A sensitive, rapid, simple and economical ultra-performance liquid chromatography–tandem mass spectro-metric method (UPLC–MS/MS) was developed and validated for simultaneous determination of imatinib, dasatinib and nilotinib in human plasma using gliquidone as internal standard (IS). Liquid-liquid extraction method with ethyl acetate was used for sample pre-treatment. The separation was performed on an Xtimate Phenyl column using isocratic mobile phase consisting of A (aqueous phase: 0.15% formic acid and 0.05% ammonium acetate)and B(organic phase:acetonitrile)(A:B=40:60,v/v).The flow rate was 0.25 mL/min and the total run time was 6 min. The multiple reaction monitoring (MRM) transitions, m/z 494.5→394.5 for imatinib, 488.7→401.5 for dasatinib, 530.7→289.5 for nilotinib and 528.5→403.4 for IS, were chosen to achieve high selectivity in the simultaneous analyses. The method exhibited great improvement in sensitivity and good linearity over the concentration range of 2.6–5250.0 ng/mL for imatinib, 2.0–490.0 ng/mL for dasatinib,and 2.4–4700.0 ng/mL for nilotinib.The method showed acceptable results on sensitivity,specificity, recovery, precision, accuracy and stability tests. This UPLC–MS/MS assay was successfully used for human plasma samples analysis and no significant differences were found in imatinib steady-state trough concentra-tions among the SLC22A5?1889T>C or SLCO1B3 699G>A genotypes(P>0.05).This validated method can provide support for clinical therapeutic drug monitoring and pharmacokinetic investigations of these three tyrosine kinase inhibitors(TKIs).

One hundred and seventy six consecutive patients with acute ischemic stroke who received intravenous recombinant tissue plasminogen activator ( rt-PA ) in 4.5 hours from symptom onset during February 2009 to July 2013 were included in the study.Modified Rankin Scale was used to evaluate the recovery of neurological functions.Patients were divided into good ( 0 -1 ) or poor ( 2 -6 ) outcome groups according modified Rankin Scale score.Univariate analysis and multivariate logistic regression analysis were used to determine the differences of clinical data between the two groups.The age of patients with good outcome was significantly lower than that of poor outcome group [ ( 61.4 ±11.5 ) vs.( 69.0 ± 13.2) years,P =0.000].Compared to patients with poor outcomes, patients with good outcome group showed lower rate of diabetes [ 13%( 12/93 ) vs.29%( 24/83 ) , P =0.009 ] , lower blood glucose level [(5.05 ±0.97) vs.(5.83 ±1.72) mmol/L,P=0.020], higher uric acid level[(404.4 ±151.7) vs.(345.6 ±107.5) μmol/L,P=0.028],shorter onset to treatment time [(1.92 ±0.94) vs.(2.30 ±1.01) h, P=0.019],lower baseline National Institute of Health Stroke Scale score [(14.0 ±5.2) vs.(16.0 ± 6.2),P=0.025],lower systolic blood pressure level at 2 h[(140.8 ±18.3) vs.(149.0 ±18.9) mmHg (1 mmHg=0.133 kPa),P=0.005]and 24 h [(137.6 ±21.9) vs.(147.1 ±17.4) mmHg,P=0.009] after thrombolysis.Logistic regression analysis showed that uric acid levels were not related to hemorrhagic transformation independently (P =0.172,OR =0.965,95%CI:0.917 -1.016), but were related to outcome independently (P=0.047,OR=0.957,95%CI:0.916-0.999).

Objective To investigate the effect of Ser473-Akt phosphorylation in the protection of atorvastatin to cerebral ischemia-re-perfusion (I/R) injury in rats. Methods Forty male Sprague-Dawley rats were randomly divided into normal group (n=10), sham group (n=10), I/R group (n=10) and intervention group (n=10). A model of cerebral ischemia-reperfusion in rats was establishied, with ischemia for 2 hours and reperfusion for 72 hours. The normal group and the sham group received no injury. I/R group was administered with normal saline only, and the intervention group received atorvastatin 10 mg/kg prepared with normal saline at palinesthesia, 24 and 48 hours after reperfu-sion. All rats were sacrificed 72 hours after reperfusion. HE staining and TUNEL staining were performed in the brain specimens. The ex-pression of Akt and Ser473-Akt in the prefrontal cortex of the brain were detected with Western blotting. Results Compared with I/R group, 72 hours after reperfusion, the damage of nerve cells significantly lessened in the intervention group;the apoptosis positive cells significant-ly reduced in the intervention group (t=-6.014, P<0.001). The expression of Ser473-Akt in prefrontal cortex was higher in I/R group than in the normal group and the sham group (t>20.327, P<0.001), and was higher in the intervention group than in I/R group (t=3.649, P=0.007). Conclusion The Ser473-Akt phosphorylation plays an important role in the protection of atorvastatin in nerve cell through anti-apoptosis of nerve cells, and reducing cerebral I/R injury.

