Objective To determine the involvement of DNA demethylation in tumor necrosis factor-α(TNF-α)-induced matrix metalloproteinase 9 ( MMP9) expression in human epidermal keratinocytes. Methods Real-time RT-PCR, Western blot, and enzyme-linked immuno sorbent assay (ELISA) were performed to determine the mRNA and protein levels of MMP9 after human keratinocyte cell line (HaCaT) cells were treated with 10 ng/ ml TNF-α or 2. 5 μmol/ L DAC/ 300 nmol/ L TSA. Bisulfite sequencing PCR ( BSP) and Methylation-sensitive high-resolution melt analysis ( Ms-HRM) were used to detect significantly differentially demethylated CpG sites in the human MMP9 promoter region in cells exposed to TNF-α. Different sites methylation constructs of promoter-luciferase reporter gene were made to detect the influences of site-specific DNA demethylation on transcription activity of MMP9 promoter. Results Compared with PBS-treated control, TNF-α significantly increased the expression of MMP9 in HaCaT cells for indicated culture duration ( P < 0. 05 ). Real time PCR, Western blot, and ELISA analysis demonstrated that the mRNA and protein levels of MMP9 were increased initially, followed by a decline with prolonged incubation time. After TNF-α treatment, varied degrees of DNA demethylation occurred at 10 CpG sites in the promoter of MMP9, and the changes at the -36 bp site were statistically significant (P<0. 05). The demethylation at the -36 bp site greatly increased the transcription activity of MMP9. Conclusion TNF-α promotes MMP9 expression in HaCaT cells through inducing -36 bp site DNA demethylation on the promoter of MMP9.

Aim To explore the hemodynamic change of elderly isolated hypertension(EISH). Methods Color doppler ultrasonography were applied to examine thirty EISH patients and twenty eight elderly healthy subjects. The left ventricular systolic function as well as hemodynamic parameters were measured. Results The indices of left ventricular systolic function in EISH group such as SV,CO,EF and CI were obviously higher than those of control group(P<0.05),Linear relevant analysis show that pulse pressure was remarkable related to SV,CO,EF and CI in EISH group,with r value 0.43,0.56,0.46,0.45 respectively.Conclusion EISH patients apparently had ventricular systolic dysfunction and hemodynamic abnormality. Pulse pressure may be associated with development of sysytolic dysfunction.

One hundred and sixteen patients with lower limb fracture were screened by Doppler ultrasound for deep vein thrombosis(DVT) of bilateral lower extremities within the first 72 h, d7 and d21 after fracture. Results showed that DVT was detected in 31 (26. 7% ) out of 116 cases within 72 h; at d7 and d21 DVT was detected in 3 and 1 patient respectively with a cumulated DVT rate of 30. 2% in 3 weeks.Serial Doppler ultrasonography is of value in screening for DVT of the lower extremities in patients with lower limb fracture at early stage.

To monitor the trans-differentiation from adult stem cells to germ cells, we analyzed the vasa expression of goat testicular tissues in different ages and constructed the germ cell specific reporting vector pVASA-EGFP. The expression of vasa was verified by RT-PCR and immunofluorescence. The vector pVASA-EGFP was constructed by molecular technology, then transfected into goat bone mesenchymal stem cells (BMSCs) by Lipofectamine 2000. Moreover, we observed the expression of the vector through green fluorescent protein (GFP). Immunofluorescence results show that Vasa was expressed in all groups of goat testicular tissues, RT-PCR results show that the levels of vasa mRNA in 3-month group and 10-month group were significantly higher than that in 10-day group. Sequencing and restriction enzyme results show that the vector was successfully constructed. After transfection and RA treatment, GFP expression was observed, which proved the validity of our reporting system. All the results proved that vasa was expressed in different ages in goat testicular tissues, and the vector pVASA-EGFP is efficient in monitoring the trans-differentiation in vitro, which paves the way for further characterization and screening of the trans-differentiation of goat BMSCs.
Animals ; Cell Transdifferentiation ; Genes, Reporter ; Genetic Vectors ; Germ Cells ; cytology ; Goats ; Green Fluorescent Proteins ; Male ; Mesenchymal Stromal Cells ; RNA, Messenger ; Testis ; metabolism ; Transfection

Objective To explore the relation ship between expression of survivin gene in leukemia cells and its clinical effects, and to study the mechanism of survivin resist-apoptosis function in leukemia.Methods Survivin expression was detected by Western blots analysis and expressions of Fas and Caspase were examined by immunohistochemistry in 18 leukemia patients.Results Thirteen cases in peripheral blood mononuclear cell survivin positive expression was detected in 18 leukemia patients(72.2%), but no survivin expression in 10 normal persons. There were significant difference of survivin expression in ALL/ANLL patients groups and different WBC groups(P

Objective:To learn the effect of personality in the course of cognitive decline.Methods:106 aged people from two districts were selected randomly.Arithmetic,digit span,picture filling,block patterns,digit symbol,visual attention span,spatial reasoning were used to learn the cognitive ability,“Kartaier's 16 personality factors test”were used to explore the personality of aged people.Results:16 personality factors test shows that old people are more deferential and self-assured.The relation between reasoning and arithmetic,rule-consciousness and arithmetic,abstractedness and digit span,reasoning and block design,dominance and digit symbol,sensitivity and special reasoning,apprehension and visual attention span,openness to change and visual attention span reach the statistical significance.Conclusion:Age is the most important reason to the cognitive impairment in aged people, education can prevent the cognitive impairment in a large degree.Personality can affect the cognitive ability although different personality factor affect different cognitive fields.

