Objective To explore the influence of Shensongyangxin capsule on plasma high-sensitivity C-reactive protein(hs-CRP) and N-terminal pro-brain natriuretic peptide(NT-proBNP) in patients with persistent atrial fibrillation after cardioversion.Methods 152 patients with persistent atrial fibrillation were randomly divided into the observation group(76 cases)and the control group (76 cases).All patients were conducted atrial cardioversion.The patients were followed up for 12 months.The plasma hs-CRP and NT-proBNP were measured before and after treatment.Results All patients with atrial fibrillation were cardioversed to sinus rhythm after cardioversion.During 12 months of follow-up,in the observation group 2 patients had recurrence of atrial fibrillation and in the control group,nine cases of recurrence,the difference between the two groups was significant (x2 =5.28,P ＜ 0.05).After treatment,the plasma hs-CRP and NT-proBNP in the two groups were significantly decreased(t =7.270,3.601,8.118,3.006,P ＜ 0.05,P ＜ 0.01),and those in the observation group were significantly lower than control group (t =4.720,2.914,all P ＜ 0.05).Conclusion Shensongyangxin capsule can significantly reduce plasma hs-CRP and NT-proBNP in patients with persistent atrial fibrillation after cardioversion.

Aim To study the effects of HSP70 on hepatic ischemia-reperfusion injury after heat preconditioning in rats.Method To establish the models of the hepatic ischemia-reperfusion injury. 42 Wistar rats were randomly divided into 6 groups, (each group had 7 rats): normal group(N); quercetin injection group(Q); ischemia-reperfusion group(I); heat preconditioning 16 hours before ischemia-reperfusion group(H+I); quercetin injection before heat preconditioning group(Q+H+I); quercetin injection before ischemia-reperfusion group(Q+I).We detected the activity of serum enzyme of ALT,AST and the pathological changes of the liver;The expressions of HSP70 of the rats were observed by Western blotting. Results The expressions of HSP70 from high to low were:group H+I,group I,group Q+H+I,group Q+I,group Q,group N; The serum levels of ALT and AST from high to low were: group Q+I,group I,group Q+H+I,group H+I,group Q,group N;All groups had visibly hepatic histological changes respectively.Conclusion The protection of heat stress pretreatment from ischemia reperfusion injury was possibly performed by inducing the expression of HSP70.

In order to investigate the effect of sequence-specific small interfering RNA on suppressing cyclin D1 expression and proliferation and cell cycle and expression of G1 phase regulators of fibroblasts derived from keloid, the plasmid expression vector of siRNA targeted against cyclin D1 was constructed and transfected into fibroblasts with LipofectamineTM 2000. The changes of cyclin D1 expression were detected by fluorescent quantitative PCR(FQ-PCR), semi-quantitative RT-PCR. The effect of sequence- specific small interfering RNA in suppressing the proliferation of fibroblasts was detected by MTT assay. Flow cytometry were used for evaluation ofceU cycle. The expression of cyclin D1, CDK4, pRb and P16 was detected by immunohistochemical method. The results showed that: (1) The sequence- specific siRNA effectively suppressed cyclin D1 expression at both mRNA levels with inhibition rate of 63.68% and 92.83% (P<0.01). (2) Significantly inhibited the proliferation of fibroblasts, and changed cell cycle in percentage of G0/G1 phase cells was increased compared with the controls groups in fibroblasts(P < 0.05). (3) 72 h after transfection, the expression of cyclin D1, CDK4 and pRb decreased, and the expression of P16 increased. It can be concluded that the plasmid expression vector for the RNAi against cyclin D1 constructed in the study can effectively and specifically suppress cyclin D1 expression, and progression of G1/S is effected by G1 phase related regulatory protein, and suppresses proliferation of fibroblasts derived fiom keloid.

