Objective To observe the short term results of emergency PTCA and stent treatment of acute myocardial infarction Methods Analyze the 52 cases of acute myocardial infarction under PTCA and stent treatment in our hospital. Results The reopen rate of infarct related vessels is 100%. One of patients was not able to implant the stent. The post operative situation, ECG and myocardial enzyme are improved obviously without complication. The mean hospitalization period is about 2 weeks. 2 D echo shows EF was normal before discharge. Conclusion The reopen rate of infarct related vessels of AMI under emergency PTCA and stent treatment can short AMI patients′ hospitalization time and improve the myocardial pump function obviously.

Objective To evaluate the effect of hydrogen on the expression of nuclear factor E2?related factor 2 ( Nrf2 ) during endotoxin?induced oxidative injury to macrophages. Methods Normally cultured Raw264.7 cells were divided into 4 groups ( n=36 each) using a random number table: control group (group C), hydrogen?rich saline group ( group H), endotoxin group ( group E) and endotoxin plus hydrogen?rich saline group (group EH). In E and EH groups, endotoxin was added with the final concentration of 1 μg∕ml. Hydrogen?rich saline ( hydrogen concentration 0. 06 mmol∕L) was added in H and EH groups. All the cells were incubated for 48 h. At 6, 24 and 48 h of incubation, cells were collect?ed to detect the activity of reactive oxygen species ( ROS) , and cells were collected and proteins extracted for determination of superoxide dismutase ( SOD) and catalase ( CAT) activities and Nrf2 expression by Western blot. Results Compared with group C, the ROS activity was significantly increased, the levels of SOD and CAT were significantly decreased, and the expression of Nrf2 was significantly up?regulated in E and EH groups (P<0.05), and no significant change was found in the parameters mentioned above in group H (P>0.05). Compared with group E, the ROS activity was significantly decreased, the levels of SOD and CAT were significantly increased, and the expression of Nrf2 was significantly up?regulated in group EH (P<0.05). Conclusion Hydrogen can attenuate endotoxin?induced oxidative injury to macro?phages, and the mechanism may be related to up?regulated expression of Nrf2.

Objective To evaluate the role of nuclear factor erythroid 2?related factor 2 ( Nrf2)∕antioxidant response element( ARE) signaling pathway in inhibition of lipopolysaccharide ( LPS)?induced inflammatory factor release from macrophages by hydrogen. Methods RAW264. 7 macrophages of mice were cultured in 6?well plates (2×106 cells∕well) and were divided into 4 groups (n=24 each) using a random number table: control group ( group C); LPS group; hydrogen?rich saline+LPS group ( group LPS+H2); Nrf2 small interference RNA (siRNA)+LPS+hydrogen?rich saline group (siRNA+LPS+H2 group) . LPS 1 μg∕ml was added in group LPS. In group LPS+H2 , LPS 1μg∕ml was added, and the cul?ture medium was then replaced with the culture medium containing 0. 6 mmol∕L hydrogen?rich saline. In group siRNA+LPS+H2 , after Nrf2?siRN was successfully transfected into the cells, the cells were continu?ously incubated for 24 h, and the culture medium was then replaced with the culture medium containing 0.6 mmol∕L hydrogen?rich saline after LPS 1 μg∕ml was added. At 24 h of incubation, the supernatant was sep?arated for determination of the lactic dehydrogenase (LDH) activity (using colorimetric method) and for detection of the concentrations of tumor necrosis factor?alpha ( TNF?α) , interleukin?1 beta ( IL?1β) , high mobility group box?1 (HMGB1) and IL?6 (by ELISA). The cells were collected for measurement of the proliferation of cells ( by methyl thiazolyl tetrazolium assay) and for determination of the expression of Nrf2 and heme oxygenase?1 ( HO?1) in cells ( by Western blot) . Results Compared with group C, the LDH activity and concentrations of TNF?α, IL?1β, IL?6 and HMGB1 in the supernatant were significantly in?creased, the proliferation of cells was significantly decreased, and the expression of HO?1 in cells was sig?nificantly up?regulated in LPS and siRNA+LPS+H2 groups, and the expression of Nrf2 in cells was signifi?cantly up?regulated in LPS and LPS+H2 groups (P<0.05). Compared with group LPS, the LDH activity and concentrations of TNF?α, IL?1β, IL?6 and HMGB1 in the supernatant were significantly decreased, the proliferation of cells was significantly increased, and the expression of Nrf2 and HO?1 in cells was sig?nificantly up?regulated in group LPS+H2 , and the expression of Nrf2 and HO?1 in cells was significantly down?regulated in group siRNA+LPS+H2 ( P<0.05) . Compared with group LPS+H2 , the LDH activity and concentrations of TNF?α, IL?1β, IL?6 and HMGB1 in the supernatant were significantly increased, the proliferation of cells was significantly decreased, and the expression of Nrf2 and HO?1 in cells was signifi?cantly down?regulated in group LPS+H2+siRNA ( P<0.05) . Conclusion The mechanism by which hydro?gen inhibits LPS?induced inflammatory factor release from macrophages is related to the activation of Nrf2∕ARE signaling pathway in mice.

