Objective To examine the role of the reactive oxygen species (ROS) and GADD 153 protein in angiotensin Ⅱ (Ang Ⅱ)-induced cardiomyocyta apeptosis in vitro.Methods Cardiomyocytes were isolated from ventricles of healthy 1-2 day old Wistar mrs of both sexes weighing 5-10 g and cultured in high glucose culture medium in an incubator filled with 5% CO2 at 37℃.The cardiomyocytes were randamiy allocated to one of 9 groups (n=5 each): group Ⅰ control (C); group Ⅱ ,Ⅲ,Ⅳ: cardiomyocytes were exposed to Ang Ⅱ 100 nmol/L for 6 h,12 h or 24 h respectively ( Ang Ⅱ 6,Ang Ⅱ12,Ang Ⅱ24) ; group Ⅴ:cardiomyocytes were preincubated with N-acetyl-L-cysteine (NAC) 5 mmol/L for 2 h before being exposed to Aag Ⅱ 100 nmol/L for 24 h (NAC + Ang Ⅱ); group Ⅵ:cardiomyocytes were exposed to NAC 5 mmol/L for 24 h (NAG); group Ⅶ:cardiomyocytes were tranfected with GADD153 antisense oligo-denxynueleotida (onti-ODN) 10 μmol/L for 24 h before being exposed to Ang Ⅱ 100 nmol/L for 24 h; group Ⅷ: cardiomyocytes were transfected with missense oligo-deoxynucleotide (mis-ODN) 10 μmol/L for 24 h before being exposed to Ang Ⅱ 100 nmol/L for 24 h and group Ⅸ: cardiomyocytes were exposed to transfection agent of same concentration for 24 h.Viability of myocytes was measured by MTT assay; ROS production was determined by spectropbotometry; apoptosis in cardiomyocytes was detected by staining with Hoechst33342; the percentage of Annexin V positive and PI negative myocytes was measured by flow cytometry as apoptosis rote and GADD153 protein expression was determined by Western blotting.Remits The viability of myocytes was significantly lower,while the ROS production,apoptosis rate and GADD153 protein expression in myocytes were significantly higher in group Ⅱ,Ⅲ,and Ⅳ(Ang Ⅱ6,Ang Ⅱ12,AngⅡ24) than in control group (P＜0.05).The cell viability was significantly higher and the ROS production,apoptosis rate and GADD153 protein expression in myocytes were significantly lower in groupⅤ(NAC+AngⅡ)and Ⅶ(anti-ODN + Ang Ⅱ) than in group Ang Ⅱ24(Ⅳ).Conclusion Angiotensin Ⅱ induces cardiomyocyte apoptosis by increasing ROS production and up-regulating GADD153 protein expression.

Mechanical ventilation is not only an important treatment method of acute respiratory distress syndrome (ARDS),but also one of the basic treatments in the intensive care unit (ICU).However,mechanical ventilation itself can cause or aggravate acute lung injury,which is called ventilator-induced lung injury (VILI).Currently,clinical pathogenesis of VILI includes four categories such as barotrauma,volutrauma,atelectrauma and hiotrauma.The pathogenesis of mechanical injury has been widely accepted,but the biological injury pathogenesis is unclear.With further research,we found that in the late stage VILI patients occured proliferation of puhnonary fibrosis,which may be formed by partial epithelial-mesenchymal transdifferentiation (EMT).Further study of specific pathogenesis of biotrauma and ARDS pulmonary fibrosis proliferation could provide new ideas for the clinical treatment of VILI.