Objective:Hyperoxia is a necessary therapy in some neonatal diseases, and long-term therapeutic hyperoxia may induce severe damaging effects on intestinal epithelial cells.The aim of this study was to investigate whether hyperoxia could promote the expression of ASK1 and P38 in intestinal epithelial cells through ROS.Methods:The human colon adenocarcinoma cell line Caco-2 cells were treated with different concentrations of H 2O 2(100 μmol/L, 200 μmol/L and 400 μmol/L)and 85% oxygen in vitro.The expression of ASK1 was detected by immunofluorescence, and the expression of P38 and p-P38 were detected by Western Blot and Real-time PCR. Results:With the increase of H 2O 2 concentration, the fluorescence intensity of ASK1 increased.The fluorescence intensity of ASK1 in the hyperoxia group was significantly stronger than that of the control group and the H 2O 2 groups.With the increase of H 2O 2 concentration(100 μmol/L、200 μmol/L、400 μmol/L), the expression of P38 protein(0.21±0.02, 0.28±0.13, 0.44±0.07)and p-P38 protein(0.09±0.02, 0.19±0.03, 0.37±0.07)gradually increased.The expression of P38 mRNA in 200 μmol/L and 400 μmol/L H 2O 2 groups(4.03±0.68、3.94±0.71)was significantly higher than that in 100 μmol/L H 2O 2 group(3.05±0.47)( P<0.01). The expressions of P38 protein, p-P38 protein and P38 mRNA in the hyperoxia group were significantly higher than those in the H 2O 2 group( P<0.01). Compared with the control group, the expressions of P38 protein, p-P38 protein and p38 mRNA in the hyperoxia group and H 2O 2 groups increased significantly( P<0.01). Conclusion:The expression of ASK1 and P38 in intestinal epithelial cells increased significantly under hyperoxia, which indicated that hyperoxia might activate ASK1 and thereby regulate the expression of downstream P38 through ROS, resulting in intestinal epithelial cells damage.

Objective To explore the role of intracellular reactive oxygen species (ROS) in the senescence of bone marrow mesenchymal stem cells(BMSCs)in patients with systemic lupus erythematosus(SLE) and the underlying mechanisms that controls the intracellular ROS levels in vitro. Methods Human bone marrow aspirates were collected from iliac of eight donors and eight SLE patients and cultured in vitro.Morphological appearance of BMSCs at different passages was examined by inverted microscope. Nuclear size was measured by fluorescence microscope. BMSCs were monitored using the senescence associated β-galacto-sidase (SAβ-gal) assay to characterize senescence in vitro. The quantification of intracellular ROS production was detected by flow cytometry. Real-time PCR technique was used to determine the gene expressions of PI3K, KRas, NRas and FoxO3 at transcription level. The expression of FoxO3, phospho-FoxO3 (p-FoxO3),AKT and phospho-AKT (p-AKT) protein were determined by Western blot analysis. Statistical analysis was conducted with t-test and Mann-Whitney rank test.Results There were no differences in morphology and nuclear size[(31.4±4.5) vs (28.2±4.8) μm, P=0.628] of BMSCs between SLE patients and normal controls.The percentage of SA β-gal positive BMSCs from SLE patients was higher than that from healthy controls [(31.8±9.0)% vs (12.4±0.7)%, P＜0.05]. Intracellular ROS levels of BMSCs from SLE patients increased more significantly than healthy donors in vitro (34600±9600 vs 17 958±5400, P＜0.05). No significant differences in the expression of PI3K, NRas, KRas and FoxO3 from SLE subjects were observed at mRNA levels compared with normal controls, though all showed a similar upward trend. The expression of p-FoxO3 and p-AKT of BMSCs from SLE patients increased significantly compared with healthy controls at protein levels.Conclusion These data suggest that BMSCs from SLE patients aged more quickly, with high SA β-gal activity and up-regulation of intracellular ROS, which is associated with up-regulation of p-FoxO3 and pAKT at protein levels. These results indicate that bone marrow mesenchymal cell senescence may be associated with the pathogcnesis of SLE by maintaining the lifespan of BMSCs.

The dielectric spectroscopy of human blood cells was measured within the frequency range of 0.01-100 MHz, and the dielectric numerical characters were determined as response to AC electric field. we measured the AC impedance of normal human blood cells with the impedance technique in the frequency domain, then the experimental data were drawn a relationship curve between the frequency of electric field and conductivity. The dielectric spectrum and the Cole-Cole plots of human blood cells were established. The characteristics of dielectric response of human blood cells were also established. The permittivity and the conductivity of human blood cells were frequency dependent, and dielectric dispersion of human blood cells had two characteristic frequencies: i.e. fc1, is 1.42 MHz, and fc2 is 3. 32 MHz.
Blood Cells ; physiology ; Electric Conductivity ; Electrochemistry ; Humans ; Spectrum Analysis ; methods

