Objective:To compare the different DNA extraction methods and establish an efficient and sensitive method of DNA extraction from bacteria for PCR and Real-time PCR.Methods:Bacillus coli DNA with the same concentration was extracted by 5 different DNA extraction methods,the Proteinase K method,the alkali-lysis method,the Chelex-100 with NP40 or Triton X-100 and the water-boiling method to determine OD260/280.Bacillus coli DNA with different concentrations wewe extracted to be amplified by PCR and Real-time PCR for compariing the sensitivity.Results:Among these extraction methods,Chelex-100 + NP40 method was with the best purity and sensitivity and offered a higher OD260/280(1.79?0.03).Neither false positive nor false negative was found in PCR and Real-time PCR,The sensitivity was at 10/ml of bacteria concentratio.Conclusion:The DNA extraction based on Chelex-100 + NP40 is effective,specific and sensitive,and it accommodate to PCR and Real-time PCR.

Severe burn is often accompanied by multiple organ damage. Acute lung injury (ALI) is one of the most common complications, and often occurs in the early stage of severe burns. If it is not treated in time, it will progress to acute respiratory distress syndrome (ARDS), which will be a serious threat to the lives of patients. At present, the treatment of ALI in patients with severe burn is still remained in some common ways, such as the liquid resuscitation, the primary wound treatment, ventilation support, and anti-infection. In recently, human umbilical cord mesenchymal stem cells (hUCMSCs) have been found having some good effects on ALI caused by various causes, but few reports on the efficacy of ALI caused by severe burns were reported. By reviewing the mechanism of stem cell therapy for ALI, therapeutic potential of hUCMSCs in the treatment of severe burns with ALI and a new approach for clinical treatment was provided.

Objective To investigate the expression levels of MiR‐96 ,HIF‐1α,Twist and Slug and other predisposing gene in cervical cancer .Methods In this study ,128 patients with cervical cancer treated in our hospital from March 2010 to May 2014 were selected as the observation group ,and 100 cases of healthy people were selected as control group .MiR‐96 ,HIF‐1α,Twist and Slug and other predisposing gene expression levels in cancer tissues were tested .(1) HIF‐1α:HIF‐1α kit was used to detect HIF‐1αmonoclonal antibodies ,the kit was prepared and stained according to the requirements ,and the positive cell rate greater than 10%under the microscope were positive .(2) MiR‐96 ,Twist and Slug:total RNA was extracted according to the instructions ,the RNA was reverse transcribed ,the relative expression values MiR‐96 ,Twist and Slug were detected by quantitative PCR .Results (1) HIF‐1αwas not expressed in normal tissues .And in tumor tissues ,the positive expression rate was much higher than that of normal tissue ,there was a significant difference(P<0 .05);(2)the relative expression values of MiR‐96 in tumor tissue were much greater than that of normal tissue (P<0 .05);(3) the relative expression rate of Twist and Slug gene in the tumor tissue were also much higher than that of normal organizations (P<0 .05) .Conclusion Compared with normal tissues ,MiR‐96 ,HIF‐1α,Twist and Slug gene expression in tumor tissues are significantly greater .

The incidence of birth defect in China is still high,which is not only affected by genetic factors,but also affected by heavy metals in surrounding that prejudices foetus' normal development.Heavy metal is a kind of inorganic pollutants with high toxicity.Excessive intake of lead,cadmium,arsenic or mercury,and the insufficient intake of zinc,calcium and copper may both lead to at least 1 kind of birth defect.The interactions between heavy metals also affect the outcomes of pregnancy.This paper reviewed different relationships between heavy metals and birth defect recording to relevant achievements such as animal experimentations and epidemiologic study made by researches from at home and abroad in recent years.It is advised to intake adequate zinc and copper and avoid being exposed to harmful metals to make sure the effective reduction of the incidences of birth defects.This paper will also point out the direction of future research about the relationship among the birth defects and heavy metal.

Objective To investigate the role of nursing intervention in the prevention of premature rupture of the membrane infection. Methods 131 patients with premature rupture of the membrane were divided into the control group(65 cases) and the experimental group(66 cases), they adopted routine nursing method and nursing intervention respectively. The duration of pregnancy, fetal condition and laboratory tests for infection were compared between two groups. Results No significant differences were found between the two groups in terms of age, duration of pregnancy, newborn weight. Significant differences were observed in Apgar score, newborn death between two groups. Conclusions Monitoring for signs of premature rupture of the membrane such as infection and laboratory tests are important for the prevention of infeclion, if infection occurs, delivery may be necessary to prevent further complications.

Objective To study the effects of N-acetylcysteine (NAC) on the proliferation, apoptosis and collagen synthesis of human lung fibroblast (HLF). Methods HLF was primarily cultured in complete medium of DMEM/F12. Different concentrations of NAC (5,10,20,40mmol/L) were administrated. Cell proliferations were tested by methylthiazolyltetrazolium (MTT) ,apoptosis and cell cycle were detected with Flow cytometer and mRNA and 40 mmoi/L )after 24 hours,the apoptosis rates were (34.38±5.80)%, (37.72±3.10)%, (44.05± 4.52) % and (59.18±5.24) % ) respectively, significantly higher than that of the controls (3.92±1.24) % pression of type Ⅰ procollagen in HLF was decreased significantly after administration of NAC at 5, 10,20, 40 mmol/L respectively (P < 0.01 ). Conclusion Administration of NAC induces apoptosis and directly inhibites the proliferation and the collagen synthesis of HLF.

