白藜芦醇通过上调脂肪变性HepG2细胞Sirt3表达改善线粒体功能

目的 探讨白藜芦醇(resveratrol,RSV)对脂肪变性肝细胞线粒体功能的影响,以及沉默信息调节因子3(sirtuin 3,Sirt3)在其中的重要作用.方法 用0.2 mmol/L棕榈酸(palmitic acid,PA)处理HepG2细胞24 h建立肝细胞脂肪变性模型,再用40 μmol/L RSV和/或50 μmol/L Sirt3抑制剂3-TYP处理24 h.实验分为对照组、PA组、PA+ RSV组、PA+ RSV+ 3-TYP组、PA+ 3-TYP组.用相关试剂盒检测甘油三酯(TG)含量、沉默调节蛋白3(sirtuin 3,Sirt3)活性、ATP合酶活性、ATP生成量以及线粒体基础呼吸率,使用DCFH-DA荧光探针检测细胞活性氧(reactive oxygen species,ROS)含量,Westernblot检测Sirt3蛋白表达.Sirt3 siRNA处理HepG2细胞后再用PA和/或RSV处理,分析ROS和甘油三酯含量.结果 相对于PA组,PA+RSV组HepG2细胞的甘油三酯、ROS含量明显降低(P＜0.05),ATP合酶活性、ATP生成量和基础呼吸率明显提高(P＜0.05).而3-TYP或Sirt3 siRNA处理后,RSV的上述作用被削弱(P＜0.05).结论 RSV通过上调脂肪变性肝细胞Sirt3的表达与活性而改善其线粒体功能.     

烧伤休克期有关补液公式的临床应用与评价

目的 评价第三军医大学烧伤休克期补液公式(简称三医大公式)在大面积烧伤患者休克防治中的应用.方法 选择2005-2007年笔者单位收治的热力烧伤患者(烧伤总面积大于或等于30%TBSA、伤后8 h内入院且无特殊疾患)共71例,分为成人组(46例)、小儿组(25例).患者入院后即按照三医大公式进行液体复苏治疗,同时监测尿量、心率、血压等指标,根据患者实际情况随时调整补液速度.记录并统计2组患者补液量、实际补液系数、尿量. 结果 71例患者均平稳度过休克期,未发生明显的因液体复苏引起的相关并发症.成人组伤后第1、2个24 h及小儿组伤后第2个24 h的实际补液量超过各自计划补液量的16%～38%.成人组第1、2个24 h的实际补液系数大于公式所要求的补液系数.2组患者第1个24 h尿量为1.1～1.2 mL·kg-1·h-1左右;第2个24 h成人组为(1.2±0.4)mL·kg-1·h-1,小儿组为(1.7±0.5)mL·kg-1·h-1. 结论 三医大公式是大面积烧伤患者休克期治疗的较好选择,在应用此公式时须强调进行个性化液体复苏治疗.     

Proteomic analysis to identify biomarker proteins in pancreatic ductal adenocarcinoma

Background: Pancreatic ductal adenocarcinoma (PDAC) is the fifth most common cause of death from cancer in Korea. PDAC is difficult to diagnose at an early stage and even more difficult to cure. Thus, there is an urgent need to identify molecular targets for early diagnosis and effective treatment. The objectives of this study were to identify differentially expressed biomarker proteins of PDAC using proteomic analysis, to validate the identified biomarker proteins associated with carcinogenesis using western blot analysis and to evaluate clinical factors influencing expression of candidate biomarker proteins. Methods: In the present study, we carried out proteomic analysis in 10 pairs of PDAC specimens with matching adjacent normal tissues to clarify the different patterns of protein expression. The proteins were separated by high‐resolution 2‐D polyacrylamide gel electrophoresis (2D PAGE) and the differentially expressed proteins were identified by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS). Differential expression of candidate biomarker proteins associated with carcinogenesis was further validated using western blot analysis. Standard statistical analysis was carried out in an attempt to establish a correlation between clinical variables and expression of candidate biomarker proteins. Results: Analysis of PDAC and the adjacent normal tissues showed reproducibly similar proteomic patterns for each group. Approximately 700 spots each were seen by silver‐stained gels from both PDAC and normal tissues. Differentially expressed protein spots were gel digested and identified by MALDI‐TOF MS. Twenty‐five proteins were identified, of which five proteins (galectin‐1, enolase‐2, α‐1‐antitrypsin, N‐myc interactor, peroxiredoxin‐4) were previously reported as being differentially expressed either at the mRNA level or protein level in human cancer. The five proteins were selected for candidate biomarker proteins related to carcinogenesis. These proteins were further validated by western blot analysis. Among the candidate biomarker proteins, galectin‐1 expression was highly correlated to histology (P = 0.019), T stage (P = 0.047), N stage (P = 0.033) and American Joint Committee on Cancer stage (P = 0.011). Conclusion: Differentially expressed 25 proteins in PDAC were identified using proteomic analysis and five proteins related to carcinogenesis were validated by western blot analysis. galectin‐1 expression was highly correlated to tumour histology and stage.     