AIM: To investigate the relationship between alteration of CHOP/GADD153 protein expression and cardiomyocyte apoptosis induced by angiotensin Ⅱ (AngⅡ), and inhibitory effects of CHOP/GADD153 antisense oligodeoxynucleotide (anti-ODN) on apoptosis in vitro.METHODS: Cultured neonatal cardiomyocytes were exposed to AngⅡ with or without preincubation of CHOP/GADD153 anti-ODN. Variability of myocytes was measured by MTT assay, the lactate dehydrogenase (LDH) release was also detected, the percentage of annexin V positive myocytes was monitored by flow cytometry as apoptosis rate, CHOP/GADD153, Bcl-2 and Bax expressions were determined by Western blotting. RESULTS: Compared with control group, the expression of CHOP/GADD153 was obviously increased from (0.20?0.02 to 0.75?0.06) in AngⅡ group (P

Objective To study the effects of selective inducible nitric oxide synthase (iNOS) inhibitor 1400W on nitric oxide (NO) exhalation and plasma nitrate concentration during endotoxemia in pigs. Methods Fourteen pigs of both sexes aged 12-16 weeks, weighing 40-60 kg were randomly divided into two groups : ( Ⅰ ) endotoxemia group ( n = 6) and ( Ⅱ ) 1400W group ( n = 8). The animals were anesthetized with pentobarbital and ketamine, intubated and mechanically ventilated. PaC02 was maintained at 35-45 mm Hg. Femoral artery and portal vein were cannulated for BP monitoring and blood sampling. E coli (LPS 20 mg L-1 ) was continuously infused iv at 4 ml h-1 for 24 h in both groups. In 1400W group, at 12 h of LPS infusion, continuous iv infusion of 1400W was started at 0.5 mg kg-1 h-1 . Blood samples were taken from artery and portal vein before LPS infusion (T0) and at 12 h (T, ) and 24 h (T2) of LPS infusion for determination of plasma nitrate concentration. Expired NO concentration was measured using NOA 280 NO analyzer. Results 1400W decreased LPS-induced increase in expired NO (P 0.05). Conclusion The selective iNOS inhibitor 1400W can inhibit iNOS activity and lead to decreased NO production.

Objective To evaluate the effects of mechanical stretch on pentraxin-3(PTX-3)mRNA and protein expression in human alveolar epithelial cells (A549 cells).Methods The human lung epithelial adenocarcinoma cells A549(A549 cells)were purchased from cell biology laboratory,Tongji Medical College,Huazhong University of Science and Technology.The cultured A549 cells were inoculated on collagen Ⅰ BioFlex plates and divided into 5 groups(n=3 wells each):group Ⅰ normal control;groupⅡsham mechanical stretch;group Ⅲ mechanical stretch;groupⅣsiRNA and groupⅤ siRNA+mechanical stretch.In group Ⅲ the cells underwent square cyclic mechanical stretch for 4 h using the Flexercell Systcm.In group Ⅳ the cells were transfected with chemosynthetic PTX-3 specific siRNA by RNAi technique.In group Ⅴ at 24 h after being transfected with PTX-3 siRNA the cells underwent mechanical stretch for 4 h.In groupⅡ mechanical stretch of the cells were prevented by Flexstep.The expression of PTX-3 mRNA in the cells was detected by real-time PCR and the expression of PTX-3 protein in the culture media was determined by Western blotting.Apoptosis of the cells was measured bv flow cytometry(Beeten-Dickinson,USA).Results PTX-3 mRNA and protein expression was signlficantly up-regulated by mechanical stretch in groupⅢand decreased by transfection with siRNA in group Ⅳand Ⅴ as compared with group Ⅰ andⅡ(P＜0.05 or 0.01).The apoptosis ratio was significantly higher in group Ⅲ and Ⅴ than in groupⅡ and was significantly lower in group Ⅴ thanin group Ⅲ(P＜0.01).Conclusion Mechanical stretch can up-regulate PTX-3 mRNA expression in A549 cells.

