DNA methylation is an important epigenetic modification,functioning under the regulation of DNA methyltransferase(DNMT),interacting with other epigenetic mechanisms and participating in the regulation of gene expression and differentiation process of cells.Many DNA methylation related changes are found in epilepsy and the emerging role of DNA methylation in epileptogenesis is becoming even more obvious.Here,we discuss the research progress of DNA methylation in the pathogenesis of epilepsy,and the perspectives of epigenetic medicine as new therapeutic strategy in epilepsies.

Objective To explore the nursing measures of teenagers with arthroscopic reduction of avulsed fracture of tibialintercondylar eminence. Methods Eighteen teenagers with avulsed fracture of tibialintercondylar eminence hospitalized from January 2013 to September 2014 were included in the study. Before operation, psychological care and operative preparation were delicately done. After operation, a plan for function training was made, the preventive measures for related complications were taken and discharge instruction was performed. Results All patients got recovered after function training 3 months since operation. The average score by Lysholm evaluation was (97.6 ± 2.3). Among them, one was Lachman test I positive, with mildly limited extension of the knee join. Conclusion The following nursing strategies inclucling psychological nursing before operation can help to reduce the incidence of complications.

Aim To study the effects of 2 ,3 ,4 ’ ,5-tet-rahydroxystilbene-2-O-β-D glucoside ( TSG ) on myo-cardial fibrosis ( MF) induced by isoproterenol ( ISO) in mice and its possible mechanism. Methods MF in mice was induced by subcutaneous injection of isoprot-erenol for 14 days. TSG (30,60,120 mg·kg-1 ) and captopril ( 40 mg · kg-1 ) were then administered by gavage to mice. The experiment was stopped 12 h after the last administration of the drugs. Hematoxylin-eosin ( HE) and Masson staining were used to estimate the extent of MF. Level of hydroxyproline in myocardial tissues was measured. Protein expressions of collagenⅠ, collagen Ⅲ and transforming growth factor-β1 (TGF-β1) in myocardial tissues were measured. Lev-els of superoxide dismutase ( SOD ) and glutathione peroxidase ( GSH-Px ) were determined. Results Compared with control mice, the level of hydroxypro-line in myocardial tissues was significantly increased in isoproterenol treated mice. Histological sections of iso-proterenol-treated hearts showed extensive myocardial fibrosis. And protein expressions of collagenⅠand col-lagen Ⅲwere markedly increased in isoproterenol trea-ted mice. However, therapy with TSG decreased the level of hydroxyproline in myocardial tissues, ameliora-ted the degree of myocardial fibrosis, and reduced the collagen expressions induced by isoproterenol adminis-tration. Moreover, treatment with TSG decreased TGF-β1 protein expression and elevated the myocardial SOD and GSH-Px activity. Conclusion TSG can inhibit MF formation induced by ISO in mice which might be due to regulating TGF-β1 protein expression and its an-tioxidant effect.

