晚期糖基化终产物对大鼠血管平滑肌细胞分泌炎症性趋化因子的影响及机制

目的 观察晚期糖基化终产物(AGE)对血管平滑肌细胞(VSMC)分泌单核细胞趋化蛋白1(MCP-1)和白细胞介素8(IL-8)的影响并初步探讨可能的细胞内信号转导机制.方法 分离、培养原代大鼠VSMC并进行鉴定.采用糖基化血清白蛋白(GSA)模拟AGE,观察不同浓度GSA(10、100和500 μg/ml)对VSMC分泌MCP-1和IL-8的影响和时间曲线;并对其进行细胞增殖率校正以消除VSMC数量增加对测定的影响;p38MAPK抑制剂(SB203580)、ERK1/2抑制剂(PD98059)及NF-κB抑制剂(PDTC)、Proteasome抑制剂(MG132)预处理后观察GSA刺激VSMC分泌MCP-1和IL-8水平的改变.结果 与空白对照组相比,100 μg/ml GSA作用24 h VSMC分泌MCP-1(13.01ng/ml±0.12ng/ml比7.02 ng/ml±0.26 ng/ml,P＜0.05)和IL-8(12.6ng/ml±0.86 ng/ml比3.07 ng/ml±0.35ng/ml,P＜0.05)水平最高.经过细胞增殖校正后,GSA仍然能够促进VSMC表达MCP-1和IL-8.MAPK抑制剂和NF-κB抑制剂预处理后发现PDTC(10 μmol/L)、SB203580(5 μmol/L)以及MG132(10 μmol/L)可以抑制GSA刺激VSMC表达MCP-1和IL-8.结论 GSA可以促进VSMC分泌致炎性趋化因子MCP-1和IL-8,这种作用独立于细胞增殖,可能是通过激活细胞内p38MAPK信号转导通路,促进核因子NF-κB的活化而实现.

Abstract：

Objective To investigate the effects of advanced glycation end products (AGEs) on the secretion of monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) in vascular smooth muscle cells (VSMCs) and explore its possible intracellular signaling mechanism. Methods Primary rat VSMCs were isolated and identified. VSMCs were treated with glycation serum albumin (GSA), an important component of AGEs, in series of concentrations and time. The role of MAPK and NF-κB inhibitors was confirmed. The levels of MCP-1 and IL-8 were determined by enzyme-linked immunosorbent assay (ELISA). Results VSMCs were treated with GSA at the doses of 10 μg/ml, 100 μg/ml and 500 μg/ml respectively. In comparison with the control group, the levels of MCP-1 ( 13.01 ng/ml ± 0.12 ng/ml vs 7. 02 ng/ml ±0. 26 ng/ml, P＜0.05) and IL-8 (12. 6 ng/ml ±0. 86 ng/ml vs 3. 07 ng/ml ±0.35 ng/ml,P＜0.05) increased in the GSA-treated group, especially at the concentration of 100 μg/ml. After adjustment for cells proliferation, the levels of MCP-1 and IL-8 were still higher in the GSA-treated group.After a pretreatment of PDTC ( 10 μmol/L), SB203580 (5 μmol/L) and MG132 ( 10 μmol/L), the levels of MCP-1 and IL-8 decreased. However, it had no change when pretreated with PD98059 (20 μmol/L).Conclusion GSA promotes the secretion of MCP-1 and IL-8 in VSMCs. Such an effect is not dependent on cellular proliferation. It may be realized through an activation of NF-κB by p38MAPK-sensitive intracellular signaling pathway.     

Antithrombin-Ⅲ without concomitant heparin improves endotoxin-induced acute lung injury rats by inhibiting the activation of mitogen-activated protein kinase

