Objective To evaluate the role of TWIK-related K+ channel 1 (TREK-1) in reduction of cerebral ischemia-reperfusion (I/R) injury by sevoflurane preconditioning in mice.Methods Sixty male Kunming mice,aged 8 weeks,weighing 21-29 g,were randomly divided into 5 groups (n=12 each) using a random number table:sham operation group (group S),group I/R,sevoflurane preconditioning group (group SP),TREK-1 small hairpin RNA (shRNA) lentivirus + sevoflurane preconditioning group (group TSP),and negative control shRNA lentivirus + sevoflurane preconditioning group (group NSP).In S,I/R and SP groups,normal saline 15 μl was injected into the lateral cerebral ventricle at 1 μl/min.In TSP and NSP groups,TREK-1 shRNA lentivirus and negative control shRNA lentivirus 15 μl were injected into the lateral cerebral ventricle,respectively,at a rate of 1 μl/min.And 14 days later,S and I/R groups inhaled 100％ oxygen for 60 min,SP,TSP and NSP groups inhaled 2.4％ sevoflurane for 60 min,followed by 15 min washout by inhaling 100％ oxygen,and then cerebral I/R was produced by occlusion of right internal carotid artery for 2 h followed by reperfusion.At 24 h of reperfusion,neurological deficit was scored (NDS).The mice were then sacrificed,and brains were removed to determine the cerebral infarct size (IS),expression of hippocampal caspase-3 and cell apoptosis in brain tissues.Apoptosis index (AI) was calculated.Results Compared with group S,the NDS,cerebral IS,expression of hippocampal caspase-3 and AI were significantly increased in I/R,SP,TSP and NSP groups.Compared with group I/R,the NDS,cerebral IS and AI were significantly decreased,and the expression of hippocampal caspase-3 was down-regulated in SP,TSP and NSP groups.Compared with group SP,the NDS,cerebral IS and AI were significantly increased,and the expression of hippocampal caspase-3 was up-regulated in group TSP,and no significant change was found in the parameters mentioned above in NSP group.Conclusion Sevoflurane preconditioning reduces cerebral I/R injury through activating TREK-1 and inhibiting apoptosis in hippocampal neurons of mice.

Objective To study the rule of callus culture and baicalin synthesis in Scutellaria baicalensis. Methods Callus was induced by plant cell culture technology and the content of baicalin was determind by HPLC. Results The optimal culture medium on the growth of callus and the synthesis of baicalin in S. baicalensis is: MS culture medium, 60 mmol/L (NH_4+∶NO_3-=1∶1), 0.5—1.5 mmol/L KH_2PO_4, 80 g/L sucrose, 0.3 mg/L IAA, 2 mg/L 6-BA, and 200 mg/L peptone. When it was cultured for 40 d, the total biomass reached 28.7 g/L and the content of baicalin was 167.4 mg/g, which was much higher than that of wild S. baicalensis. Conclusion The growth of S. baicalensis callus and the accumulation of baicalin are not underway simultaneously; the callus grows first and then its secondary metabolic products synthesize. It is obvious for sucrose to regulate the baicalin synthesis. When the concentration of sucrose is less than 3%, it could only promote the callus growth; when between 3% and 8%, it could greatly increase not only the callus growth but also the baicalin synthesis, when 8%, both of them arrive to the maximum content.

Objective To investigate the role of phosphatase and tensin homologue deleted on chromosome 10 (PTEN) protein in the spinal cord neurons in diabetic neuropathic pain (DNP) in rats.Methods Male Sprague-Dawley rats,aged 2 months,weighing 180-200 g,were studied.Diabetes mellitus was induced by single intraperitoneal injection of streptozotocin (STZ) 60 mg/kg.Sixteen rats with DNP were randomly divided into 2 groups (n =8 each) using a random number table:DNP group and DNP+PTEN inhibitor bpv (pic) group (DPN-bpv group).Another 16 rats were equally and randomly divided into either control group (group C) or bpv group.In DNP-bpv and bpv groups,bpv (pic) 0.2mg/kg was injected intraperitoneally once a day within 14-28 days after injection of STZ.Before STZ injection (T1),and at 2,7,14,21 and 28 days after STZ injection (T2-6),the mechanical paw withdrawal threshold (MWT) was measured.After measurement of MWT,the rats were sacrificed,and the lumbar segments of the spinal cord (L4.5) were removed for determination of PTEN protein activity (by ELISA) and Akt (s473) phosphorylation (by Western blot).Results Compared with group C,the MWT was significantly decreased at T4-6,and the PTEN protein activity and Akt (s473) phosphorylation were significantly increased in DNP and DNP-bpv groups (P＜0.05 or 0.01).Compared with group DNP,the MWT was significantly increased at T6,and the PTEN protein activity and Akt (s473) phosphorylation were significantly decreased in group DNP-bpv (P＜0.05).Conclusion PTEN protein in the spinal cord neurons is involved in the maintenance of DNP in rats.