Objective To investigate the oxidative stress and inflammation in trophoblast cells stimulated by different chain length fatty acids.MethodsSerum-free trophoblast cells cultured in vitro were divided into five groups,which were incubated with DMEM medium without free fatty acid (F-FFA),short chain fatty acids (SC-FFA),medium chain fatty acids (MC-FFA),long chain fatty acids (LC-FFA),very long chain fatty acids (VLC-FFA).Then cells in each group were stimulated by DMEM medium,reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor (apocynin) and p38 mitogen-activated protein kinases (p38MAPK) inhibitor (SB203580) and were subdivided as each FFA plus-DMEM group, plus-NADPH-Ⅰ and plus-p38MAPK-Ⅰ groups.Expressions of mRNA and protein of p38MAPK and cyclooxygenase 2 (COX-2) in trophoblast cells were detected by real-time PCR and western blot.Results (1) The mRNA expression of p38MAPK in LC-FFA + DMEM,VLC-FFA + DMEM,LC-FFA + NADPH-Ⅰ,LC-FFA + p38MAPK-Ⅰ,VLC-FFA + NADPH-Ⅰ,VLC-FFA + p38MAPK-Ⅰ group were 4.56 ±0.28,22.65 ±2.40,0.87 ±0.06,1.02 ±0.15,19.87 ± 1.93,10.22 ±0.75 separately,and the protein expressions were 0.79 ± 0.02,0.93 ± 0.10,0.43 ± 0.06,0.44 ± 0.19,0.79 ± 0.10,0.81 ±0.14.Compared with other groups,the mRNA and protein expressions of p38MAPK in LC-FFA + DMEM,VLC-FFA + DMEM group were increased ( P ＜ 0.05 ).Compared with LC-FFA + DMEM group,mRNA and protein expressions of p38MAPK in LC-FFA + NADPH-Ⅰ and LC-FFA + p38MAPK-Ⅰ group were significantly decreased (P ＜ 0.05 ).Compared with VLC-FFA + DMEM group,mRNA and protein expressions of p38MAPK had no difference in VLC-FFA + NADPH-Ⅰ group (P ＞ 0.05 ),mRNA expression of p38MAPK in VLC-FFA + p38MAPK-Ⅰ group was significantly decreased (P ＜ 0.05 ),but there was no difference in protein expression ( P ＞ 0.05).(2) The mRNA expression of COX-2 in LC-FFA + DMEM,VLC-FFA +DMEM,LC-FFA + NADPH-Ⅰ,LC-FFA + p38MAPK-Ⅰ,VLC-FFA + NADPH-Ⅰ,VLC-FFA + p38MAPK-Ⅰ group were 3.97 ±0.03,39.08 ±0.63,0.99 ±0.13,0.98 ±0.18,20.93 ±3.70,13.46 ± 2.31 separately,and the protein expressions were 1.32 ± 0.20,1.33 ± 0.25,0.59 ± 0.13,0.58 ± 0.30,0.88 ± 0.18,0.91 ± 0.24.Compared with other groups,mRNA and protein expressions of COX-2 in LC-FFA + DMEM and VLC-FFA + DMEM group were significantly increased ( P ＜ 0.05 ).Compared with LC-FFA + DMEM group,mRNA and protein expressions of COX-2 in LC-FFA + NADPH-Ⅰ and LC-FFA +p38MAPK-Ⅰ group were decreased ( P ＜ 0.05 ).Compared with VLC-FFA + DMEM group,mRNA and protein expressions of COX-2 in VLC-FFA + NADPH-Ⅰ and VLC-FFA + p38MAPK-Ⅰ group were all decreased ( P ＜ 0.05 ).( 3 ) The correlation analysis showed that there were significantly positive correlations between the mRNA and protein expressions of p38MAPK and COX-2 in LC-FFA group ( P ＜ 0.05 ).There were significantly positive correlations in protein expression ( P ＜ 0.05 ),but no conrelation in the mRNA expression between p38MAPK and COX-2 in the F-FFA,SC-FFA,MC-FFA,VLC-FFA groups (P ＞ 0.05).ConclusionsThe oxidative stress and inflammation may exist in trophoblast cells which were stimulated by LC-FFA and VLC-FFA.p38MAPK signal transduction pathway may contributed in this process.

