Objective To study the role of caspase 3 in ischemic brain damage of rats, and further understand the molecular mechanisms of ischemic cerebral vascular disease.Methods Rat models of the left middle cerebral artery (MCA) occlusion/reperfusion were made using a modification of the intraluminal sature method of Longa established by Belayev, infarct zones were confirmed by 2,3,5 triphenyltetrazolium chloride (TTC) staining, and caspase 3 expression on brain sections at the mRNA and active protein level was detected with in situ hybridization and immunohistochemistry technique, respectively.Results After 2 hours of left MCA ischemia followed by 24 hours of reperfusion, obvious infarct in the MCA dominate regions was confirmed by TTC staining; low levels of caspase 3 mRNA, and fewer of its active protein expression was found in normal brains, sham brains and contralateral brains of MCAO rats; both caspase 3 mRNA and activated protein expression in ipsilateral region were increased after 24 hours of recirculation, and even higher levels were detected at 48 hours of reperfusion.Conclusion Apoptotic mechanism might involve in delayed neuronal death after cerebral ischemia, and caspase 3 might play an important role in ischemic neuronal injury.

Objective To investigate the efficacy and safety of different strategy of antithrombotic therapy in elderly patients with atrial fibrillation (AF) and coronary disease.Methods 224 inpatients and outpatients with AF and coronary disease whose mean follow-up time was 2 years,were divided into warfarin group,warfarin combined with antiplatelet drug(aspirin or clopidogrel) group,antiplatelet drug(aspirin or clopidogrel) group and control group.The thromboembolic event rate and adverse events incidence were observed in different groups.Results The risk of thromboembolic events,particular emerging risk of cerebral infarction in the control group(12.50 ％) was significantly higher than other groups (P＜ 0.05).Thrombotic event rates in warfarin + single joint group(2.78％) was lower than other groups(warfarin group 4.29 ％,single joint group 5.81％),but there was no significant statistical difference.The overall risk of bleeding in the control group (3.13 ％) was significantly lower than other antithrombotic group (P＜0.05).The incidence of skin bruising in single joint group was lower than other antithrombotic groups,the overall bleeding risk,especially gasfrointestinal bleeding in warfarin+ single joint group was significantly higher than other groups (P＜ 0.05).Conclusion Elderly patients with coronary heart disease and atrial fibrillation need antithrombotic therapy to prevent thrombotic events.Using warfarin alone or warfarin combined antiplatelet drug does not lead to serious bleeding events,but generally bleeding risk should be attracted more attention.

Objective To explore the conditions, safety and efifcacy of exchange transfusion for extremely severe hyper-bilirubinemia in term newborns requiring mechanical ventilation. Methods Ten full-term newborns of extremely severe hyper-bilirubinemia requiring mechanical ventilation were selected from January 2010 to March 2013 in the department of neonatology. After stable vital sign was achieved by the use of conventional life support, peripheral arterial and venous synchronous exchange transfusion was performed. The bilirubin, platelets, blood calcium, blood coagulation and blood gas were monitored before and after exchange transfusion. The effects and adverse events of exchange transfusion in newborns on mechanical ventilation were observed. Results There was no death in the study, and the replacement rate of total bilirubin was 50.5%. The main adverse event was thrombocytopenia (80%). There was a signiifcant difference in platelet counting before and after exchange transfusion (P<0.05). There was no signiifcant difference in blood coagulation, pH/HCO3-of the blood gas, prothrombin time (PT) and acti-vated partial thromboplastin time (APTT) before and after exchange transfusion. Conclusions It is relatively safe to implement an exchange transfusion in the full-term newborns with severe hyperbilirubinemia in newborns requiring mechanical ventilation, but the use of conventional life support is prerequisite because it stabilizes vital signs. The relatively ideal replacement rate of bili-rubin and the low incidence of adverse reactions are expected. However long-term prognosis of neural system is still unknown.

