Objective To investigate the correlation between asymmetric dimethylarginine (ADMA) and endothelial dysfunction in patients with type 2 diabetic nephropathy.Methods The 81 patients with type 2 diabetic nephropathy were selected.According to the urinary albumin excretion rate (UAER),the patients were divided into micro-albuminuria group (DN1 group,UAER 21-199 mg/24 h,38 cases) and macro-albuminuria group (DN2 group,UAER ≥ 200 mg/24 h,43 cases).The 20 healthy people were defined as control group.Intimal-media thickness and endothelial dysfunction of the radial artery of right forearm were detected by color Doppler ultrasound.The serum level of ADMA was determined by enzyme-linked immunosorbent assay.Results There was no significant difference in radial artery inner diameter intimal-media thickness among the 3 groups (P ＞ 0.05).The Endothelial dependent diastolic function (EDD) and endothelial independent diastolic function (EID) in DN1 group and DN2 group were significantly lower than those in control group [(10.45 ± 2.58)％ and (7.56 ± 2.17)％ vs.(15.72 ± 3.05)％,(15.42 ± 2.71)％ and (15.37 ± 2.92)％ vs.(19.31 ± 3.76)％,P ＜ 0.05],and the EDD in DN2 group was significantly lower than that in DN1 group (P＜ 0.05).The serum ADMA in DN1 group and DN2 group was significantly higher than that in control group [(0.63 ± 0.08) and (0.92 ± 0.12) μ mol/L vs.(0.39 ± 0.05)μmol/L,P ＜0.05],and in DN2 group it was significantly higher than that in DN1 group (P ＜0.05).In patients with type 2 diabetic nephropathy,the serum ADMA and EDD had negative correlation (r =-0.81,P =0.020),but the serum ADMA and C reactive protein had positive correlation (r =0.75,P =0.034).Conclusions The serum level of ADMA is significantly increased in patients with type 2 diabetic nephropathy.There is a close correlation between ADMA and endothelial dysfunction of artery.

Objective To investigate the effects of curcumin on mRNA expression of cytokines related to Toll-like receptor (TLR) 4 signaling in THP-1 cells.Methods After pretreatment with different concentrations (50,25,12.5 mg/L) of curcumin or dexamethasone for 12 hours,THP-1 cells were stimulated by lipopolysaccharide (LPS.1 mg/L) for 4 hours followed by the collection of cells.Then total RNA was isolated from these cells and subjected to reverse transcription-polymerase chain reaction (RT-PCR) for the detection of mRNA expression of tumor necrosis factor receptor-associated factor 6 (TRAF6),interleukin-1 receptorassociated kinase (IRAK1) and nuclear factor (NF)-κB.THP-1 cells without pretreatment or stimulation served as negative control,and those only stimulated with LPS served as LPS group.Results After stimulation with LPS (1 mg/L) for 4 hours,the mRNA expressions of TRAF6,IRAK1 and NF-κB were significantly upregulated in THP-1 cells compared with negative control cells (f=38.69,39.13,23.99,all P<0.01).Curcumin of 50 mg/L and 25 mg/L significantly inhibited the mRNA expressions of TRAF6.IRAK 1 and NF-κB upregulated by LPS with an inhibition rate of more than 50% (all P<0.0 1).Conclusions Certain concentrations of curcumin can inhibit the mRNA expressions of TRAF6.IRAK1 and NF-κB.which demonstrates the regulatory effect of curcumin on the mRNA expressions of TLR4 signaling pathway-associated cytokines.