Objective To investigate the clinical significance of YMDD mutation during Lamivudine therapy on chronic hepatitis B.Methods Fluorometric analysis PCR, ELISA were used to estimate the YMDD mutation, HBVDNA quantative level and HBeAg for HBV of 72 cases with chronic hepatitis B before therapy (0 month), and after Lamivudine therapy for 9,12,18 months.Results The YMDD mutation was not observed in these cases before Lamivudine therapy. The mutation was found in 8 cases (11.1%), 17 cases (23.6%) and 28 cases (38.9%) at 9, 12, 18 month for therapy. The YMDD mutation rate rose with the therapy time lasting (P＜0.05). Moreover, the YmDD mutation rate in the patients with HBVDNA quantity higher than 108 copies/ml was significantly higher than that in the patients with HBVDNA quantity lower than 108 copies/ml (P＜0.005). The YMDD variation rate in patients with HBeAg positive and in patients with HBeAg negative showed no significant difference (P＞0.05). The HBeAg negative conversion rate was significantly higher in non-mutation group than that in mutation group (P＜0.05).Conclusion The serum virus quantity may be regard as an early estimate indication of the development of YMDD mutation during Lamivudine therapy.

Objective To understand the status and pathogenic characteristics of healthcare-associated infection (HAI)in cardiovascular surgical intensive care unit (ICU)patients undergoing heart transplantation.Methods Pa-tients who underwent heart transplantation in a hospital between July 1 ,2013 and June 30,2014 were performed targeted monitoring.Results Of 66 patients undergoing heart transplantation,16 developed 18 times of HAI,inci-dence of HAI was 24.24%.Incidence of HAI and ventilator-associated pneumonia(VAP)in patients undergoing heart transplantation were both higher than non-transplantation patients (24.24% vs 6.24%,χ2 =33.718;7.58%vs 1 .72%,χ2 =12.199,respectively,both P <0.001 ).The infection tyes were as follows:lower respiratory tract infection(n=7),VAP (n=6),bacteremia (n=3),superficial incision infection (n =1 ),as well as skin and soft tissue infection (n = 1 ). The isolated pathogens were fungus (n = 8 ),Klebsiella pneumoniae (n = 3 ), Staphylococcus aureus (n=2),Acinetobacter baumannii (n=2),Enterobacter cloacae (n=1 ),Acinetobacter hae-molyticus (n=1 ),and Citrobacter freundii (n =1 ).Conclusion Incidence of HAI is high in patients undergoing heart transplantation,the main infection type is lower respiratory tract infection,the major pathogen is fungus.

Objective To detect nuclear DNA degradation of bone marrows and brains in rat cadavers at different temperatures,and develop a new parameter for estimating early postmortem interval(PMI).Methods The brain and bone marrow were taken out for every 4h,during 0~40h after death at 10℃ and 20℃,respectively.And the single cell gel electrophoresis(SCGE) was carried out to detect the nuclear DNA degradation.Linear regression analysis was used to assay the relationship of the comet parameter HeadDNA%,Tail Length(TL) and Olive TailMoment(TM) with PMI.Results Different decline degrees of comet HeadDNA% were found in both brain cells and bone marrow cells after death,the decline of HeadDNA% in brain cells at 20℃ was faster.Compared with degradation in marrow cells,the linear relation between degradation of brain cells and PMI was better.Conclusion with that of comet parameters TL and TM,the perfect linear relationship between HeadDNA% and PMI was also observed.Conclusion Brain tissues are more suitable for PMI estimation by detecting degradation of DNA with SCGE.The HeadDNA% is more valuable for PMI estimation than TL and TM.

Background and purpose:Studies have shown that renin-angiotensin system (RAS) is closely associated with tumor progress. angiotensinⅡ (AngⅡ) is the most important component of RAS. This study aimed to investigate the possible mechanism by which AngⅡ affected the cell proliferation in human breast cancer cell line MCF-7.Methods:CCK-8 was used to investigate the cell proliferation alteration of MCF-7 cells after treatment of AngⅡ at different dose and time. The inlfuence of losartan (an AT1R inhibitor) and PD98059 (a MAPK inhibitor) in AngⅡ-enhanced cell proliferation was detected by CCK-8. Protein expression was analyzed by Western blot.Results:AngⅡ stimulated the growth of breast cancer cells in a dose- and time-dependent manner. The maximal proliferation effect on MCF-7 cells was obtained with 10-7 mol/L AngⅡ and 24 h, respectively (P<0.000 1). Losartan signiifcantly decreased the level of AngⅡ-induced proliferative effects (P<0.05). Western blot showed that AngⅡ caused rapid activation of p-ERK. In addition, PD98059 could signiifcantly suppress AngⅡ-promoted cell proliferation.Conclusion:AngⅡ can promote MCF-7 cell proliferation through AT1R/ERK/MAPK pathway activation, which could be reversed by losartan or PD98059. Therefore, targeting AngⅡ/AT1R/MAPK signaling could be a novel therapeutic for breast cancer.