Objective:To establish a rapid method to detect drug-resistance genotypes of extended spectrum-β-Lactamases (ESBLs) produced by gram negative bacillus using the real -time fluorescence quantitative PCR. Methods: According to clinical common genotypes of ESBLs, SHV, TEM.CTX-M.OXA and their homology, 9 pairs of specific primers were designed including SHV, TEM, CTX-M-1, CTX-M-2, CTX-M-8, CTX-M-9, OXA-1, OXA-2 and OXA-10. To extract DNA template by boiling assay, and then establish and grade up SYBR GREEN I real-time fluorescence quantitative PCR reaction system, finally definite real-time fluorescence quantitative PCR method. Its precision and range of linearity were tested. With established assay 51 multi- drug resistant ESBLs- E. coli K. pneumoniae were detected and compared with improved three dimensional extract tests. Results: Except OXA-2, 8 genotypes SHV, TEM, CTX-M-1, CTX-M-2, CTX-M-8, CTX-M-9, OXA-1 and OXA-10 were amplified by quantitative PCR from 39 ESBLs+ and 51 multi-drug resistant ESBLs-E. coli K. pneumoniae and confirmed by sequence testing. The range of linearity was 3×10~3-3×10~8 copies/mL, r =-0.994 7. Repetitive experiments showed that the average coefficient of variation between -runs was 9.6%. Comparing with three dimensional extract test, there was no significant difference (χ2 = 1.125,P> 0.05). Conclusion: Testing drug-resistance genotypes of ESBLs with SYBR GREEN I real-time fluorescence quantitative PCR is a rapid,specific and sensitive method, which is capable of inspecting genotypes of ESBLs from clinical strains.

The regulations of unregistered herbal medicine had important changes in the UK after the EU Directive 2004/24/EC promulgation. British Drugs Act was amended to allow the sale of unregistered herbal medicine by registered practitioners, however, the regulations about the register of parishioners has not been issued yet. At the same time, the unregistered herbal medicine can be sold during the transitional period. But as problems about unregistered herbal medicine security appeared frequently, MHRA decided that the deadline for transitional period was April 30, 2014 through the consulting from all sectors of society. Therefore, the TCM practitioners in the UK cannot sell, and even have no right to sell unregistered herbal medicine after this date, which will bring a huge obstacle on the TCM development in the UK.

Traditional Chinese medicine (TCM) developed rapidly in the world, but the TCM international standard system has not been established, affecting the healthy development of TCM. To develop TCM international standardization strategy must master the current technical legislation and regulation status related to TCM. Through TM/CAM legislative investigation of the 4 Participating Countries of ISO/TC249 in western Pacific region, the author found that:Member states already have national policy on TM/CAM; laws and regulations on herbal product production, marketing, regulatory and management practitioners have already been enacted;TM/CAM practitioners in larger quantities, a higher education was required for practitioners and standardized management. This indicated that there is a good environment for TCM development in western Pacific region.

The widespread application of traditional medicine has attracted increasing attention. To promote the security of traditional medicine, WHO has established a series of global policies and strategies with consistency since 1960s. It also formulated guidelines for harvesting, production and operation, and quality control of botanical medicine according to the development of traditional medicine in different times. Thanks to the technical support of WHO, member states have established technical regulations for traditional medicine according to their own situations, and made significant achievements in the promotion of traditional Chinese security.