Background Antithrombin-Ⅲ (AT-Ⅲ), the major inhibitor of thrombin in plasma, also has anti-inflammation property and might have positive effect on sepsis. The present study aimed to investigate the effects of AT-Ⅲ on inflammatory reaction and pulmonary protection in endotoxin-induced acute lung injury (ALI) rat.Methods Sixty male Sprague-Dawley rats were randomly assigned equally to normal control group, ALl group, AT-Ⅲ treatment group, AT-Ⅲ+heparin treatment group, and heparin treatment group. The pulmonary vascular permeability index (PVPI) was measured by single nuclide tracer technique. The activity of AT-Ⅲ in plasma was determined by the method of synthetic chromogenic substrata. Tumor necrosis factor-a (TNF-a) and interleukin-6 (IL-6) levels in serum were determined by enzyme-linked immunosorbent assay. The expressions of lung tissue mitogen-activated protein kinases (ERK1/2, P38 and JNK MAPK) were determined by Western blotting.Results Rats had significantly improved lung histopathology in the AT-Ⅲ treatment group and heparin treatment group compared with the ALI group. The PVPI of the ALI group was 0.38±0.04, significantly higher than that of the normal control group (0.20±0.02, P <0.01), AT-Ⅲ treatment group (0.30±0.04, P <0.01) and heparin treatment group (0.28±0.04,P <0.01) respectively. There were no significant differences of PVPI in the ALl group and AT-Ⅲ+heparin treatment group.The activity of AT-Ⅲ in plasma in the ALl group was (76±8)%, significantly lower than that of the normal control group ((96±11)%, P <0.05) and AT-Ill treatment group ((105±17)%, P <0.05) respectively. The serum levels of TNF-α and IL-6 of the ALI group were (2.770±0.373) pg/L and (1.615±0.128) ng/ml respectively, significantly higher than those of the normal control group ((0.506±0.093) pg/L and (0.233±0.047) ng/ml respectively, all P <0.01), AT-Ⅲ treatment group ((1.774±0.218) μg/L and (1.140±0145) ng/ml respectively, all P <0.01) and heparin treatment group ((1.924i±0.349) μg/L and (1.223±0.127) ng/ml respectively, all P <0.01). The lung tissue levels of phospho-ERK1/2 and phospho-P38 MAPK expressions were markedly higher in the ALI group than in the normal control group, AT-Ⅲ treatment group and heparin treatment group respectively.Conclusions AT-Ⅲ without concomitant heparin inhibited the activation of ERK1/2 and P38 MAPK, down-regulated the levels of downstream cytokines TNF-α and IL-6, relieved endothelial permeability, and improved the ALl in endotoxin-induced rats. It might be helpful to administrate AT-Ⅲ alone, not with concomitant heparin, to those patients with ALI and sepsis.     

焦痂切除对大面积烧伤患者血栓并发症的预防

目的观察焦痂切除对大面积烧伤患者血栓并发症的预防。方法选取2013年4月至2014年12月我院收治的大面积烧伤患者60例为研究对象,采用随机数表法分为观察组和对照组各30例,对照组入院后立即进行深静脉置管,并给予乌司他丁及生理、心理干预,观察组在此基础上实施焦痂切除术,记录两组创面愈合时间、住院时间、补液量,分析凝血酶原片段1+2(F_(1+2))、纤维蛋白肽(FPA)及抗凝血酶活性(AT)、蛋白C活性(PC),并观察并发症发生率。结果观察组创面愈合时间(25.18±1.44)d、住院时间(8.73±1.05)d与对照组比较明显较短(P0.05),观察组补液量(2.14±0.55)L低于对照组(P0.05);观察组F1+2(1.03±0.14)nmol/L、FPA(2.34±0.57)μg/L、TNF-α(0.14±0.06)ng/ml、IL-6(0.64±0.05)ng/ml较对照组低(P0.05),观察组AT活性(71.63±1.21)%、PC活性(82.31±1.25)%明显高于对照组(P0.05);观察组并发症发生率6.7%低于对照组26.7%(P0.05),两组血栓并发症发生率3.3%、20.0%比较亦有统计学意义(P0.05)。结论焦痂切除可有效预防大面积烧伤患者血栓并发症,改善其凝血功能,值得在临床推广应用。     

鞘内注射雷帕霉素对小鼠神经病理性痛行为的影响

目的 探讨鞘内注射雷帕霉素对小鼠神经病理性痛行为的影响.方法 选择鞘内置管成功无运动障碍及严重体重减轻的成年雄性C57/BL6小鼠48只,随机分为两组:假手术组(sham组,n=24)和慢性坐骨神经损伤模型组(CCI组,n=24).建立模型后将各组随机分为3组,组-不做任何处理,组二在术后1～6 d鞘内注射雷帕霉素1μg/5μl,组三鞘内给予溶媒20％ DMS0 5μl(sham,sham+R,sham+V,CCI,CCI+R,CCI+V,n=8).在手术前1d,手术后1d,3d,5d,7d,10d,14 d,17d,21d,28 d检测小鼠双侧足底机械缩足阈值( Mechanical Withdraw Threshold,WMT)和热辐射缩足潜伏期(Thermal Withdraw Latency,TWL).结果 与sham组相比,CCI组小鼠手术侧足底MWT和TWL均明显降低[第7天,MWT:( 1.02±0.12)g,(0.42±0.12)g,F=51.01,P＜0.05;TWL:(7.03±0.71)s,(3.26±0.66)s,F=38.27,P＜0.05].而CCI后1-6d鞘内注射雷帕霉素1μg/5μL,使小鼠的MWT明显升高[第7天,(0.42±0.18)g,(0.86±0.25)g,F=6.56,P＜0.05],并且至少持续至术后10d.与之相对的,雷帕霉素对CCI小鼠的热痛觉过敏也有缓解趋势,但差异无统计学意义.sham组和对侧足底痛行为未见明显改变(P＞0.05).结论 鞘内注射雷帕霉素可以明显缓解CCI小鼠的机械痛觉异常,但对热痛觉过敏无显著作用.     