BACKGROUND: Type Ⅰ collagen is a specific collagen secreted by in vitro cultured osteoblast, and the formed network is the basis of bone mineralization, which also reflects the ability of osteoblast bone formation. Studies have shown astragalus root increased osteoblast proliferation. However, the effect of astragalus root on improving type Ⅰ collagen expression of osteoblast remains poorly understood.OBJECTIVE: To evaluate the effect of astragalus root injection on the abilities of rat cranium-derived osteoblast proliferation and type Ⅰ collagen expression.METHODS: Rat osteoblast was cultured in vitro and divided into control group (MEM culture solution containing calf serum) and astragalus root groups (different concentrations). The effect on osteoblast proliferation was evaluated on days 1, 3, 5, 7, and 9 by MTT method. Moreover, the expression of type Ⅰ collagen protein was observed after 6 hours of treatment with astragalus root injection using in cell western-blot method. In addition, the gene expression of COLLal was investigated by real-time PCR method.RESULTS AND CONCLUSION: From days 3 to 9, the different concentrations of astragalus root injection improved osteoblast proliferation, respectively compared with control group (P ＜ 0.05), and this ascending trend peaked on day 7. Different concentretions of astragalus root injection improved COLLol mRNA expression, especially 15% astragalus root injection was the most effective. The type Ⅰ collagen protein expression of 15% and 10% astragalus root injection were significantly greater compared with the control group (P ＜ 0.05). Astragalus root injection improved in vitro cultured osteoblast proliferation and type Ⅰ collagen secretion in a certain dose-effect manner.

Objective To develop a dual real-time fluorescent RT-PCR method for rapid detection of enterovirus(EV)and en terovirus type 71(EV71).Methods Specific primers and probes were designed and the dual real-time fluorescent RT-PCR reaction system was established.The quantitative standard curve was drawn;its sensitivity and precision were evaluated.Feces and throat swab specimens of 109 clinical patients with hand foot and mouth disease were collected and tested by using this method.Then the obtained results were compared with those detected by commercial EV71 PCR kit.Results The relative coefficient(2)of EV and EV71 standard curve established by the dual real-time fluorescent RT-PCR method were both 0.998.Its sensitivity reached 0.5 TCID50/mL for detecting EV and 0.05 TCID50/mL for detecting EV71.The within-run precision for detecting EV and EV71 was ＜3％ and total precision≤4％.The results showed good specificity for the detection of enterovirus and non-enterovirus.In 109 detected clinical samples,84 cases of EV positive samples were detected,in which 56 cases were EV71 positive with the total positive rate of 51.4 ％,which was consistent with the result of simple fluorescent RT-PCR commercialization kit(P=1.000).Conclusion The established dual real-time fluorescent RT-PCR method has high sensitivity and good stability,which has an important significance for early high throughput rapid diagnosis of hand foot and mouth disease.

Objective To explore the types and drug resistance of pathogens in patients with type Ⅰ incision surgical site infection in orthopedics department.Methods Patients with type Ⅰ incision surgical site infection in orthopedics department at Peking University Third Hospital from January 2005 to December 2013 were retrospectively collected.Clinical characteristics of patients,distribution and drug resistance of pathogens were analyzed.Results A total of 58.2 thousands patients with type Ⅰ incision surgical site were hospitalized from January 2005 to December 2013 in orthopedics department,and among them 442 patients had infection in the type Ⅰ incision surgical site.The infection rate was 0.8％.Infection was mainly observed in elderly patients.The most common diseases were lumbar canal stenosis (21.7％),cervical spondylosis (20.6％) and lumbar intervertebral disc herniation (14.0％).A total of 453 pathogenic strains were detected,of which 52.9％ were gram-positive bacteria,45.5％ were gramnegative bacteria and 1.6 ％ were fungi.The common pathogens were Staphylococcus aureus (25.2 ％),Staphylococcus epidermidis (14.1 ％),Escherichia coli (11.5 ％),Enterobacter cloacae (7.3 ％),Pseudomonas aeruginosa (6.2 ％) and Acinetobacter baumannii (6.0 ％).The percentage of Meticillinresistant Staphylococcus aureus (MRSA) was 23.7％ and the percentage of Meticillin-resistant Staphylococcus epidermidis (MRSE) was 43.8％.Vancomycin or linezolid-resistant Staphylococcus aureus or Staphylococcus epidermidis were not detected.Proportion of extended-spectrum beta-lactamases (ESBL) producing strains in Escherichia coli was 53.8％,and proportion of ESBL-producing strains in Klebesiella pneumonia was 50.0％.The resistance rates to impenem and meropenem of the three different species in Enterobacteriaceae,including Escherichia coli,Enterobacter cloacae and Klebsiella pneumonia,were 0.Resistance rates of Pseudomonas aeruginosa to cefoperazone-sulbactam,piperacillin-tazobactam were less than 10 ％.Resistance rate of Acinetobacter baumannii to minocyline was 11.1％ and resistance rates of it to other drugs were more than 20％.Conclusions The rate of type Ⅰ incision surgical site infection in orthopedics department is low.Gram-positive and gram-negative bacteria each account for half of the pathogens.The proportion of resistant pathogens is high and empirical treatment is needed to cover these pathogens.

