Six compounds, syringaresinol (Ⅰ), 2,5－dimothoxy－1,4－benzoquinone(Ⅱ), ?－sitost erol (Ⅲ)verticine (Ⅳ), verticiaone (Ⅴ) and solnidine(Ⅵ) were isolated from the aerial parts of Fritillaria thunbergii Miq. Their structures were determined by spectral data and chemical,evidences Ⅰ and Ⅱ were obtained from the Fritillaria L. for the first time.

Objective To evaluate the effect of remifentanil on Toll-like receptor 2 (TLR2) mRNA expression during renal ischemia-reperfusion (Ⅰ/R) in rats.Methods Fifty-four male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 3 groups (n=18 each) using a random number table:sham operation group (group S),Ⅰ/R group and remifentanil group (group R).Renal Ⅰ/R injury was produced by clamping the bilateral renal arteries for 45 min followed by reperfusion in Ⅰ/R and R groups.Bilateral renal arteries were only exposed but not clamped in group S.Remifentanil 1.0 μg · kg-1 · min-1 was infused via the tail vein starting from 15 min before ischemia until 30 min of reperfusion in group R,while the equal volume of normal saline was given instead in S and Ⅰ/R groups.The animals were sacrificed at 15 min before ischemia and 6 and 24 h of reperfusion,and the renal specimens were obtained for examination of the pathological changes (with light microscope) and for determination of the contents of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) (by ELISA) and expression of TLR2 mRNA (by RT-PCR) and cell apoptosis (by double staining and flow eytometry).The apoptotic rate was calculated.Results Compared with group S,TLR2 mRNA expression was significantly up-regulated,and the contents of TNF-α and IL-6 and apoptotic rate were increased at 6 and 24 h of reperfusion in Ⅰ/R and R groups.Compared with group Ⅰ/R,TLR2 mRNA expression was significantly down-regulated,and the contents of TNF-α and IL-6 and apoptotic rate were decreased at 6 and 24 h of reperfusion in group R.The pathological changes were significantly attenuated in group R as compared with group Ⅰ/R.Conclusion The mechanism by which remifentanil reduces renal Ⅰ/R injury is related to down-regulation of TLR2 expression and decrease in TLR2 activity and inhibition of inflammatory responses in renal tissues and cell apoptosis in rats.

Objective To investigate the effect of remifentanil on protein kinase C (PKC) activity during renal ischemia-reperfusion (I/R) in rats.Methods Seventy-five male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 5 groups (n=15 each) using a random number table:sham operation group (group S),I/R group,remifentanil group (group R),naloxone group (group N),and naloxone + remifentanil group (group NR).Renal ischemia was induced by clamping the bilateral renal arteries for 45 min using an atraumatic clamp followed by reperfusion.In R and NR groups,remifentanil 1.0 μg · kg-1 · min-1was infused via the caudal vein starting from 15 min before ischemia until 30 min of reperfusion.In N and NR groups,naloxone 0.3 mg/kg was injected via the caudal vein at 20 min before ischemia and 35 min of ischemia,respectively.The rats were sacrificed at 24 h of reperfusion and the kidneys were removed for determination of the ultrastructure of the renal tubular epithelial cells (using transmission electron microscope),activity of PKC in renal tissues (by ELISA),and expression of the PKC in renal tissues (by immuno-histochemistry).Results Compared with group S,the activity of PKC in renal tissues was significantly increased in the other four groups,and the expression of the PKC in renal tissues was up-regulated in group R.Compared with group I/R,the activity of PKC in renal tissues was significantlyincreased,the expression of PKC in renal tissues was up-regulated,and the pathological changes were attenuated in group R.Compared with group R,the activity of PKC in renal tissues was significantly decreased,the expression of PKC in renal tissues was down-regulated,and the pathological changes were aggravated in N and NR groups.Conclusion The mechanism by which remifentanil attenuates renal I/R injury may be related to up-regulation of PKC expression and increase in PKC activity through activating opioid receptors in rats.

