Objective:To investigate the regulation of PTEN with RNAi on NR2B after OGD.Methods:In the OGD model with anaerobic gas mixture and deoxygenated,glucose-free extracellular solution,shRNAspten-GFP and unrelated control plasmid PshGFP transfection were done respectively.The quantities of NR2B in the neuronal membrane were quantitatively measured with colorimetric assay,and the expression of NR2B mRNA were analyzed by RT-PCR.The neuronal activity was analyzed by AlamarBlue andintracellular free calcium concentration were measured with Fura-2 AM by laser confocal microscope(LCM).Results:shRNAspten-GFP plasmid can been expressed successfully in cultured neurons.The study demonstrated that there are numerous NR2B subunit expression on neuronal membrane by colorimetric assay(OPD)assay and RT-PCR.The expression and quantity of neuronal NR2B with shRNAspten-GFP treatment was significantly decrease after oxygen and glucose deprivation(OGD)and reperfusion(P

Objective: To investigate the regulation of PTEN on apoptosis and the active roles in blocking Ca2+ influx and neuroprotection.Methods: In the OGD model with anaerobic gas mixture and deoxygenated,glucose-free extracellular solution,shRNAspten-GFP and unrelated control plasmid PshGFP transfection were done respectively.Intracellular free calcium concentration were measured with Fura-2 AM fluorescent indicator,the expression of PTEN were analyzed by immunoblotting(blot) and neuron apoptosis were detected by AO/EB and Flow Cytometry.Results: Apoptosis rate and calcium concentration after OGD was significantly higher than that of normal group(P

Objective To investigate the time and distribution characteristics of neuron apoptosis occurred after global cerebral ischemia in rats,so as to provide some helpful experimental basis both for the early interference of injured neuron and the prevention and treatment of brain secondary damage.Methods In the global cerebral ischemia model with modified 4-vessel occlusion method,the amount and distribution of apoptotic neuron in hippocampus and cortex after ischemia and reperfusion were detected with TTC staining,TUNEL labeling and AO/EB fluorescence examination techniques.Intracellular free calcium concentration was measured with Fura-2 AM fluorescent indicator.Results TUNEL showed that apoptotic neurons scattered in hippocampus 3 h after reperfusion,then spread around to cortex.The amount of apoptotic cell reached to the peak at 24-48 h.The died cells appeared later and mainly dispersed around the apoptotic cells.There was a significant increase of the amount of died cells 72 h after ischemia and reperfusion.AO/EB fluorescence examination showed that the total amount of the apoptotic neurons in hippocampus,frontal cortex and parietal cortex reached their peak at 24 h and 72 h respectively,and they were significantly higher than that in control group(P

Objective:To observe clinical therapeutic effect of recombinant human erythropoietin (rhEPO)on pa-tients with heart failure complicated chronic kidney disease.Methods:A total of 84 patients with heart failure com-plicated renal insufficiency were selected.They were divided into routine treatment group (n=42)and rhEPO group (n=42,received rhEPO based on routine treatment)according to random number table method.Echocardiographic examination results and renal function were compared between two groups after 12-week treatment.Results:Com-pared with routine treatment group,there were significant rise in left ventricular ejection fraction [LVEF,(37.2± 10.3)% vs.(45.4 ± 11.4)%]and left ventricular fractional shortening [LVFS,(19.6 ± 4.3)% vs.(24.5 ± 3.8)%],and significant reductions in left ventricular end-diastolic diameter [LVEDd,(6.12±0.67)mm vs.(5.01 ±0.54)mm],24h urinary protein [(0.76±0.1)g vs.(0.24±0.09)g],24h urine microalbumin [(319.6±39.6) mg vs.(107.3±26.7)mg],blood urea nitrogen [(10.3±1.9)mmol/L vs.(6.2±1.5)mmol/L]and serum creati-nine [(97.2±16.8)μmol/L vs.(79.3±15.7)μmol/L]in rhEPO group,P<0.01 all.Conclusion:Recombinant human erythropoietin could significantly improve heart,renal function in patients with heart failure complicated re-nal insufficiency.

