Human herpes simplex virus 1 (HSV-1) causes facial,ocular,and encephalitic disease and is associated with latent infection and cancer.Here,we developed a means of studying the pathogenesis of HSV-1 infection at the protein level by using the SELDI Protein Chip to detect changes of protein expression in Vero cells cultured in vitro.After infection with HSV-1 and culture for 12,24 or 48 h,cells were harvested and lysed.IMAC3 arrays were applied to SELDI-TOF-MS to detect proteomic differences before and after infection.The chip detected a series of differentially expressed protein peaks.Interestingly,both peaks at 16 912 Da and 17 581 Da corresponded precisely with the molecular mass of ISG 15,which may participate in antiviral activity during the process of infection.Thus,the results we obtained can serve as a basis to study the pathogenesis of HSV-1 and the interaction between the virus and its host.In addition,they can help in the discovery of new therapeutic targets for treatment of HSV-1 infection.

Objective To explore the feasibility of culturing dermal papillae cells (DPC) of hu- man hair in a serum-flee medium,and to observe the growth characteristics of these cells.Methods Cell culture flasks (plates) were pretreated with fibronectin,and DPC (2nd passage) were incubated with Williams E serum-flee medium supplemented with insulin-transferrin-selenite (ITS).Cells were observed by an inverted phase-contrast microscope.Proliferation of DPC was evaluated with 3-(4,5-dimethylthia- zol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by their growth curve.Results In a serum-free medium,2nd passage DPC adhered to the flask surface within two to four hours of incubation; two to three days later,confluence,of the cells was observed,without noticeable proliferation.Four days later,cell connection was interrupted,isolated cells or cell clusters were seen,and detachment of some cells from the flask surface was observed.One to two weeks later,most cells had died.After incubation with 4% bovine serum for ten hours,cell proliferation was observed surrounding the remaining viable cell colonies. DPC growth curve showed stagnant phase and slow growth phase;however,log growth phase was not ob- served.Conclusion DPC could be successfully cultured in serum-free medium.However,the culture con- dition needs to be further optimized.

Objective To explore the gene expression of interleukin-13 receptor (IL-13R)?2 and its relationship to proliferation activity of human gliomas.Methods The gene expression of IL-13R?in 50 hu- man gliomas,2 malignant human glioma cell lines and 6 normal brain tissues were studied by RT-PCR.Ki-67 labeling index (Ki267 LI) of all sample were detecteded by immunohistochemical staining.Results Only one normal brain tissues expressed very low IL-13R?2 mRNA,whereas 35 (70%) of 50 human brain tumors expressed 1L-13R?2 mRNA.The positive rate and expression level of IL-13R?2 mRNA were increased with the ascending of WHO tumor grade.(former:rs=0.87;letter:rs=0.69,P＜0.01).The difference of posi- tive rate and expression level of IL-13R 2?mRNA between the low grade and high grade tumors was statistical- ly significant,the proliferation activity of gliomas evaluated by Ki-67LI (Ki-67 Labeling Index,Ki-67LI) was positively correlated with IL-13R?2 gene expression and the tumor grade.Conclusion In human cerebral gliomas,IL-13R?2 genes may play an role in the malignant progression.The expression level of malignancy in molecular level and selecting the target of gene therapy.

OBJECTIVETo observe the dynamic expression of calcium-sensing receptor(CaSR) in myocardium of diabetic rats.

METHODSThirty male Wistar rats were randomly divided into 3 groups including control, diabetic-4 week and diabetic-8 week groups(n = 10). The type 2 diabetes mellitus models were established by intraperitoneal injection of streptozotocin (STZ, 30 mg/kg) after high-fat and high-sugar diet for one month. The cardiac morphology was observed by electron microscope. Western blot analyzed the expression of CaSR, phospholamban (PLN), a calcium handling regulator, and Ca+-ATPase(SERCA) in cardiac tissues.

RESULTSCompared with control group, the expressions of CaSR and SERCA were decreased, while the expression of PLN was significantly increased in a time-dependent manner in diabetic groups. Meanwhile diabetic rats displayed abnormal cardiac structure.

CONCLUSIONThese results indicate that the CaSR expression of myocardium is reduced in the progression of DCM, and its potential mechanism may be related to the imnaired intracellular calcium homeostasis.

Animals ; Calcium-Binding Proteins ; metabolism ; Diabetes Mellitus, Experimental ; complications ; Diabetes Mellitus, Type 2 ; Diabetic Cardiomyopathies ; metabolism ; physiopathology ; Disease Progression ; Heart ; physiopathology ; Male ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Wistar ; Receptors, Calcium-Sensing ; metabolism ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; metabolism ; Streptozocin

BACKGROUNDCoronary computed tomographic angiography (CCTA) has been widely used in patients who are at intermediate risk for having stable coronary artery disease (SCAD), and 2013 European Society of Cardiology Guidelines on the Management of SCAD (2013G) recommended the appropriate application of CCTA. However, 2013G has not been subjected to systematic analyses for subsequent impact on clinical practice.

