased, trend of granulocyte directional differentiation emerged, and differentiation degree was increasd(P <0. 05) ;expression level of c-my and VEGF mRNA was down-regulated (P < 0. 05), but expression level of HIF-1α mRNA was up-regulated (P < 0. 05). VEGF protein level in HL-60 cells culture supernatant was decreased by blocking the expression of MMP-9 or MMP-2(P <0. 05).Conclusion VEGF expression has positive correlation with c-myc expression,but has negative correlation with HIF-1α expression. MMP-9 and MMP-2 may be the main factors regulating VEGF secretion in HL-60 cells.

Objective To study the molecular mechanism of different sensitivities to apoptosis induced by low concentration of As2O3 in PML-RARα negative HL-60 cells and PML-RARα positive NB4 cells. Meth-ods NB4 and HL-60 cells were cultured with As2O3 for 1 to 4 days; cell proliferation were detected by MTT method; the apoptosis was detected by flow cytometry,Bcl-2,Bax and Fas mRNA were determined by RT-PCR. Results The proliferation of NB4 cells was inhibited obviously by As2O3(1.0 μmol/L)with the induction of apoptosis( P <0.05) ,which was accompanied by the down-regulation of Bcl-2 mRNA expression( P <0.05)and the ratio of Bcl-2/Bax(P <0.05), but there was no obvious variation of Bax and Fas expression( P >0.05). Inhibition of proliferation and apoptosis were not obvious in PML-RARα negative HL-60 cells induced by low concentration As2O3 ( P >0.05), and there was no obvious variation of Bcl-2, Bax, Fas mRNA expres-sion or Bcl/Bax ratio( P >0.05). Conclusion The ratio of Bcl-2/Bax is contributed to the different sensitiv-ities of PML-RARα negative HL-60 cells and positive NB4 cells induced by low concentration of As2O3.

The prognosis of hepatocellular carcinoma (HCC) is closely associated microvascular invasion (MVI), and in recent years, more and more clinical studies have been conducted on MVI, but there are few articles on the development of basic pathology and related controversies. As the basis of clinical research, it is of great significance to understand the definition of MVI and the controversy over pathological grading. With reference to related articles, this article reviews the development history of the definition of MVI, analyzes the current status of the research on the criteria for pathological grading of MVI, and assesses the value of different criteria in predicting postoperative recurrence of HCC, so as to provide a reference for the clinical research on MVI.

OBJECTIVEThe effect of triptolide on the DNA methylation level of MMP-9 gene and the mRNA expression of tissue inhibitors of met-alloproteinases (TIMPs) were examined in human fibrosarcoma HT-1080 cells to explore the molecular mechanisms involved in the anticancer activity of triptolide.

METHODHT-1080 cells were cultured in MEM containing 10% newborn calf serum and 1% penicillin-streptomycin. Triptolide was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 1 goL-1 and stored at -20 degrees C. Triptolide was freshly diluted with culture medium perior to use and directly added to cell cultures at the indicated concentration, and incubated for 72 hours at 37 degrees C in a humidified atmosphere with 5% CO2, with changes of reagents every 24 hours. Methylation specific PCR (MSP)was applied to assess the methylation status of MMP-9 gene promoter, and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was employed to measure the mRNA expression of tissue inhibitors of metalloproteinases (TIMPs) in human fibrosarcoma HT-1080 cells after 72 hours of treatment with 6 nmol x L(-1), 12 nmol x L(-1) or 18 nmol x L(-1) triptolide, respectively.

RESULTSThe methylation index of MMP-9 gene promoter was statistically elevated in HT-1080 cells after 72 hours of treatment with 18 nmol L(-1) triptolide, compared with those in controls (0.61 +/- 0.10 vs 0.39 +/- 0.10, P < 0.05), while no significant difference was noted between 6 nmol x L(-1) or 12 nmol x L(-1) triptolide treated HT-1080 cells and controls (0.40 +/- 0.15 vs 0.39 +/- 0.10, 0.46 +/- 0.20 vs 0.39 +/- 0.10, respectively, both P > 0.05). The mRNA expression of TIMP-1, -2, -3 or -4 was not significantly changed in HT-1080 cells after 72 hours of treatment with the indicated concentrations of triptolide, respectively compared with those in controls (all P > 0.05).

CONCLUSIONThe results demonstrated that triptolide upregulates the methylation level of MMP-9 gene in HT-1080 cells in vitro.

Antineoplastic Agents, Alkylating ; pharmacology ; Cell Line, Tumor ; DNA Methylation ; drug effects ; Diterpenes ; pharmacology ; Epoxy Compounds ; pharmacology ; Fibrosarcoma ; drug therapy ; genetics ; metabolism ; Gene Expression Regulation, Enzymologic ; drug effects ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Phenanthrenes ; pharmacology ; Tissue Inhibitor of Metalloproteinases ; genetics ; metabolism

Objective To evaluate the applicationof the double-disk synergy test(DDST) and combined disk test(CDT) in clini cal metal enzyme phenotype deteetion.To evaluate the value of multiplex PCR in detecting the metallo-β-1actamase in clinical.Methods 56 strains of metallo-β-1actamase-positive strains were identified [NDM-1(n=9),VIM(n=32),IMP(n=15)]for appraise the two methods.By optimizing the design of PCR primers,3 pairs of primers were designed and detected (IMP,VIM,NDM-1) in one tube for evaluate the method.Results The DDST was 80.36％(45/56),and 100.00％(56/56) in the CDT.The accuracy of multiplex PCR was 100.00 ％ (56/56),and the size of the amplified fragment was used to distinguish three types of metallo-β-lactamase.Conclusion The CDT is more suitable for clinical application than DDST.Multiplex PCR has the characteristics of simple,rapid and accurate in detection of metallo-β-1actamase.It is suitable for daily use of clinical microbiological laboratory,which will help the clinical timely and effective administration.

Through summarizing and analyzing the characteristics of elderly infertile people,the problems emer-ging in the assisted reproduction process and the possible ethical problems emerging in the assisted reproduction treatment of elderly patients,this paper explored how to build the ethical path which aimed at elderly pregnancy -assisted people and suitable for Reproductive Center in the First Affiliated Hospital of Xinjiang Medical University. And aiming at the possible ethical problems emerging in the process of assisted reproduction treatment of elderly pa-tients,this paper put forward that it should establish normative ethical working path,to be more convenient to fully conduct ethical supervision and examination in the process of assisted reproduction treatment of elderly patients.