Objective To observe the effect of Wnt/β-catenin pathway on proliferation of human umbilical vein endothelial cell (HUVEC) using the glycogen synthase kinase 3β (GSK-3β)-targeting RNAi recombinant adenovirus vector. Methods Homologous recombination and cloning techniques were used to construct RNAi recombinant adenoviral expressive vectors specific to GSK-3β. Then, the adenovirus plasmids was transfected into HEK 293A cells to produce adenovirus and amplify the adenoviral stock. Plaque forming assay was used to titer the adenoviral stock. The GSK-3β and β-catenin protein expressions were detected by Western blot and immunohistochemistry. The proliferation of HUVEC was detected with MTr assay. Results The RNAi adenovirus vectors specific to GSK-3β were successfully produced with high titer. The expression of GSK-3β protein in HUVEC could be down-regulated efficiently by the RNAi adenovirus, along with increased β-catenin protein expression. The proliferation of HUVEC was significantly increased (P < 0.05 or P < 0.01) after infected with GSK-3β RNAi recombinant adenovirus for 3, 5, 7 days. Conclusion RNAi adenovirus is an important tool that can inhibit the expression of GSK-3β efficiently, along with increased β-catenin protein expression. Up-regulating of the Wnt/β-catenin pathway might play an important role in the proliferation of HUVEC.

Objective To understand the functional exercise situation of the total hip arthroplasty(THA)patients in hospital. Methods Using self-designed functional exercise in patients with TKA outline the structure of observation, observation and collection of Orthopedic Surgery,71 cases (91 hips)received THA in patients with relevant information. Results After the former 3d,22.5% of patients did not exercise;After 14 d continuous passive motion (CPM) ,hip joint exercises were separately accounted for 71.8% and 81.7% ,and static muscle contraction,straight leg raising were accounted for 40.8 percent and 81.7 percent. The four exercise started lately, training methods were not standardized the patients, insisted on a shorter time;single-and double-hip replacement hip replacement in patients with Shimoji routing practice started time respectively, after (6.51 ± 2.90 ) d and ( 10.30 ± 3.21 ) d. In the functional exercise process,CPM was assisted by nurse,in other exercises was assisted by retaining workers and patients families. Conclusion THA patients hospitalized during the actual exercise of the number,frequency,timing and methods vary, nursing staff should be further explored how to ensure that patients really effective rehabilitation exercise to further enhance the effect of patients with rehabilitation exercises.

OBJECTIVE:To provide reference for avoiding the recurrence of medical safety(adverse)events or inducing more severe medical accident. METHODS:The medical safety(adverse)events reported by a class A grade 3 hospital from July 2013 to March 2016 were studied retrospectively,and excluded purely adverse drug reactions and transfusion reactions to analyze the rea-son and characteristics. RESULTS:Of 747 events,there were 578 medical,nursing and drug-related medical safety (adverse) events,accounting for 77.38%;among 175 medical related medical safety(adverse)events,top 3 reasons were basic skills defi-ciencies,defects in the system processes,inadequate communication and education, accounting for 32.00%,24.00%,19.43%. The occurrence of medical safety (adverse) events were closely related to working life and qualifications;among 578 medical, nursing and drug-related medical safety (adverse) events,the incidence of adverse events was 71.11% in 5-year working life group,and that was 73.18% in junior titles group. CONCLUSIONS:Various professionals should strictly implement procedure and regulations,strengthen communication and education in daily work,and improve professional knowledge and skills so as to prevent the occurrence of adverse event completely.

OBJECTIVE:To establish the quality standard of Flemingia philippinensis. METHODS:The properties and micro-scopic characteristics were observed;TLC was adopted for qualitative identification of genistin and genistein;the moisture,total ash and extract were detected;HPLC was adopted for contents determination of genistin and genistein:the column was Thermo BE-TASIL C18 with mobile phase of Acetonitrile-0.5% glacial acetic acid(gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 261 nm,column temperature was 25℃,and the injection volume was 10μl. RESULTS:The properties and micro-scopic characteristics of F. philippinensis and F. macrophylla showed strong specificity. TLC spots of genistin and genistein we- re clear and well separated,with no interference in negative control. The moisture was 3.69%-8.37%,total ash was 1.72%-6.74%,and extract was 5.89%-19.65%. The linear range was 0.012 9-2.588 μg for genistin(r=0.999 9)and 0.004 6-0.923 2 μg for genistein (r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 3.0%;recoveries were 99.63%-101.87%(RSD=0.82%,n=6)and 97.19%-100.34%(RSD=1.23%,n=6). CONCLUSIONS:The method is simple,accurate and specif-ic,and can be used for the quality control of F. philippinensis.