METHODSA total of 5320 patients suspected with SCAD were enrolled and scheduled for CCTA from March 2013 to September 2014. For each patient, pretest probability of SCAD was calculated according to updated Diamond-Forrester model (UDFM). Appropriate CCTA or appropriate stress test was determined as described in the 2013G. A generalized estimating equation model was used to determine the trends in the half-monthly rate of appropriate CCTA.

RESULTSOverall, only 61.37% of patients received appropriate CCTA, and there was insignificant change over time (P = 0.8701). The application of CCTA in patients who should have had a stress test accounted for most of the inappropriate CCTA before (22.29%) or after (19.98%) the publication of the 2013G. In all patients or any subgroup, no significant change in the adjusted half-monthly rate of appropriate CCTA was found after the publication of the 2013G (odds ratio, 1.002; 95% confidence interval, 0.982-1.021; P = 0.8678).

CONCLUSIONSThese findings suggest that the 2013G have not, to date, been fully incorporated into clinical practice, and the clinical utilization of CCTA remains unreasonable to some extent.

Aged ; Coronary Angiography ; methods ; Coronary Artery Disease ; diagnosis ; Female ; Humans ; Male ; Middle Aged ; Odds Ratio

OBJECTIVETo establish and evaluate a rat model of type 2 diabetes mellitus (T2DM) associated with depression for further elaborating the disease.

METHODSTwenty-four Wistar rats were randomly divided into three groups: normal group (group N), T2DM group (group T) and T2DM with depression group (group T + D), with 8 rats in each group. The T2DM rat model was induced by high fat diet and low dose of Streptozotocin (STZ) injection, and in addition, the T2DM rats were made restraint stress for 21 days. After the model was established, the insulin tolerance test (ITT) and oral glucose tolerance test (OGTT) were performed. Then the rat depression level was analyzed by open field test, and the concentration of 5-hydroxytryptamine (5-HT) and dopamine (DA)was determined by ELISA to confirm the model identity.

RESULTSThe blood glucose level in group T and group T + D didn't return to the normal level at 180 minutes in the ITT and OGTT test; Compared with the group N, the max movement distance, retaining time in the central zone and the retaining frequency within 5 minutes in the group T + D decreased; 5-HT and DA level in the serum of rats in. group T + D was reduced.

CONCLUSIONA rat model of type 2 diabetes mellitus associated with depression has been successfully established by high fat diet and injection of low dose streptozotocin in combination with restraint stress for 21 days. This rat model is useful for further relevant studies.

Animals ; Depression ; complications ; Diabetes Mellitus, Experimental ; complications ; Diabetes Mellitus, Type 2 ; Diet, High-Fat ; Glucose Tolerance Test ; Rats ; Rats, Wistar ; Restraint, Physical ; Serotonin ; Streptozocin ; Stress, Psychological

OBJECTIVETo study the mechanism involved in the control of glioma cell proliferation with transfection of connexin (Cx) 43 gene.

METHODSC6 rat glioma and TJ905 human glioblastoma cell lines without Cx43 gene expression were transfected with Cx43cDNA mediated by lipofectamine. Northern blot, in situ hybridization and immunohistochemical technology were used to detect the expression of Cx43mRNA and its protein with MTT assay and silver colloid stain for the detection of cell proliferation, TUNEL method for determination of cell apoptosis, scrape loading dye transfer (SLDT) for GJIC, Western blot and immunohistochemical technology for bFGF, PDGF, EGFR, IGF-I and IGFBP3 expression.

RESULTSCx 43 gene transfected glioma cells showed decreased proliferation, restored GJIC and decreased bFGF, PDGF, IGFBP3, except EGFR expression and cell apoptosis which showed no change.

CONCLUSIONThe mechanism of Cx 43 gene inhibiting gliomas cell proliferation is the restoration of GJIC and decreased autocrine growth factors.

Animals ; Apoptosis ; Cell Division ; physiology ; Connexin 43 ; genetics ; physiology ; DNA, Complementary ; genetics ; Glioma ; pathology ; Rats ; Transfection ; Tumor Cells, Cultured

OBJECTIVETo explore the inhibitory effect of arsenic trioxide (A(s2)O3) on the growth of rat C6 glioma cells (C6 cells) as well as finding out the feasibility of using As2O3 as chemotherapy of gliomas.