Objective To studythe efficacy of levosimendan in treating refractory heart failure complicating severe renal in sufficiency.Methods Sixty-seven cases of refractory heart failure complicating severe renal insufficiency in the internal medicine department of our hospital were randomly divided into the levosimendan treatment group(L group,n=33)and dopamine treatment group(D group,n=34).The changes of N-terminal pro-B-type natriuretic peptide(NT-pro-BNP),left ventricular ejection fraction (LVEF)and glomerular filtration rate(GFR)before treatment and on 1,3,7,30 d after treatment were compared between the two groups and analyzed.Results Before the treatment,there were no statistically significant differences in the baseline indicators between the two groups(P＞0.05).The group L:the NT-pro-BNP level on 1,3,7 d after treatment was decreased significantly(P＜ 0.05),LVEF on 3,7 d was significantly increased compared with th baseline(P＜0.05)and GFR on 1,3,7,30 d was significantly increased compared with the baseline(P＜0.05).The group D:the NT-pro-BNP level on 7 d of treatment was significantly decreased(P＜0.05),LVEF on 7 d of treatment was significantly increased compared with the baseline(P＜0.05),and no statistically significant changes were observed in GFR on 1,3,7,30 d(P＞0.05).After treatment,NT-pro-BNP,LVEF and GFR significant level values in the group L were better than those in the group D.Conclusion Levosimendan is superior to dopamine in improving heart and renal function for the patients with refractory heart failure complicating severe renal insufficiency.

Objective: To study caspase-3 activities and the neuroprotective effect of caspase-3 inhibitor Ac-DEVD-CMK in primary cultures of rat hippocampal neurons during hypoxia/reoxygenation (H/R). Methods: After pretreated with Ac-DEVD-CMK or the vector DMSO fo 1 h, primary cultures of rat hippocampal neurons were induced to hypoxia for 3 h, followed by 12 to 48 h of reoxygenation. The neuronal viability was estimated by MTT assay, and caspase-3 cellular activities were measured by a colorimetric method using Ac-DEVD-pNA as a substrate. Results: During H/R, the cultured rat hippocampal neuronal viability gradually decreased, and caspase-3 activities in the primary cultures of rat hippocampal neurons were significantly increased, and peaked at 24 h of reoxygenation. Caspase-3 activities were decreased in the neurons pretreated with Ac-DEVD-CMK in a dose dependent manner. Conclusion: Delayed neuronal death is detected in cultured hippocampal neurons during H/R, and apoptosis mediated by caspase-3 may play an important role in it, and caspase-3 inhibitor has a neuroprotective effect on them.

OBJECTIVE To search a method for identifying Clostridium perfringens and genotyping their toxin for gene diagnosis by multiplex PCR.METHODS The mutiplex PCR was developed with three sets of primers(designed) based on the sequences of three C.perfringens toxin genes(CP?,CP? and CPE) published in GenBank to identify C.perfringens and genotype their three toxin genes.RESULTS Three expected(sequences) were (obtained) successfully by multiplex PCR and identified by electrophoresis.CONCLUSIONS The(specific) sequences of C.perfringens could be amplified and their three genes of toxins could be identified by this multiplex PCR(system).Such method should be helpful for developing gene diagnosis well.

OBJECTIVE To discuss a highly effective method to immobilize probe on the surfaces of piezoelectric DNA sensors.METHODS Pseudomonas aeruginosa probe was immobilized on the gold surface of gene sensor(array) with routine self-assembly method(SAM)(non-reduction method) and SAM with deoxidized probe((reduction) method),respectively.The changes in frequency and time-cost were compared in reactions with(different) concentrations of probe.RESULTS Reduction method had the advantage of more probe immobilization;less time consumed in testing and higher changes in frequency during the reaction than non-reduction method.CONCLUSIONS Reduction method has a better ability to immobilize probe on the surfaces of piezoelectric DNA sensors.

Objective To evaluate the diagnostic value of high frequency echocardiography on mice with doxorubicin-induced cardiotoxicity and find out the most valuable monitoring indexes for early doxorubicin-induced myocardial damage.Methods Forty mice were divided into doxorubicin-injection group (ADM group) and control group randomly.Twenty mice in each group were injected with doxorubicin and saline at the same dose for 5 weeks continuously.The parameters of cardic function of ADM group and contol group were observed by high-frequency echocardiography after infection.Brain natriuretic peptide (BNP) tested by euzymelinked immunosorbent assay(ELISA) and pathological findings were used as the golden standard.Results ①The left ventricular long axis view and the apical four chamber view could be displayed clearly by transthoracic high-frequency echocardiography.②After 5 weeks intervention,weight,heart rate,left ventricular ejection fraction (LVEF),left ventricular fractional shortening (LVFS),E/A,E wave deceleration time(EDT) and other parameters showed marked difference between ADM group and control group(P ＜0.05).③The decrease of diastolic function was earlier than systolic function in ADM group.④LVEF and E/A presented good negative linear correlation to BNP(r =-0.622,P ＜0.05; r =-0.593,P ＜0.01).EDT showed positive linear correlation to BNP(r =0.501,P ＜0.05).Conclusions Cardic function of mice with doxorubicin-induced myocardial damage could be early evaluated accurately by high-frequency echocardiography,which contained good relativity with BNP.

