Objective To study the differentiation of bone marrow mesenchymal stem cells in artery tissue under different calcification environments.Methods We made a vascular calcification model using warfarin,vitamin K1 and vitamin D3.After the model was successfully made,we took artery tissue of normal SD rat arteries and calcified arteries co-cultured with MSC,which were divided into three groups.The normal group included normal artery tissue with MSC; calcified inducers group included calcified inducers (dexamethasone with β-glycerophosphate and ascorbic acid),normal arterial tissue and MSC; calcification group included calcified artery tissue and MSC.Each group was cultured for 3 weeks.On the 10th day of the experiment,osteoprotegerin (OPG) protein secretion was detected by ELISA.After three weeks,changes of cell morphology were observed using inverted microscope,and total protein content and alkaline phosphatase (ALP) activity were detected by ELISA.In additional,the Ror2 (receptor tyrosine kinase-like orphan receptor 2) mRNA expression was detected by using RT-PCR method.Results MSC in calcification group spontaneously proliferated and differentiated to osteoblast-like cells.Compared with normal group,calcification group showed that the total protein content,ALP activity of bone metabolism markers,OPG were significantly elevated,while Ror2 mRNA expression was significantly decreased.MSCs in calcified inducers group did not differentiate to osteoblast-like cells,and the total protein content and OPG were increased,while ALP activity had no significant difference as compared with the normal group.However,Ror2 mRNA expression was lower in calcified inducers group than in normal group,while was higher than that in calcification group.Conclusions MSCs proliferate into bone-like differentiation in vascular calcification environment and aggravate the vascular calcification.And in normal vascular with calcified inducers environment,MSCs proliferate into smooth muscle cell differentiation and rehabilitate the vascular calcification.These phenomenons may be related to the Ror2 expression in artery.

Objective According to principle of forming primer dimer,a new cloning gene method was established. Methods Two and more oligonucleotide with complementary partnership in the 3′ end were synthesized by artifical method,oligonucleotide,Taq DNA polymerase and dNTP were mixed in the proportion of reasonable,then expanded and extended by PCR,result was identifed by agarose gel electrophoresis.Results A clear special band was appeared in the agarose gel. Conclusion Short fragment target gene less than 500 bp may be synthesized by this method.

Objective To explore the connotation of patient safety competency of nurses, and to provide references for the safety of education and patients' safety competency. Methods The connotation of patient safety competence of nurses was discussed through semi-structured interview and expert meeting method. Results Patient safety competency of nurses contained 7 aspects, including patient-centered care,team cooperation, safety risk management,information evidence-based application, patient safety culture, clinical safety and continuous quality improvement. Conclusions Patient safety competency of nurses is based on the knowledge, attitude and skill, and includes comprehensive abilities in all other aspects.

Objective To explore the correlation between nurses′ preventive behavior against needle stick injuries and perception of risks, hospital safety climate as well as provide some references for strategy making. Methods In accordance with a multi-stage sampling method, questionnaires were used to collect data from 1 012 nurses in 8 level three class A general hospitals. Results The average scores of nurses′ preventive actions toward needle stick injuries and hospital safety climate were (3. 71 ± 0. 50) and (3. 91 ± 0. 53) respectively. Nurses with high perception of risks accounted for 67. 1%. In addition, the Spearman correlation analysis suggested that nurses′preventive behavior was negatively correlated with perception of risks (r= -0. 093,P<0. 01), while it positively correlated with hospital safety climate (r=0. 463,P<0. 01). Conclusions Clinical nurses should have further regulations for protective behavior against needle injuries. We can take measurements actively to reduce nurses perception of risks and create safety work climate that can be benefits for nurses taking favorable protective behavior and reducing the incidence of needle injuries.

Objective To develop a Patient Safety Nurse Competency Evaluation Scale, and evaluate its reliability and validity. Methods The Patient Safety Nurse Competency Evaluation Scale was developed through literature review, semi-constructed interview and experts consultation. A total of 590 nurses in 5 Class ⅢGrade A hospitals in Beijing were selected by convenient sampling methods and investigated by self-evaluation and peer evaluation. Results The final instrument included 35 items, organized into 6 composites by exploratory factor analysis, with the cumulative contribution of 65.983%. The factor analysis confirmatory showed that χ2/df was 2.742, and RMSEA was 0.064. Correlation coefficients between composites and total scale ranged from 0.694 to 0.812, and correlation coefficients between composites ranged from 0.367 to 0.647, with statistical significance (P< 0.05). The content validity index (S-CVI) was 0.933. The total scale internal consistency reliability was 0.942. The retest reliability coefficient of the general scale was 0.832. The correlation coefficients between self-evaluation and peer evaluation ranged from 0.342 to 0.517 in medium correlation. Conclusions The Patient Safety Nurse Competency Evaluation Scale demonstrates adequate reliability and validity. It is culturally appropriate for nurses self-evaluation and peer evaluation on patient safety competency.