1,25-Dihydroxyvitamin D3 pretreatment enhances the efficacy of allergen immunotherapy in a mouse allergic asthma model

Background  Allergen-specific immunotherapy can induce immune tolerance to specific allergens by regulating immune status of individuals. However, its clinical application is limited due to individual differences in efficacy among patients and un-confirmed safety. 1,25 Dihydroxyvitamin D₃ (1,25(OH)₂D₃) has been shown to be involved in a variety of physiological processes, including immune response regulation. In the present study we explored the role of 1,25(OH)₂D₃ pretreatment for immunotherapy.

Methods  Seventy-five BALB/c mice were randomly divided into five groups (15 mice per group). The mouse allergic asthma model was established by intra-peritoneal injection of ovalbumin (OVA, 10 μg) and aluminium hydroxide (2 mg) as an adjuvant. Intra-peritoneal injection of 50 ng of 1,25(OH)₂D₃ served as a pretreatment, subcutaneous injection of OVA (100 μg) as an immunotherapy, and 1% OVA inhalation as a challenge. Histopathological analysis was performed on four mice per group. The number of cells and their classification in bronchoalvolar lavage (BAL) fluid were assayed. Levels of serum OVA-specific immunoglobulin E (sIgE) and IFN-γ, IL-4, IL-5 and IL-10 in BAL fluid were measured by ELISA.

Results  After 1,25(OH)₂D₃ pretreatment, immunotherapy could significantly inhibit the infiltration of inflammatory cells into lung tissues and BAL fluid of mice with allergic asthma when compared with un-treated animals (eosinophils: (7.46±1.34)×10⁴/ml vs. (13.41±1.67)×10⁴/ml, P <0.05). In addition, levels of IL-4 ((36.91±7.87) pg/ml vs. (43.70±6.42) pg/ml, P >0.05) and IL-5 ((41.97±7.93) pg/ml vs. (60.14±8.35) pg/ml, P <0.05) in BAL fluid and serum sIgE ((0.42±0.05) vs. (0.75±0.06) OD units, P <0.05) were profoundly reduced. However, the IL-10 level in BAL fluid was significantly increased ((67.74±6.57) pg/ml vs. (44.62±8.81) pg/ml, P <0.05).

Conclusions  These results indicated that 1,25(OH)₂D₃ pretreatment enhanced the inhibitory effects of immunotherapy on allergic airway inflammation. In the treatment of allergic diseases, 1,25(OH)₂D₃ pretreatment may be beneficial for improving the efficacy of immunotherapy. 

     

Antagonistic Effects of Tranilast on Proliferation and Collagen Synthesis Induced by TGF-β2 in Cultured Human Trabecular Meshwork Cells

