[ ABSTRACT] AIM:To investigate the effects of BARF1 down-regulation on EBV-positive gastric carcinoma cell apoptosis, and the molecular mechanisms by BARF1 silencing-mediated apoptosis.METHODS: After NUGC3 and SNU719 cells were transfected with NCsiRNA and siRNA, respectively, the protein levels of BARF1, Bcl-2, Bax, cyto-chrome C, caspase 3 and capase 9 were detected by Western blot, and the mRNA expression of BARF1, Bcl-2 and Bax was determined by RT-PCR.The cell viability was measured by the method of Trypan blue exclusion and the cell apoptosis was analyzed by flow cytometry analysis with Annexin V-FITC/PI staining.The expression of the apoptosis-related proteins in the cells transfected with siRNA and NCsiRNA was examined by human apoptosis antibody arrays.Mitochondrial mem-brane potential was determined by flow cytometry.The interaction between Apaf-1 and caspase 9 was confirmed by immuno-precipitation.RESULTS: Compared with untreated and NCsiRNA groups, BARF1 gene silencing significantly inhibited the cell viability, induced apoptosis, and reduced the mitochondrial membrane potential in the NUGC3 and SNU719 cells transfected with siRNA.BARF1 gene silencing up-regulated the expression of pro-apoptotic proteins and down-regulated the expression of anti-apoptotic proteins, and the Bcl-2/Bax ratio was significantly decreased.In BARF1 gene silencing cells, the caspase inhibitor z-VAD-fmk inhibited BARF1 silencing-mediated apoptosis, and significantly increased the levels of cleaved caspase 3 and caspase 9.The concentration of cytochrome C significantly increased as compared with NCsiRNA group, and Apaf-1 interacted with caspase 9 in the cytoplasm.CONCLUSION:BARF1 silencing induces apoptosis via the mitochondrial pathway through regulating the expression of Bcl-2 and Bax proteins in a caspase-dependent manner in the NUGC3 and SNU719 cells.

Objective: To study the effects of adrenomedullin (ADM) and proadrenomedullin N terminal 20 peptide (PAMP) alone or in combinations on the isolated rat hearts as well as the possible signaling pathways involved in their actions. Methods: In isolated rat hearts, the left ventricular pressure (LVP), LVP?dp/dtmax, coronary fluid (CF) and heart rate(HR) of the hearts infused at different concentrations of ADM and/or PAMP were determined by a 4 cannal physiological recorder, then the cAMP contents were assayed in myocardium. Results: After being infused with ADM from 10 -11 to 10 -8 mol?L -1 or PAMP from 10 -11 to 10 -8 mol?L -1 , the LVP and LVP?dp/dtmax of the isolated hearts decreased gradually in a concentration dependent manner, and at the same concentration, the effects of PAMP were more potent than those of the ADM. When ADM and PAMP were co administrated with both concentrations as low as from 10 -11 to 10 -10 mol?L -1 , the cardiac parameters were decreased more than either ADM or PAMP administrated alone. However, the inhibitory effects of ADM and PAMP were attenuated when they were in combination at higher concentrations as from 10 -9 to 10 -8 mol?L -1 . When the rat hearts were infused with ADM, PAMP,and ADM plus PAMP, the CF were always higher than those of the controls and decreased when co administrated with L NAME, an inhibitor of NOS, but the decreaseddegree of LVP and LVP?dp/dtmax were attenuated by L NAME.The cAMP contents in the left cardiac ventricle were increased significantly by ADM infusions but not changed obviously by PAMP, and were of no statistical difference in rat hearts with ADM administrated alone from those combinated with ADM and PAMP. Conclusion: These results showed that ADM and PAMP infused alone or in combinations inhibited the function of rat hearts in vitro, which might be partly involved with the NOS/NO pathway.

