Objective To study the chemical constitients of Polygonum cuspidatum.Methods The compounds were isolated through column chromatography and their structures were elucidated through physicochemical and spectral analyses.Results Seven compounds were obtained from the diethyl ether fraction of ethanol extract and identified as physcion(Ⅰ),emodin(Ⅱ),ambrettolide(Ⅲ),?-sitosterol(Ⅳ),oleanolic acid(Ⅴ),coumarin(Ⅵ),and 2-ethoxy-8-acetyl-1,4-naphthoquinone(Ⅶ).Conclusion Compound Ⅶ is a new compound named as cuspidatumin A,and compound Ⅲ is obtained from P.cuspidatum for the first time.

Objective To explore the effect of bcl 2 in vascular smooth muscle cells(VSMC) apoptosis induced by nitric oxide(NO),we investigated the effect of NO on Bcl 2 protein expression in cultured vascular smooth muscle cells(VSMCs),and effect of Bcl 2 over expression on apoptosis is(VSMCs) induced by NO. Methods Cultured rat VSMCs at passage 5 to 8 were used for experiments,S nitroso N acetypenicillamine(SNAP) was used as NO donor.Cells were preincubated for 24-48 hours in special culture dish with conditioned medium to reach quiescent,and then incubated in conditioned medium containing 0 5 mmol/L SNAP.After 8 to 10 hours,the cells were fixed and Bcl 2 expression was analyzed by ACAS 570 through fluorescent immunochemistry.Vascular smooth muscle cells were transfected with bcl 2 gene through retrovirus to allow overexpression of Bcl 2 in VSMCs.Finally,we observed the apoptosis in normal VSMCs and VSMCs infected with bcl 2. Results SNAP downregulated Bcl 2 expression in VSMCs.PCR and Southern hybridization confirmed the integration of the bcl 2 retrovirus vector into vascular smooth muscle cells genomic DNA.And fluorescence immunohistochemistry confirmed that Bcl 2 protein was overexpressed in VSMCs infected with bcl 2.And ratio of apoptosis become lower in VSMCs infected with bcl 2. Conclusion bcl 2 can inhibit apoptosis of VSMCs induced by SNAP.It is suggested that downregulation of Bcl 2 protein is one of the ways in which SNAP induces VSMCs apoptosis,and apoptosis in VSMCs induced by SNAP is regulated by bcl 2.;

Objective To investigate the influence of recombinant neuregulin-1 beta (rhNRG-1β) on neural stem cell proliferation through extracellular regulated protein kinases (ERK) signaling pathway in oxygen and glucose deprivation (OGD) environment.Methods Neural stem cells were obtained from embryonic brain of mice pregnant for 14-17 d,cultured and identified by immunochemical staining through detection of the indicator nestin using the SABC-FITC (POD)double standard kit.Neural stem cells were divided into three experiment groups (OGD group,control group and OGD + rhNRG-1β group).Control group:identified neural stem cells,2 × 107,were cultured for 3 h in the 24-hole culture plate with DMEM/F12 complete culture medium;OGD group:neural stem cells,2 × 107,were cultured in the 24-hole culture plate deprived glucose DMEM/F12 in a wet airtight container (37 ℃ constant temperature),cells were cultured with mixed gas of nitrogen (950 ml/L) and oxygen (50 ml/L) for 1 h,and then the culture medium was replaced with complete culture medium and cultured for 3 h;OGD + rhNRG-1β group:before OGD intervention,100 μg/L rhNRG-1β was given for 3 h.Neurospheres formation:the three groups of stem cells were dispersed into single cells,1 × 106/ml cells were inoculated to culture plates containing cover slips coated with poly lysine,and cultivated for 7 d,and neurospheres formation of the 3 groups of neural stem cells was observed under microscope,which was aimed to record neural stem cells proliferation changes.Colony formation:the three groups of stem cells,vaccinated in 60 mm in a petri dish,2 × 107 in number,were cultivated in complete culture medium for 24 h.The colony formation of the three groups of cells was observed under microscope,and neural stem cells proliferation changes were observed.Western blotting:the change of phosphorylation ERK (pERK) protein of the three groups of stem cells was determined,and the effect of pERK protein expression regulated by rhNRG-1β in mice neural stem cells proliferation through ERK signaling pathway was observed.Results Microscopically the primary cultured stem cells grew in single or in pairs,in a round shape;neural stem cell proliferated in clumps or colony;neural stem cells expressed the specificity of nestin protein markers with fluorescent yellow-green color.The differences in the aspects of the average diameter of neurospheres and neurospheres quantity in the neural stem cells between groups were statistically significant (F =693.66,1 002.09,all P ＜ 0.01),and among them the neuropheres formation of OGD group was significantly suppressed.Formation quantity and average diameter in OGD group [(88.78 ± 7.14) numbers,(62.12 ± 2.52) μm] were significantly lower than those in the control group [(246.34 ± 8.67) numbers,(128.45 ± 2.33) μm] and those in OGD + rhNRG-1β group [(237.87 ± 6.61) numbers,(118.37 ± 2.71) μm,all P ＜ 0.01].The difference of colony formation rate of neural stem cell was statistically significant (F =132.03,P ＜ 0.01),and among them colony formation of OGD group significantly suppressed.Formation rate in OGD group [(11.65 ± 0.94)％] was significantly lower than that in the control group [(33.23 ± 2.93)％] and that in OGD + rhNRG-1β group [(31.42 ± 2.61)％,all P ＜ 0.01].Western blotting showed that the difference of pERK protein expression of neural stem cells between groups was statistically significant (F =63.76,P ＜ 0.01).Relative expression of the pERK protein in OGD group (0.487 ± 0.072) was significantly lower than that in the control group (1.013 ± 0.112) and that in OGD + rh-NRG-1β group (1.752 ± 0.278,all P ＜ 0.01).Conclusion rhNRG-1β preserves neural stem cell proliferation with phosphorylation ERK protein expression up-regulated in oxygen and glucose deprivation environment.