Whether tranilast had antagonistic effect on proliferation inhibition and collagen synthesis promotion induced by TGF-β2 in cultured human trabecular meshwork cells was investigated. Suspension of 1 × 104 cultured human trabecular meshwork cells of 3-5 passage was distributed in each well of a 96-well disk and divided into control group and experimental group. After 24 h, 0 μg/ml (control), 12.5 μg/ml, 25 μg/ml, 50 μg/ml tranilast with 3.2 ng/ml TGF-β2 were added into the incubation medium. Another 24 h later, proliferation and collagen synthesis in cultured human trabecular meshwork cells were examined respectively by using tetrazolium-based semiautomated colormetric (MTT) assay and 3 H-proline incorporation with liquid scintillation technique. The results showed absorbance (A) values of the experimental groups were 0. 9036 ± 0. 3017, 1.1361 ±0.1352, 1.2457 ±0.1524 according to the different concentrations of tranilast, and 0. 8956 ±0. 1903 of the control group. In comparison with the control group, 25 μg/ml (q′= 3. 23, P＜0.05), 50 μg/ml (q=4.70, P＜0.01) tranilast significantly antagonized the decrease of the A values induced by TGF-β2 in the cultured human trabecular meshwork cells. In comparison with the control group [817.37±124.21 cpm/104 cells], 12.5 μg/ml (620.33±80.46 cpm/104 cells, q′=4.26, P＜0.05),25 μg/ml (594. 58±88.13 cpm/104 cells, q′=4. 81, P＜0.01), 50 μg/ml (418. 64±67.90 cpm/104 cells, q′=8.62, P＜0.01) tranilast significantly inhibited the incorporation of 3 H-proline into the cultured human trabecular meshwork cells promoted by TGF-β2 in a dose-dependent manner. It was concluded that tranilast had the antagonistic effect on the proliferation inhibition and collagen synthesis promotion induced by TGF-β2 in the cultured human trabecular meshwork cells.     

鞘内注射吗啡和舒芬太尼超前镇痛的效果比较

目的 观察鞘内注射吗啡、舒芬太尼超前镇痛对术后疼痛的影响及不良反应情况,同时观察阿片类药物鞘内注射超前镇痛与传统硬膜外给药在术后镇痛方面的区别.方法 选择60例全子宫切除术患者,随机分成3组,每组20例,均实施腰-硬联合麻醉.采用双盲法鞘内注药:吗啡组(M组):罗哌卡因20 mg+吗啡0.3 mg;舒芬太尼组(S组):罗哌卡因20 mg+舒芬太尼8 μg;对照组(R组):罗哌卡因20 mg,3组均加10%葡萄糖至3 ml.患者自控硬膜外镇痛泵(PCEA)配方:各组均为0.16%罗哌卡因+0.0025%氟哌利多+0.9%氯化钠溶液至100 ml,其中R组在PCEA中添加吗啡6 mg(浓度为0.006%).镇痛泵设置:背景剂量2 ml/h,自控镇痛1 ml,锁定时间15 min,每小时限量6 ml,镇痛时间24 h.随访记录术后2、4、6、8、24、36、48 h时患者的镇痛情况,采用视觉模拟评分(VAS)和普瑞斯-亨利(P-H)评分进行评价;比较3组患者PCEA总按压次数和有效按压次数,并记录围术期不良反应.结果 各组VAS及P-H评分呈现一致性,M组于各时点VAS分别为(10±7)、(13±8)、(11±8)、(16±8)、(10±8)、(6±6)、(3±4)分,均明显低于R组各时点VAS:(25±13)、(32±10)、(40±9)、(39±12)、(28±10)、(20±9)、(13±11)分(P＜0.05);S组各时点评分为(2±2)、(9±11)、(19±15)、(25±9)、(19±8)、(12±6)、(8±7)分,均明显低于R组(P＜0.05);S组VAS于术后2 h低于M组(P＜0.05),后5个时点VAS M组低于S组(P＜0.05).PCEA泵的使用情况:M组与S组总按压次数和有效按压次数在各时间点与R组相比,差异均有统计学意义(P＜0.05).围术期不良反应:M组、S组及R组发生恶心者分别为17、2、17例;发生呕吐者分别为11、1、8例;出现瘙痒者分别为6、12、5例.3组不良反应发生率间差异有统计学意义(P＜0.05).结论 鞘内注入吗啡0.3 mg或舒芬太尼8 μg用于超前镇痛的效果明显优于传统镇痛方式.其中鞘内注射吗啡的镇痛作用强而持久但不良反应多,可用于中长手术的术后镇痛;鞘内注射舒芬太尼的镇痛时效短、作用强,且副作用少,可应用于剖宫产术等时间较短手术的术后镇痛.     

