Objective To assess effects of cerebral ischemic postconditioning(IPost)on neuronal apoptosis and phosphated glycogen synthase kinase-3β(GSK-3β)after cerebral ischemic/reperfusion.Methods The experiment was conducted at the center for animal experiment of Renmin Hospital,Wuhan University.Thirty male Wistar rats were randomly divided into three groups:sham operation(S),ischemic/reperfusion(I/R)and ischemic postconditioning(IPost).Each group contained ten rats.Global brain ischemia was produced with four-VO method.Animals were killed after two days.Apoptosis in neurons in the cortex region were detected by TUNEL assay; infarct areas were detected by TTC ; activity of p-GSK-3β was detected by spectrum assay; Statistical software SPSS13.0 was applicated to perform one-way ANOVA Student-Newman-Keul test; correlation was detected by linear regression.Results Compared with group S,TUNEL-positive cells and infarct areas increased(P ＜0.01),the activity of p-GSK-3β decreased in I/R and IPost groups(P ＜ 0.01).Compared with group I/R,TUNEL-positive cells and infarct areas significantly decreased(P ＜ 0.01),the activity of p-GSK-3β increased in group IPost(P ＜ 0.01).The activity of p-GSK-3β and TUNEL-positive cells had highly correlation,so as infarct areas(P ＜ 0.01).Conclusions IPost can lessen the ischemic/reperfusion injury of Cortex,through increas the activity of p-GSK-3[β and decreasing neuronal apoptosis.

Objective To investigate the effect of radix paeoniae rubra (RPR) on expressions of p38MAPK, NF-κB and iNOS in ltpopolysaccharide (LPS)-induced acute lung injury (ALI) and explore its protective mechanism. Methods Forty Wistar rats with ALI were divided randomly into five groups: saline control group (Group A) , LPS group (Group B), RPR for treatment group (Group C), RPR for prevention group (Group D) and SB203580 group (Group E). The effects of RPR on protein content and the ratio of neutrophiles in bronchoalveolar lavage fluid (BALF), contents of malondidehyde (MDA) and serum NO in lung tissue were observed. Arterial blood was collected for blood gas analysis. Expressions of p38MAPK, NF-κB and iNOS in rat lung tissues were detected. Results Compared with Group A, expressions of p38MAPK, NF-κB and iNOS were significantly increased (P <0.01 orP< 0.05) , the protein content and the ratio of neutrophiles in BALF, contents of MDA and serum NO were significantly higher (P<0.01 or P<0.05) in Groups B, C, D and E. There was a significant decrease in the level of arterial bicarbonate and partial pressure of oxygen in Groups B, C, D and E (P<0.01 or P<0.05). Compared with Group B, the expressions of p38MAPK, NF-κB and iNOS were significantly lower, and the protein content and the ratio of neutrophiles in BALF, contents of MDA and serum NO were significantly decreased, while the levels of arterial bicarbonate and partial pressure of oxygen were significantly higher in Groups C, D and E (P<0.05 or P<0.01). Compared with Group B, the lunginjury of Groups C, D and E was significantly alleviated, while there was no statistical difference among Groups C, D and E in the indices of lung injury. Conclusion Protective effect of RPR on LPS-induced ALI is closely related to the inhibited expressions of p38MAPK, NF-κB and iNOS.

Objective To investigate the role of p38 mitogan-activated protein kinase signal transduction pathway in lipopolysaccha-ricle-indaced acute lung injury (ALI) in rats. Methods 48 wistar rats were randomly divided into 3 groups, saline control group (group A), lipopolysaccharide (LPS) group(group B) and SB203580 group(group C). Models of lipepelysaccharide-imluced ALI were used to oh-serve the expression of p38MAPK in rat lung, protein content, the ratio of neutrophiles in bronchoalveolar lavage fluid (BALF) , pulmonary MDA content and the activities of serum NO. After LPS dripping for 6 hours, arterial blood was drawn for analysis and lung tissue was detec-ted. Results Compared with those in group A, expression of p38MAPK were markedly increased in group B and C (P<0.01). But in group C, expression of P38MAPK was significantly lower than that in group B (P<0.05). The protein content, the ratio of neutrophiles in bronchoalveolar lavage fluid (BALF), content of pulmonary MDA and the activities of serum NO in group B, C were significantly higher than those in group A (P<0.01). There was a significant decrease in the level of arterial bicarbonate and partial pressure of oxygen in group B and C (P<0.01). Compared with those in group B, these indexes of lung injury were significantly lower while the level of arterial bicar-bonate and partial pressure of oxygen was increased in group C(P <0. 05 orP <0. 01). Under light microscope, the pathologic changes in-duced by LPS were significantly attenuated by SB2035g0. Conclusion The activation of P38MAPK play an important role in the mechanism of lipopolysaccharide-induced ALI.