Objective To investigate the effect of hydrogen on endotoxin-induced expression of zonula occludens-1 (ZO-1) in human colon epithelial cells (Caco-2 cells).Methods Caco-2 cells were cultured routinely,seeded in Transwell chambers or wells,and randomly divided into 4 groups (n =45 each) using a random number table:control group (group C);hydrogen-rich culture medium group (group H);endotoxin group (group E);hydrogen-rich culture medium + endotoxin group (group HE).The cells were cultured in high-glucose DMEM culture medium in group C.The cells were incubated in hydrogen-rich culture medium containing hydrogen 0.6 mmol/L in group H.The cells were incubated in highglucose DMEM culture medium containing 50 μg/ml lipopolysaccharide in group E.The cells were incubated in hydrogen-rich culture medium containing 50 μg/ml lipopolysaccharide and 0.6 mmol/L hydrogen in group HE.Transepithelial electrical resistance (TEER) was measured before incubation or culture,and at 6,12 and 24 h of incubation or culture.The viability of Caco-2 cells was measured by methyl thiazolyl tetrazolium assay at 24 h of incubation or culture.The expression of ZO-1 mRNA in Caco-2 cells was determined using real-time reverse transcriptase polymerase chain reaction at 6,12 and 24 h of incubation or culture.The distribution of ZO-1 in Caco-2 cells was observed by immunofluorescence at 24 of incubation or culture.Results Compared with group C,TEER was significantly decreased at 6,12 and 24 h of incubation or culture,and the expression of ZO-1 mRNA was significantly down-regulated in E and HE groups (P＜0.05),and no significant change was found in the parameters mentioned above in group H (P＞0.05).Compared with group E,TEER was significantly increased at 6,12 and 24 h of incubation or culture,and the expression of ZO-1 mRNA was significantly up-regulated in group HE (P＜0.05).The distribution of ZO-1 protein in cell membrane became discontinuous,and the distribution of ZO-1 protein in cytoplasm was significantly increased in group E.Compared with group E,the distribution of ZO-1 protein in cell membrane was significantly increased and gradually became continuous,and the distribution of ZO-1 protein in cytoplasm was significantly decreased in group HE.Conclusion The mechanism by which hydrogen reduces the damage to human colon epithelial cell barrier is related to up-regulation of ZO-1 expression and improvement in the redistribution of ZO-1 protein.

Objective To investigate the role of Rho kinase (ROCK) in the protective effects of hydrogen on intestinal epithelial barrier function in sepsis. Methods Caco-2 cells were cultured routinely, and divided into 6 groups randomly (n=3):control group (C group), hydrogen-rich medium group (H group), lipopolysaccharide (LPS)-treatment group (L group), hy?drogen+LPS-treatment group (HL group), Rho kinase inhibitor (Y-37632) treatment group (Y group) and Rho kinase inhibi?tor Y-27632+LPS-treatment group (YL group). H group was treated with 0.6 mmol/L hydrogen-rich media. The concentra?tion of LPS and Y-27632 were 50 mg/L and 25μmol/L separately. After the Caco-2 monolayer model was established, the transepithelial electrical resistance (TEER) values were measured regularly. When the TEER value reached 800Ω·cm2, the treatment was administered. Then TEER values were measured at 6 h, 12 h and 24 h, and FITC-dextran permeability was de?tected at 24 h. Cells were seeded on 6-well plates. After cell density reached 80%-90%, treatments were given randomly. The real time-polymerase chain reaction (RT-PCR) was conducted to assess mRNA levels of ZO-1 and ROCK mRNA. ZO-1 and ROCK protein expression levels were detected by Western blot assay. Results Compared with C group, TEER values were elevated in 12 h and 24 h in H group (P<0.05). There were no statistical significances in FITC-dextran permeability,
protein expression levels of ZO-1 and ROCK between C group and H group (P>0.05). TEER values were elevated at 6 h, 12 h and 24 h in Y group (P<0.05). There was no significant difference in FITC-dextran permeability between C group and Y group (P > 0.05). The mRNA expression of ZO-1 increased and mRNA expression of ROCK decreased in Y group (P <0.05). The TEER values reduced at 6 h, 12 h and 24 h in L group. The FITC-dextran permeability increased significantly, mRNA and protein expressions of ZO-1 significantly decreased, mRNA and protein expressions of ROCK significantly in?creased in L group (all P<0.05). Compared with L group, TEER values increased significantly at 6 h, 12 h and 24 h in YL group, FITC-dextran permeability decreased, mRNA expressions of ZO-1 increased, mRNA expressions of ROCK de?creased in YL group (P<0.05). Compared with L group, TEER values increased at 6 h, 12 h and 24 h in HL group, FITC-dextran permeability reduced markedly, protein expressions of ZO-1 increased at each time point, protein expressions of ROCK decreased at each time point in HL group (P<0.05). Conclusion Hydrogen can protect intestinal barrier function against sepsis, ameliorate the integrity and permeability of intestinal epithelium and increase the expressions of intercellular tight junction proteins. The suppression of Rho kinase over-expression induced by LPS may be involved in these protective effects of hydrogen.