Objective:To determine the effects of cytochrome P450 1A1 (CYP1A1) on nitric oxide (NO) production in lipopolysaccharide (LPS)-induced macrophages and the underlying mechanism.Methods:The peritoneal macrophages (PMs) were isolated from healthy C57BL/6 mice and stimulated with 10 mg/L LPS to establish inflammatory response model. The CYP1A1 mRNA and protein expressions in the cells were determined. The mouse macrophages RAW264.7 cell line with CYP1A1 overexpression (CYP1A1/RAW) were cultured in vitro, and they were stimulated by 10 mg/L LPS at logarithmic phase. The negative control-expressed RAW264.7 cells (NC/RAW) were established. The protein and mRNA expressions of activator protein-1 (AP-1) and inducible nitric oxide synthase (iNOS) in the cells as well as the content of NO in the cell supernatant were determined. The RAW264.7 cells were cultured in vitro, and they were stimulated by 10 mg/L LPS and 100 nmol/L 12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE] only or in combination at logarithmic phase. The blank control group was set up. The expression of iNOS mRNA in the cells and NO content in the cell supernatant were determined to observe whether the effect of CYP1A1 on LPS induced NO production in macrophages was related to 12(S)-HETE produced by metabolism. The RAW264.7 cells with CYP1A1 overexpression and hydroxylase activity mutation (CYP1A1m/RAW) were cultured in vitro, and they were stimulated by 10 mg/L LPS at logarithmic phase. The CYP1A1/RAW cell control group was set up. The iNOS mRNA expression in the cells and NO content in the cell supernatant were determined to observe the effect of hydroxylase activity of CYP1A1 in regulating NO production in macrophages. Results:Compared with the phosphate buffered saline (PBS) control group, the CYP1A1 mRNA expressions were elevated significantly from 2 hours after LPS stimulation and reached a peak at 12 hours [CYP1A1 mRNA (2 -ΔΔCt): 6.41±0.98 vs. 1.00±0.00, P < 0.05], while CYP1A1 protein expressions were increased from 6 hours after LPS stimulation and reached a peak at 24 hours, suggesting that CYP1A1 expression might be involved in LPS-induced macrophage over-activation. Compared with NC/RAW+LPS group, the iNOS mRNA expressions and NO contents both increased in CYP1A1/RAW+LPS group and reached a peak after 12 hours and 24 hours, respectively [12-hour iNOS mRNA (2 -ΔΔCt): 54.42±8.21 vs. 24.22±3.89, 24-hour NO (μmol/L): 66.52±4.09 vs. 41.42±2.09, both P < 0.05], while the iNOS protein expression and AP-1 phosphorylation also enhanced, suggesting that CYP1A1 might increase NO production by promoting AP-1 activation and iNOS expression. LPS and 12(S)-HETE stimulation only or in combination had no effect on iNOS mRNA expression and NO production, and no significant difference was found between the 12 (S)-HETE+LPS group and LPS group [12-hour iNOS mRNA (2 -ΔΔCt): 34.24±4.07 vs. 34.35±4.01, 24-hour NO (μmol/L): 44.02±3.14 vs. 44.56±3.21, both P > 0.05], suggesting that the regulation of CYP1A1 on NO production might not be induced by 12 (S)-HETE. There was no significant difference in the iNOS mRNA expressions or NO content between the CYP1A1m/RAW+LPS group and CYP1A1/RAW+LPS group [12-hour iNOS mRNA (2 -ΔΔCt): 52.11±6.84 vs. 50.21±5.19, 24-hour NO (μmol/L): 60.42±4.14 vs. 52.01±5.12, both P > 0.05], suggesting that CYP1A1 hydroxylase activity deficiency showed no effect on NO production. Conclusions:LPS stimulation significantly increases CYP1A1 expression in macrophages. CYP1A1 overexpression promotes NO production by activated macrophages through AP-1/iNOS pathway, while hydroxylase-deficiency or 12(S)-HETE has no effect on this regulation.

Severe trauma or massive deep burn can cause significant immunosuppression associated with sepsis and multiple organ failure. Dendritic cell (DC), as the professional antigen presenting cells and activating factor of immune response, plays an extraordinary role in initiating and regulating congenital and adaptive immune response. The quantity, functional changes, relevant molecular mechanisms and reverse measures of DC after trauma/burn were reviewed in order to intensively study the changes of DC after trauma/burn and provide a reference for exploring effective intervention measures for trauma/burn.

Objective To probe into the correlation among pricking wound related knowledge, attitude and behavior of clinical nurses, and provide reference for preventive strategy. Methods Multistage sampling method was used to choose 1 126 ward nurses in 8 level three general hospitals, and self designed pricking wound KAP scale was utilized to investigate. Results The correct rate of clinical nurses′pricking wound related knowledge was 86. 67%, the median of attitude score 4. 20 and average score of related behavior (3. 71 ± 0.50). The pricking wound related knowledge and attitude presented positive correlation with behavior (r =0. 210,0. 251;P <0. 01). Conclusions Clinical nurses master pricking wound related knowledge and have positive attitude, but preventive behavior is not ideal.

