AIM: To investigate the molecular mechanisms of apoptosis and to elucidate the apoptosis signaling pathway triggered by etoposide in Jurkat human leukemia cells. METHODS: Apoptosis was detected using annexin V - FITC and propidium iodide (PI) staining, respectively, and annexin V - FITC positive cells and hypodiploid cells were analyzed by flow cytometry. Mitochondrial membrane potential (△Ψm) was detected using 3,3' - dihexyloxycarbocyanine iodide [ DiOC6 (3)] staining and △Ψm low cells were analyzed by flow cytometry. Preparation of cytosolic extracts and isolation of mitochondria were completed by centrifugation. Western blotting analysis was used to evaluate the level of cytochrome c, caspase - 3, and poly ( ADP - ribose) polymerase (PARP) expression. RESULTS: Etoposide induced apoptosis showing phosphatidylserine externalization and DNA fragmentation in a time - dependent manner and the apoptosis could be inhibited by a broad caspase inhibitor benzyloxycarbonyl - Val - Ala - Asp - fluoromethylketone ( zVAD. fmk). Collapse of △Ψm induced by etoposide preceded DNA fragmentation and phosphatidylserine externalization. In contrast, it was not blocked by zVAD. fmk. Etoposide caused cytochrome c release from mitochondria into cytosol, subsequent activation of caspase-3 (32 kD) presented with an intermediate (20 kD) and its active product (17kD), and cleavage of full- length PARP (116 kD) into the so- called apoptotic 85 kD fragment. CONCLUSION:Etoposide - induced Jurkat cell apoptosis is initiated through mitochondria signaling pathway with cytochrome c release into cytoplasm and caspase is the ultimate executioner of cell apoptosis.

Objective To evaluate the effects of traditional Chinese medicine likeDansen and western medicine in treating traumatic brain edema (TBE). Methods A total of 130 Wistar rats were divided randomly into six groups: Group A (normal contrast group), Group B (operation group), Group C (western medicine treatment group, WM group), Group D (western medicine and salvia injection), Group E (western medicine and Honghua injection) and Group F (western medicine and Sanqi injection). The model of brain edema was established by liquid nitrogen treatment to assay whole blood viscosity, water content and Even blues (EB) in brain tissues and to observe the content and pathologic changes of endothlin (ET) in plasma. Results Whole blood viscosity of Groups D, E and F was lower than that of Group B (P0.05). Water content and EB in brain tissues and ET of plasma in Groups C, D, E and F were lower significantly than those in Group B (P

Objective To investigate the clinical effect and security of spraying Zhikang Capsules guided by gastroscopy in treatment elderly upper gastrointestinal bleeding.Methods Eighty-four elderly gastric duodenal ulcer bleeding patients were randomly assigned to group A,B and C (n =28 for each group).Patients in group A were used noradrenalin to spray with gastroscopy,in group B were used thrombin and in group C were used Zhikang Capsules.The total effective,the immediate homeostasis rate,the incidence of rebleeding,the emergency operation rate and the incidence of adverse reaction were recorded and analyzed.Results Total effective rate of group C was 96.4％ (27/28),significantly higher than that in group A (71.4％,20/28),B (75.0％,21/28) and the difference was significant(P＜0.05).The incidence of rebleeding in group A,B,C were 28.6％ (8/28),17.9％ (6/28),3.6％ (1/28) respectively,and the difference was significant (P ＜0.05).The same trend was seen regarding of the incidence of adverse reaction among three groups (25.0％,7/28;10.7％,3/28;0,0/28;P＜0.05).Conclusion Spraying Zhikang Capsules guided by gastroscopy is showed with good therapeutic effect in treating elderly upper gastrointestinal bleeding and high safety,and with less adverse reaction.

Objective To screen the differential expression proteins in the children with neuroblastoma (NB) by proteomics tools.Methods Three specimens were collected from the patients diagnosed Ⅳstage NB by biopsy at Department of Pediatric Surgery of Chinese PLA General Hospital in Beijing from July to December,2011.Another three specimens were acquired from the same patients underwent tumor excision.Average age was 3.17 years.Proteins in neuroblastoma before and after chemotherapy were separated by two dimensional gel electrophoresis,analyzed by high performance liquid chromatography-eleetrospray tandem MS (Nano-UPLC-ESIMS/MS).Results After two dimensional gel electrophoresis,we obtained the maps about tissues before and after chemotherapy.There were 7 differential protein spots identified by using the Image Master two dimensional gel electrophoresis software,in which 2 were up-regulated,including Nm23 protein and neuropolypeptide h3,5were down-regulated after chemotherapy,including stathminl,heat shock protein 27,mitochondrial short-chain enoyl-coenzyme A,peroxiredoxin 1 and peroxiredoxin 3.Conclusion The differential expression proteins of children neuroblastoma before and after chemotherapy were successfully identified by two dimensional gel electrophoresis and Nano-UPLC-ESI-MS/MS.