The oxidative stress, inflammatory cytokines and apoptosis have been strongly implicated in the pathogenesis of multiple diseases. Recently, more and more research findings have demonstrated that hydrogen-rich saline (HRS) has the anti-oxidant, anti-inflammatory and anti-apoptotic effects in vivo and in vitro, and can be used to treat multiple diseases, such as ischemia/reperfusion injury, stroke, neurodegeneration, sepsis, neuropathic pain and multiple organ dysfunction syn-drome diseases. This article reviews the possible mechanism of HRS for the treatment of diseases.

ABSTRACT:Objective To study the effects of propofol on the metastasis of tumor cells related PI3K/Akt signaling pathway.Methods The breast cancer model was established by transplanting human derived breast cancer cell lines into immunodeficient mice with naked gene.The mice,inoculated successfully,were randomly divided into 4 groups:control group (C group,n =6),propofol group (P group,n =6),propofol+PI3K inhibitor (BYL71 9)group (P+B group,n =6),and PI3K inhibitor group (BYL71 9)(B group,n =6).The expressions of PI3K,p-Akt and Akt were examined by Western blot at week 4 after administration;the gene levels of PI3KR1, Akt1 and Akt2 were detected by RT-PCR at week 4 after administration;the number of metastatic lung nodules from both lungs was also observed at week 4 after administration.Results Compared with those in C group,the expressions of PI3K and p-Akt were significantly higher in P group (P <0.05),the level of PI3KR1 mRNA but not Akt1 and Akt2 mRNA was significantly increased(P < 0.05 ),and metastatic lung nodules significantly decreased (P <0.05).In B group,the expressions of PI3K and p-Akt were significantly decreased (P <0.05 ),the levels of PI3KR1,Akt1 and Akt2 mRNA were not significantly increased (P >0.05),but metastatic lung nodules significantly increased (P < 0.05 ).Compared with those in B group,in P+ B group the expressions of PI3K and p-Akt were markedly higher (P <0.05),the level of PI3KR1 mRNA but not Akt1 and Akt2 mRNA was significantly increased (P <0.05),and metastatic lung nodules significantly decreased (P <0.05).Conclusion Propofol can inhibit the metastasis of tumor cells through the upregulated and activated PI3K/Akt signaling pathway.

Objective To evaluate the role of autophagy in activation of astrocytes in the spinal cord of rats with neuropathic pain (NP).Methods One hundred thirty-six male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were randomly divided into 4 groups (n =34 each) using a random number table: sham operation group (group S), group NP, autophagy inhibitor 3-methyladenine (3-MA) group (group MA), and autophagy activator rapamycin group (group R).NP was induced by chronic constriction injury.In 3-MA and R groups, 3-MA 15 mg/kg and rapacymin 10 mg/kg were injected intraperitoneally, respectively, at 1 h before NP.Ten rats were randomly selected, and the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before NP and at 1, 3, 7 and 14 days after NP.Before NP and at 1, 3 and 7 days after NP, 6 rats were randomly sacrificed, the L4-6 segments of the spinal cord was harvested to detect microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ), Beclin-1 and glial fibrillary acidic protein (GFAP) expression by Western blot.Results Compared with group S, the MWT was significantly decreased, the TWL was shortened, and the expression of LC3 Ⅱ , Beclin-1 and GFAP was up-regulated at each time point after NP in group NP (P＜ 0.05).Compared with group NP, the MWT was significantly decreased, the TWL was shortened, the expression of LC3 Ⅱ and Beclin-1 was down-regulated, and the expression of GFAP was up-regulated at each time point after NP in group MA, and the MWT was significantly increased, the TWL was prolonged, the expression of LC3 Ⅱ and Beclin-1 was up-regulated, and the expression of GFAP was down-regulated at each time point after NP in group R (P＜0.05).Conclusion Autophagy is involved in the development and maintenance of NP through promoting the activation of astrocytes in the spinal cord of rats.