Objective To investigate the effect of adrenomedullin on rat renal tubular epithelial cell line (NRK-52E) apoptosis induced by hypoxia-reoxygenation (HR) injury and its mechanism.Methods NRK-52E cells were cultured and randomly allotted to the following 4 groups:control group,HR group,empty plasmid + HR group,ADM + HR group.NRK-52E cells were transfected with pcDNA3.1-myc-his B empty vector or pcDNA3.1-ADM by transfection complex comprising optimal proportion of plasmid and Fugene HD reagents.Cells were counted by trypanblau and cell survival rate was computed.The concentration of lactate dehydrogenase (LDH) was detected by spectrophotometric method to evaluate cell vitality.The apoptotic rate of NRK-52E cells was measured by flow cytometry.The transfer efficiency was detected by immunocytochemistry.The mRNA expressions of ADM,Bax,Bcl-2 and Fas were determined by Semi-quantitative RT-PCR,and active caspase-3,8,9 protein expression were examined by Western blotting.Results Compared with control group,the expression of ADM significantly increased in HR group (P ＜ 0.05).The expression of ADM significantly increased in ADM + HR group than that in empty plasmid + HR group.Compared with control group,in HR group,the living cell counts and cell survival rate significantly decreased; the LDH concentration in media,apoptotic rate and the levels of Bax,Bcl-2,Bax/Bcl-2,Fas,active Caspase -3,8,9 significantly increased (all P ＜ 0.05).Compared with HR group,in ADM + HR group,the living cell counts and cell survival rate significantly increased; the LDH concentration in media,the cell apoptotic rate and the levels of Bax,Bax/Bcl-2,Fas,active Caspase-3,8,9 significantly decreased,while Bcl-2 was promoted (all P ＜ 0.05).The above indexes had no differences between empty plasmid + HR group and HR group (all P ＞ 0.05).Conclusion The increased expression of ADM can inhibit NRK-52E apoptosis induced by HR,and the mechanism might be achieved by inhibiting mitochondrial and death receptor-mediated apoptotic pathways.

OBJECTIVE　To develope an HPLC assay for the determination of tanshinoneⅡA.METHOD　C18ODS column was used.The mobile phase was consisted of MeOH-water(70∶30).The detection wavelength was at 269 nm.RESULT　The linear regression equation was Y=9725X-2584,r=0.9997.The average recovery was 98.29% with RSD=1.86%.(n=5).CONCLUSION　The method may be used for quality control.

Objective:To study the apoptosis of T lymphocyte subsets of peripheral blood in systemic lupus erythematosus(SLE) and the correlated pathogenesis.Methods:The percentages of T lymphocyte subsets,the positive expression rates of Fas/FasL on T lymphocyte subsets,and the apoptosis of T lymphocyte subsets were determined by three-colorflow cytometry.The level of serum IL-10 was analyzed by ABC-ELISA.IL-10 antibody and FasL antibody were added respectively into the 48 hours' culture of 10 SLE patients' PBMCs in vitro,whose serum IL-10 significantly increased,and then,the changes of T lymphocyte subsets of SLE PBMCs were determined by three-colorflow cytometry.Results:1) In SLE,mainly in hyperactive SLE,the apoptosis of CD4+ T lymphocytes increased significantly(P

Objective: To study the apoptosis of T lymphocyte subsets of peripheral blood in systemic lupus erythematosus(SLE)and the correlated pathogenesis.Methods: The percentages of T lymphocyte subsets,the positive expression rates of Fas/FasL on T lymphocyte subsets,and the apoptosis of T lymphocyte subsets were determined by three-colorflow cytometry.The level of serum IL-10 was analyzed by ABCELISA.IL-10 antibody and FasL antibody were added respectively into the 48 hours' culture of 10 SLE patients' PBMCs in vitro,whose serum IL-10 significantly increased, and then, the changes of T lymphocyte subsets of SLE PBMCs were determined by three-colodlow cytometry.Results: 1)In SLE, mainly in hyperactive SLE, the apoptesis of CD4~+ T lymphocytes increased significantly(P＜0.05),and the ratio of CD4~+/CD8~+ decreased compared with normal controls,and the abnormal apoptesis of CD4~+ T lymphocytes had a positive correlation with the increased expression rates of Fas/FasL on CD4~+ and CD8~+ T lymphocytes(P＜0.05).2)The serum IL-10 level increased significantly in SLE (P＜0.01), expecially in hyperactive SLE, and serum IL-10 had a positive correlation with the level of serum anti-dsDNA antibodies, CD4~+/CD8 + ratio, and the positive expression rate of FasL on CD4 * T lymphocytes(P＜0.05).3)According to a tracking experiment on two different groups of SLE patients,we found that:with the stabilization of the illness,the level of serum IL-10 decreased significantly, the positive expression rate of Fas/FasL on T lymphocytes,particularly on CD4~+ T lymphocytes, reduced evidently,and the apoptosis of CD4~+ T lymphocytes decreased(P＜0.05).Conclusion: The abnormal activities of T lymphocytes, particularly of CD4~+ T lymphocytes from patients with SLE,which lead to AICD,cause the percentage of CD4~+ T lymphocytes and the ratio of CD4~+/CD8~+ to decrease,disorder immune response, and accelerate the process of SLE.As a key factor in modulating the positive expression rate of Fas/FasL on CD4~+ T lymphocytes,IL-10 plays an important role in causing SLE.