After discussing the reason for insufficiency of committee member in current ethical review , and considering the requirement for improving the function of Ethics Review Committee , this paper proposes that Ethics Review Committee should appoint more non -affiliated scientific members to ensure the independence of ethical re-view , thus protecting the rights and beneficence of subjects , and to secure the justice in ethical review of protocols , the paper also discussed the resolutions to address the challenge brought by appointing non -affiliated members .

Objective:To explore the relationship between sleep structure changes and state or trait anxiety in patients with insomnia.Methods:Investigations were conducted with the State-trait Anxiety Inventory (STAI) and whole-night ploysomnography (PSG) recorded in 31 patients with insomnia and 20 normal controls.Repeated examinations were conducted in return visits 3～4 months after discharge.Results:Compared to control group,the patients with insomnia had decreased sleep time[(333.71±84.33) min vs.(403.65±19.29) min] and sleep efficiency[(70.41±17.35) % vs.(83.45±4.42) %],and increased sleep latency[(39.48±24.24) min vs.(19.65±8.57) min],REM sleep latency[(106.60±42.89) min vs.(86.80±12.25) min],proportion of S_1 sleep time[(25.36±14.22) % vs.(8.86±1.77) %]and awakening times[(4.45±2.51)vs.(1.75±1.07)].The proportions of S_(3+4) time[(7.38±9.70) % vs.(13.78±4.24) %] and REM sleep time reduced[(14.54±5.61) % vs.(19.18±2.14)%] (Ps<0.05).According to the STAI,scores of both state anxiety[(47.94±8.96) vs.(39.15±4.51)] and trait anxiety[(49.94±8.90) vs.(42.05±7.13)] were significantly higher in insomnia group than in control group (Ps<0.05).State anxiety was positively correlated with sleep latency (r=0.42),REM sleep latency (r=0.25),awakening times (r=0.44) and proportion of S_1 sleep time (r=0.34) (Ps<0.05),and negatively correlated with proportion of REM sleep time (r=-0.41,P<0.01).Trait anxiety was positively correlated with sleep latency (r=0.37,P<0.01) and wakefulness frequency (r=0.29,P<0.05).In return visits,the sleep structure improved,state anxiety score reduced,and trait anxiety had no significant changes.Conclusion:Patients with insomnia have significant sleep structural changes and higher state and trait anxiety.Their sleep structural changes are possilly related to state and trait anxiety.

Objective To explore the changes of plasma fibrinogen ,plasminogen and D-dimmer before and after thrombolytic therapy in patients with acute myocardial infarction (AMI) ,to provide the evidence for treatment and prognosis AMI .Methods 93 cases of patients with AMI were divided into twe groups ,reperfusion group of 77 cases ,obstract group of 16 cases .At the same pe-riod ,30 cases myocardial infarction of patients with non thrombolytic therapy were selected as control group .The levels of plasma Fib ,Plg and D-dimer in each group were detected at different times .Results The levels of Fib and D-D in AMI patients were higher than those in the control group before thrombolysis therapy (P<0 .05) .Compared with before thrombolysis therapy ,the Fib and Plg levels of AMI patients were decreased significantly after thrombolytic therapy 6 h(P<0 .01) ,and returned to the normal level after 48 h .The D-dimmer level of AMI patients was increased significantly after thrombolytic therapy 6 h(P<0 .01) ,and returned to the normal level after 7 d .After thrombolytic therapy 6 h ,the levels of Fib and Plg of reperfusion group were decreased signifi-cantly(P<0 .05) ,the level of D-dimmer was increased significantly (P<0 .05) .Conclusion The levels of plasma fibrinogen ,plas-minogen and D-dimmer have important reference value for monitor the thrombolytic therapy effect in patients with AMI .