Objective To explore the effects of the neonatal handling and enriched environmental stimulation on brain damage in neonatal rats with hypoglycemia.Methods Thirty-six neonatal rats were randomly divided into the normal group,hypoglycemia intervention group and the hypoglycemia non-intervention group.Those rats in hypoglycemia intervention and hypoglycemia non-intervention groups were weaned for 12 h,then the blood sugar of both groups were monitored.After neonatal rat models with hyperglycemia were prepared,the rats in hypoglycemia intervention group received the neonatal handling for 14 d and then were kept in an enriched stimulation environment for another 14 d.Rats in normal group and hypoglycemia non-intervention group were fed in the routine way.Neonatal handling was done when the rats were born for 24 h.The rat was rubbed with the brush from head to tail softly.Rats in the hypoglycemia non-intervention group was not handled.The enriched environment stimulation was used after 15 d when the rats were born.Rats in the hypoglycemia intervention group was put into the enriched environment for 1 h per day until 28 d when the rats were born,and rats in the hypoglycemia non-intervention group was put into the normal environment.Then the body weight was scaled at 0 d,7 d,14 d,21 d and 28 d when the rats were born.Space learning and memory were tested with Morris earter at their third month's age.After that,changes of pathology was observed in their occipital cortex.Results The weight increase,the ability of space learning,memory and the number of survival pyramid neurons of occipital cortex in normal group were better than those in hypoglycemia intervention and hypoglycemia non-intervention group(Pa

HSP90, which is the biomarker of cell stress and endogenous protective protein, functions as a molecular chaperone. Many client proteins of HSP90, including EGFR, Met, Raf-1, IKK and p53, play important roles in the occurrence and development of tumor. Binding of HSP90 inhibitors triggers the deactivation of HSP90, resulting in client protein degradation, and hence inhibits the tumor growth by blocking multiple targets involved in signaling of tumor proliferation. This review summarizes recent development of small molecule inhibitors bound to N-terminal of HSP90.

The paper is to report the establishment of a method for quickly evaluating compatibility of Chinese medicines injections. Isothermal titration calorimetry (ITC) was used to evaluate compatibility of Yiqifumai (YQFM) and vitamin C injection (Vc)/5% glucose injection (5% GS). The diversification of Gibbs free energy (deltaG), enthalpy (deltaH) and entropy (deltaS) were used to decide reaction types of colliquefaction procedures of different injections. The reactive profiles were used to determine signs and quantity of heat. And high performance liquid chromatography (HPLC) was used as a supportive method for ITC. Then, feasible binding sites were analyzed based on the information of spatial structures of major compositions. During the colliquefaction procedure of YQFM and Vc, [deltaH] > T[deltaS], so, the reaction is enthalpy-driving. And the reactive profile showed that a big deal of heat was given out during the procedure. Obviously, chemical reactions happened and the major compositions changed. On the other side, the reaction of YQFM mixed with 5% GS was entropy-driving, because [deltaH] < T[deltaS]. The reactive profile showed there was a little heat gave out. So, non-chemical reactions happened, and the major compositions did not change. The conformity existed between the results of ITC and HPLC. ITC could be used to evaluate the compatibility of Chinese medicines injections because of the advantages of ITC, such as real time, fast, sensitive and having more parameters.

Objective To explore the expression of Bcl-6 mRNA in the Tfh cells of HIV/AIDS patients and the relationship between Bcl-6 mRNA and the progression of HIV/AIDS.Methods This experiment chose 60 patients who were confirmed by HIV antibody test positive,during May 2014 to November from AIDS Research Institute in the First Affiliated Hospi-tal,Henan College of Traditional Chinese Medicine.According to the amount of CD4+T cells,the patients with HIV/AIDS were divided into A(CD4<200 cells/μl),B(200 cells/μl500 cells/μl)three groups.The Tfh cells expressed CD4+ CXCR5 + ICOS+ and lymphocyte subpopulation including T,B and NK cells were detected by flow cytometry (FCM).The expression of Bcl-6 mRNA levels were detected by reverse transcription-polymerase chain reaction (RT-PCR).The association between Tfh cells and Bcl-6 mRNA,B cells were analyzed.Results The expression of Bcl-6 mRNA from patients of A,B and C three groups was increased than in healthy individuals (2.94±0.91,2.27±0.62,2.15± 0.351,P <0.05).The percent of Tfh cells from group A patients was higher than group C patients (5.88±3.01 vs 1.26± 0.87,P =0.032)and also the healthy control group (5.88±3.01 vs 0.78±0.42,P =0.004).The percentage of NK cells and B cells in HIV/AIDS patients was no statistically significant.Correlation analysis showed that there was positive corre-lation between the proportion of Tfh cells and Bcl-6 mRNA (r=0.799,P =0.000).And Tfh cells were observed no relation-ship with B cells (r=-0.083,P =0.657).Conclusion The changes of Bcl-6 mRNA and Tfh cells may be related to the progression of HIV/AIDS.