Objective:To explore the potent effects of denatonium benzoate(DB)on Mas-related G protein-coupled receptor-B2(MrgprB2)-mediated rat mast cell degranulation.Methods:RBL-2H3 cells were treated with DB overnight,before challenged with MrgprB2 ligands substance P(SP).The release of β-hex from MrgprB2-activated RBL-2H3 was detected by substrate method.Detec-tion of LTC4,IL-6,TNF-α and cPLA2 activity were performed by ELISA.The Ca2+influx and the expression of RBL-2H3 MrgprB2 re-ceptors were measured by fluorescence assay.Results:The results showed 10 μmol/L,50 μmol/L,80 μmol/L,100 μmol/L DB treat-ments promoted β-hex and LTC4 releases from activated RBL-2H3,accompanied by increased Ca2+mobilization and cPLA2 activa-tion.DB treatments did not affect IL-6 and TNF-α LTC4 releases in MrgprB2-activated RBL-2H3,as well as the levels of MrgprB2 ex-pression in mast cells.Conclusion:Taken together,DB enhanced the MrgprB2-mediated RBL-2H3 degranulation in vitro,probably by up-regulating Ca2+mobilization in activated cells.

Objective To evaluate the protective effect of pterostilbene against ultraviolet B (UVB)-induced acute damage in HaCaT cells,and to explore related mechanisms.Methods The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazo1ium (MTS) assay and flow cytometry were performed to estimate the proliferative activity and the apoptosis and necrosis rate of HaCaT cells treated with different concentrations of pterostilbene respectively,so as to screen the non-toxic concentration of pterostilbene.HaCaT cells were randomly divided into several groups:normal control group receiving no treatment,UVB group irradiated with 57 mJ/cm2 UVB,3 pterostilbene groups treated with 2.44,4.88 and 9.75 μmol/L pterostilbene respectively for 24 hours,3 pterostilbene + UVB groups treated with 2.44,4.88 and 9.75 μmol/L pterostilbene respectively for 24 hours followed by UVB radiation.Western blot analysis was conducted to detect changes of the transcription factor NF-E2-related factor 2 (Nrf2) expression in cell nuclei and cytoplasm before and after the treatment with pterostilbene and UVB,quantitative PCR to determine the mRNA expression of catalase and superoxide dismutase in the HaCaT cells,and enzyme-linked immunosorbent assay (ELISA) to evaluate the activity of catalase and superoxide dismutase.Results MTS assay and flow cytometry showed that 2.44,4.88 and 9.75 μmol/L pterostilbene had non-toxic effect on HaCaT cells.The protein expression of Nrf2 in the nuclei and cytoplasm in the normal control group was 1.03 ± 0.08 and 1.04 ± 0.11 respectively.Compared with the normal control group,the protein expression of Nrf2 in the nuclei and cytoplasm experienced no significant changes in the 2.44-,4.88-and 9.75-μmol/L pterostilbene groups,and the UVB group showed similar protein expression of Nrf2 in the cytoplasm,but significantly increased protein expression of Nrf2 in the nuclei (1.77 ± 0.08,q =17.24,P ＜ 0.01).Compared with the normal control group and UVB group,the 2.44-,4.88-and 9.75-μmol/L pterostilbene + UVB groups all showed significantly lower protein expression of Nrf2 in the cytoplasm (0.86 ± 0.10,0.87 ± 0.11 and 0.46 ± 0.11 respectively,all P ＜ 0.05),but significantly higher protein expression of Nrf2 in the nuclei (2.38 ± 0.11,2.57 ± 0.11 and 2.07 ± 0.13,all P ＜ 0.01).As qPCR showed,UVB radiation could significantly inhibit the mRNA expression of CAT (P ＜ 0.05),but had no obvious effect on the mRNA expression of SOD (P ＞ 0.05).The mRNA expression of CAT and SOD experienced no significant changes in the 2.44-,4.88-and 9.75-μmol/L pterostilbene groups compared with the normal control group (P ＞ 0.05).However,2.44,4.88 and 9.75 μmol/L pterostilbene could significantly reduce the inhibitory effect of UVB radiation on the mRNA expression of CAT (P ＜ 0.05) and up-regulate the mRNA expression of SOD in the pterostilbene + UVB groups (P ＜ 0.05).ELISA revealed that UVB radiation could inhibit the activity of CAT and SOD in the HaCaT cells (both P ＜ 0.001),while 2.44,4.88 and 9.75 μmol/L pterostilbene could reduce the inhibitory effect of UVB radiation on the activity of CAT and SOD (all P ＜ 0.05).However,the activity of CAT and SOD were still lower in the 2.44-,4.88-and 9.75-μmol/L pterostilbene + UVB groups than in the normal control group (P ＜ 0.05).Conclusion Pterostilbene can prevent UVB-induced acute damage in HaCaT cells by activating the Nrf2 pathway and up-regulating the expression of the downstream antioxidant enzymes CAT and SOD.