Objective To investigate the inhibitory effect of icariin(ICA) on the xenograft tumors growth of esophageal car‐cinoma and to preliminarily investigate its mechanism .Methods The MTT assay and Giemsa staining were applied to detect and observe the in vitro inhibitory effect of ICA on esophagus cancer cell lines Eca‐109 and TE‐13 .The xenograft tumor model of nude mouse esophagus cancer cell was constructed and divided into 3 groups ,6 cases in each group .Each mouse in the experimental group was intraperitoneally injected by ICA 50 mg/kg ;while the control group was injected by the same volume of normal saline and the positive control group was injected by cis‐platinum 2 mg/kg ,once every 2 days ,a total of 14 days .The tumor volume was measured once per 3 d .After experiment ,the tumor weight was measured;the TUNEL staining was used to observe the morphological chan‐ges and cell apoptosis of tumor tissue in each group .The changes of Fas and FasL protein expression in tumor tissues were analyzed by immunohistochemistry .The FasL and IFN‐γlevels in peripheral blood were tested by the ELISA assay .Results ICA exerted no obvious inhibitory effect on the proliferation of Eca‐109 and TE‐13 cell in vitro .The average volume and weight of xenografts tumor had statistical difference between the experimental group and the positive control group (P<0 .05) .The TUNEL staining results showed that the tumor tissues had obvious apoptosis ,the number of apoptosis cells was significantly increased compared with the control group(P<0 .05) .The immunohistochemistry experimental results showed that the expression of Fas and FasL was signifi‐cantly increased(P<0 .05) .The ELISA experimental results demonstrated that the FasL and IFN‐γlevels of peripheral blood in the experimental group were significantly increased(P<0 .05) .Conclusion ICA had no obvious inhibitory effect on esophageal cancer cell proliferation in vitro ,but could induce in vivo apoptosis through the Fas expression and secretion of FasL and IFN‐γ,thus plays the role of anti‐esophageal cancer .

Aim To study the effect of 2,3,4′,5-tetrahydroxystilbene-2-O-?-D glucoside(TSG) on vascular smooth muscle cells(VSMCs) proliferation induced by fetal bovine serum and explore the underlying mechanisms.Methods VSMCs were cultured from rat aorta using explant technique.VSMCs proliferation was analyzed by 5-bromo-2-deoxyuridine(Brdu) incorporation assay.Cell cycle phase distributions were determined by flow cytometry.The expression of proliferating cell nuclear antigen(PCNA) was evaluated by Western blot analysis.The level of intracellular reactive oxygen species(ROS) was estimated through fluorescence assay.The intracellular activities of superoxide dismutase(SOD),catalase(CAT),and glutathi one peroxidase(GSH-px) were measured using biochemical method.Results TSG in the experimental concentration 0.1～100 ?mol?L~-1 exhibited no apparent cytotoxocity.10 ?mol?L~-1 and 100 ?mol?L~-1 TSG significantly inhibited the proliferation of VSMCs induced by serum,cell cycle transition from G0/G1phase to S phase,PCNA expression in nucleus of VSMCs and level of intracellular ROS.100 ?mol?L~-1 TSG markedly increased the activities of SOD and GSH-px.Conclusions Certain concentration of TSG can prevent VSMCs proliferation effect mediated by serum;the mechanism about TSG effect may be associated with arresting G0/G1 to S progression,decreasing PCNA expression and improving the antioxidation of VSMCs.