OBJECTIVE:To establish a method for the simultaneous determination of costunolide and dehydrocostus lactone in Haoweilai soft capsule. METHODS:HPLC was performed on the column of Dikma C18 with mobile phase of acetonitrile- water (V/V,55∶45) at a flow rate of 1.0 ml/min,detection wavelength was 225 nm,column temperature was 25 ℃,and the injection volume was 10 μl. RESULTS:The linear range were 24.00-108.00 μg/ml(r=0.999 7) for costunolide and 20.88-93.98 μg/ml for dehydrocostus lactone (r=0.999 8);RSDs of precision,stability and reproducibility tests were lower than 1%;recoveries were 98.71%-100.00%(RSD=0.25%,n=6)and 96.88%-99.18%(RSD=0.40%,n=6). CONCLUSIONS:The method is simple with good stability and reproducibility,and can be used for the simultaneous determination of costunolide and dehydrocostus lactone in Haoweilai soft capsule.

Coxsackievirns A16(CVA16),together with enterovirus type 71(EV71),is responsible for most cases of hand,foot and mouth disease(HFMD) worldwide.Recent findings suggest that the recombination between CVA16 and EV71,and the co-circulation of these two viruses may have contributed to the increase of HFMD cases in China over the past few years.It is therefore important to further understand the virology,epidemlology,virus-host interactions and host pathogeuesis of CVA16.In this study,we describe the viral kinetics of CVAI6 in human rhabdomyosarcoma(RD) cells by analyzing the cytopathic effect(CPE),viral RNA replication,viral protein expression,viral RNA package and viral particle secretion in RD cells.We show that CVA16 appears to first attach,uncoat and enter into the host cell after adsorption for 1 h.Later on,CVA16 undergoes rapid replication from 3 to 6 h at MOI 1 and until 9 h at MOI 0.1.At MOI 0.1,CVA16 initiates a secondary infection as the virions were secreted before 9 h p.i.CPE was observed after 12 h p.i.,and viral antigen was first detected at 6 h p.i.at MOI 1 and at 9 h p.i.at MOI 0.1.Thus,our study provides important information for further investigation of CVA16 in order to better understand and ultimately control infections with this virus.

Objective To study the expression of tumor stem cell marker aldehyde dehydrogenase 1 (ALDH1)in invasive bladder cancer tissue and to clarify its relationship with the biological behavior of bladder cancer. Methods The ALDH1 expression in 109 cases of primary invasive carcinomas specimens (case group)and 20 cases of normal bladder tissue surrounding cancer (control group)was detected by immunohistochemistry. At the same time,the ALDH1 expression in 6 cases of metastatic pelvic lymph node tissue and 20 cases of non-metastatic pelvic lymph node tissue was detected. The relationship between the ALDH1 expression and the chinicopathological charateristics of invasive bladder cancer and its influence in the survival rate and disease-free survival were analyzed. Results The positive rates of ALDH1 expression in bladder cancer tissue and normal bladder tissue were 33.94%(37/109)and 5.00% (1/20),respectively,there was significant different between them (P<0.01);they were 19.05% (8/42)and 43.28% (29/67)in the cases with non muscle invasive and nmuscle invasive bladder cancer, respectively,there was significant difference (P<0.01);they were 13.04% (3/23)and 39.53% (34/86)in the cases of bladder cancer with low grade and high grade,respectively,there was significant difference (P<0.05);they were 50.00% (3/6)and 12.90% (4/31)in the tissue of bladder cancer with metastatic lymph nodes and non metastatic ones,respectively,there was significant difference (P<0.05);they were 50.00% (3/6)and 0.00%(0/20)in the metastatic lymph nodes and non metastatic ones,respectively,there was significant difference (P<0.01).The overall survival rate in the patients with positive ALDH1 expression was 64.9% while it was 84.7% in negative ones,there was significant difference (P<0.05);the disease-free Survival was 51.4% and 75% in the patients with positive and negative ALDH1 groups,respectively,there was significant difference (P<0.05). Conclusion The high expression of tumor stem cell marker ALDH1 is associated with staging, grading and prognosis of invasive bladder cancer.ALDH1 may play a role in the tumorigenesis,progression and metastasis of bladder cancer.