Objective To investigate the role of autophagy in the lung injury in the septic mice.Methods Thirty-six male C57BL/6 mice, aged 6 weeks, weighing 20-25 g, were randomly divided into 3 groups (n=12 each) using a random number table: sham operation group (group S);cecal ligation and puncture (CLP) group;CLP + autophagy inhibitor 3-methyladenine (3-MA) group (group CLP+3-MA).Sepsis was produced by CLP.In group CLP+3-MA, 3-MA 10 mg/kg was injected intraperitoneal at 1 h after operation.Arterial blood samples were taken at 24 h after operation for blood gas analysis, and the oxygenation index was calculated.The lungs were removed for microscopic examination of pathologic changes which were scored, and for determination of wet/dry lung weight ratio (W/D ratio) , myeloperoxidase (MPO) activity (using colorimetric method) and the expression of autophagy protein microtubule-associated protein 1 light chain 3 Ⅱ] (LC3 Ⅱ), Beclin-1 and lysosomes-associated protein Rab7 and lysosome-associated membrane protein-2 (LAMP2) (by Western blot).The lung was lavaged, and broncho-alveolar lavage fluid (BALF) was collected for determination of the total cell count and polymorphonuclear leukocyte (PMN) count.Results Compared with group S, the pathological score, W/D ratio, MPO activity, and the total cell and PMN counts in BALF were significantly increased, and oxygenation index was decreased in CLP and CLP +3-MA groups, and the expression of LC3 Ⅱ , Beclin-1, LAMP2 and Rab7 was up-regulated in group CLP (P＜0.05).Compared with group CLP, the pathological score, W/D ratio, MPO activity, and the total cell and PMN counts in BALF were significantly increased, the oxygenation index was decreased, and the expression of LC3 Ⅱ , Beclin-1, LAMP2 and Rab7 was down-regulated in group CLP+ 3-MA (P＜0.05).Conclusion Autophagy is involved in the endogenous protective mechanism of acute lung injury in the septic mice.

Objective To investigate whether hydrogen-rich saline could ameliorate the expressions of cytokines in a rat model of inflammatory pain through activating heme oxygenase-1 (HO-1).Methods Eighty male Sprague-Dawley (SD) rats,weighing 180 ～ 220 g,were randomly divided into five groups (n =16 in each group):Control group (Con),inflammation pain group (CFA),inflammation pain + hydrogen-rich saline group (CFA + H2),inflammation pain + HO-1 inhibitor Znpp-Ⅸ group (CFA + Znpp-Ⅸ),and inflammation pain + hydrogen-rich saline + HO-1inhibitor Znpp-Ⅸ group (CFA + H2 + Znpp-Ⅸ).The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were tested on days 1 (T1),2 (T2),3 (T3),5 (T4),7 (T5),and 14 (T6) after inflammation pain.The expressions of spinal HO-1 mRNA and protein were measured with real-time quantitative polymerase chain reaction (RT-PCR) and Western blot,and spinal inflammatory cytokines were measured with enzyme-linked immunosorbent assay (ELISA) on day 7 after inflammatory pain.Results Compared to Con group,MWT and TWL were significantly reduced;the spinal HO-1 mRNA level,protein expression and activity were increased;and the levels of tumor necrosis factor-α (TNF-α),interleukin (IL)-1β,IL-6 and IL-10 in spinal tissues were also increased in CFA group (P ＜ 0.05).Compared to CFA group,MWT and TWL were significantly increased;the spinal HO-1 mRNA level,protein expression and activity were further increased;and the levels of TNF-α,IL-1β and IL-6 were decreased,while IL-10 was further increased in CFA + H2group (P ＜ 0.05).Compared to CFA + H2 group,MWT and TWL were decreased;the spinal HO-1 mR-NA level,protein expression and activity were decreased;and the levels of TNF-α,IL-1β and IL-6 in spinal tissue were significantly increased,while IL-10 was decreased in CFA + H2 + Znpp-Ⅸ group (P ＜0.05).Conclusions Hydrogen-rich saline can ameliorate the mechanical and thermal allodynia in a rat model of inflammatory pain,and reduce the release of inflammatory cytokines via activating HO-1.