ObjectiveTo investigate the effect of sivelestat sodium on the prognosis in patients with acute lung injury (ALI) and acute respiratory distress syndrome (ARDS).Methods Databases including PubMed, EBSCO, Springer, Ovid, Wanfang data, CNKI and China Biology Medicine (CBM) were searched to identify randomized controlled trials (RCTs) regarding sivelestat sodium treatment for ALI/ARDS published from 1985 to December 2014. The patients in treatment group received intravenous infusion of sivelestat sodium, and those in control group received normal saline. The items for analysis were 28-day mortality, duration of mechanical ventilation, length of intensive care unit (ICU) stay, and oxygenation index on day 3. According to the evaluation method of Cochrane system, data extraction and quality assessment from the literature were carried out. Meta-analysis was performed using RevMan 5.3. The publication bias was analyzed with funnel plot.Results Five RCTs with a total of 780 participants were included, with 389 patients in sivelestat sodium group, and 391 in control group. Meta analysis showed: compared with control group, sivelestat sodium could not lower the 28-day mortality [odds ratio (OR) = 0.91, 95% confidence interval (95%CI) =0.66-1.26,P = 0.58], or shorten the duration of mechanical ventilation or length of ICU stay [duration of mechanical ventilation: mean difference (MD) = -0.02, 95%CI = -0.29 to 0.24,P = 0.87; length of ICU stay:MD = -9.63, 95%CI =-23.34 to 4.08,P = 0.17], but it could improve oxygenation index on day 3 (MD = 0.88, 95%CI = 0.39 to 1.36, P = 0.000 4). Heterogeneity was not significant for the main analysis and no publication bias was shown on funnel plot. Conclusion Sivelestat sodium gave rise to a better oxygenation on day 3, but did not change the length of mechanical ventilation and ICU stay, and it did not improve 28-day mortality in ALI and ARDS.

Objective To investigate the effect of exogenous protectin DX (PDX) on acute lung injury in septic mice.Methods Thirty male C57BL/6 mice,aged 6-8 weeks,weighing 20-25 g,were randomly divided into 3 groups (n =10 each) using a random number table:sham operation group (Sham group),sepsis group (S group) and PDX group.Sepsis was produced by cecum ligation and puncture (CLP) in the mice anesthetized with pentobarbital sodium.At 1 h after CLP,PDX 300 ng was injected intraperitoneally in PDX group,and the equal volume of normal saline was given in Sham and S groups.At 24 h after CLP,the mice were sacrificed,and the broncho-alveolar lavage fluid (BALF) was collected for determination of interleukin-1beta (IL-1β),tumor necrosis factor-alpha (TNF-α),IL-6 and IL-10 concentrations,and the lungs were removed for microscopic examination and for determination of the myeloperoxidase (MPO) activity,wet/dry lung weight ratio (W/D ratio) and phosphorylation of nuclear factor-kappa B (NF-κB) p65.Lung injury scores were calculated.Results Compared with Sham group,the lung injury score,MPO activity,W/D ratio,phosphorylation of NF-κB p65,and concentrations of protein and inflammatory factors in BALF were significantly increased in S and PDX groups (P＜0.05).Compared with S group,the lung injury score,MPO activity,W/D ratio,phosphorylation of NF-κB p65,and concentrations of protein,IL-1β,TNF-α and IL-6 in BALF were significantly decreased,and the concentration of IL-10 in BALF was significantly increased in PDX group (P＜0.05).Conclusion Exogenous PDX can alleviate acute lung injury through inhibiting NF-κB activity in the lung tissues of septic mice.

