OBJECTIVE:To study the effects of Akt inhibitor MK-2206 on the proliferation and apoptosis of lung adenocarcino-ma A549 cells. METHODS:The optical density of A549 cells was detected by MTT assay after treated with 0(blank control),0.5, 1,2.5,5,10,20 and 30 μmol/L MK-2206 for 24 h;after pretreatment with 0(blank control),5,10 and 20 μmol/L MK-2206 for 24 h,morphological changes of A549 cells were observed with inverted microscope. Cell cycle and apoptosis rate were analyzed by flow cytometry. Western blot was used to detect the expression of cell cycle-related proteins Cyclin D1,p21 and p27 and apopto-sis-related protein PARP(cf-PARP),cf-caspase-3,Bcl-2 and Bax. RESULTS:Compared with blank control,the optical density of A549 cells decreased,cells shrank and presented vesicular state after treatment of MK-2206;A549 cells arrested in G0/G1 stage, the protein expression of p21 and p27 strengthened while that of Cyclin D1 decreased;the apoptotic rate of cells increased,the ex-pression of cf-PARP,cf-caspase-3 and Bax in cells increased while that of Bcl-2 decreased. All reponse were in concentration-de-pendant manner (P<0.05 or P<0.01). CONCLUSIONS:MK-2206 can inhibit the proliferation of A549 cells,and induce the apoptosis of A549 cells by adjusting the expression of activating caspase-3,down-regulating Bcl-2 and up-regulating Bax.

Objective To investigate the reverse effect and mechanism of the extracted solution of chinese medicine MBC on the drug-resistance of K562/ADM in vitro. Methods MTT assay was used to determine the IC50 of ADM on K562/ADM cell line and the reverse multiples of the drug-resistance before and after MBC treatment. Flow cytometry(FCM) was applied to assess the intracellular concentration of Rh123. Results 8?10 -1 ?g/ml MBC increased cytotoxic effect of ADM on 562/ADM cells, the reverse multiple was 6 28, and MBC could increase the intracellular concentration of Rh123. Conclusion MBC could increase killing effect of ADM on the drug-resistant K562/ADM cell and partly reverse the drug-resistance of K562/ADM cell.

Objective To analyze the effect of clinical pathway of nursing on health education in patients undergoing tonsil-lectomy.Methods Participants included 180 patients undergoing tonsillectomy in our hospital from September 2012 to September 2013 were randomly divided into the clinical pathway of nursing group ( experimental group) and the traditional nursing group ( control group) with 90 cases in each group.The sleep quality, heart rate, blood pressure, body weight, hemorrhage, health education com-pliance rate and nursing service satisfaction were recorded and compared between the two groups.Results The sleep quality, health education compliance rate and nursing service satisfaction in the experimental group was better than that in the control group;The inci-dence of heart rate speed up, blood pressure increase, loss weight, the standard rate of health education, hemorrhage and the satisfac-tion of nursing quality was lower than that in the control group.There was statistical significance in the difference between the two groups (p<0.05).Conclusion Health education with the clinical pathway of nursing can improve the quality of nursing, promote the rehabilitation and increase the nursing service satisfaction .

Objective To construct simulation medical care comprehensive training and cultivate stu-dents'nursing professional competence.Methods By setting medical simulation scenarios,comprehensive training of medical care based on admission,hospitalization,discharge and re-admission process for chronic medical patients was constructed.Self-designed questionnaire was adopted to investigate and evaluate the effect of teaching by systematic sampling from students and teachers in two dimensions.Results Compared with the control group,students'nursing skills,clinical comprehensive thinking,self-learning ability,communication skills and other professional competence of the experimental group were significantly increased.Conclusions Construction and implementation of comprehensive medical care with simulated clin-ical scenarios training and inpatient demand-oriented in vocational nursing students achieves a seamless con-nection on teaching and clinic and the nursing professional ability of students can also be improved.