Objective To observe the effects of exogenous sodium phosphocreatine (PCr) on cerebral repeffusion injury of rats after ischemia in order to explore the potential mechanism. Method Thirty-six healthy adult male Wistar rats with body weight 200- 220 g were randomly (random number) divided into sham operation group, ischemic reperfusion (I/R) group and PCr treatment group. The I/R model was established by using electro-cauterizing bilateral vertebral arteries and occluding bilateral common carotid arteries with atraumatic carotid clasps for 10 min, and then the clasps were released for 48 hours reperfusion. In sham operation group, bilateral common carotid arteries were exposed without occlusion. In PCr treatnent group, PCr in dose of 150 mg/kg was administered intravenously 60 min before the occlusion of bilateral common carotid arteries. Normal saline was administered intravenously instead of PCr into rats of I/R group. After reporfusion for 48 hours, the rats were sacrificed and brains removed for detections of neuron apoptosis by using TUNEL, malondialdebyde (MDA) level by using chromtometry and calmodulin (CaM) activity by using ELISA. Results Compared with sham operation group, TUNEL-positive cells, MDA level and CaM activity increased in I/R group and PGr treatment group ( P ＜0.01). Compared with I/R group, TUNEL-positive cells, MDA level and CaM activity were lower significantly in PCr treatment group ( P ＜ 0.01). Conclusions PCr can lessen cerebral ischemic reperfusion injury and neuron apoptosis, the mechanism maybe relates to the attenuation of abnormalities in calcium balance and reduction of oxygen free radicals by PCr.

OBJECTIVE:To explore the new method for clinical pharmacists training,and provide reference for improving the quality of clinical pharmacists training. METHODS:Based on the full-awareness of Definition and characteristics of stage-progres-sive learning(SPL)and problem-based learning(PBL)teaching method,SPL-PBL teaching method was used in the teaching of the-oretical and clinical practice in the process of students training in clinical pharmacist training base. The teaching course of theoreti-cal knowledge was mainly SPL in first half and PBL in another half. While in the clinical practice teaching of ward rounds,check-ing doctor's advice,participation in the development of treatment programs and case discussion,PBL was mainly used interspersed by SPL;SPL was mainly in inquiry,pharmaceutical care and medication education interspersed by PBL. RESULTS:SPL-PBL teaching method had not only effectively aroused the enthusiasm of the students'learning,enhanced the sense of responsibility and improved their clinical professional knowledge and practice ability,but also strengthened the cultivation of students communication ability,document retrieval ability,writing communication ability and self-learning ability(four abilities) to make students more quickly into the role and improve their subjective initiative. CONCLUSIONS:Practice has proved that the SPL-PBL teaching meth-od not only gives consideratin to the inndividual differences among studengts,but also mobilizes the enthusiam,and not only gives consideration to the teaching about basic theoretical knowledge,but also strengthens the training for clinical thingkig andfour abili-ties,wich can be used for the clinical pharmacists training. Meanwhile,SPL has expanded the aplication surface of PBL beacuse that SPL has reduced the requirements of PBL for overall qulity. However,it needs to be improved because of the short time appli-cation.

Objective To investigate the role of gamma-aminobutyric acid (GABA) A receptor (GABAAR) in lung tissues in lipopolysaccharide (LPS)-induced acute lung injury in rats.Methods Thirty-two male Wistar rats,aged 8 weeks,weighing 200-230 g,were randomly divided into 4 groups (n =8 each) ∶ control group (group C),group LPS,GABA pretreatment + LPS group (group GABA) and GABAAR antagonist bicuculline pretreatment + LPS group (group BIC).Acute lung injury was induced by intravenous LPS 5 mg/kg in groups LPS,GABA and BIC,while the equal volume of normal saline was given in group C.GABA 50 mg/kg and bicuculline 10 μmol/kg were injected intraperitoneally 30 min before LPS injection in GABA and BIC groups,respectively.Arterial blood samples were collected at 6 h after LPS injection for measurement of arterial partial pressure of oxygen (PaO2).The animals were then sacrificed and lungs removed for determination of W/D lung weight ratio,GABAAR expression,contents of interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α) and malondialdehyde (MDA) and superoxide dismutase (SOD) activity in lung tissues and for microscopic examination.Results Compared with group C,PaO2 was significantly decreased in the other three groups,and W/D lung weight ratio,TNF-α,IL-6 and MDA contents were significantly increased,GABAAR expression was up-regulated,and SOD activity was decreased in groups LPS and GABA (P ＜ 0.05).Compared with group LPS,W/D lung weight ratio,TNF-α,IL-6 and MDA contents were significantly increased,GABAAR expression was up-regulated,and SOD activity was decreased in group GABA (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group BIC and in PaO2 in groups GABA and BIC (P ＞ 0.05).Conclusion GABAA R in lung tissues is involved in the development of acute lung injury induced by LPS.

