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 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 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.

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 clinical features and management strategies of re-pregnant women with chronic hepatitis B virus infection after implementation of two-child policy.Methods A total of 173 women with chronic hepatitis B virus infection in a subsequent pregnancy were enrolled from Hangzhou Xixi Hospital, among whom 91 cases were before implementation of the policy (from January to August 2015, control group), and 82 cases were after policy implementation (from March to December 2016, study group).The clinical data including basic information, pregnancy complications, postpartum complications, delivery mode and perinatal complications were retrospectively analyzed,t test or χ2 test was used for comparison between the two groups.Results There were significant differences in proportion of age>35 years (31.71% vs.16.48%,χ2=5.528), frequency of abortion(3.2±0.3 vs.1.3±0.4,t=5.723), fertility interval with first pregancy[(57.8±21.4)months vs.(40.4±16.6)months,t=6.691],rate of assisted reproduction(14.63% vs.5.49%,χ2=4.066),proportion of abnormal liver function(13.41% vs.4.40%,χ2=4.618),antiviral treatment during pregnancy (34.14% vs.19.78%,χ2=4.561) and proportion of HBV DNA loads >105IU/mL(35.36% vs.20.88%,χ2=4.515)between study group and control group (all P<0.05).The incidences of gestational hypertension(12.20% vs.3.30%), gestational diabetes mellitus(14.63% vs.5.49%),placenta previa(9.76% vs.2.20%),placental implantation(3.66% vs.0), hydramnio (12.20% vs.3.30%), fetal distress (14.63% vs.5.49%) and preterm birth(14.63% vs.5.49%)were also statistically significant between two groups (χ2=4.914, 4.066, 4.524, 3.838, 4.914, 4.066 and 4.066,all P<0.05).The incidence of cesarean section again(50.00%), postpartum hemorrhage(14.63%)and postpartum anemia(19.51%) were significantly higher in the study group than that in the control group(35.16%,4.40% and 8.79%,χ2=3.892, 5.387 and 4.149,all P<0.05).Compared with the control group, the incidence of low birth weight(14.63% vs.5.49%), neonatal asphyxia(19.51% vs.6.59%)and neonatal HBV infection rate(15.85% vs.6.59%)were higher in the study group (χ2=4.066, 6.486 and 3.883,all P<0.05).Conclusions The clinical characteristics of re-pregnant women with chronic hepatitis B virus infection are more complex and suffer greater risk of pregnancy, so antenatal care should be further reinforced after the comprehensive implementation of two-child policy.

Objective To observe the efficacy of asarone injection combined with inhalation on the treatment of COPD acute phase.MethodsAccording to chronic obstructive pulmonary disease diagnosis and treatment guidelinesin the diagnostic criteria for chronic obstructive pulmonary disease in acute phase, 81 patients were randomized double-blindly divided into observation group of 40 cases and a control group 41 cases.Administering inhalation nebulizer patients in the control group, asarone injection combined with inhalation treatment in the observation group.Serum levels of interleukin-8 (IL-8), interferon-γ (IFN-γ), tumor necrosis factor (TNF-α) levels before and after treatment, using a blood gas analyzer patients arterial oxygen pressure (PaO2) and arterial carbon dioxide tension (PaCO2) level, measured using a spirometer patients accounted for one second forced expiratory percentage of predicted value (FEV1% pred) and total airway resistance, and a separate determination combination therapy regimen based on the above indicators Comparative efficacy.Results①IL-8, IFN-γ, TNF-α: After treatment, the observation group was significantly lower than the control group (t=11.498;t=10.279;t=11.576), the differences were statistically significant (P<0.05).②blood gas PaO2 and PaCO2: post-treatment observation group than the control group (t=11.021;t =8.868), the differences were statistically significant (P<0.05).③lung function FEV1% pred and total airway resistance: After treatment, the observation group than the control group (t=7.182;t=6.341), the differences were statistically significant (P<0.05).The total effective rate was significantly higher (χ2 =4.742) after ④observation group, the difference was significant (P<0.05).ConclusionAsarone injection combined with inhalation can reduce chronic inflammation in the body in patients with acute obstructive pulmonary disease, the body adjust blood gas, improve lung function, improve the treatment of patients with acute COPD.

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.

Objective To investigate the adherence mechanism of Candida albicans to host cells in molecular level. Methods and Results Yeast cell wall protein of Candida albicans were extracted, and purified fibronectin (Fn) adhesin was obtained from yeast cell wall extract. Fn adhesin was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE), the molecular weights of Fn adhesin were 60kd and 105kd. In order to find out the effect of the protein on adherence to host cells, human buccal epithelial cells and Candida albicans cells were incubated at 37℃ for 2 hours on a rotator, and yeasts adhering to the surfaces of epithelial cells were assayed microscopically. The adherence rate of C.albicans to buccal epithelial cells was 44 7?6 28%, and after treating the epithelial cells with Fn adhesin, the adherence rate reduced to 28 3?6 18%, P

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.