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.

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 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.

AIM: To evaluate the antihistamine effects of domestic azelastine hydrochloride. METHODS: Histamine increased skin vascular permeability and induced shock model in guinea pigs. In isolated guinea pig ileal rings, the contraction of smooth muscle induced by histamine was determined. RESULTS: Pretreament with azelastine hydrochloride( 0.05, 0.15,and 0.45 mg?kg -1,ig) significantly inhibited the increase in skin vascular permeability induced by histamine in a dose-dependent manner in guinea pigs. Pretreament with azelastine hydrochloride( 0.05, 0.1,and 0.2 mg?kg -1,ig)produced a significant improvement of shock, as shown by a decrease in reactivity degree, mortality and a prolongation of latency. In isolated guinea pig ileal rings, azelastine hydrochloride (10 -8,3?10 -8,10 -7,3? 10 -7 mol?L -1)significantly inhibited contraction of smooth muscle induced by histamine in a concentration-dependent manner and caused a parallel right shift of the dose-effect curve of histamine (pA 2= 8.55). CONCLUSION: Domestic azelastine hydrochloride shows significant the antihistamine effects.

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

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.

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.