Objective To investigate the mechanism of myocardial ischemia and reperfusion-induced acute lung injury (ALl) and protective effect of ischemic postconditioning.Methods Forty SD rats were allocated to sham group,myocardial ischemia/reperfusion group (reperfusion group),ischemic postconditioning group (postconditioning group),and ischemic postconditioning + phosphatase and tensin homolog deleted on chromosome ten (PTEN) inhibiting group (inhibitor group) according to the random number table,with 10 rats per group.Myocardial ischemia/reperfusion was induced by left anterior descending coronary artery occlusion.Postconditioning was performed within 1 minute before reperfusion consisting of 3 10 s cycles of reperfusion followed by 10 s occlusion.Lung was immediately removed 120 minutes after reperfusion for HE stain,immunohistochemical detection of inflammatory factors and apoptosis factors,TUNEL assay of cell apoptosis,and Western blot of protein kinase B (Akt),phospho-Akt (p-Akt),glycogen synthase kinase-3β (GSK-3β),and phospho-GSK-3β (p-GSK-3β).Results Down-regulated B-cell lymphoma-2 (Bcl-2) and IL-10 and up-regulated Bcl-2 associated X protein (Bax),cysteinyl aspartate specific proteinase-3 (Caspase-3),IL-6 as well as IL-8 were observed in other 3 groups compared with sham group (P ＜0.01).Moreover,down-regulated Bax,Caspase-3,IL-6,IL-8 as well as TUNEL and up-regulated Bcl-2 as well as IL-10 were observed in reperfusion group compared to postconditioning group and tensor group (P ＜ 0.01).No statistical differences were found among the four groups in levels of Akt,p-Akt,and GSK-3β,but level of p-GSK-3β was significantly down-regulated in reperfusion group compared to other 3 groups(P ＜ 0.01).Conclusion Development of ALI may relate to down-regulation of p-GSK-3β evoked directly by the release of inflammation factors in early period of myocardial ischemia/reperfusion and ischemic postconditioning may attenuate the condition.

AIM: To investigate effect of Wujibaifeng Pills (WJBFP) on osteoporosis of ovariectonmized (OVX) rat. METHODS: Ovariectonmized (OVX) rat model was established to evaluate osteoporosis of which parameters investigated included bone gla protein (BGP), alkaline phosphatase (ALP), bone minera density (BMD), Serum phosphorus and serum total calcium. RESULTS: WJBFP(1.0g/kg,2.0g/kg,4.0g/kg) could enhance the contents of serum estradiol and calcitonin, decrease serum BGP level in OVX rats; It had no effect on serum total calcium and ALP activities but increase level of serum phosphorus; It could enhance BMD, prevent OVX rat from decreasing bone loss without raising body weight; furthermore, it could inhibit both the uterus and adrenal gland atrophy. CONCLUSION: WJBFP might have better prevention on osteoporosis of ovariectionmized rats.

AIM　To investigate the effect of 4-aminopyridine (4-AP) on ion channels of myocytes. METHODS　L-type calcium channel and sodium channel currents were recorded in guinea pig single ventricular myocyte using whole-cell patch-clamp techniques. RESULTS　4-AP, 0.1, 0.5 and 1.0 mmol*L-1 were shown to inhibit L-type calcium channel currents (ICa,L) and sodium channel currents (INa) concentration-dependently. The percentage of inhibition were (11.6±1.7)%, (37.5±8.3)% and (54.5±6.9)% (P<0.01) respectively for ICa,L, and (22.1±14.3)% (P<0.05), (39.4±8.8)% and (62.3±6.8)% (P<0.01) respectively for INa. 4-AP 0.5 mmol*L-1 shifted the I-V curves of ICa,L and INa upwardly. CONCLUSION　4-AP blocked L-type calcium channel and sodium channels in guinea-pig ventricular myocytes concentration-dependently.

Objective To investigate the effect of penehyclidine hydrochloride on the cell apoptosis in lung tissues in a rat model of traumatic acute lung injury (ALI) .Methods Fifty-four SD rats weighing 225-275 kg were randomly divided into 3 groups ( n = 18 each) : control group (group C) , ALI group, penehyclidine hydrochloride group ( group P) . Traumatic ALI was induced by dropping a self-made impact device on the chest of anesthetized rats according to the technique described by Raghavendran et al. Intraperitoneal penehyclidine hydrochloride 2 mg/kg was injected immediately after blunt chest trauma and at 12 h after blunt chest trauma in group P. Six rats in each group were sacrificed at 3, 12 and 24 h after blunt chest trauma and the lung tissues collected for microscopic examination and determination of cell apoptosis (by TUNEL) and expression of Bax and Bcl-2 (by immuno-histochemical staining) . The apoptosis index was calculated. Results The apoptosis index and expression of Bax and Bcl-2 were significantly higher, while the ratio of Bcl-2/Bax was significantly lower at each time point in groups ALI and P than in group C ( P ＜ 0.05) . The apoptosis index and Bax expression were significantly lower,while the Bcl-2 expression and ratio of Bcl-2/ Bax higher at each time point in group P than in group ALI.The microscopic examination showed that penehyclidine hydrochloride injection significantly attenuated the pathologic changes. Conclusion Penehyclidine hydrochloride can reduce the traumatic ALI through inhibiting the cell apoptosis in rat lung tissues.