OBJECTIVE:To discuss clinical efficacy and safety of leuprolide acetate in the treatment of endometriosis. METH-ODS:66 endometriosis patients were divided into observation group(37 cases)and control group(29 cases)according to random number table method. Observation group was treated with Leuprolide acetate microsphere for injection,3.75 mg/time,on the upper arm,abdomen or buttocks since the first-fifth day of menstrual cycle(administration time was available at hysterectomized patients disposal),and then every 4 weeks;one administration was recognized as a treatment course. Control group was treated with Ethi-nyl estradiol cyproterone tablet orally,1 tablets per day for 3 weeks,one week drug withdrawal,as a treatment course. 2 groups were given 3-5 courses of treatment according to their tolerance or the improvement of clinical symptoms. The improvement of clini-cal symptoms were compared between 2 groups,and the levels of FSH,LH and E2 were compared before and after treatment;the occurrence of ADR and the time of the return of menses were recorded. RESULTS:2 patients in observation group and 1 patient in control group withdrew from the test. After treatment,total effective rate was 97.14% in observation group,which was significant-ly higher than 75.00% in control group,with statistical significance (P<0.05). Before treatment,there was no statistical signifi-cance in the levels of FSH,LH and E2 between 2 groups (P>0.05). After treatment,the levels of FSH,LH and E2 were de-creased significantly,and the observation group was more significant than the control group,with statistical significance (P<0.05). Total incidence of ADR was 20.00% in observation group,compared to control group (21.42%),there was no statistical significance (P>0.05). The time of the return of menses in observation group was (89.75 ± 3.34) d after drug withdrawal,com-pared to control group [(88.46±2.94)d],there was no statistical significance(P>0.05). CONCLUSIONS:Leuprolide acetate can effectively improve clinical symptoms of patients with endometriosis,and reduce the levels of FSH,LH and E2 with slight ADR, and it doesn’t influence the time of the return of menses.

Objective To evaluate the effect of dexmedetomidine and small dose of ketamine on the expression of P2X4 receptor (P2X4 R) mRNA and P2X7 receptor (P2X7R) mRNA in the dorsal root ganglion of rats with neuropathic pain.Methods Ninety male Sprague-Dawley rats,aged 6-9 weeks,weighing 180-220 g,were randomly divided into 5 groups (n =18 each):sham group (group S),chronic constrictive injury group (group CCI),dexmedetomidine group (group D),ketamine group (group K) and dexmedetomidine + ketamine group (group DK).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 400 mg/kg.Neuropathic pain was induced by CCI in CCI,D,K and DK groups.The sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1mmintervals with 4-0 silk thread.In group S,the sciatic nerves were only exposed but not ligated.In D,K and DK groups,dexmedetomidine 50μg/kg,ketamine 10 mg/kg and dexmedetomidine 25μg/kg + ketamine 5 mg/kg were injected intraperitoneally,respectively,while the equal volume of normal saline was injected in S and CCI groups,once a day for 14 consecutive days after CCI.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before CCI,and 3,7 and 14 days after CCI.Six animals were sacrificed after measurement of pain threshold at 3,7 and 14 days after CCI and the lumbar segments (L4-6) of the dorsal root ganglion were removed for determination of P2X4 R mRNA and P2X7 R mRNA expression by RT-PCR.Results Compared with group S,MWT and TWL were significantly decreased at 3,7 and 14 days after CCI in groups CCI,D,K and DK,the expression of P2X4R mRNA and P2X7R mRNA was up-regulated at 3,7 and 14 days after CCI in groups CCI,D and K,and the expression of P2X4 R mRNA and P2X7 R mRNA was up-regulated at 3 and 7 days after CCI in group DK (P ＜ 0.05).Compared with group CCI,TWL and MWT were significantly increased and the expression of P2X4 R mRNA and P2X7 R mRNA was down-regulated at 3,7 and 14 days after CCI in groups D,K and DK (P ＜ 0.05).Compared with D and K groups,TWL and MWT were significantly increased and the expression of P2X4 R mRNA and P2X7 R mRNA was down-regulated at 3,7 and 14 days after CCI in group DK (P ＜ 0.05).Conclusion The mechanism by which the combination of dexmedetomidine and small dose of ketamine produces a synergistic antinociception in rats with neuropathic pain may be related to down-regulation of the expression of P2X4 R mRNA and P2X7 R mRNA.