ObjectiveTo explore the influence of clinical symptoms and applicability of discontiguous naikan cognitive therapy(DNCT) among convalescent schizophrenic patients.MethodsApplying DNCT,100 convalescent paranoid schizophrenic patients with convalescent clinical state were consecutively recruited.All the patients were randomly divided into DNCT group and control group and were pretreated with antipsychotic agent therapy,40 patients in DNCT group and 49 patients in control group entered the statistic analysis,11 lost.In DNCT group,the patients received DNCT for successive 28 days.In control group,the patients only received antipsychotic agent therapy.Positive and Negative Syndrome Scale (PANSS),Nurses'Observation Scale for Inpatient Evaluation (NOSIE) were administered to all subjects pre- and post-treatment.ResultsAfter treatment,in the study group,total PANSS scales ( (54.00 ± 10.19 ) vs (45.05 ± 5.28 ),t =5.430,P ＜ 0.01 ),the positive symptom item ((11.00±3.33) vs (9.53 ±1.85),t=3.670,P=0.01),negative symptoms item((12.15 ±4.38) vs (9.40± 2.15 ),t =4.371,P ＜ 0.01 ),general psychopathology item ( (26.90 ± 5.66) vs (22.65 ± 3.07 ) 分,t =4.494,P＜0.01 ) scored lower than before,The difference was statistically significant.PANSS study group after treatment,total scores( (45.05 ±5.28 ) vs (52.04 ± 10.36),t=-3.876 P＜0.01 ),negative symptom item score( t =- 3.789,P ＜ 0.01 ),composite item ( t =2.251,P =0.027 ),the general psychopathology item ( t =- 3.336,P =0.01 ),score significantly lower than the control group.After twelve weeks follow-up study,in the study group,PANSS total scores ( t =4.764,P ＜ 0.01 ),item score of positive symptoms ( t =2.335,P =0.025 ),negative symptoms item score( t =3.083,P =0.004) ),genial psychopathology item score ( t =4.325,P ＜ 0.01 ) was still significantly lower than before treatment,the difference was statistically significant.In study group,after treatment,NOSIE Scale total negative factors scores( t =3.083,P =0.004) were significantly lower than before,total positive factors( t =-2.446,P=0.019),the total estimated factor in the disease scores ( t =-4.730,P ＜ 0.001 )were significantly higher than before treatment.After treatment,in the study group,negative factors ( t =-3.953,P=0.000) were significantly lower than the control group,twelve weeks follow-up,study group total negative factors of NOSIE scale score( t =2.126,P =0.040) was still lower than before treatment,the difference was statistically significant,total positive factor( t =- 2.054,P =0.047 ) still higher than before treatment,the difference was statistically significant.ConclusionDNCT can possibly improve part clinical symptoms of patients with convalescent schizophrenia to a certain extent,especially negative symptom,and the impact remained to the twelve weeks,but need to further prove the effect of naikan cognitive therapy.

OBJECTIVE To set-up safe alternatives to ethidium bromide(EB).METHODS Comparative analysis was performed on the DNA staining efficiencies of 4 fluorescent dyes including SYBR Gold,SYBR Green Ⅰ,(GoldView) and EB in preparative agarose gel electrophoresis.RESULTS Although both SYBR Gold and SYBR Green Ⅰ altered electrophoretic mobility and thus DNA size estimates,they were cost-effective alternatives to EB.SYBR Gold was more sensitive than SYBR Green Ⅰ at detecting short fragments,but 50 bp bands were clearly(visible) using either dye when visualized with a long integration time.CONCLUSIONS SYBR Gold or SYBR Green Ⅰ are sensitive and relatively safe alternatives to EB.