Objective To investigate Danhong in improving the movement function of reperfused rat after stroke. Methods Rats to make middle cerebral artery occlusion model,after 24 hours of reperfusion were divided into control group,high dose and low dose of Danhong groups and Venorruton group randomly. Rats in Control group were injected saline. High and low dose group were injected DanHong according to their weight,high dose as 14. 4 mg/kg,low dose as 3. 6 mg/kg. Venorruton group were injected Venorruton as 0. 04 g/kg. The infarcted square of rat brain was measured,the rats were tested with walking wood bar and two fore limb grasp strength. Results Rat infarcted squre in 24 hours and 3 days,control groups is the biggest,compared with others,there are statistically difference(P<0. 05). Score in each group of walking wood bar in 3rd days are (1. 30 ± 0. 91),(3. 78 ± 1. 72),(4. 18 ± 2. 05),(4. 63 ± 2. 45). Compared between control and other druge groups,there are statistically differnec(P<0. 05). There are statistically differences between control and other groups in two fore limb grasp testing(P<0. 05). Conclusion Danhong has the same effect which can improve the movement function to stroke rat as Venorruton.

Objective To investigate the efficacy and safety of different doses of dezocine at different administration time on analgesia after laparoscopic cholecystectomy.Methods 80 patients elected from ASA Ⅰ-Ⅱ grade laparoscopic cholecystectomy were randomly divided into four groups,20 cases in each group.Group Ⅰ was anesthetized by intravenously injected dezocine 0.10mg/kg before the surgery,group Ⅱ was anesthetized by dezocine 0.15mg/kg before the surgery,group Ⅲ was anesthetized by intravenously injected dezocine 0.10mg/kg after stopping anesthetic surgery,and group Ⅳ was anesthetized by intravenously injected dezocine 0.15mg/kg after stopping anesthetic surgery.The postoperative pain scores were observed 1,6,12,24 hours after operation in the four groups by using visual analog scale (VAS),comfort score (BCS),and anesthesia recovery score (modified Aldrete score).Results TheVAS of the four groups 1 hour after surgery:group Ⅰ and group Ⅲ was significantly different(t =2.308,P =0.036),group Ⅰ and group Ⅳ was significantly different (t =2.106,P =0.042),group Ⅱ and group Ⅲ was significantly different (t =2.711,P =0.014),group Ⅱ and group Ⅳ was significantly different (t =2.317,P =0.037).The BCS 1 hour after surgery:group Ⅰ and group Ⅲ was significantly different(t =2.108,P =0.042),group Ⅰ and group Ⅳ was significantly different(t =2.069,P =0.048),group Ⅱ and group Ⅲ was significantly different (t =2.353,P =0.033),group Ⅱ and group Ⅳ was significantly different (t =2.361,P =0.036).The VAS 6 hours after surgery:group Ⅰ and group Ⅲ was significantly different (t =2.084,P =0.045),group Ⅱ and group Ⅲ was significantly different(t =2.309,P =0.038),group Ⅱ and group Ⅳ was significantly different(t =2.303,P =0.040).The BCS 6 hours after surgery:group Ⅰ and group Ⅲ was significantly different (t =2.294,P =0.041),group Ⅱ and group Ⅲ was significantly different(t =2.322,P =0.035),group Ⅱ and group Ⅳ was significantly different (t =2.070,P =0.048).The BCS 12 hours after surgery:group Ⅱ and group Ⅲ was significantly different(t =2.518,P =0.047).VAS and BCS scores at other time points had no significant difference (P ＞ 0.05).Conclusion The analgesic after laparoscopic gallbladder surgery using dezocine 0.10mg/kg-0.15mg/kg,especially 0.15mg/kg administered anesthesia before surgery,can effectively relieve postoperative pain and improve postoperative comfort,reduce postoperative analgesic(pain pump) and has less adverse reactions,which is worthy of promotion.