Objective To investigate the memory monitoring ability in patients with hepatolenticular degeneration(HLD) and explore the mechanism of their memory impairment.Methods The feeling-of-knowing (FOK) paradigm of episodic memory(EM) and semantic memory(SM)were established and subsequently applied to 30 HLD patients and 30 healthy control(HC) participants who were matched in age and educational level.Results Compared with healthy control group (FOK-EM recall (64.17 ± 29.21) ％ ; FOK-SM recall (84.72 ± 11.44)％),the FOK-EM recall((26.55±20.92)％) and FOK-SM recall((53.93±28.42)％) in HLD patients were significantly lower(t=-5.140,P＜0.01 ; t=-5.123,P＜0.01).The positive judgment and correct recognition of FOK-EM ((50.64±29.43) ％) and the negative judgment and correct recognition of FOK-EM((12.80± 18.32) ％) in the HLD group were significantly different from the HC group (the positive judgment and correct recognition of FOK-EM:(75.15±31.73)％ and the false judgment and correct recognition of FOK-EM:(1.81±5.41)％; t=-2.693,P＜0.05 ; t=3.026,P＜0.01).Most importantly,the stroop effect was positively correlated with the negative judgment and correct recognition of FOK-EM in HLD group(r=0.601,P＜0.01).Conclusion The results show that the HLD group underestimate their memory performance on episodic FOK,and the impairment of memory monitoring is positively correlated with the deficit of executive function,indicating that the prefrontal impairment can be an influential factor of memory disorder in HLD,whereas the unimpaired semantic metamemory FOK indicates the episodic and semantic metamemory monitoring may depend on different neural network.

The changes of plasma myostatin levels in patients with type 2 diabetes mellitus (T2D) and their clinical correlation were investigated. We recruited 43 T2D patients and 20 age-matched healthy subjects. Plasma myostatin, lipid and glucose, and serum insulin were determined. T2D patients showed significantly higher fasting plasma glucose (FPG), serum insulin and triglyceride levels, and lower high-density lipoprotein levels than normal control subjects (P<0.01). Mean plasma myostatin level in T2D patients and health controls was (66.5±17.8) and (46.2±13.8) ng/mL, respectively. An unpaired t test showed that the increase of myostatin in the T2D patients was significant (P<0.001). In both healthy control and T2D groups, the female subjects showed higher myostatin levels than the male subjects. In the T2D patients, plasma level of myostatin was negatively correlated with body mass index (BMI, r=-0.42, P<0.01) and FPG (r=-0.51, P[Symbol: see text]0.01), but positively correlated with insulin resistance index (HOMA-IR, r=0.48, P<0.01). Up-regulation of plasma myostatin in the T2D patients and its correlation with BMI, FPG and blood insulin sensitivity suggests that plasma myostatin may be implicated in the pathogenesis of T2D and thus presented as a therapeutic target for treating the disease. Furthermore, circulating myostatin levels may be used as a biomarker for the disease.

Objective To explore β-sodium aescinate on vascular endothelial function ( FMD ) , homocysteine ( Hcy ) and hypersensitive C-reactive protein ( hs-CRP) and clinical efficacy in patients with acute cerebral infarction.Methods 198 acute cerebral infarction patients from March 2013 to April 2015 were randomly divided into observation group (n=100) and control group (n=98).Control group were treated according to the condition of the disease, observation group were treated by β-sodium aescinate base on control group, 20mg was added to 250mL saline for intravenous drip,one times per day.Continuous used 14d for one treatment courses.Compared the change of vascular endothelial function, Hcy and hs-CRP and clinical efficacy.Results The total effective rate of observation group was 90.00%, which was significantly higher than that of 71.42% in control group (χ2 =11.01,P<0.05).Post-treatment the value of FMD significantly increased, Hcy and hs CRP were significantly decreased both in observation group and control group respectively, which the difference had a statistically significant as compared with Pre-treatment (P<0.05);but, the value of FMD was significantly higher, Hcy and hs CRP was significantly lower in observation group than that of control group (P<0.05).Conclusion It has a significant β-sodium aescinate clinical effect in treatment of acute cerebral infarction, and FMD are significantly higher, Hcy and hs-CRP are significantly decrease.