Objective To reveal the potential different longitudinal motion characteristics and differences of left and right muscular band (L-MB,R-MB)of interventricular septum (IVS) during systole and diastole in normal subjects.Methods Dual Doppler tissue imaging was applied for the longitudinal velocity curves of L-MB and R-MB of anterior and posterior interventricular septum(P-IVS,A-IVS) at basal,mid,and apical levels in eighty-two healthy subjects.Peak isovolumetric contraction (VIVC),systolic (Vs),early(Ve) and late (Va) diastolic velocities were measured,and myocardial velocity gradient(MVG)and Ve/Va ratio of each segment of L-MB and R-MB were calculated.Results ①Peak velocity of MB:the VrvC,Vs,Ve and Va of R-MB of both A-IVS and P-IVS were higher than those of L-MB; the VrvC,Vs,Ve and Va were gradually decreased from the base to apex of both A-IVS and P-IVS;the VIVC and Vs of A-IVS were higher than those of P-IVS; the Ve and Va of A-IVS were lower than those of P-IVS.②Correlation and MVG analyses of peak velocity of MB:the systolic and diastolic velocities between L-MB and R-MB of both A-IVS and P-IVS have shown a very strong positive correlation;the isovolumetric systolic MVG1 of L-MBof A-IVS,systolic MVG1 of R-MB of P-IVS and the diastolic MVG1 of both A-IVS and P-IVS were higher than those of MVG2.Conclusions The heterogeneity of longitudinal peak systolic and diastolic velocity and regional myocardial compliance,the consistency of moving direction and positive correlation of longitudinal peak systolic and diastolic velocity between L-MB and R-MB of both A-IVS and P-IVS have been demonstrated in normal subjects.

Objective To investigate and analyze the perioperative nursing of patients with breast cancer, and explore the effectiveness of the application of mind map in perioperative nursing care of patients with breast cancer. Methods The cluster sampling method was employed, and 265 patients with breast cancer receiving radical mastectomy treatment from department of general surgery of the First Affiliated Hospital of Liaoning Medical University were gathered according to the inclusion and exclusion criteria. 137 cases were assigned to test group (May 2014 to April 2015), and 128 cases were assigned to control group (May 2013 to April 2014 ). Nursing work was improved by establishing research group, discussing the design of perioperative nursing mind map and applying mind maps. Effect was evaluated by evaluating the incidence of postoperative complications in patients with breast cancer, the quality of patient′s life and the patient′s satisfaction. Results The incidence of bleeding, seroma, flap necrosis, upper extremity swelling and other postoperative complications of patients with breast cancer in test group was 26.28%(36/128),and that in control group was 73.44%(94/137). The differences between the two groups were of statistical significance (χ2=58.891, P<0.01). The perioperative survival quality scores of physiology, society/family, emotions, function, additions and total in the test group were (2.43 ± 1.09), (2.64±0.97), (2.85±0.97), (2.43±1.09), (2.22±1.12),(2.34±1.13), and that in the control group were (1.89± 1.09), (1.89 ± 1.06), (1.78 ± 0.92), (1.89 ± 1.09), (1.76 ± 1.05), (2.00 ± 1.09)respectively. The differences between the two groups were statistically significant (t=4.664-10.829,P<0.01). The proportion of patient satisfaction in test group was 94.89%(130/137),and that in control group was 82.03%(105/128), respectively. The differences between the two groups were statistically significant (χ2=10.900, P < 0.01). Conclusions Application of mind map in nursing intervention is able to help nurses to grasp the key content and direction of perioperative nursing on patients with breast cancer. More importantly, not only can it improved patients′quality of life and reduce the incidence of postoperative complications, but also increases patients′satisfaction.

Objective To explore the clinical expression of P53, Livin and PARP in the epithelial ovarian cancer and its correlation with the chemotherapy resistance and clinical prognosis.Methods 74 specimen of epithelial ovarian cancer confirmed from January 2009 to June 2011 in our gynecology department were selected.During the follow-up visit, the subjects were divided into chemotherapy sensitivity group and chemotherapy resistance group according to the recurrence cases, the clinical expression and survival rate for two groups were compared, the influence factors of survival time were analyzed.Results The positive rate of P53, Livin and PARP for chemotherapy sensitivity group was 47.1%, 56.9%and 52.9%;the positive rate for chemotherapy resistance group was 73.9%, 95.7% and 95.7%,the diyforences were significant(P<0.05).After 1, 3 and 5 years of treatment, the survival rate for chemotherapy sensitivity group was 100.0%, 82.4% and 66.7%,The survival rate for chemotherapy resistance group was 87.0%, 26.1% and 8.7%,the diyforences were significant(P<0.05).Based on the Cox regression model, the influence factors of the patient's age, pathological differentiation degree, clinical staging and chemotherapy sensitivity were introduced.It was known that the patient's survival time was greatly influenced by clinical staging and chemotherapy sensitivity (P<0.05).Conclusion For patients with epithelial ovarian cancer, the expression of P53, Livin and PARP is correlated with chemotherapy resistance.Therefore, the clinical effect is predictable, for patients with higher expression, the personalized therapy can improve the patient's prognosis.