Objective To explore effect of irregular chemotatic factor fractalkine(FKN),phosphatidyl inositol 3 kinase and nuclear factor?κB(NF?κB)on interleukin?6(IL?6)expression in peripheral blood monocytes and the effect of valsartan intervention,so as to research the signal conduc?tive mechanism of FKN impacting on IL?6. Methods Peripheral blood monocytes were isolated from fresh blood of healthy volunteers by the densi?ty gradient centrifugation. The extractive peripheral blood monocytes were divided into seven groups:the control group,the FKN group,the LY294002 group(PI3K inhibitors),the PDTC group(NF?κB inhibitors),the FKN+valsartan group,the FKN+LY294002 group,and the FKN+PDTC group,the latter two were pretreated by LY294002 and PDTC respectively before FKN inducing PBMC cells. The IL?6 expression in cell me?dium was measured in each group by ELISA at 12 hours and 24 hours after PBMC treatment. Results After 12 hours of culture,compared with the control group,the expression of IL?6 in the FKN group was decreased(P<0.05),while LY294002 and PDTC groups had no significant difference (P>0.05). Compared with the FKN group,the expression of IL?6 was decreased in the FKN+valsartan group(P<0.05),increased in the FKN+LY294002 group(P<0.05),and was decreased in the FKN+PDTC group(P<0.05). After 24 hours of culture,the IL?6 expression in each group had no significant difference(P>0.05). Conclusion FKN can adjust the expression of peripheral blood PBMC IL?6 in a two?way pattern, inhibiting the expression of IL?6 by PI3K pathway and promoting the expression of IL?6 by NF?κB pathway,overall,FKN can inhibit the expression of IL?6. Valsartan can increase FKN to inhibit the expression of IL?6.

Objective To investigate the effects of vagus nerve electrostimulation (VNS) on the brain damage of rat middle cerebral artery occlusion (MCAO)/reperfusion model and its mechanism. Methods Twenty four adult male SD rats were randomly divided into two groups (12 each): MCAO/reperfusion group (MCAO group) and MCAO/reperfusion+VNS group (MCAO+VNS group). Subsequently, the neurological function deficit was determined by neurological scoring according to Zea Long scoring method 24h after MCAO/reperfusion. The cerebral infarction volume was determined by TTC assay. The cell apoptosis in brain damage zone was determined by TUNEL assay. Then, the effect of VNS on cAMP response element binding protein (CREB) and p-CREB protein expression was determined by Western blotting. The effect of VNS on Bcl-2 and Bax expression was determined by immunohistochemistry assay. Results Compared with MCAO group, VNS significantly inhibited MCAO-induced neurological deficit (P<0.01), decreased brain infarct volume (P<0.01) and cell apoptosis (P<0.01), increased the expression of p-CREB protein (P<0.01) and the number of Bcl-2-position cells (P<0.01) together with decreasing the number of Bax-position cells (P<0.01). However, VNS did not affect the expression of CREB protein (P>0.05). Conclusion VNS may ameliorate MCAO-induced neurological deficit and decrease brain infarct volume, which may be related to the promotion of p-CREB protein expression level.

OBJECTIVE:To strengthen the supervision of clinical antibiotics use through establishing electronic consultation system. METHODS:Based on hospital information system(HIS)and antibiotic management system,consultation system for anti-biotics of special use(ASU)was established,and the effects of the system after used(Jan. 2014)were also evaluated. RESULTS:Real-time consultation of ASU medical orders and related consultation results inquiry and statistics could be achieved through de-signing ASU consultation application form,establishing consultation experts and confirming their working content,adding statistics function,etc. The consultation have been finished 8275 times from 2014 to 2015. The inspection rate for pathogenic microbes in ASU patients rose from 78.68% in 2013 to 83.90% in 2015. CONCLUSIONS:The establishment and application of ASU consulta-tion system can improve consultation efficiency and quality,so as to promote rational use of antibiotics in the clinic.