Objective To know the influence of psychological intervention on delivery results of par-turient women, and then reference to certain effective psychological nursing cares for clinical field. Methods Divided 226 partreient women of spontaneous labor into the experimental group and the control group randomly, there were 113 eases in each group. Traditional routine nursing cares was used in the control group, psychological intervention was used in the experimental group in addition. Compared the condi-tion of birth process and the incidence rate of labor-related complications between the two groups. Re-sults There were significant differences between the two groups about all the indexes which had indicated the delivery. Conclusions Correct psychoanalysis and proper nursing intervention and effective release the pains for parturient women, and then decrease the incidence rate of medical negligence.

Hepatitis c virus (HCV) infection has become one of the global public health problem,while there is no vaccine to prevent HCV infection, the so-called "cocktail" therapy that use a combination of drugs targeting multiple steps in the HCV infection cycle could achieve better curative effect. the process of HCV entering into host cell is the important step of drug intervention, in which HCV envelope protein El and E2, Host cell factors including Heparan sulfate(HS), CD81, scavenger receptor class B type I (SR-BI), Occludin (OCLD), Claudin (CLDN), low densitity lipoprotein receptor (LDLR), dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN), Liver/lymph node specific ICAM-3-grabbing integrin(L-SIGN), trans- ferrin receptor 1 (TfR1) and so on play a important role. The virus and the host factors can be used as targets of hcv entry inhibitors many studies have shown that as novel and promising compounds, HCV entry inhibitors combinating with other drugs can be more effective in the treatment of HCV, this paper have re- viewed targets and inhibitors of HCV enterring into host cell since 1990s.
Animals ; Antiviral Agents ; pharmacology ; Hepacivirus ; drug effects ; physiology ; Hepatitis C ; genetics ; metabolism ; virology ; Humans ; Receptors, Virus ; genetics ; metabolism ; Viral Envelope Proteins ; genetics ; metabolism ; Virus Internalization ; drug effects