Objective To evaluate the effects of dexmedetomidine on the activity of cAMP response element binding protein (CREB) and c-fos in the spinal dorsal horn in a rat model of neuropathic pain.Methods Fifty-four adult male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly divided into 3 groups (n =18 each):sham operation group (group S),chronic neuropathic pain group (group C) and dexmedetomidine group (group D).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 350 mg/kg.The sciatic nerve was exposed and 4 ligatures were placed on the right sciatic nerve at 1 mm intervals with 4-0 silk thread in C and D groups.In group D,dexmedetomidine 50 μg/kg was injected intraperitoneally once a day starting from the end of operation until 1 day before the animals were sacrificed,while the equal volme of normal saline was injected instead of dexmedetomidine in S and C groups.Paw withdrawal threshold to mechanical stimulation with yon Frey filament (MWT) and paw withdrawal latency to thermal stimulation (TWL) were measured on 1 day before operation and 3,7 and 14 days after operation.The animals were sacrificed after measurement of MWT and TWL.Their lumbar segments (L4-6) of the spinal cord were removed for measurement of the expression of phosphorylated CREB (pCREB) and c-fos by immunohistochemistry.Results Compared with group S,MWT was significantly decreased,TWL was shortened,and the expression of pCREB and c-fos was up-regulated on 3,7 and 14 days after operation in C and D groups (P ＜ 0.05).Compared with group C,MWT was significantly increased,TWL was prolonged,and the expression of pCREB and c-fos was down-regulated on 3,7 and 14 days after operation in group D (P ＜ 0.05).MWT was significantly lower,and TWL was shorter on 3,7 and 14 days after operation than on 1 day before operation in C and D groups (P ＜ 0.05).MWT was significantly lower,TWL was shorter,and the expression of pCREB and c-fos was higher on 7 and 14 days after operation than on 3 days after operation in C and D groups (P ＜ 0.05).MWT was significantly higher,TWL was longer,and the expression of pCREB and c-fos was lower on 14 days after operation than on 7 days after operation in C and D groups (P ＜ 0.05).Conclusion The mechanism by which dexmedetomidine reduces neuropathic pain is related to inhibition of the activity of CREB and c-fos in the spinal dorsal horn of rats.

Objective To investigate the effects of Sulfotanshinone Sodium Injection (SSI) on neuropathic pain in rats.Methods One hundred and eight adult male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly divided into 3 groups (n =36 each):sham operation group (group S) ; chronic constrictive injury (CCI)group; group SSI.The animals were anesthetized with intraperitoneal 10% chloral hydrate 350 mg/kg.In groups CCI and SSI,4 ligatures were placed on the right sciatic nerve at 1 mm intervals with 4-0 silk thread according to the method described by Bennett et al.In group S,the right sciatic nerves were exposed,but not ligated.In group SSI,SSI 25 mg/kg was injected intraperitoneally once a day starting from the end of operation until one day before the animals were sacrificed,while the rats received the equal volume of normal saline (5 ml/kg) instead of SSI in groups S and CCI.Twelve animals in each group were chosen at 1 day before operation and 3,7 and 14 days after CCI (T1-4) to measure mechanical paw withdrawal threshold to yon Frey stimuli (MWT) and paw withdrawal latency to thermal nociceptive stimulus (TWL).Six rats in each group were sacrificed at T2-4 after measurement of pain threshold,and their lumbar segnents (L4-6) of the spinal cord were immediately removed for determination of Bcl2 and caspase-3 expression in spinal dorsal horn (by immune-histochemistry),and MDA content and SOD activity (by spectrophotometry) in spinal cord.Results Compared with group S,PWT was significantly decreased,PWL was shortened,the expression of Bcl-2 and caspase-3 was up-regulated,MDA content was increased and SOD activity was decreased at T2-4 in groups CCI and SSI (P ＜ 0.05).Compared with group CNP,PWT was significantly increased,PWL was prolonged,the expression of Bcl-2 was up-regulated,the expression of caspase-3 was downregulated,MDA content was decreased and SOD activity was increased at T2-4 in group SSI (P ＜ 0.05).Conclusion SSI can mitigate neuropathic pain in rats and inhibition of oxidative stress in spinal cord tissues and reduction of apoptosis in spinal dorsal horn neurons are involved in the mechanism.

Objective To evaluate the effect of dexmedetornidine on the expression of Toll-like receptor 4 (TLR4) and nuclear factor kappa B (NF-κB) in the spinal cord in rats with neuropathic pain (NP).Methods One hundred and eight male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly assigned into 3 groups (n =36 each):sham operation group (group S),NP group and dexmedetomidine group (group D).NP was induced by chronic constrictive injury in anesthetized rats.Sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals with 4-0 silk thread.In group S,the right sciatic nerves were exposed,but not ligated.Dexmedetomidine 50 μg/kg was injected intraperitoneally once a day from the onset of operation to one day before the rats were sacrificed in group D,while the equal volume of normal saline was injected in groups S and NP.Mechanical withdrawal threshold (MWT) and thermal pain threshold (TPT) were measured on the day before operation (T0) and 3,7,and 14 days after operation (T1-3).After measurement of pain threshold at T1,T2 and T3 after operation,the L4-6 segments of the spinal cord were removed for determination of the expres-sion of TLR4 and NF-κB mRNA (by RT-PCR) and the expression of TLR4 and NF-κB in spinal dorsal horn (by immuno-histochemistry).Results Compared with group S,MWT and TPT were significantly decreased and the expression of TLR4,NF-κB and TLR4 and NF-κB mRNA was up-regulated after operation in groups NP and D (P ＜ 0.05).Compared with group NP,TPT and MWT were significantly increased and the expression of TLR4,NF-κB,TLR4 mRNA and NF-κB mRNA was significantly down-regulated after operation in group D (P ＜ 0.05).Conclusion The mechanism by which dexmedetomidine attenuates NP in rats is related to inhibition of the expression of TLR4 and NF-κB in rat spinal cord.