METHODC6 cells were treated by different dose of As2O3 (1, 2, 4, 6 and 8 micromol L(-1)). MTT assay and staining for PCNA were used for cell proliferation. Cell apoptosis was determined by TUNEL method and Bcl-2 expression was studied by Western blot. Parental rat C6 cells (5 x 10(5)/15 microL) were implanted into right caudate nucleus of male SD rats as control group. Rats bearing cerebral C6 gliomas as treated group were treated with 1 mmol x L(-1) As2O3. The general manifestation, survival time, MRI dynamic scanning and histopathological changes of all rats were observed.

RESULTAll the treated cells showed decreased proliferation in vitro as detected by MTT method (P < 0.01) and staining for PCNA. In situ labeling apoptotic DNA fragment of the treated cells demonatrated that the cell apoptosis significantly increased following treatment with As2O3 (P < 0.01). Western blot showed that the expression of Bcl-2 protein was decreased. All rats in control group died of cerebral gliomas within 3 weeks after implantation of C6 cells (17.8 +/- 0.92) d. Eight out of 10 rats in treated group died within 24-36 days (32.1 +/- 1.35) d and other 2 ones kept alive beyond 120 days with one treated rat being totally disappear of the tumor foci and another having a little residue of tumor.

CONCLUSIONThe result demonstrates the potential efficacy of As2O3 in the treatment of gliomas. It also suggests that As2O3 may be a good candidate for chemotherapy of human gliomas.

Animals ; Arsenicals ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Glioma ; drug therapy ; Male ; Oxides ; pharmacology ; Rats ; Rats, Sprague-Dawley

Objective To explore the influence of social network on the quality of life of elderly stroke patients in communities. Methods The general situation questionnaire, the stroke impact scale ( SIS) and social network scale for the elderly were used to investigate the community elderly stroke patients. Results The total score of social network was(39. 82±5. 87). The scores of social participation,social con-tact and social support were(11. 79±1. 61),(14. 90±3. 31) and (13. 13±3. 19). Results of univariate a-nalysis showed there were statistically significant differences ( P<0. 05) in quality of life among stroke pa-tients of different ages (F=7. 803),educational level (F=2. 664),family income (F=4. 754),smoking (F=5. 467),alcohol consumption (F=7. 058),physical exercise (F=10. 393),body mass index (t=4. 841) and hypertension (t=-2. 862). Multiple linear regression results showed that 3 times or more exercises per week ( B=5. 689),normal body mass index (B=6. 613),high degree of social support (B=0. 710),social partici-pation (B=1. 421) and social contact (B=1. 372) were protective factors for the quality of life of stroke pa-tients. Age 80 years and above (B=-10. 426),frequent smoking (B=-9. 005),frequent drinking (B=-8. 419) and hypertension ( B=-5. 224) were risk factors for quality of life. Conclusion The elderly stroke patients should quit smoking and alcohol,strengthen exercise,maintain a normal body mass index and avoid overweight and obesity in daily life. While promoting health-related behaviors,more attention should be paid to the promotion effect of social network on treatment,rehabilitation and overall quality of life of stroke patients.

Objective To investigate the clinical factors influencing survival prognosis in patients with supratentorial malignant glioma (MG). Methods A total of 194 patients with MG,admitted to our hospital from January 2004 to December 2009, were chosen in our study; anaplastic astrocytoma (AA) was noted in 120 patients and glioblastoma multiforme (GB) was noted in 74 patients.Their survival status was followed-up.Survival related statistics were calculated using the Kaplan-Meier method,and differences between survival curves were analyzed by the log-rank test.The interaction of each prognostic factor and their effect on overall survival (OS) and progression-free survival (PFS) were analyzed with the Cox proportional hazards model. Results The median PFS and OS in AA patients were 18 and 21 months,and those in GB patients were 10 and 12 months.When PFS and OS were used as endpoints,AA patients lived longer than GB ones, while patients ≤ 40 years old lived longer than those ＞40 years old; patients with Karnofsky performance status (KPS) scored 80-100 had longer survival than those with KPS scored 60-70; patients with preoperative seizure enjoyed the same situation as compared with those without preoperative seizure. Both univariate and multivariate Cox analyses confirmed that independent influencing factors were age,KPS scores,grade of pathology and appearance of preoperative seizure; patients with young age, high KPS scores, low grade of pathology and preoperative seizure had longer PFS and OS than those without. Conclusion Young age,high KPS scores,low grade of pathology and preoperative seizure are protective prognostic factors in patients with MG,while gender,tumor size,tumor location,and extent of resection have no prognostic significance; the prognostic significance of contrast - enhancing is uncertain.