Objective To determine whether field triage would reduce median contact-to-device ( C2D ) time in patients with ST-segment elevation acute myocardial infarction ( STEMI ) . Methods Consecutive patients with STEMI underwent primary percutaneous coronary intervention( PCI) from March 2010 to February 2014 in Shanghai Pudong Gongli Hospital were analyzed. Patients were divided into two groups. A total of 121 patients were admitted by field triage and 101 patients by non-field triage. The primary study point was C2D time and the study points secondary included ( door-to-balloor, D2B) time, peak Troponin I ( TnI) levels, hospital mortality and 30 days follow-up mortality. Results Baseline and procedural characteristics between the two groups were comparable. Comparing to non-field triage group, the C2D time was reduced [(92. 0 ± 56. 0)min vs. (131. 0 ± 61. 0)min,P﹤0. 01]. The D2B time was lower in the field triage group vs. the non-field triage group [(55. 0 ±26. 0)min vs. (96. 0 ±31. 0)min,P﹤0. 01]. The percentage of patients with C2D time less than 90 minutes increased significantly from 85. 1% to 98. 3%( P﹤0. 01 ) in the field triage group. Peak TnI level was significantly reduced in the field triage group [(23. 5 ±22. 0) μg/L vs. (43. 5 ± 39. 0) μg/L,P﹤0. 01]. In-hospital mortality and 30 days follow-up mortality did not significantly differ between the 2 groups (3. 3% and 3. 0%, P=0. 885;3. 3% and 5. 0%, P=0. 544, respectively). Conclusions In STEMI patients, field triage was associated with significantly reduced C2D and D2B times.

Objective To evaluate the effects of intrathecal TRESK gene recombinant adenovirus on inflammatory responses mediated by chemokine in the spinal cord of rats with neuropathic pain ( NP ) . Methods Thirty?six male Sprague?Dawley rats, weighing 200-250 g, were randomly divided into 6 groups (n=6 each) using a random number table: control group (group C); sham operation group (group S);NP group; TRESK?overexpressed adenovirus group ( group TRESK ); negative adenovirus group ( group Virus); normal saline group ( group NS) . Spinal nerve injury was produced by exposing the sciatic nerve and its branches and ligation and transection of tibial nerve and common fibular nerve in anesthetized rats. In TRESK, Virus and NS groups, pAd∕CMV∕V5?DEST?TRESK 25 μl (109IU∕ml), negative adenovirus 25 μl and normal saline 25 μl were intrathecally injected, respectively. At 1 day before operation ( base?line, T0 ) and 1, 3, 7 and 14 days after operation ( T1-4 ) , the mechanical paw withdrawal threshold ( MWT) and thermal paw withdrawal latency were measured. Six rats in each group were sacrificed after measurement of pain threshold at T3 . The L4,5 segments of the spinal cords were removed for determination of monocyte chemotactic protein?1 ( MCP?1) , MIP?2, tumor necrosis factor?alpha ( TNF?α) , interleukin?1 beta ( IL?1β) and IL?6 mRNA expression by real?time PCR. Results There was no significant difference in thermal paw withdrawal latency at each time point between groups. Compared with C and S groups, MWT at T1-4 in NP and TRESK groups and at T1-3 in Virus and NS groups were significantly decreased, and the expression of MCP?1, MIP?2, TNF?α, IL?1βand IL?6 mRNA was up?regulated in NP, TRESK, Virus and NS groups. Compared with group NP, MWT was significantly increased at T1-4, and the expres?sion of MCP?1, MIP?2, TNF?α, IL?1β and IL?6 mRNA was down?regulated in group TRESK. Conclusion The mechanism by which intrathecal TRESK gene recombinant adenovirus reduces NP is re?lated to inhibition of inflammatory responses mediated by chemokine in the spinal cord of rats.

Objective To observe the effect of adenosine monophosphate activated protein kinase (AMPK) on attenuating inflammation in fibrosis induced hy acute ischemia reperfusion injury (IRI) in mice.Methods Forty eight male C57BL/6 mice were randomly divided into four groups:sham operation group (sham group),IRI group,AMPK inhibitor+IRI group (AMPK/IRI group) and normal saline+IRI group (NS/IRI group),12 mice each group.The mice with renal IRI were occluded for 30 min through clipping bilateral renal pedicle,then released renal perfusion.Mice in sham group were performed the separation of renal pedicle without clipping.Mice in AMPK/IRI group and NS/IRI group were respectively intraperitoneal injected AMPK inhibitor and normal saline before IRI.At the 2 d after operation,6 randomly-selected mice from each group were blooded by extraction eyeball to detect BUN and Scr.The renal histopathological changes were observed through HE staining.The mRNA expression of IL-1β,IL-6 and TNF-α was detected by real time PCR,and the level of AMPK phosphorylation was detected by Western blotting.At the 14 d after operation,Collagen 1 (COL1),α-SMA and fibronectin (FN) were detected by immunofluorescence and Western blotting in 6 remained mice from each group.The degree of kidney fibrosis was observed through sirus red staining.Results Compared with those in sham group,tubular interstitial damage was aggravated (P ＜ 0.05),BUN and Scr were increased (P ＜ 0.05),the mRNA expression of IL-1β,IL-6 and TNF-α was increased at the 2 d after operation (all P ＜ 0.05),and the level of AMPK phosphorylation was activated in IRI group and NS/IRI group (all P ＜ 0.05);the degree of kidney fibrosis and the expression of COL1,α-SMA and FN were increased obviously at the 14 d (all P ＜ 0.05).Compared with those in IRI group,in AMPK/IRI group tubular interstitial damage was aggravated (P ＜ 0.05),BUN and Scr were increased (all P ＜ 0.05),the mRNA expression of IL-1β,IL-6 and TNF-α was increased at the 2 d (all P ＜ 0.05),and the level of AMPK phosphorylation was decreased (P ＜ 0.05).Moreover,the degree of kidney fibrosis and the expression of COLI,α-SMA and FN were increased obviously at the 14 d in AMPK/IRI group (all P ＜0.05).Conclusions AMPK can ameliorate the acute renal ischemia reperfusion injury induce fibrosis in mice,and the mechanism may be related to the decrease of inflammatory reaction.