Objective:To establish the quality standard system of Gui Erbai gel based on a method ofa system to multiple evalu-ation and discuss the feasibility of the method used for the quality standard for traditional Chinese medicine. Methods:TLC identifi-cation of Gui Erbai gel was established by one thin layer system. An HPLC method was used to detect 6 active components in Gui Erbai gel. Results:Five active components in the gel could be identified by one thin layers system simultaneously with clear spots and good separation. Six active components in the gel could be determined by the same HPLC system with high accuracy. The average content of podophylotoxin,quercetin,kaempferol,imperatorin,dictamnine and rutin is as follows 0. 154,0. 052,0. 138,0. 051,0. 060,0. 048 mg· g-1 . RSD<3％. Conclusion:The established method based on a system to multiple evaluation can be used for the quality standard establishment for Gui Erbai gel with the properties of promising feasibility, simple operation, low cost, high accuracy and good stabili-ty.

Objective To discuss the coping methods of pediatric nurses with adverse psychology and behavior of parents with terminal children. Methods 198 parents who were assessed with adverse psychology and behavior with terminal children were chosen and randomly divided into the observation group(100 cases) and the control group(98 cases). The control group was given conventional care, while in the observation group, nursing intervention of humanistic care and reinforced health education were adopted in addition. The changes of adverse psychology and behavior of the two groups were observed. Χ2 test was used. Results The adverse psychology and behavior in the observation group was reduced significantly than that of the control group after nursing intervention. Conclusions Nursing intervention can effec-tively mitigate the adverse psychology and behavior in parents with terminal children. It plays an active role beth on the terminal children and their parents.

Objective To investigate the effects of 13-hexyl-berbefine hydroehlofide (HB-13) and 13-hexyl-paimatine hydrochloride (HP-13) on the activation of nuclear factor-kappa B (NF-kB) and phosphorylation of p38 mitogen-activated protein kinase (MAPK) in a human keratinocyte cell line, HaCaT stimulated by tumor necrosis factor alpha (TNF-α). Methods HaCaT cells were cultured in the presence of various concentrations (0.39, 0.78, 1.56 μg/mL) of HB-13 or HP-13 for 120 minutes followed by the stimulation with recombinant human TNF-α for 120 minutes (in phosphorylatEd-IkB-α test) or 15 minutes (in phosphorylated-p38 test). Then, HaCaT cells were disrupted, total protein was extracted, and the expressions of phosphorylated I B-α and phosphorylated p38 were detected with Western blot. HaCaT cells receiving neither pretreatment nor stimulation served as blank control, untreated HaCaT cells stimulated by rhTNF-α as stimulator control, and HaCaT cells pretreated with turmeric root tuber and stimulated by rhTNF-α as positive control. Results From 0.39 to 1.56 μg/mL, both HB-13 and HP-13 significantly inhibited the expression of p-IkB-α in HaCaT cells stimulated by rhTNF-α, and a nonsignificant dose-dependent trend was observed for their inhibitory effect, with the ICo value being 0.441 μg/mL for I-IB-13 (r = -0.990, n = 3, P > 0.05) and 0.832 μg/mL for HP-13 (r = -0.992, n = 3, P > 0.05). In contrast, neither 1-113-13 nor HP-13 within the experiment concentration range had a significant effect on the expression of p-p38 in HaCaT cells stimulated by rhTNF-α (P > 0.05). Conclusions Within the experimental concentration range, both HB-13 and HP-13 can inhibit the activation of NF-kB in HaCaT cells induced by TNF-α signal, but neither of them suppress the phosphorylation of p38MAPK induced by TNF-α signal in HaCaT cells.