不同剂量缬沙坦联合氨氯地平治疗老年高血压的临床效果评价简

目的 研究不同剂量缬沙坦联合氨氯地平治疗老年高血压的临床效果.方法 将2011年1月～2012年12月临安市人民医院收治的160例老年高血压患者纳入研究,随机分为给予不同剂量缬沙坦联合氨氯地平治疗的观察组和对照组,分别于治疗前和治疗后3个月时检测血压情况、肾功能情况,并分析血压与肾功能的相关性.结果 ①治疗后,观察组与对照组患者的24 h平均收缩压（24 h SBP）[（121.8±15.9）mmHg比（122.6±14.7）mmHg,t=1.038,P＞0.05,1 mm Hg=0.133 kPa]、24 h平均舒张压（24 h DBP）[（72.8±9.8）mm Hg比（73.1±13.1）mm Hg,t=0.938、P＞0.05]差异无统计学意义,24 h平均收缩压变异度（24 h SBPV）[（8.4±1.4）mm Hg比（13.8±1.9）mm Hg,t=6.832,P＜0.05]、24 h平均舒张压变异度（24 h DBPV）[（6.9±1.9）mm Hg比（10.8±2.4）mmHg,t=6.384,P＜0.05]均低于对照组;②治疗后,观察组患者的血肌酐[（82.8±11.3）μmol/L比（121.8±15.3）μmol/L,t=5.893,P＜0.05]、血尿素氮[（4.3±0.9）mmol/L比（6.5±0.8）mmol/L,t=5.374,P＜0.05]、24 h尿蛋白[（0.23±0.03）g比（0.48±0.06）g,t=6.842,P＜ 0.05]水平均低于对照组,内生肌酐清除率[（99.5±14.5）mL/min比（89.5±13.8）mL/min,t=4.982,P＜ 0.05]高于对照组;③收缩压和舒张压水平均与血肌酐、血尿素氮、24 h尿蛋白水水平呈正相关关系,与内生肌酐清除率呈负相关关系.结论 加倍剂量缬沙坦联合氨氯地平治疗有助于减小血压昼夜变异度,改善肾功能状况,对于老年高血压的治疗具有积极价值.     

转化生长因子、碱性成纤维细胞生长因子与大鼠膀胱出口梗阻后逼尿肌收缩功能障碍的关系

目的 探讨转化生长因子β1(TGFβ1)、碱性成纤维细胞生长因子(bFGF)与大鼠膀胱出口梗阻(BOO)后逼尿肌收缩功能障碍的关系.方法 建立Wistar大鼠BOO模型(BOO 2周组11只;BOO 6周组10只),假手术组(8只)作为对照,测定离体逼尿肌肌条M受体激动剂刺激下收缩力,RT-PCR方法测定逼尿肌中TGFB1 mRNA、bFGF mRNA的表达,ELISA测定尿液中TGFβ1、bFGF 的水平.结果 在1×10-4、1×10-3 mmo/L氯化氨基甲酰胆碱(Carbachol)浓度下BOO 2周组的最大逼尿肌收缩力[(0.96±0.11)、(1.98±0.21)g]大于对照组[(0.85±0.18)、(1.82 ±0.19)g,均P＜0.05];大鼠在1×10-5、1×10-4、1×10-3和1 × 10-2 mmol/L Carbachol浓度下BOO 6周组的最大逼尿肌收缩力[(0.19 ±0.02)、(0.65 ±0.06)、(1.12 ±0.08)和(1.40 ±0.19)g]均小于BOO 2周组[(0.24±0.03)、(0.96 ±0.11)、(1.98 ±0.21)和(2.16 ±0.21)g,均P＜0.05]和对照组[(0.23 ±0.04)、(0.85 ±0.18)、(1.82 ±0.19)和(2.12 ±0.26)g,均P＜0.05].TGFβ1 mRNA在对照组、BOO 2周组、BOO 6周组逼尿肌的表达分别为0.32 ±0.01、0.34 ±0.10和0.72 ±0.21,后两组之间差异有统计学意义(P＜0.01);bFGF mRNA表达分别为0.10±0.05、0.21±0.07和0.38 ± 0.13,组间两两比较,差异均有统计学意义(均P＜0.05).离体逼尿肌收缩力与其TGFβ1 mRNA、bFGF mRNA表达水平呈负相关(均P＜0.05).BOO 6周组尿中TGFβ1表达[(606±216)μg/mol肌酐]明显高于BOO 2周组[(131 ±49)μg/mol肌酐]和对照组[(107 ±22)μg/mol肌酐,均P＜0.05].结论 逼尿肌bFGF mRNA表达随BOO进展持续升高,TGFβ1 mRNA在失代偿阶段表达才明显升高;尿液TGFβ1在大鼠BOO 6周时呈强表达,有可能成为预测BOO后逼尿肌收缩功能的指标.     