Objective To study whether bellidifolin has rehabilitation effect on injured sciatic nerve rats,and whether ciliary neurotrophic factor (CNTF) is involved in this mechanism.Methods 225 male wistar rats were made to be sciatic nerve injured models,with right sciatic nerve being cut and sewed under microscopy,left sciatic being sham.Rats were randomly divided into control group,bellidifolin 25 mg,50 mg,75 mg groups and mecobalamin group,with 45 rats in each group.Rats in control group were just injected sodium chloride,others were intra-peritonealiy injected different doses of bellidifolin and mecobalamin after operation.Results Three weeks after operation,SFl of control group,mecobalaming group,bellidifolin 25 mg,50 mg and 75 mg group were-84.35± 4.87,-45.20±2.30,-70.42±4.21,-57.73±3.46 and-64.38±4.38 respectively.Compared with control group,others showed significant differences (P＜0.05).There were statistically differences between bellidifolin groups and mecobalaming group(P＜0.05).Within bellidifolin groups,50 mg group showed difference compared with 25 mg and 75 rg groups(P=0.031).TSW results also showed differences among bellidifolin groups,control group and mecobalaming group.There were statistical differences among bellidifolin groups(P＜0.05).Each groups with immunohistochemistry analysis,CNTF expression showed statistically differences among bellidifolin 50 mg group and 25 mg,70 rg groups,Bellidifolin 50 mg group was higher than others(P＜0.05).Conclusion Bellidifolin can promote the recovery of injured sciatic nerve,especially the concentration of 50 mg bellidifolin,and CNTF is involved in the rehablitation process.

Objective To study the rehabilitation effect of Bellidifolin for injuried sciatic nerve,and to explore whether ciliary neurotro-phic factor ( CNTF) is involved in this mechanism. Methods The right sciatic nerver of 225 male wistar rats was cut and sewed under mi-croscopy. Rats were devided into 5 groups,as control group,Bellidifolin 25 mg group,50 mg group、75 mg group and Mecobalamin group. The control group were injected sodium chloride,other groups were injected different dose of Bellidifolin and Mecobalamin. 1,3 and 5 weeks later, the motor nerve conduction velocity( MNVC) and gastrocnemius muscle cross-sectional area were detected,CNTF positive area were analysed by immunohistochemical method. Results There were differences among bellidifolin groups,control group and mecobalamin group in Nerve conduction velocity. Within Bellidifolin groups,50 mg group compared with 25 mg and 75 mg groups,there were statistically differences( P=0. 025). Three weeks after operation,gastrocnemius muscle cross-sectional area of control group,mecobalaming grop and Bellidifolin 25 mg group,50 mg group,and 75 mg group were(455. 06 ± 29. 38),(679. 03 ± 81. 48),(465. 31 ± 71. 55),(670. 24 ± 91. 26) and (669. 28 ± 78. 54) respectively,compared with control group and Bellidifolin 25 mg group,others had a significant difference(P<0. 05). CNTF expres-sion showed billidifolin 50 mg group are higher than others(P<0. 05). Conclusion Bellidifolin can improve the rehabilitation of injured sciatic nerve. CNTF is involved in this mechnism.