Posterior tibial artery-based multilobar combined flap free transfer for repair of complex soft tissue defects

In this report, the posterior tibial artery (PTA) based multilobar combined flap is introduced for the repair of complex soft tissue defects. The flap was designed based on the perforatoring branches of PTA in the anterior soleus muscle septum, which supply the skin over the medial side of the calf and the entire soleus muscle. The saphenous nerve was included in one perforator flap of the combined flap for reinnervation. The tibial artery was repaired with a vein graft after harvest of flap. From October 2005 to February 2007, eight patients (6 males, 2 females) underwent PTA-based multilobar combined flap transfer for coverage of soft tissue defects involving the foot (three cases), hand (two cases), and calf (three cases). Each combined flap composed of two to three perforator flaps, and the size of the perforator flaps ranged from 4 x 2 cm to 10 x 8 cm. With an average follow-up of 6 months, all flaps survived without complications and injured extremities showed a good functional recovery with restoration of the partial protective sensation on the flap with reinnervation. This clinical report has shown that a reliable multilobar combined flap can be designed based on the perforators of the posterior tibial artery and used for coverage of the complex wound.     

Effect of internal biliary drainage on plasma levels of endotoxin,cytokines, and C-reactive protein in patients with obstructive jaundice

Preoperative biliary drainage may improve the cytokine and acute-phase response derangements observed in patients with obstructive jaundice. We conducted a prospective longitudinal, before-after trial in our 600-bed teaching hospital. Twenty-four patients with obstructive jaundice were investigated, 11 with benign obstruction and 13 with malignant disease. Endoscopic internal biliary drainage was performed in all patients (7 by papillotomy and 17 by endoprostheses). Endotoxin, tumor necrosis factor alpha (TNF-a), interleukin-6 (IL-6), nitric oxide production, and C-reactive protein (CRP) were determined at admission and on days 2 and 7 after internal biliary drainage was accomplished. Bile cultures were obtained before and at the time of drainage. Endotoxin, IL-6, TNF-a, and CRP were significantly higher in patients with cancer. After internal drainage, endotoxin (11.4 vs. 2 EU/L; p <0.05), TNF-a (87.5 vs. 48 pg/ml; p = 0.03), and IL-6 (324 vs. 232 pg/ml; p <0.05) plasma levels decreased significantly in the early postdrainage period in patients with cancer. Endotoxin, cytokines, as well as the CRP plasma values, however, increased again on day 7 after drainage. This trend was less marked in patients with benign obstruction. Patients with positive bile cultures after drainage displayed higher levels of CRP (115 vs. 62 mg/L; p = 0.03), IL-6 (598 vs. 330 pg/ml; p = 0.04), and endotoxin (10.6 vs. 4.8 EU/L; p = 0.02) than those with negative bile cultures. Biliary tract obstruction is associated with an increase in endotoxin levels, a positive acute-phase response, and plasma cytokine elevation. After biliary drainage a transitory improvement of these alterations was observed, although values remained high 1 week postdrainage. These findings were associated with positive bile cultures.     

High glucose increases inducible NO production in cultured rat mesangial cells. Possible role in fibronectin production.

BACKGROUND/AIM: Increased nitric oxide (NO) generation and action have been suggested to be associated with glomerular hyperfiltration and increased vascular permeability early in diabetes. However, previous studies have primarily focused on the constitutive nitric oxide synthase (cNOS) pathway present in endothelial cells, and the role of the inducible NOS (iNOS) pathway in diabetic nephropathy has remained unclear. This study examined whether high glucose modulates NO synthesis by the iNOS pathway in rat mesangial cells. In addition, the effect of inhibition of the iNOS pathway on fibronectin production was determined to examine the role of the iNOS pathway in high glucose-induced extracellular expansion by mesangial cells. METHODS: NO synthesis by the iNOS pathway was evaluated by nitrite and iNOS mRNA and protein productions. The effects of protein kinase C (PKC) inhibitor and aldose reductase inhibitor on the iNOS mRNA expression and aminoguanidine, a relatively specific inhibitor of the iNOS on fibronectin protein production were examined. RESULTS: High 30 mM glucose concentration led to significant increases in nitrite production of rat mesangial cells upon stimulation with lipopolysaccharide (LPS) plus interferon-gamma (IFN-gamma) compared with control 5.6 mM glucose concentration. Mesangial iNOS mRNA expression and protein production also increased significantly in response to high glucose. The addition of calphostin C, a PKC inhibitor, and 6-bromo-1,3-dioxo-1H-benz[d,e]isoquinoline-2(3H)-acetic acid, an aldose reductase inhibitor, significantly suppressed the enhancement of iNOS mRNA expression in high glucose concentration. High glucose also significantly increased fibronectin protein production of mesangial cells upon stimulation with LPS plus IFN-gamma compared to control glucose. Aminoguanidine reversed this high glucose-induced fibronectin production at dose inhibiting iNOS mRNA expression. CONCLUSIONS: These results indicate that high glucose enhances cytokine-induced NO production by rat mesangial cells, and that the activation of PKC and aldose reductase pathway may play a role in this enhancement. In addition, high glucose-induced NO production by the iNOS pathway may promote extracellular matrix accumulation by mesangial cells under certain condition.     