Objective To investigate the efficacy and safety of balloon-assisted catheter directed thrombolysis (CDT) for acute lower extremity deep vein thrombosis (DVT).Methods From September 2008 to February 2011,94 patients with acute lower extremity DVT were admitted.The cases in early stage were treated by CDT (Group A,n =50),and the cases in late stage were treated by balloon-assisted CDT ( Group B,n =44).The clinical data of these patients were retrospectively analyzed.The circumference difference between normal and affected limbs,scores of venous patency,and rates of venous patency were recorded for judging the efficacy.The total dose of urokinase and retention time of infusion catheter was compared between the two groups.The incidence of pulmonary embolism and bleeding were used to judge the safety of treatment.The venous patency was followed up by ultrasound or/and venography.Measurement data with normal distribution was described by mean + standard,and was analyzed using T test.Measurement data with non-normal distribution was described by M ( QL,QU ),QL =P25,QU =P75,and was analyzed using Wilcoxon' s test.Categorical variable data was analyzed using Chi-Square test Results The prior treatment circunfference difference between normal and affectéd limbs were (5.37 ±1.97) cm (thigh) & (4.14 ± 1.57) cm (calf) in Group A and (5.41±2.22) cm (thigh) & (4.05 ±1.61) cm (calf) in Group B ; and the difference between the groups was insignificant ( thigh:t =- 0.113,P=0.910; calf:t =0.288,P =0.774).The post treatment correspondences were:(2.96 ± 1.10) cm (thigh) & ( 1.93 ± 0.84 ) cm (calf) in Group A and ( 1.78 ± 1.40) cm ( thigh ) & ( 1.41± 1.17 ) cm (calf) in Group B; the difference between the groups was significant (thigh:t =4.66,P ＜0.0001; calf:t =2.548,P =0.012 ).The prior treatment score of venous patency was 9 (8,10) in Group A and 8.3(7,10) in Group B without significant difference (Z =- 1.5172,P =0.1292).The post treatment score of venous patency was 3.5 ( 2,5 ) in Group A and 0 ( 0,1) in Group B with significant difference ( Z =-5.7702,P ＜0.01).The rate of venous patency after the treatment was 55.0％ (42.3％,72.4％ ) in Group A and 100％ (88.5％,100％ ) in Group B,with significant difference ( Z =4.9148,P ＜ 0.01).The total dose of urokinase used in the treatment was 5.950 ( 5.525,7.225 ) × 106U in Group A and 4.100 (3.600,5.050) × 106U in Group B with significant difference (Z =-6.0133,P ＜ 0.01).The retention time of perfusion catheter was 10 (9,12) d in Group A and 6 (5,7) d in Group B with significant difference ( Z =- 8.0358,P ＜ 0.01).No symptomatic pulmonary embolism occurred in both groups during the treatment and follow-up period.The rate of bleeding complication was 38.0％ (19/50) in Group A and 22.3％ (10/44) in Group B,without significant difference (x2 =2.5590,P =0.1097 ).The removal rate of optional filter was 88.37％ (38/43) in Group A and 100％ (39/39) in Group B,with significant difference ( x2 =4.829,P =0.028 ).The rate of venous patency at the last follow-up point was 50.0％ (44.4％,59.2％ ) in Group A,and 95.4％ (83.6％,100％ ) in Group B,with significant difference (Z =- 3.2721,P =0.0011).Conclusions Balloon-assisted CDT was a promising treatment for acute lower-extremity DVT.It improved the effect of thrombolysis and reduced the dosage of urokinase,and did not increase the risk of pulmonary embolism.