脂多糖致大鼠急性肺损伤早期Ⅰ型前胶原羧基端肽的表达

目的：观察脂多糖（LPS）致大鼠急性肺损伤（ALI）早期是否存在肺纤维增生,探讨其在ALI发病机制中的作用。方法：将27只SD大鼠随机分成3组,其中对照组为气管内滴注NS（生理盐水）12h后取材,LPS1组、LPS2组为气管内滴注LPS 1.0ml/kg（浓度为100μg/ml）后分别于12h、24h取材。用酶联免疫吸附试验（ELISA）方法测定血清及肺泡灌洗液（BALF）中Ⅰ型前胶原羧基端肽（PⅠCP）的浓度。结果：LPS2组大鼠血清PⅠCP浓度为（64.75±11.97）μg/L显著高于对照组（12.38±3.59）μg/L和LPS1组（13.96±4.30）μg/L（P〈0.05）,而LPS1组和对照组间差异无显著性（P〉0.05）;LPS2组大鼠BALF中PⅠCP浓度为（35.26±3.32）μg/L显著高于对照组（6.90±1.99）μg/L和LPS1组（6.92±2.50）μg/L（P〈0.05）,而对照组和LPS1组间差异无显著性（P〉0.05）。结论：LPS致大鼠ALI后24h血清及BALF中PⅠCP浓度显著增高提示ALI早期肺纤维增生存在。     

硫化氢对急性肺损伤大鼠肺组织核转录因子κB及细胞间黏附分子1表达的影响

目的 探讨硫化氢(H2S)对油酸诱导的急性肺损伤(ALI)大鼠肺组织核转录因子κB(NF-κB)与细胞间黏附分子1(ICAM-1)表达的影响.方法 雄性SD大鼠42只,采用随机数方法分为对照组(6只)、油酸组及油酸+硫氢化钠(NaHS)组,后2组大鼠分别在2、4、6h3个时间点进行观察,每个时间点大鼠为6只.复制大鼠ALI模型,对照组大鼠经过尾静脉注射0.1 ml/kg生理盐水;油酸组大鼠通过尾静脉注射油酸0.1 ml/kg;油酸+NaHS组大鼠先腹腔注射NaHS 56 μmol/kg(溶于0.5 ml生理盐水),30 min后经大鼠尾静脉注射油酸0.1 ml/kg.对支气管肺泡灌洗液沉渣行瑞士染色进行白细胞分类计数;对大鼠肺组织病变进行半定量肺损伤评分;测定肺组织中H2S的含量;应用免疫组织化学法对肺泡上皮细胞中ICAM-1表达进行定位及半定量分析,并对NF-κB核转位进行半定量分析.结果 油酸组大鼠2、4、6h支气管肺泡灌洗液多形核白细胞(PMN)比例[(74.5±3.0)％、(80.2±2.0)％和(87.2±2.7)％]及肺损伤评分(5.2±0.8、6.4±0.6和6.8±0.8)均明显高于对照组[(3.1±1.6)％和0.4±0.6,均P＜0.01];肺组织中H2S含量均明显低于对照组[ (21.20 ±0.38) μmol/g、( 20.80±0.53)μmol/g、(18.92±0.75)μmol/g比(26.81±0.65)μmol/g,均P＜0.01];肺泡上皮细胞中NF-κB核表达和ICAM-1胞膜表达则均明显高于对照组(均P＜0.05).油酸+NaHS组支气管肺泡灌洗液PMN比例及肺损伤评分在2、4和6h3个时间点均明显低于油酸组(均P＜0.05);肺组织中H2S含量在4和6h明显高于油酸组(均P＜0.05);肺泡上皮细胞中NF-κB核表达和ICAM-1胞膜表达在4和6h则明显低于油酸组相应时间点(均P＜0.05).结论 H2S可能通过抑制ALI大鼠肺部炎症反应发挥保护效应,其抗炎效应与其抑制大鼠肺泡上皮细胞NF-κB的表达有关.     

代谢综合征患者葡萄糖代谢率与血浆脂联素及瘦素水平变化的关系

目的 探讨代谢综合征(MS)患者葡萄糖代谢率与血浆脂联素、瘦素水平的关系.方法 筛选30例MS患者和20名健康者为对照组.所有受试者均行高胰岛素-正常血糖钳夹试验;检测血浆脂联素与瘦素浓度及空腹胰岛素(FTNS)、血糖、血脂等生化指标;测定血压、腰围、身高、体重等人体参数.结果 (1)高胰岛素-正常血糖钳夹试验稳态时(120～150 min),MS组的葡萄糖代谢率显著低于对照组[(8.33±1.59)与(4.13±1.34)mg·kg-1·min-1],差异有统计学意义(P＜0.01).(2)MS组的血浆脂联素水平显著低于对照组[(5.15±2.54)mg/L比(10.28±5.50)mg/L,P＜0.01],瘦素水平显著高于对照组[(189±90)μg/L比(127±73)μg/L,P＜0.01].(3)在MS组,腰围、体质指数、甘油三酯、高密度脂蛋白胆固醇、FINS、脂联素、瘦素等与葡萄糖代谢率相关(均P＜0.05).结论 提示MS患者的胰岛素抵抗可能与血浆脂联素降低及瘦素升高有关.