Objective To investigate the effects of miR-335-5p on the proliferation and apoptosis of osteoblasts which were exposed to high glucose condition, and explore its possible molecular mechanisms. Methods MC3T3-E1 osteoblasts were divided into four groups:control group(5. 5 mmol/L glucose), high glucose group(HG group, 22. 0 mmol/L glucose), agomir-335-5p group(transfected with agomir-335-5p and exposed to 22. 0 mmol/L glucose) , and agomir negative control group( agomir NC group, transfected with agomir negative control and exposed to 22. 0 mmol/L glucose), cultured for 7 days. Cell proliferaton, cell apoptosis, expressions of miR-335-5p and dickkopfhomolog1(DKK1)mRNA,proteinlevelsofDKK1andcysteinylaspartate-specificproteinase-3(caspase-3) were detected using MTT, flow cytometry, quantitative realtime PCR and western blot, respectively. Results Compared with control group, the expression of miR-335-5p mRNA and cell proliferation in HG group were significantly decreased(P<0. 05), while cell apoptosis and the protein levels of DKK1 and caspase-3 were increased significantly(P<0. 05), the expression of DKK1 mRNA did not change (P>0. 05). The miR-335-5p mRNA expression and cell proliferation in agomir-335-5p group were higher than those in HG group and agomir NC group(P<0. 05). However, Cell apoptosis and the protein levels of DKK1 and caspase-3 in agomir-335-5p group were lower than those in HG group(P<0. 05). Conclusion High glucose inhibits the proliferation and induce the apoptosis of MC3T3-E1 osteoblast through decreasing the expression of miR-335-5p and subsequently increasing the DKK1 expression.

Objective To establish the preferable puberty pathological aggression animal model.Methods The experimental models were established towards puberty rats by frustration test of non-reward and instigation test.Rat models were tested for specificity using open-field test,saccharine test,elevated plus-maze(EPM)and olfactory sensibility.Results ①Compared with normal aggression(52.5±5.36)and control group(8.83±1.34)in total aggressive times,the pathological aggression group(101.17±2.85)increased significantly(P<0.01);②Some behavior items of each model group were moderately or hishly correlated to total score(r=0.379～0.929);③There was a significant difference between pathological aggression group and normal aggression group in the numbers of attack toward vulnerable-body regions,persistence attack after intruder displayed submissive and hish attack/threat ratios(P<0.01);④The pathological aggression group did not show depression,anxiety and olfactory disorder,except for space cognitive function(P>0.05).However,the normal aggression group displayed obvious depressive mood.Conclusion Each behavioral index matches the criteria of pathological aggression model.Meanwhile,it also excludes other factors of disturbing the specificity of the model.It suggests this model may be preferable puberty pathological aggression animal model.

Objective To investigate the effect of endoplasmic reticulum stress in renal damage caused by hyperlipidemia. Methods Forty male Wistar rats were randomly divided into two groups: normal control group (NC, n=20) and high-fat group (HF, n=20). Rats in NC group were fed with normal diet while those in HF group were fed with high-fat diet. Five rats in each group were randomly chosen in week 4, 8, 12 and 18. Serum lipid, urine protein in 24 hours and the pathological changes of renal tissues were observed; the apoptosis of renal cells was detected by TUNEL staining; the expression of GRP78 protein in the kidney was examined by immunohistochemistry. The expression of GRP78 mRNA and CHOP mRNA was examined by RT-PCR. Results Compared with those in NC group, serum lipid as well as the expression of GRP78 mRNA and protein in the kidney were increased in week 4, 8, 12 and 18 in HF group(P<0.05). In contrast, urine protein in 24 hours, the apoptosis index of renal cells and the expression of CHOP mRNA were increased in week 8, 12 and 18 (P<0.05). Conclusion CHOP pathway of endoplasmic reticulum stress is involved in renal damage caused by hyperlipidemia.