Objective To discuss the problem of upper respiratory tract infection presenting various clinical manifestations in adults thereby likely making misdiagnosis and to put forth the patients with the symptom of acute abdominal pain as examples in order to caution emergency physicians to avoid misdiagnosis and mistreatment.Methods Data of 58 adult patients with upper respiratory tract infection were collected from January 2010 to December 2011.The chief complain of these patients was acute abdominal pain.After analysis of data including clinical history,symptoms and signs,laboratory findings and medication,patients were divided into two groups post hoc:correctly diagnosed group (group A) and misdiagnosed group (group B).In group A,anti-chill medicine such as tylenol cold tablets (a compositus of paracetamol,pseudoephedrine hydrochloride,dextromethorphan and chlorpheniramine) was given to patients,while in group B,anisodamine (muscarine cholinergic blocker) and antibiotics were given instead of anti-chill medicine.The anti-chill medicine such as tylenol cold tablets would be given to patients of group B when abdominal pain was not released 8-48 hours after treatment.The course of treatment and outcome of patients in the two groups were observed.Results Of 28 patients in group A,27 had abdominal pain relieved or disappeared 2-3 hours after treatment and only 1 patient had abdominal pain lasted 12 hours after treatment.Of 30 patients in the group B,only 2 patients presenting the symptom of acute gastroenteritis got totally relived 2-3 hours after treatment,while the rest of 28 patients did not get rid of abdominal pain,or only had brief period of painlessness after anisodamine injection.Alternatively,8-48 hours later as Tylenol Cold Tablets was given to patients of group B,the abdominal pain was relieved or disappeared after 2-3 hours without recurrence in 2-48 hours.Conclusions Upper respiratory tract infection manifests diversely in adults,and acute abdominal pain could be a main clinical manifestation.Anti-chill medicine,such as tylenol cold tablets,has a noticeably therapeutic effect for acute abdominal pain caused by upper respiratory tract infection in adults,while anisodamine injection makes poor efficacy.

Objective To analyze right ventrieular diastolic dysfunction in essential hypertension by quantitative tissue velocity imaging(QTVI). Methods Twenty normotensives and forty-five hypertensives underwent standard Doppler echocardiography and QTVI of right ventricular lateral tricuspid annulus. By QTVI,diastolic measurements were obtained. According to WHO standard, the patients with hypertension were classified into three groups, grade Ⅰ (group Ⅰ) 15 cases, grade Ⅱ(group Ⅱ) 15 cases, grade Ⅲ (group Ⅲ) 15 cases. Results Pulsed Doppler eehocardiography showed impaired Doppler diastolic indexes in hypertensives, without changes among the eases groups. In all hypertension groups, QTVI showed reduction of right ventricular Ea, increase of Aa and Ea/Aa<1 comparison with controls. IVRT of Ⅱ, Ⅲgroups was longer than normal. Conclusions Right ventricular diastolic function with essential hypertension decreased according to the degree of staging. QTVI has a greater predictive value than conventional imaging, and is useful for evaluating right ventricular function in patients with essential hypertension.

Objective To observe the association between homocysteine (Hcy)level and low limb vascular disease(LLVD) in newly-diagnosed type 2 diabetic patients. Methods The study subjects were divided into three groups : control group (NC), newly-diagnosed type 2 diabetes group without peripheral artery disease (T2DM) and peripheral artery disease with newly-diagnosed type 2 diabetes group (T2DM+LLVD).Both Hcy and the ankle-brachial index(ABI) were measured in three groups. Results Compared with NC group, clinical parameters of body mass index,waist-to-hip ratio,systolic blood pressure, total cholesterol(TC),triglycerides(TG),low density lipoprotein-cholesterol(LDL-C),and glycated hemoglobin A1C (HbA1c) increased markedly,and the content of high density lipoprotein-cholesterol (HDL-C) was markedly lower in both T2DM and T2DM+LLVD groups.In T2DM+LLVD group the contents of Hcy,LDL-C and HbA1c were markedly higher than in T2DM group,but the content of HDL-C decreased significantly.Hcy was independently associated with the development of peripheral artery disease in newly-diagnosed type 2 diabetic patients by logisitic regression analysis. Conclusions Increased Hcy is an independent risk factor for peripheral artery disease in newly-diagnosed type 2 diabetes.