Expression conditions of induction strategies for the cytoplasmic inclusion bodies (IBs) production of liver targeted interferon IFN-CSP by recombinant Escherichia coli (E. coli) BL21 (DE3) were optimized in shake-flask cultures in this study. The factors of the optimized protocol included in the present study were pH, inducer IPTG (isopropyl beta-D-thiogalactoside) concentration, culture growth temperature, incubation time and induction point. The effects of those factors were investigated by 'single variable at a time' method, aimed to analyze characterization of the recombinant strain. Orthogonal experimental design was further used to optimize the above critical factors for IFN-CSP production. According to the expression optimization result, it was confirmed that the main influence factors were cell density and induction temperature. The IFN-CSP gene expression optimized conditions were: pH value of the culture medium was 6.0, culture temperature 37 degrees C, adding IPTG to final concentration 0.4 mmol/L when the recombinant strain growth density OD600 achieved 0.8 and induction time 4 h. At this point, the IBs represented 74.3% of the total cellular protein. Compared with the non-optimized condition, IFN-CSP production obtained in optimized induction strategies were increased by approx. 1.2-fold. The optimized induction strategy yielded 688.8 mg/L of IFN-CSP, providing experimental data to study the biology activity and productive technology of IFN-CSP.
Biotechnology ; methods ; Cell Culture Techniques ; methods ; Culture Media ; chemistry ; Escherichia coli ; metabolism ; Gene Expression ; Interferons ; biosynthesis ; Liver ; Temperature

Objective To evaluate the immunotoxicity effect of Liver targeting interferon (IFN -CSP) on mice.Methods Mice were randomly divided into five groups:low, middle and high dose of IFN-CSP, solvent control group(saline) and Positive control group (cyclophosphamide).They were injected subcutaneously for 2 weeks.Delayed hypersensitivity test was used to determine the cell immunefunction and plaque forming cell assay was used to determine the humoral immune function.Results There was no significant difference of the the index of immune organ and the ear swelling degree between IFN-CSP groups and control group.There was also no significant difference on hemolytic plaque test between them.Conclusion IFN-CSP has no significant effect on both cellular immunity function and humoral immune function of mice, this results will provides the basis for further safety evaluation.

Objective To design a kind of aseptic drainage device with safe practice and accurate measurement for clinical nurses. Methods The experiment group included 63 cases using drainage device with accurate measurement,and control group included 63 cases using disposal drainage bag with scale of visual measurement.The accuracy of drainage liquid measurement was compared between two groups,and bacterial culture was made for the drainage liquid of two groups.Results The measurement error of experiment group and control group was(3.31±1.8)mL and(56.0±5.8)mL,respectively.The difference was significant(t=-4.593,P<0.01).The bacterial culture results showed no obvious difference between two groups(positive rate were 15.9%vs 17.5%,χ2=0.057,P>0.05). Conclusion The accurately measured aseptic drainage device is convenient for nurses to practice in clinics,and its application can reduce workload of nurses and decrease the occurrence of pollution.

Objective To evaluate clinical value of MGFA classification and QMG score on pre-dicting late extubation after thymectomy for myasthenia gravis(MG).Methods Total of 61 patients with MG received extended thymectomy from January 2007 to February 2012 were enrolled.Patients were divided into two groups:normal extubation group contained the other 47 patients without pro-longed postoperative mechanical ventilation and delayed extubation group included 14 patients with prolonged postoperative mechanical ventilation.The following factors were evaluated:gender,age, weight,MGFA classification,QMG score,history of steroid hormones or anticholinesterase drugs be-fore operation,the function of liver and kidney before operation,preoperative electrolyte,preoperative hemoglobin content,etc.Receiver operator characteristic curve (ROC)was plotted,and the predictive value, sensitivity and specificity of preoperative MGFA clinical classification and QMG score predicting postoperative ventilation in MG were calculated.Results Fourteen patients(22.95%)de-veloped breathing support after the anaesthetic or endotracheal intubation again in 48 hours.the area under ROC curve(AUC)for preoperative MGFA clinical classification predicting postoperative ventila-tion was 0.723 in MG,it had the sensitivity of 78.5% and specificity of 63.8%.The AUC for QMG score predicting postoperative ventilation was 0.866,the QMG score threshold value of 8.5 had the sensitivity of 78.6% and specificity of 87.2%.Conclusion MGFA classification and QMG score can predict late extubation after thymectomy in patients with myasthenia gravis.

Objective To investigate the incidence of thyroid space occupying lesion by ultrasound imaging in patients with breast cancer.Methods From 2009 to 2012,150 patients with breast cancer as experimental group and another 150 patients with breast benign diseases as control group underwent thyroid ultrasound examination.Ultrasound results of the two groups were analyzed to find the differences.Restult More obvious space occupying lesions were found in experimental group than those in control group (P＜0.05).Conclusion The incidence rate of thyroid space occupying lesion in the patients with breast cancer is higher than that in the patients with no breast cancer.