Objective To investigate the curative effect of sodium nitroprusside in the treatment of EV71-related pneumoedema/pneumorrhagia and hypotension in this study. Methods This was a retrospective study of a total 10 children with EV71-related pneumoedema/pneumorrhagia treated with sodium nitroprusside using case-control methods. The clinical manifestations and outcomes of the 10 children (present cohort) were compared with those of 8 children (past cohort) who had EV71-related pneumoedema/pneumorrhagia without sodium nitroprusside. Results Among these 10 patients,all were at the appearance of pneumoedema/pneumorrhagia, cardiac arrhythmia and hypotension. Two patients showed severe cardiac arrhythmia, two patients showed cardiac arrest for many times. All 10 patients were treated with mechanical ventilation and other conventional treatments. At the first time of the manifest of hypotension,sodium nitroprusside were put into practice in all 10 patients. Ten patients were treated with intravenously sodium nitroprusside in the stage of hypotension by maxi30 min ～ 1 h,the patients showed an improvement in vultus,pulse and peripheral circulation and decrease of heart rate and elevation of blood pressure after 30 min ～2 h,but at least 2～6 h later,the blood pressure tended to stabilize at normal standard. Conclusion Hypotension is the intensive stage in EV71-related hand,foot and mouth disease ,and the shock syndrome caused by acute left ventricular disorder is related to sympathetic nerve activity. Sodium nitroprusside can effectively reduce the cardiac afterload,and correct shock and improve the prognosis.

Objective To investigate the expression of patterned matrix vasculogenic mimicry in hepatocellular carcinoma (HCC) and its prognostic significance. Methods HCC tissues from 151 patients undergoing curative resection in Anhui Provincial Hospital from January 2003 to December 2008 were studied.Postoperative follow-up and clinicopathologic data were reviewed.Immunohistochemical staining of laminin,CD34 and transmission electron microscopy were used to identify patterned matrix VM in HCC.The relations between clinicopathologic features, prognosis and patterned matrix VM were analyzed.Results Patterned matrix VM was positive in 31 out of 151 cases (20.5％ ).The expression of patterned matrix VM was positively correlated with tumor size ( x2 =4.132,P =0.042),vascular invasion ( x2 =5.825,P=0.016),high Edmondson grade (x2 =5.256,P=0.022),and late pTNM stage (x2 =6.218,P =0.013).Kaplan-Meier survival analysis revealed that cases of the VM positive group had poor overall 1,3 and 5 year survival (OS) and disease-free survival (DFS) than that of the VM negative group (67.7％,34.6％,11.5％ and 86.7％,64.7％,40.3％,respectively,x2 =14.852,P ＜ 0.001 ; and 41.9％,19.4％,6.5％ and 63.3％,40.6％,19.6％,respectively,x2 =10.065,P =0.002).Univariate and multivariate analyses revealed that multiple tumor nodules,vascular invasion and VM were independent prognostic factors for overall survival,while vascular invasion and VM were independent prognostic factors for disease-free survival. Conclusions Patterned matrix VM exists in HCC. The expression of patternedmatrix VM is associated with tumor size,Edmondson grade,pTNM stage and vascular invasion,and it might serve as an unfavorable prognostic factor for HCC patients.