Objective To evaluate the role of pretreatment with docosahexaenoic acid (DHA) in the protective effect of angiopoietin-1 (Ang-1) on rat brain microvascular endothelial cells (BMVECs) during oxygen glucose deprivation (OGD).Methods BMVECs were sub-cultured in vitro and divided with a random number table into 7 groups:normal control group,normal control + Ang-1 group,OGD group,OGD + Ang-1 group,OGD + DHA group,OGD + DHA + Ang-1 group,and OGD + DHA + GW9662 + Ang-1 group.The normal control and normal control + Ang-1 groups were cultured in DMEM containing serum,5 mmol/L glucose,and 1.25 mmol/L pyruvate;OGD groups were cultured in glucose-and serum-free DMEM.DHA (40 μmol/L) was added to OGD + DHA,OGD + DHA + Ang-1,and OGD + DHA + GW9662 + Ang-1 groups,and 5 μmol/LGW9662 [inhibitor of peroxisome proliferator-activated receptor gamma (PPAR-γ)] to OGD + DHA + GW9662 +Ang-1 group,before pretreatment for 1 hour in 5％ CO2 and 95％ air.After the pretreatment,Ang-1 (250 ng/ml)was added to normal control + Ang-1,OGD + Ang-1,OGD + DHA + Ang-1,and OGD + DHA + GW9662 +Ang-1 groups.The cells were cultured in 94％ N2∶ 5％ CO2∶ 1％ O2 for 24 hours,except for normal control and normal control + Ang-1 groups,which were cultured in 5％ CO2 and 95％ air instead.The cell apoptosis rate was detected with flow cytometry,expressions of Bax,Bcl-2,caspase-3,Tie-1,Tie-2,pTie-2,pAkt,ZO-1proteins with Western blot.Results Compared with normal control group,the cell apoptosis rate in OGD,OGD + Ang-1,OGD + DHA,OGD + DHA + Ang-1,and OGD + DHA + GW9662 + Ang-1 groups were significantly increased (P =0.000,0.000,0.000,0.004,0.000);the expression levels of Bax,caspase-3,and Tie-1 were significantly enhanced (Bax:0.62 ± 0.03,0.38 ± 0.03,0.45 ± 0.03,0.26 ± 0.02,0.33 ± 0.02vs.0.16 ±0.01;caspase-3:0.76 ±0.05,0.42 ±0.04,0.52 ±0.02,0.32 ±0.02,0.40 ±0.02 vs.0.15 ±0.01;Tie-1:0.51 ±0.03,0.25 ±0.01,0.33 ±0.02,0.16±0.01,0.22±0.02 vs.0.12 ±0.01;all P=0.000);the expression levels of Bcl-2,Bcl-2/Bax,Tie-2,and pTie-2 were significantly decreased [Bcl-2:0.09±0.01,0.20±0.01,0.16±0.02,0.31±0.01,0.22±0.01 vs.0.34±0.01;Bcl-2/Bax:(14.93±1.86)％,(68.03±5.56)％,(36.93 ±2.22)％,(119.1 ±13.3)％,(64.23 ±6.07)％ vs.(208.33 ±7.37)％;Tie-2:0.07±0.01,0.16±0.02,0.11 ±0.01,0.21±0.01,0.18±0.01 vs.0.26±0.01;pTie-2:0.05±0.01,0.15 ±0.01,0.07 ±0.01,0.22 ±0.02,0.16±0.01 vs.0.27 ±0.01;allP=0.000],in addition,pAkt and ZO-1 in OGD,OGD + Ang-1,OGD + DHA,and OGD + DHA + GW9662 +Ang-1 groups were also significantly reduced (pAkt:0.13 ±0.01,0.26 ±0.01,0.14 ±0.01,0.28 ±0.02vs.0.39±0.02;ZO-1:0.08±0.01,0.18±0.01,0.10±0.01,0.19±0.01vs.0.23±0.02;allP=0.000).Compared with the OGD group,OGD + Ang-1 and OGD + DHA + Ang-1 groups demonstrated significantly reduced cell apoptosis (both P =0.000).Compared with normal control + Ang-1 group,the expression levels of pTie-2,pAkt,and ZO-1 were significantly decreased in OGD + Ang-1 and OGD + DHA +Ang-1 groups (pTie-2:0.15 ±0.01,0.22 ±0.02 vs.0.52 ±0.02;pAkt:0.26 ±0.01,0.37 ±0.04 vs.0.67 ± 0.05;ZO-1:0.18 ± 0.01,0.24 ± 0.02 vs.0.39 ± 0.05;all P =0.000).Compared with OGD +Ang-1 group,OGD + DHA + Ang-1 group had significantly decreased cell apoptosis and expression levels of Bax,caspase-3 and Tie-1,and significantly increased expressions of Bcl-2,Bcl-2/Bax,Tie-2,pTie-2,pAkt,and ZO-1 (all P =0.000),while OGD + DHA + GW9662 + Ang-1 group showed no significant differences in these indexes (P =0.202,0.770,0.382,0.448,0.233,0.736,0.143,0.526,0.495,0.670).Conclusion Pretreatment with DHA may enhance the protective effect of Ang-1 on rat BMVECs under the condition of OGD,possibly via activating PPAR-γ,suppressing Tie-1 expression,and hence amplifying the activation of Tie-2.

OBJECTIVETo investigate the role of endogenous hydrogen sulfide (H2S) in erectile dysfunction (ED) induced by androgen deficiency.

METHODSWe randomly divided 30 eight-week-old healthy male SD rats into six groups: 2-week control (A), 4-week control (B), 2-week castration (C), 4-week castration (D), 2-week castration + androgen replacement (E), and 4-week castration + androgen replacement (F), those in groups E and F subcutaneously injected with testosterone propionate (TP) at the physiological dose of 3 mg/kg per day after castration, while those in the other groups with isodose oil instead. At 2 and 4 weeks after operation, we determined the level of serum testosterone (T) , intracavernous pressure (ICP) , mean carotid arterial pressure (MAP) of the rats, measured the concentration of H2S in the plasma and corpus cavernosum tissue, and detected the expressions of cystathionine-P3-synthase (CBS) and cystathionine-gamma-lyase (CSE) by immunohistochemistry and Western blot.

