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.

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 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.

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 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 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 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 study the efficacy and immunoregulation of pidotimod combined with routine treatment in the children with recurrent respiratory tract infections. Methods:Totally 114 cases of children with recurrent respiratory tract infections were divid-ed into the control group and the treatment group according to the sequence of hospitalization. The 58 patients in the control group re-ceived the conventional treatment, while the 56 patients in the treatment group were given pidotimod additionally. After two-month treatment, all the children were given 1 -6-month follow-up, and the symptom disappearance time, clinical efficacy, recurrence, physiological indices and change of immunological function in the two groups were observed and compared. Results:The disappearance time of symptom and lung physical signs in the observation group was much shorter than that in the control group (P<0. 05);the total effective rate of the observation group was 92. 86%, which was significantly higher than that of the control group (67. 24%, P <0. 001). The number of upper and lower respiratory tract infections in the observation group was significantly lower than that in the con-trol group (P<0. 05). After the treatment, the immune parameters in the observation group were significantly improved (P<0. 01), which were significantly higher than those in the control group (P<0. 05 or P<0. 01). The immune parameters before and after the treatment in the control group showed no statistically significant difference (P>0. 05). Conclusion:Pidotimod combined with routine treatment in the children with recurrent respiratory tract infections exhibits promising efficacy, which can improve immunity effectively.