Objective To investigate the distribution, clinical diagnosis and treatment methods of the extrapulmo-nary complications in children with mycoplasma pneumoniae (MP). Methods The clinical data of 1 100 patients confirmed the diagnosis of mycoplasma pneumonia and with the positive serum MP-IgM test were collected in this study. The distribu-tion and clinical characteristics and MP-DNA detection rates were compared between 417 patients with extrapulmonary com-plications and 683 cases without complications. The occurrence of various complications in a four-year period was analyzed. Clinical data were compared between fiberoptic bronchoscopy lavage group and non-surgical group. Results The MP-DNA detection rate and the length of hospital stay were higher in patients with pulmonary complications than those of patients without complications. The most common types of extrapulmonary complications were liver damage, skin rashes and gastrointestinal reactions , but less severe. Encephalitis, nephritis and myocarditis were rare complications, but severe and occult. The fatal hemophagocytic lymphohistiocytosis (HLH) was also visible in patients. Bronchoscopy lavage was conducive to the recovery of the disease. Conclusion MP pneumonia showed high incidence and risks of extrapulmonary complica-tions, which required careful clinical observation and inspection, the dynamic monitoring laboratory markers and comprehen-sive treatment as well.

The changes of plasma myostatin levels in patients with type 2 diabetes mellitus (T2D) and their clinical correlation were investigated. We recruited 43 T2D patients and 20 age-matched healthy subjects. Plasma myostatin, lipid and glucose, and serum insulin were determined. T2D patients showed significantly higher fasting plasma glucose (FPG), serum insulin and triglyceride levels, and lower high-density lipoprotein levels than normal control subjects (P<0.01). Mean plasma myostatin level in T2D patients and health controls was (66.5±17.8) and (46.2±13.8) ng/mL, respectively. An unpaired t test showed that the increase of myostatin in the T2D patients was significant (P<0.001). In both healthy control and T2D groups, the female subjects showed higher myostatin levels than the male subjects. In the T2D patients, plasma level of myostatin was negatively correlated with body mass index (BMI, r=-0.42, P<0.01) and FPG (r=-0.51, P[Symbol: see text]0.01), but positively correlated with insulin resistance index (HOMA-IR, r=0.48, P<0.01). Up-regulation of plasma myostatin in the T2D patients and its correlation with BMI, FPG and blood insulin sensitivity suggests that plasma myostatin may be implicated in the pathogenesis of T2D and thus presented as a therapeutic target for treating the disease. Furthermore, circulating myostatin levels may be used as a biomarker for the disease.
Blood Glucose ; Diabetes Mellitus, Type 2 ; blood ; Female ; Humans ; Insulin ; blood ; Lipids ; blood ; Male ; Middle Aged ; Myostatin ; blood

OBJECTIVETo illuminate the regulating effect of all-trans retinoic acid (ATRA) on gap junctional intercellular communication (GJIC) and connexin 43 (Cx43) gene expression in glioma cells, which is tissue- and organ-specific.

METHODRat C6 glioma cells were exposed to ATRA at a concentration of 1, 10, 100 micromol/L and the GJIC function of the cells was examined with scrape-loading dye transfer assay 24 hours, 48 hours and 72 hours after ATRA treatment. The effect of ATRA on Cx43 gene expression was measured with semiquantitative reverse transcription polymerase chain reaction (RT-PCR) 24 hours after ATRA exposure.

RESULTSThe GJIC function of C6 glioma cells was significantly increased by ATRA at each concentration applied. The dye passed 4 to 5 rows of cells from the scraping edge in ATRA treated cells, but only 1 or 2 rows in the control. The augment effect was observed 24 hours after each concentration ATRA treatment, and lasted till 72 hours after treatment with 1 micromol/L and 10 micromol/L ATRA. Forty-eight hours after exposed to 100 micromol/L ATRA, the enhancement of GJIC was less obvious. There was no significant increase induced by ATRA on the transcription of Cx43 gene, as demonstrated by semiquantitative RT-PCR.

CONCLUSIONATRA turned out to be a potent enhancer on GJIC function in C6 glioma cells, andthe enhancement effect was most probable at post-transcriptional level.

Animals ; Antineoplastic Agents ; pharmacology ; Brain Neoplasms ; metabolism ; pathology ; Connexin 43 ; biosynthesis ; genetics ; Gap Junctions ; physiology ; Gene Expression ; Glioma ; metabolism ; pathology ; RNA, Messenger ; genetics ; Rats ; Tretinoin ; pharmacology ; Tumor Cells, Cultured