Objective High expression of multi-resistant transporter ATP-binding cassette super family G member 2 (ABCG2) is a major cause of drug resistance and chemotherapeutic failure of cancer .This study was to investigate the significance of ABCG2 expression in adrenocortical cancer cells after cyclophosphamide ( CTX) intervention in vivo . Methods Ten male and fe-male BALB/C-nu mice were randomly divided into a cyclophosphamide ( CTX) group and a control of equal number .SW-13 cells were subcutaneously injected into the nude mice to establish a model of subcutaneous transplantation tumor , followed by intraperitoneal injec-tion of CTX and isotonic saline solution into the two groups of mice , respectively .Then the expression of ABCG 2 in tumor tissue and primarily cultured cells was detected by immunohistochemistry and flow cytometry . Results The expression of ABCG 2 in the tumor tissue was significantly higher in the CTX than in the control group ([69.1 ±1.83]%vs [53.4 ±1.65]%, P<0.05), and so was that in the primarily cultured cells ([97.89 ±1.36]% vs [81.88 ±8.31]%, P<0.05). Conclusion The ABCG2 gene is in-volved in the drug resistance of adrenocortical carcinoma and may be a therapeutic target of the malignancy .

Objective To evaluate the clinical outcome in the treatment of humerus shaft comminuted fractures using limited open reduction and internal fixation combined with an external fixator.Methods Data of 80 patients with comminuted humerus shaft fractures treated from January 2005 to January 2013 were analysed retrospectively.All the patients underwent limited open reduction and internal fixation combined with an external fixator (treatment group) and open reduction and plate fixation (control group) according to the random number table.In the treatment group,there were 40 patients (28 males,12 females),at mean age of 33.5 years (range,21-54 years),with causes of injury including traffic accidents in five patients,falls in nine,crashes in seven and others in six.There were seven patients with open fractures and 33 with closed fractures.In the control group,there were 40 patients (25 males,15 females),at mean age of 32.9 years (range,19-55 years),with causes of injury including traffic accidents in 16 patients,tumbling in seven,crush in seven and others in ten.There were eight patients with open fractures and 32 with closed fractures.The operation time,intraoperative blood loss,bone union time and complications in both groups were recorded.Clinical efficacy was evaluated using the Stewart and Hundley standard.Results Mean follow-up was 19 months (range,15-24 months).Treatment and control groups showed significant differences in operation time [(55.5 ± 10.3) minutes vs.(120.5 ± 15.3) minutes],intraoperative blood loss [(120.4 ± 20.7) ml vs.(245.4 ± 26.7) ml] and bone union time [(11.6 ± 1.3) weeks vs.(14.9 ± 2.3) weeks] (P ＜ 0.05).Rate of incision infection was 8％ (3/40) in treatment group and 10％ (4/40) in control group (P ＞ 0.05).In treatment group the results were excellent in 31 patients and good in nine.In control group the results were excellent in 27 patients,good in nine,fair in one and poor in three.One patient with radial nerve injury after a second surgery for implant removal and two patients with osteomyelitis or bone nonunion were noted in control group.Conclusion Limited open reduction and internal fixation in combination with an external fixator is associated with small trauma,easy operation,short operation time,few bleeding,rigid fixation,early functional exercises and reduced bone nonunion for treatment of comminuted humerus shaft fractures,which exhibits great clinical value.

Objective To explore the effect of adriamycin on the characteristics of colony derived from human adrenal cortical carcinoma cells (ACC) SW-13.Methods Treatment with Adriamycin (ADM) was used in BALB/c-nude mouse tumor xenograft model established using the ACC cell line SW-13.The characteristic of colony was assessed for the formation rates,the percentagc of three colony types and growth curve of single cell.Hoechst33342 dyeing test was used to test drug resistance.Results The Single-cell colony formation rate of experimental group were significantly higher than control group (P ＜ 0.05),and the holoclone percentage of experimental group were significantly higher than control group (P ＜ 0.05).In the Hoechst33342 dyeing tcst,the fluorescence intensity of control was higher than experimental group.Conclusion The treatment of ADM in vivo is beneficial for the colony formations of ACC cell and the formations rate of holoclone,and can improve the ability of drug resistance of ACC cell SW-13.