ObjectiveTo study the effects of 6-hydroxydopamine (6-OHDA),and inhibitors for vesicular monoamine transporter (VMAT) on 5-hydroxytryptamine (5-HT),norepinephrine (NE) and dopamine (DA) and the expressions of tryptophan hydroxylase (TpH) mRNA,dopamine-beta-hydroxylase (DβH) mRNA and tyrosine hydroxylase (TH) mRNA in PC12 cells.Methods The cell viability was determined using MTT assay, the density of 5-HT, NE and DA was detected using enzyme-linked immunosorbent assay,and the expressions of TpHmRNA,DβHmRNA and THmRNA were detected using RT-PCR in PC12 cells at different time points (0,12,24,36,48 h )after exposure to different concentrations of 6-OHDA(25,50,100,200 μmol/L),and VMAT inhibitors,reserpine (50,100,400,1600 nmol/L),which combined with 6-OHDA( 100 μmol/L).Results (1)The cell viability declined with the increasing concentration of 6-OHDA which showed time dependence.The cell viability in PC12 cell which treated with reserpine decreased significantly in the responding group.The density of 5-HT in PC12 cell did not decrease with the increasing concentration of 6-OHDA,but the change had the time dependence,and the density of 5-HT was lowest at 36 h.The density of NE decreased with the increasing concentration of 6-OHDA which showed time dependence. The density of DA in PC12 cell decreased with the increasing concentration of 6-OHDA,but the change did not have the time dependence.The density of 5-HT,NE and DA in PC12 cell which treated with reserpine decreased significantly in the responding group. (2) The expressions of TpHmRNA, DβHmRNA and THmRNA in PC12 cell decreased with the increasing concentration of 6-OHDA which showed time dependence.The expressions of TpHmRNA(0.006 ± 0.001,0.003 ± 0.000,0.003 ± 0.000,0.002 ± 0.000) ; DβHmRNA (0.005 ± 0.002,0.003 ± 0.001,0.002 ±0.001,0.001 ± 0.000) and THm RNA (0.005 ± 0.002,0.003 ± 0.001,0.002 ± 0.001,0.001 ± 0.000) in PC12 cell which treated with reserpine decreased significantly in the responding group(F =13.336,9.000,9.393,all P =0.000).Conclusions6-OHDA can decrease the cell viability in PC12 cell,reduce the density of 5-HT,NE and DA and decrease the expressions of TpHmRNA,DβHmRNA and THmRNA,and the effects have dose and time dependence.Reserpine can aggravate this damage.

Objective To study the effect of glutamate on the expression of vascular endothelial growth factor (VEGF) mRNA and protein in cultured rat astrocytes. Methods Cultured rat astrocytes were randomly divided into 6 groups: control group (C), glutamate group (G), QA group (Q), DCG-IV group (D), L-AP4 group (L) and glutanmte-FMCPG gronp (G+M). Cells were cultured under nomoxic condition (95% air, 5% CO2). RT-PCR and ELISA methods were used to detect the expression of VEGF mRNA and protein in cultured astrocytes, respectively. G+ M group was preincubated with lmM MCPG for 30 min prior to the stimulation with glutamate. There were 7 time points at 0,4,8,12,16,24 and 48 h in each group except G+M group. Results The expression of VEGF mRNA and protein did not differ significantly among D group, L group and C group. Different from that in C group, the expression of VEGF mRNA and protein could be enhanced both in a dose-dependent and time-dependent manner in G group and Q group. Meanwhile, the enhanced expression of VEGF mRNA and protein in G group was completely suppressed by MCPG after 24 h. Conclusion Glutamate can increase the expression of VEGF mRNA and protein in cultured astrocytes, which may be due to the activation of group I metabotropic glutamate receptors in astrocytes.

Objective To investigate the properties of proliferation and differentiation of neonatal rat retinal progenitor cells (RPCs) in vitro. Methods RPCs were isolated from neonatal SD rats neural retina and cultured in DMEM/F12+N2 with EGF and bFGF (suspension medium )or 10%FBS without EGF and bFGF (differentiation medium). The cells grew as suspended spheres or adherent monolayers, depending on different culture conditions. The neural stem cells or retinal progenitors, neurons, astrocytes, retinal ganglion cells, rod photoreceptors and the proliferating cells were evaluated with immunofluorescence analysis by Nestin or Pax6, Map2, GFAP, Thy-1, Rhodopsin and BrdU antibodies respectively. Results RPCs could propagate and differentiate in suspension or differentiation medium and express the markers of Nestin (92.86%) or Pax6 (86.75%), Map2 (38.54%), GFAP (20.93%), Thy-1 (27.66%) and Rhodopsin(13.33%)in suspension medium; however, Nestin (60.27%), Pax6 (52%), Map2 (34.94%), GFAP (38.17%), Thy-1(30.84%) and Rhodopsin (34.67%) in differentiation medium. 96.4% of the population in the neurospheres was BrdU-positive cells. The cells could spontaneously adherent forming some subspheres and retinal specific cell types. Conclusion Neonatal rat RPCs possess the high degree of proliferation and can differentiate into neurons, astrocytes, retinal ganglion cells and rod photoreceptors in vitro. There are different proportions for RPCs to differentiate into specific cell types.