Abstract：

Objective To investigate the relationship of glucose metabolic rate (GMR) and plasma levels of adiponectin and leptin in patients with metabolic syndrome (MS). Methods A total of 30 MS subjects aged 36 -60 years old were selected as MS group. And 20 normal adults were selected as control group. The GMR was evaluated by the technique of hyperinsulinemic euglycemia clamp. The plasma concentrations of adiponectin and leptin were detected by enzyme-linked immunosorbent assay (ELISA). Blood pressure, waist circumference (WC), body weight and body height were measured.Results ( 1 ) During the steady state ( last 30 min), the GMR was significantly lower in MS group than that in control Group [ (4. 13 ± 1.34) mg· kg- 1 · min -1 vs ( 8.33 ± 1.59 ) mg · kg- 1 · min -1, P ＜ 0. 01 ].(2) The plasma level of adiponectin was significantly lower in MS group than that in control group [ (5. 15 ±2. 54) μg/ml vs ( 10. 28 ±5.50 ) μg/ml, P ＜0. 01 ]. The plasma level of leptin were significantly higher in MS group than that in control group [(189.37 ±90.48) nng/ml vs(126.55 ±72.70) ng/ml, P ＜0.01].(3) In MS group, glucose metabolic rate was associated with WC, BMI, TG, HDL-C FINS, leptin, and adiponectin, ( all P ＜ 0. 05 ). Conclusion The technique of hyperinsulinemic euglycemic clamp shows that the BMR of MS patients significantly decreases. It may be associated with their lowered plasma levels of adiponectin and leptin.     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

类风湿关节炎患者TH17细胞及其相关细胞因子的表达及临床意义

目的 探讨类风湿关节炎(RA)患者TH17细胞及其相关细胞因子的表达及临床意义.方法 收集大连医科大学附属第一医院门诊RA初治活动期患者的外周血标本32例为活动期RA组,健康献血者30例为健康对照组,所有受试者清晨空腹坐位肘静脉采血.应用酶联免疫吸附法(ELISA)及反转录PCR(RT-PCR)法,分别检测外周血血清中IL-17水平和外周血单个核细胞(PBMC)中IL-17mRNA表达水平;将PBMC一组不加刺激,另一组用佛波酯(50 ng/mL)和离子霉素(1μg/mL)刺激4h后,流式细胞仪检测TH17细胞数量.结果 活动期RA患者血清IL-17水平(15.76 ±0.62) pg/mL与健康对照组(8.67±0.12) pg/mL比较,差异有显著性意义(P＜0.05).活动期RA患者PBMC的IL-17mRNA表达水平(1.94±0.46)与健康对照组(0.84±0.16)比较,差异有显著性意义(P＜0.05).RA组血清IL-17水平与mRNA表达水平呈正相关(r=0.70,P＜0.05),且与疾病活动程度(DAS28)评分呈正相关(r=0.99,P＜0.05).活动期RA患者外周血TH17细胞比例(2.50±0.54)％与健康对照组(0.03±0.00)％比较,差异有显著性意义(P＜0.05).但刺激组(3.23±0.05)％与无刺激组(2.50±0.54)％比较,差异无显著性意义(P＞0.05).RA患者组TH17细胞数量与疾病活动程度(DAS28)评分呈正相关(r=0.96,P＜0.05).结论 活动期RA患者TH17细胞及其主要效应性细胞因子IL-17在细胞、蛋白和基因表达水平上均明显升高并与疾病活动程度呈正相关,对临床制定治疗方案、判断疗效有指导意义.     