Objective To investigate the effects of adenovirus containing human beta-nerve growth factor (Ad-hNGFβ) gene on substance P (SP) and calcitonin gene-related peptide (CGRP) content of the spinal cord in a rat model of neuropathic pain by chronic constrictive injury (CCI). Methods Forty-eight male SD rats weighing 200-250 g were randomly divided into 3 groups (n=16 each) : group Ⅰ sham operation; group Ⅱ CCI and group Ⅲ CCI + Ad-hNGFβ gene IT. The animals were anesthetized with intraperitoneal choral hydrate 300-350 mg/kg. The right sciatic nerve was exposed and 4 ligatures were placed on the sciatic nerve at 1-2 nun intervals as described by Bennet and Xie[5]. In sham operation group, right sciatic nerve was exposed but not ligated. In group Ⅰ and Ⅱ artificial cerebrnspinal fluid was injected IT instead of Ad-hNGFβ gene. The behavior score and the paw-withdrawal latency (PWL) to radiant heat and mechanical stimulus were measured one day before operation and every 4 days within the 28 days after gene transfection. Four animals were killed at 4, 7, 14 and 28 day after IT gene transfection in each group and lumbar segment (L4-6 ) of the spinal cord was removed for determination of SP and CGRP content by immunohistochemistry. Results The behavior scores were significantly higher and PWL to radiant heat and mechanical stimulus were significantly lower in group Ⅱ and Ⅲ than in group Ⅰ. There was no significant difference in the behavior score and PWL to mechanical stimulus between group Ⅱ and Ⅲ while the PWL to radiant heat was significantly higher in group Ⅲ than in group Ⅱ. After operation SP and CGRP content were significantly higher in group Ⅱ and group Ⅲ than in group Ⅰ , and significangly lower in group Ⅲ than in group Ⅱ 7-28 days after operation. Conclusion The recomhinant Ad-hNGFβ gene transfection can attenuate heat hyperalgesia by reducing SP and CGRP content of the spinal cord in a rat model of neuropathic pain induced by CCI.

Objective To determine the half-effective target plasma concentration(EC50)of remifentanil inhibiting airway response when combined with propofol by TCI in patients undergoing fiberoptic bronchoscopy.Methods Forty ASAⅠorⅡ patients,aged 20-64 yr, undergoing elective fiberoptic bronchoscopy, were randomly divided into 2 groups (n = 20 each).Anesthesia was performed with TCI of remifentanil and propofol in both groups. The target effect site concentration of propofol was 3 μg/ml.The target effect site concentration of remifentanil was determined by up-and-down sequential hial. The target effect site concentration of remifentanil was set at 5 μg/ml in the first patient and the ratio of the target concentrations between the two consecutive patients was 1.1. BIS ≤ 60 was defined as the suitable depth of anesthesia in group A.The airway response≤grade Ⅱ was defined as the suitable depth of anesthesia in group B. The EC50 and 95% confidence interval (CI) required to inhibit the airway response were calculated in the two groups.Results The EC50 and 95% CI of remifentanil required to inhibit the airway response were 4.50μg/L (95% CI 3.88-5.36 μg/L)in group A and 4.10μg/L(95% GI 3.31-5.00 μg/L) in group B. The EC50 of remifentanil inhibiting airway response was significantly higher in group A than in group B. Conclusion The EC50 of remifentanil inhibiting airway response is 4.10 μg/L when combined with propofol by TGI (effect site concentration 3μg/ml)in patients undergoing bronchoscopy. BIS is not a suitable index assessing the depth of propofol-remifentanil anesthesia.

Objective To evaluate the changes in the expression of diplopore potassium ion channel TRESK mRNA in dorsal root ganlion (DRG) in rats with neuropathic pain (NP) .Methods Thirty-two male SD rats weighing 220-250 g were randomly divided into 2 groups ( n = 16 each) : group sham operation (group S) and group NP. NP was induced by ligation and severance of left tibial and common fibular nerves according to the technique described by Decosterd. Eight rats in each group were sacrificed 1 day before and 14 day after operation and their L4,5 DRGs in the operated side were isolated for determination of TRESK mRNA expression by RT-PCR. In the remaining 8 rats in each group paw withdrawal threshold to mechanical stimuli ( MWT) and paw withdrawal latency to a thermal nociceptive stimulus (TWL) were measured at 1 day before (baseline) and 1, 3, 5, 7, 14 day after operation. Results MWT was significantly lower in group NP than in group S. The TRESK mRNA expression in L4,5 DRGs in the operated side was significantly decreased after operation as compared with the baseline before operation in group NP and was significantly lower in group NP than in group S. Conclusion The development and maintenance of NP may be closely related with down-regulation of TRESK mRNA.