Objective To observe effect of intra-and post-operative mild hypothermia using an ice blanket on patients with severe traumatic brain injury (sTBI).Methods Twenty sTBI patients with Glasgow Coma Scale (GCS) of 3-8 points were included and were assigned to either ice bag cooling (Bag group) or ice blanket cooling (Blanket group) (n =10 each) according to random number table.Patients in Bag group had temperature reduction by placing ice bag over great vessels,whereas in Blanket group an ice blanket (temperature was set as the nasopharyngeal temperature of 33℃-34℃) was employed to have temperature reduction.Hypothermia therapy in the two group groups was initiated from the beginning of operation and continued for 48 hours after operation.Intracranial pressure,cerebral perfusion pressure (CPP) and GCS in both groups were recorded respectively at 10 minutes before operation (T0) and at 8,12,24,48 and 72 hours after operation (T1,T2,T3,T4 and T5).Venous blood of the two groups was harvested to assay the serum concentration of neuronspecific enolase (NSE),myelin basic protein (MBP)and S-100β at T0,T3,T4,-Ts and at 96 hours after operation (T6) by ELISA method.Glasgow Outcome Scale (GOS) was evaluated at postoperative six months.Results In Bag group,body temperature (T1-T5) of the patients had no significant decrease (P ＞ 0.05) and NSE (T3-T6),S-100β (T3-T6) and MBP (T4-T6) were increased (P ＜ 0.05 or 0.01) when compared with those in T0 ; intracranial pressure (T2-T5) was increased (P ＜ 0.05) and CPP (T3-T5) was lowered (P ＜ 0.05) when compared with those in T1.In Blanket group,body temperature (T1-T6) of the patients presented was decreased significantly (P ＜ 0.01) and NSE (T3-T6),MBP (T5-T6) and S-100β (T4-T6) were increased (P ＜ 0.05 or 0.01) when compared those in T0 ; intracranial pressure (T2-T6) was increased (P ＜ 0.05) and CPP had no significant changes (P ＞0.05) when compared with those in T1.By contrast with those in the same time points in Bag group,lower body temperature (T1-T5) (P ＜ 0.001),lower intracranial pressure (T2-T5),higher CPP (T3-T5) as well as lower NSE (T4-T6),MBP (T4-T6) and S-100β(T6)were observed in Blanket group (P ＜0.05 or 0.01).Changes of GCS and GOS in the two groups were no significance (P ＞0.05).Conclusion Intraoperative and postoperative mild hypothermia therapy using an ice blanket may alleviate the degree of brain injury in sTBI patients.

Objective To observe whether bone marrow mesenchymal stem cell transplantation can improve vasospastic rats sense and motor function.Methods Rats grouped with randomized number method as Control group,Subarachnoid hemorrhage group.Stem cell culture media group and Stem cell transplantation group.Subarachnoid hemorrhage model were made with tail artery blood twice injection,2 days after 2' nd injection.Bone marrow mesenchymal stem cell were transplanted to lateral cistern.Subarachnoid hemorrhage(SAH) group didn' t transplant stem cell.Stem cell culture media group injected DMEM media as DMEM group.Stem cell transplantation group injected 30μl Bone Marrow mesenchymal stem cell suspension,so called BMSCs group.Neurofunctional score and learning memory expression were detected with morris mazer and Neurofunctional Score Scale in each group.Results After transplantation for 7 d,functional score of Control,SAH,DMEM and Stem cell group were 3.95 ±2.51,7.20 ± 1.03,7.23 ± 1.79 and 5.81 ± 1.11 respectively.Compared with others groups,Stem cell group score was significantly decrease(P=0.017).After transplanting stem cell for 14 d,the mean spanning plate time in Control group,SAH group,DMEM group and Stem cell group were 7.38 ± 1.73,4.52 ± 0.90,5.11 ± 1.93 and 7.32 ± 2.16 respectively,SAH and DMEM group vs other 2 groups,there were clearly statistically differences (P =0.009),while between control group and stem cell group,there were no statistically differences (P =0.14).Conclusion SAH rat transplant stem cell can improve sense,motor and learning expression in certain level.

ObjectiveTo study the risk factors and pathogenic characteristics of ventilator associated pneumonia(YAP). MethodsRisk factors of VAP,pathogens and drug resistance in 118 patients in ICU were analyzed retrospectively. ResultsThe incidence of VAP was 44.9％, and age, the mechanical ventilation time, state of consciousness,tracheotomy,the antibiotic combination are risk factors of incidence of VAP. 53cases of VAP(68.2％ )were infected by Gram negative bacilli,25.9％ were by Gram positive cocci,and 5.9％ were by fungi. Drug resistance was observed obviously. ConclusionThe occurrence of VAP was related with multiple factors. The gram negative bacteria are the major pathogens of VAP, and the rate of drug resistance was high. The occurrence of VAP could severely affect patients' prognosis.