Objective To investigate the apoptosis of bone marrow mesenchymal stem cells(BMSCs)from systemic lupus erythematosus(SLE)patients and the expression of apoptotic molecules.Methods BMSCs were isolated from bone marrow of SLE patients and normal controls by density eentrifugation and adhesive culture in vitro.The apoptosis of BMSCs were evaluated by TUNEL assay.The expressions of Fas.Bcl-2 and the activity of Caspase 8 were detected by flow cytometry.Real-time PCR technique was used to determine the gene expressions of Fas,Bcl-2,Bax,Bcl-w,Caspase 8 and Apaf-1.Meanwhile,cytochrome C was detected by immunocytochemistry.Statistical analysis was conducted with t-test and Mann-Whitney rank test.Results The percentage of apoptotic BMSCs increased in SLE patients compared with healthy donors[(64±10)%vs [14±9)%,U=0,P<0.05 ].The expression of Bcl-2 in BMSCs of SLE patients was lower than the normal controls at protein leve[(11±9)%vs(56±18)%,U=0,P<0.05 ],and mRNA level(0.2±0.2vs 2.4±0.7,U=24,P<0.05).More cytochrome C positive pellets in the cytosolic fraction could be detected in BMSCs from SLE patients compared with healthy controls [(56±21)%vs (16±16)%,U=1,P<0.05].The activity of Caspase 8 was enhanced[(49±14)%vs(16±12)%,U=0.P<0.05 ],although with no significant difierence at mRNA Ievel.Both groups expressed Fas but with no significant difference (U=19,P>0.05).Conclusion BMSCs from SLE patients undergo more apoptosis,the mechanisms may be associated with the down regulation of Bcl-2,up-regulation of Cytoehrome C in cytoplasm and the activation of Caspase 8,which directs the intrinsic and extrinsic apoptosis pathways.

To investigate the effects of gambognic acid (GA) on TRAIL-induced apoptosis of cancer cells, human colon HT-29 cancer cells were treated with GA to promote apoptosis. Inhibition of the cell proliferation was measured with MTT assay and cell apoptosis was detected with formation of DNA ladders in agarose gel electrophoresis, and activation of caspase activity. The content of cytosolic reactive oxygen species (ROS) was measured with flow cytometry. The activities of Caspase-3, -8, -9 were detected using spectrophotometric assay. The levels of c-FLIP, CHOP, DR4 and DR5 in cells were tested by Western blot. Combination of GA (1 µg · mL(-1)) and TRAIL (40 ng · mL(-1)) significantly reduced proliferation and increased apoptosis of HT-29 cells over those induced by each agent alone. Percentage of apoptotic cells was increased to 45.5%. GA markedly enhanced the intracellular ROS generation. Expression of CHOP, DR4 and DR5 was up-regulated to 7.38, 5.41, and 4.85 times of the control group, respectively. GA promoted activation of Caspase-3, -8, and -9 by TRAIL (P<0.05). Furthermore, the expression of anti-apoptotic protein c-FLIP was down-regulated to 0.22 ± 0.08 times of the control group. In conclusion, GA sensitizes HT-29 cells to TRAIL-induced apoptosis by promoting ROS-activated ERS pathways, up-regulating of DR4 and DR5, and inhibiting c-FLIP expression.

OBJECTIVE:To explore the preventive effect and safety of levetiracetam combined with sodium valproate of diaze-pam on recurrent febrile seizures(FS). METHODS:A total of 90 children with recurrent FS were randomly divided into observa-tion group and control group with 45 cases in each group. Control group was treated with sodium valproate or diazepam orally. On the basis of control group,observation group additionally received levetiracetam orally,with initial dose of 15 mg/kg,bid,for 7 d,and then decreasing gradually;decreasing to 10 mg/kg on 8th-12th day,bid;decreasing to 5 mg/kg on 13th-15th day,bid;drug withdrawal on 16th day. The children of 2 groups were followed up for 1 years,and received routine test every 2 months. The times of fever,the rate of recurrent convulsion,the conversion of epilepsia and the incidence of ADR were recorded in 2 groups during follow-up period. The serum levels of NSE and S-100β protein were determined in 2 groups before treatment and 6 months after treatment. The intelligence and behavior ability of 2 groups were scored by Chinese Modified Wechsler Intelligence Scale for Children and Children’s Adaptive Behavior Rating Scale. RESULTS:3 children of observation group and 2 of control group were failure in follow-up. During the follow-up period,fever times and the rate of recurrent convulsion in observation group were signifi-cantly lower than in control group,with statistical significance(P<0.05). There was no statistical significance in the rate of epilep-sia conversion and the incidence of ADR between 2 groups (P>0.05). After treatment,serum levels of NSE and S-100β in 2 groups were decreased significantly,and the observation group was significantly lower than the control group,with statistical signif-icance(P<0.05). To the end of follow-up,verbal IQ,performance IQ and total IQ score of observation group were significantly higher than those of control group,and the cognition factor,social factor and behavior ability scores of observation group were sig-nificantly higher than those of control group,with statistical significance (P<0.05). CONCLUSIONS:Levetiracetam combined with sodium valproate or diazepam can prevent the occurrence of recurrent FS,relieve cerebral injury and improve the intelligence and behavior ability of the children,so as to improve the life quality of Children.