Objective The incidence of alcoholic fatty liver increases year by year in recent years .The aim of this study was to establish an animal model of AFL to investigate the pathogenesis of hepatic fibrosis . Methods This study involved 40 male Japa-nese rabbits aged (17.01 ±1.54) d and weighing 1.00-1.52 kg, which were equally randomized to an experimental group and a control group.The animals in the former group received lavage of 10 mL of 50%ethanol twice a day, with normal feedstuff and water, while those in the control group received normal feedstuff and water only .We performed ultrasonography for dynamic liver presentation before and at 12, 16, and 20 weeks after feeding, followed by pathological observation of the livers . Results After 12 weeks of eth-anol garage , fatty liver was observed in 18 of the rabbits and it deteriorated with the prolonged time of administration . The body weight was significantly decreased in the experimental rabbits as com-pared with the controls at 16 weeks ([2.48 ±0.30] vs [2.78 ± 0.15] kg, P<0.05) and 20 weeks ([2.61 ±0.44] vs [3.10 ± 0.13] kg, P<0.05).Ultrasound and pathological grading showed 1 mild, 3 moderate, and 13 severe cases of fatty liver in the experimen-tal group, but none in the control , and pathological examination re-
vealed similar results (1 mild, 4 moderate, and 12 severe cases of fatty liver) in the former group.At 20 weeks, alcoholic fatty liver was found mainly in the S3-S4 stage. Conclusion Alcoholic fatty liver models could be successfully established in rabbits by etha-nol garage and ultrasonography is useful for monitoring the development and progression of the condition .

Objective To evaluate the efficacy and safety of uterine artery chemoembolization(UACE)in conjunction with dilata-tion and curettage in the treatment of cervical pregnancy.Methods UACE was performed in 7 patients with cervical pregnancy,after bilateral uterine artery perfusion of methotrexate,using gelatin sponge for embolization.Dilatation and curettage was carried out within 1 week after the procedure.Results The symptom of vaginal bleeding was effectively controlled after UACE,then dilatation and curettage performed in 6 within 1 week,the blood loss during curettage procedure was 10-30 mL(mean 21 mL).The serumβ-HCG decreased to normal range within 1 week in 1 and 3 weeks in 6 after intervention,average 1 6.4 ± 6.9 days.No serious com-plications occurred and follow up did not show symptoms of premature ovarian failure.Conclusion UACE combined with dilatation and curettage significantly improve the efficacy and safety of the treatment of cervical pregnancy,and avoid serious incident,such as massive hemorrhage and unnecessary resection,the clinical application is safe and effective.

Objective Folate receptors ( FRs) , overexpressed on the surface of a variety of tumor cells , are potential targets for tumor targeting therapy .This study aimed to prepare an FR-mediated drug nanocarrier with folate conjugated pullulan acetate ( FPA) to target chemotherapeutic agents to FR-overexpressed tumor cells and investigate its tumor-suppressing effect in vitro. Methods The cytotoxicity of epirubicin-loaded FPA nanoparticles ( FPA/EPI NP) against HepG2 and Hela cells was evaluated by MTS assay.The HepG2 and Hela cells were divided into five groups to be treated with NPs (NP control), chlorpromazine, chloro-quine, amiloride, and folate, respectively, followed by detection of the fluorescence intensity by flow cytometry . Results FPA/EPI NP was successfully formulated into NPs , with the mean particle size of (268.5 ±12.0) nm, by dialysis with an almost spherical shape . The drug-loading rate and entrapment rate of FPA/EPI NP were (6.45 ±1.04) and (72.45 ±11.50) %, respectively.The survival rates of the HepG2 and Hela cells were both >95%after 24 hours of incubation with FPA NP at 5, 40, 200, 400 and 1000μg/mL and 90.0%after 72 hours.The survival rates of the HepG2 cells treated with 5, 40, 200, 400 and 1000μg/mL FPA/EPI NP for 24 hours were (92.3 ±5.2), (70.4 ±4.6), (50.0 ±4.0), (41.1 ±4.1) and (27.0 ±3.6) %, respectively.Compared with the NP control group, the Hela cells of the chlorpromazine , amiloride and folate groups showed a significantly lower rate of NP uptake (P<0.05), and so did the HepG2 cells pretreated with chlorpromazine or amilo-ride (P<0.05).At 72 hours, the half maximal inhibitory concentrations (IC50) of FPA/EPI NP against HepG2 and Hela cells were 168 and 105μg/mL, respectively . Conclusion Clathrin-mediated endocytosis and macropinocytosis are involved in the internaliza-tion of FPA/EPI NP in HepG2 cells, while clathrin-and FR-mediated endocytosis in that of Hela cells .FPA NP may serve as a new drug carrier for tumor-targeted therapy .