MicroRNA-687 Induced by Hypoxia-Inducible Factor-1 Targets Phosphatase and Tensin Homolog in Renal Ischemia-Reperfusion Injury

Ischemia-reperfusion injury contributes to tissue damage and organ failure in clinical settings, but the underlying mechanism remains elusive and effective therapies are still lacking. Here, we identified microRNA 687 (miR-687) as a key regulator and therapeutic target in renal ischemia-reperfusion injury. We show that miR-687 is markedly upregulated in the kidney during renal ischemia-reperfusion in mice and in cultured kidney cells during hypoxia. MiR-687 induction under these conditions was mediated by hypoxia-inducible factor-1 (HIF-1). Upon induction in vitro, miR-687 repressed the expression of phosphatase and tensin homolog (PTEN) and facilitated cell cycle progression and apoptosis. Blockade of miR-687 preserved PTEN expression and attenuated cell cycle activation and renal apoptosis, resulting in protection against kidney injury in mice. Collectively, these results unveil a novel HIF-1/miR-687/PTEN signaling pathway in ischemia-reperfusion injury that may be targeted for therapy.     

不停跳冠状动脉旁路移植术中应用MostCare/PRAM监测系统进行血流动力学监测的临床研究

目的探究Most Care/PRAM系统监测下不停跳冠状动脉旁路移植术(OPCABG)患者术中血流动力学变化情况和预后分析。方法纳入2016年10月至2017年1月安贞医院89例OPCABG患者,其中男53例、女36例,年龄(60.50±8.40)岁。记录术中血流动力学变化情况。按是否发生心肌梗死、低心排血量等严重循环不良事件,分为平稳组和严重循环不良事件组,进行相关分析。结果手术全程监测完整血流动力学数据患者65例,开胸前和关胸后被动抬高试验的每搏量(SV)升高均值分别为23.00%±3.20%和29.40%±3.70%。麻醉、开胸、应用肝素时、搭桥中、应用鱼精蛋白时、关胸和术毕7个时间段,SV明显下降,外周血管阻力指数(SVRI)持续显著增加,最大压力梯度(d P/d T)和心脏循环效率(CCE)在麻醉后明显下降,搭桥时下降到最低,其后逐渐升高;每搏量变异率(SVV)和脉压变异率(PPV)在麻醉后下降,开胸后一直升高。89例患者发生严重循环不良事件共9例,其中4例死亡。严重循环不良事件组术中基础SVRI、SVV和PPV均显著高于平稳组(P<0.05),CCE、d P/d T和SV差异无统计学意义(P>0.05)。术中基础SVRI、CCE、d P/d T、SVV、PPV和SV均值与预后指标均无明显相关性。结论OPCABG术中易出现血流动力学的改变,因此,OPCABG术中宜应用Most Care/PRAM仪进行血流动力学监测,并及时纠正血流动力学异常。     

Osteopontin inhibits macrophage nitric oxide synthesis to enhance tumor proliferation

BACKGROUND: Interactions between tumor cells and their host environment can play a major role in regulating survival programs required for tumor progression. Osteopontin (OPN) is a glycophosphoprotein overexpressed by tumors, and is a key molecule for tumor progression and metastasis. OPN also inhibits expression of autocrine and paracrine inducible nitric oxide synthase (iNOS). Given the cytotoxic effects of macrophage NO expression, we hypothesized that tumor-derived OPN inhibits expression of local macrophage iNOS to potentiate tumor survival. METHODS: We used a coculture system of murine CT26 colorectal cancer cells with RAW264.7 murine macrophage cells. CT26 expresses OPN at high levels. RNA interference was utilized to produce long-term specific silencing of OPN in CT26. RESULTS: Inhibition of constitutive OPN synthesis in CT26 upregulates local NO production with inhibition of CT26 proliferation and promotion of CT26 apoptosis. Macrophage iNOS expression is accompanied by increased binding activity of nuclear factor-kappaB DNA. When the CT26 culture media were examined for a panel of proinflammatory cytokines, elevated concentrations of granulocyte colony-stimulating factor (G-CSF) were found. Subsequently, in CT26 cells treated with antisense-G-CSF, NO levels in CT26-RAW cocultures were significantly decreased. CONCLUSION: In our system of CT26-RAW264.7 coculture, we conclude that inhibition of OPN synthesis in CT26 results in G-CSF-mediated induction of macrophage iNOS expression with resultant inhibition of CT26 proliferation via increased apoptosis. Our results suggest that tumor-derived OPN may enhance tumor survival by down regulating expression of NO in the local microenvironment. This is one mechanism by which OPN may potentiate cancer survival and progression.