RESULTSThe serum T level was significantly lower in group C ([0.63 +/- 0.15] nmol/L) than in A ( [ 16.55 +/- 4.17] nmol/L) and E ( [ 18.99 +/- 4.62] nmol/L) (P <0.05), as well as in group D ([0.70 +/-0.22] nmol/L) than in B ([15.44 +/-5.18] nmol/L) and F ([20.99 +/-6.41] nmol/L) (P <0. 05) , and so were ICP/MAP after 5 and 7 V electrical stimulation of the pelvic ganglia (P <0. 05) , H2 S concentration (P <0.05), and the expressions of CBS and CSE (P <0.05). The expressions of CBS and CSE proteins were also significantly decreased in group C as compared with D (P <0.05).

CONCLUSIONThe reduced expressions of CBS and CSE may inhibit the H2 S signaling pathway, which might be one of the mechanisms underlying androgen deficiency-induced ED in rats.

Androgens ; deficiency ; Animals ; Cystathionine beta-Synthase ; metabolism ; Cystathionine gamma-Lyase ; metabolism ; Erectile Dysfunction ; metabolism ; Hydrogen Sulfide ; metabolism ; Male ; Orchiectomy ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To investigate current status of knowledge-attitude-behavior about unplanned extubation (UEX) of central venous catheter among infusion nurses in Guangdong Province.Methods The nurses who attended the 2015 intravenous therapy nursing training provided by Guangdong Nursing Association were selected as participants.A self-designed questionnaire about nurses' knowledge-attitude-behavior about UEX of central venous catheter was used to investigate 450 nurses.Results The proportion of nurses who reached good level in knowledge were 41.1％,nurses who had positive attitude were 71.0％,nurses who had positive behaviors were 30.0％.There were significant differences among nurses with different educational background,different titles and different hospital ranks(P＜0.05).Conclusion The knowledge-attitude-behavior of nurses are not satisfactory,especially those who are junior or have lower education or work in non-tertiary hospital.

Objective To investigate the diagnosis and treatment of visceral artery aneurysms (VAAs).Methods From June 2008 to December 2013,20 patients (11 male and 9 female) with visceral artery aneurysms were treated in our hospital.The clinical data and treatment outcomes were retrospectively analyzed.Results There were 20 patients,including 11 males and 9 females.The site of aneurysmal disease was splenic artery in 12 cases,mesenteric artery in 2 cases,hepatic artery in 2 cases,renal artery in 2 cases,celiac artery in 1 case and gastroduodenal artery 1 case.10 patients were treated by open surgery,and other 10 patients treated by endovascular.1 patient died,and others were discharged.All patients were followed up 28.5 months (ranged from 10 to 76 months).Two patients died due to cirrhosis of the liver,and the remaining patients were alive,no treatment-related complications,no serious complications,such as aneurysm recurrence occurs.Conclusions Both traditional surgical and endovascular treatment of visceral artery aneurysms may obtain a satisfactory outcome.

Inflammatory bowel disease is thought to be regulated by the balance between Th1 and Th2 cytokines secreted by T cells, and NF-kappaB p65 also plays a predominant role in the intestinal inflammation. We evaluated the potency of oxymatrine, one of active components of Sophora Root, in inhibiting the immune responses and inflammation in 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. The inflammation was markedly ameliorated in the oxymatrine-treated rats. The level of IL-2 was increased and that of IL-10 was decreased in colon tissue in the rat model, which was reversed by the treatment of oxymatrine. Moreover, the elevated expression of NF-kappaB p65 in colon tissue in the model was also improved by oxymatrine treatment. Our results suggest that oxymatrine might be beneficial for the abnormal immune responses and inflammation by regulating the unbalance of Th1 and Th2 cytokines secretion and inhibiting the expression of NF-kappaB p65 in colon tissue.

Objective To investigate the applications of antibacterial agents to outpatients in primary hos -pitals in Hefei City of Anhui Province,and to provide reference for rational use of antibacterial agents.Methods In 2011, fourty-five primary hospitals in Hefei City were selected randomly ,including urban community health service centers (Group A) and township hospitals(Group B),and thirty or fourty outpatient prescriptions were analyzed monthly . Results In Group A, the percentage and intensity of antimicrobial usage , the proportion of the combination and injectable formulation were ( 45.36 ±20.02 )%, ( 89.73 ±25.50 ) DDDs · ( 100 cases ) -1 · d-1 , 13.34%, 23.16%,respectively,and the data in Group B were (61.36 ±17.18)%,(108.46 ±32.27)DDDs· (100 cases) -1 · d-1,29.13%,46.39%,respectively,which the former were significantly lower than the latter.Conclusion In primary hospitals,the applications of antibacterial agents to outpatiants are not rataional ,including high percentages of usage and unreasonable selection of species ,and more supervision and training need to be given to the medical staff , especially in township hospitals .