Objective To investigate the effects of triptolide on the expression of a series of proteins associated with interferon-γ (IFN-γ)signaling in HaCaT keratinocytes.Methods After pretreatment with difrerent dosages of triptolide(10-10-10-7 mol/L),HaCaT cells were stimulated by recombinant human IFN-γ(rhIFN-γ,500 U/mL)for various periods followed by the collection of cells.Then,total protein was extracted from these cells and subjected to Western blotting for the detection of expression of interferon-γ receptor α(IFN-γRα),phosphorylated Janus kinase 2(pJAK2)and suppressor of cytokine signaling (SOCS1).Results Triptolide at the concentrations of 10-8 mol/L and 10-7 mol/L significantly inhibited the IFN-γRα expression upregulated by rhIFN-γ(both P＜0.05).The expression of pJAK2 induced bv rhIFN-γ was also suppressed by triptolide at the concentrations of 10-9 moI/L and 10-8 mol/L(both P＜0.05).The inhibition of triptolide on IFN-γRα and pJAK2 expression was dose-dependent and the 50%inhibitory concentrations(IC50 value)were 1.37×10-8 mol/L and 2.83×10-9 mol/L,respectively.On the contrary,triptolide upregulated the expression of SOCS1 stimulated by rhIFN-γ at the concentrations of 10-10,10-9 and 10-8 mol/L(P＜0.05,0.05,0.01,respectively)with the 50%effective dosage(ED50 value)at 3.32 × 10-11 mol/L.Conclusions By inhibiting the expression of IFN-γRα as well as phosphorylation of JAK2 and upregulating the expression of SOCS1,triptolide inhibits the phosphorylation of STAT-1,resulting in the inhibition of genetic transcription of multiple inflammatory factors induced by IFN-γ signaling in HaCaT keratinocytes,and the inhibition probably contributes to the efficacy of triptolide in the treatment of IFN-γ-dependent inflammatory skin disorders,such as psoriasis.

Objective To investigate the role of protein kinase C (PKC) in induction of vascular endothelial growth factor (VEGF) secretion by isoflurane in primary cultured rat cardiomyocytes. Methods Primary cultured neonatal rat cardiomyocytes were randomly divided into 6 groups ( n = 6 each): control group (group C), 3 different concentration isoflurane groups (group Ⅰ1-3 ), PKC inhibitor calphostin C group (group P), and PKC inhibitor + isoflurane group (group PI). The cells were exposed to 0.7%, 1.4% and 2.1% isoflurane for6 h in group Ⅰ1-3 respectivly. Calphostin C was added to the culture medium with a final concentration of 50 nmol/L in group P. Calphostin C was added to the culture medium with a final concentration of 50 nmol/L, then the cells were exposed to 1.4% isoflurane for 6 h in group PI. VEGF concentrations and expression of PKC isoforms were determined by ELISA and Western blot respectively. Results Compared with group C, the VEGF concentration was significantly increased in group Ⅰ2 and Ⅰ3, and PKCε expression was down-regulated in the cytoplasm while upregulated in the cytomembrane in group Ⅰ2 ( P ＜ 0.01 ), but no significant change was found in the parameters mentioned above in group Ⅰ2 ( P ＞ 0.05). PKCα, PKCδ and PKCζ expression was significantly higher in the cytoplasm than in the cytomembrane in group C and Ⅰ2. VEGF concentrations were gradually increased with the increase in isoflurane concentrations ( P ＜ 0.05). VEGF concentrations were significantly lower in group PI than in Ⅰ2 ( P ＜0.05) .Conclusion Isoflurane induces VEGF secretion in primary cultured rat cardiomyocytes through translocation of PKCε from the cytoplasm to the cytomembrane, suggesting that it is a mechanism of the cardioprotective effects of isoflurane.