三级战备状态对官兵心理和部分生化指标的影响及干预效果

目的 探讨三级战备状态对官兵心理、部分生化指标的影响及心理行为干预的效果.方法 以进入三级战备状态的某摩步营189名官兵为研究对象,以班为单位随机分为研究组和对照组.战备期间2组进行统一的军事训练和思想政治教育,研究组增加心理行为干预.在进入三级战备状态当天(战备前)和结束次日(战备后),2组分别进行抑郁自评量表(SDS)、焦虑自评量表(SAS)、简易应对方式问卷(SCSQ)评定,超氧化物歧化酶(SOD)、丙二醛(MDA)、皮质醇(COR)和醛固酮(ALD)含量测定.结果 研究组战备后SDS总分(42.1±9.3)分、SAS总分(43.8±7.2)分、MDA(2.6±0.51)μmol/L、COR(252.5±52.4)ng/ml、ALD(97.5±24.4)pg/ml低于对照组,积极应对(21.2±6.4)分和SOD(1551±354)U/gHb高于对照组(均P＜0.01或0.05).对照组战备后SDS总分(49.2±10.3)分、SAS总分(50.6±10.2)分、MDA(2.9±0.35)μmol/L、COR(333.8±62.6)ng/ml、ALD(123.8±29.6)pg/ml高于战备前,积极应对(18.2±5.4)分和SOD(1302±352)U/gHb低于战备前(均P＜0.01).SDS、SAS总分与SOD显著负相关(r=-0.142、-0.119,P＜0.01或0.05),与COR、ALD显著正相关(r=0.156,0.151,0.159,0.156,P＜0.01或0.05);SCSQ的积极应对分与SOD显著正相关(r=0.141,P＜0.01),与COR和ALD显著负相关(r=-0.152,-0.155,P＜0.01).结论 三级战备状态下官兵产生了明显的心身应激反应,心理行为干预可有效改善应激反应水平.     

氧自由基在创伤性大鼠脑损伤后肠道细菌移位中的作用

目的 探讨氧自由基在创伤性脑损伤大鼠肠黏膜屏障应激性变化机制中的作用.方法 雄性Wistar大鼠64只,随机分为2组:创伤性脑损伤(TBI)组(32只)和对照组(32只),再分别按术后6、12、24、48 h时相点分为4个亚组(n=8).测定肠黏膜组织丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽(GSH)水平,检测肠系膜淋巴结等多器官的组织匀浆中标记大肠杆菌的检出率.结果 TBI组多脏器荧光标记大肠杆菌检出率明显高于对照组(6、12、24、48 h组分别为:8.3%比2.1%、25.0%比2.1%.27.1%比2.1%、12.5%比0.0%,均P＜0.05);同时,该组各时相点肠黏膜组织匀浆中MDA含量明显高于对照组(nmol/mg pro,6、12、24、48 h组分别为:4.9±0.7比2.6±0.3、6.1±0.8比2.8±0.5、5.9±0.4 比 2.6±0.3、5.3±0.5比2.7±0.4,均P＜0.05);而GSH(mg/gpro,6、12、24、48 h组分别为:287±36比408±53、192±25比421±46、160±48比432±35、241±31比394±51,均P＜0.05)、SOD(U/mg pro,6、12、24、48 h组分别为:19.5±3.3比21.1±1.9、11.8±2.6比20.7±5.3、13.3±3.3比20.0±3.3、15.1±1.5比21.2±3.2,均P＜0.05)则明显低于对照组.结论 创伤性脑损伤模型大鼠伤后早期肠黏膜通透性升高,导致细菌移位增加,氧自由基在这一病生理过程中起重要作用.     

老年重症患者术中应用自由基清除剂依达拉奉的多中心疗效分析

目的 观察老年重症患者手术中应用依达拉奉对预后的影响.方法 2008年7月1日至9月30日在北京朝阳医院、北京同仁医院、北京安贞医院和卫生部北京医院4家医院的400例老年重症手术患者按随机数字表法分为试验组与对照组,各200例.麻醉开始前试验组患者静脉泵入依达拉奉(60mg/40 ml)直至手术结束,对照组以等量生理盐水代替.分别于桡动脉穿刺后、手术开始后1 h及缝皮前检测超氧化物歧化酶(SOD)与丙二醛水平,记录术中情况及术后病死率、总住院日、重症监护病房(ICU)停留时间、术后机械通气时间与术后并发症情况.其中行不停跳冠状动脉旁路移植术患者于术前和术后24 h检查肌钙蛋白Ⅰ(cTn Ⅰ)与左室射血分数(LVEF).结果 试验组手术开始后1 h与缝皮前SOD水平均高于对照组[(87±14)U/ml比(78±14)U/ml,(83±13)U/ml比(77±14)U/ml,P＜0.01、＜0.05];丙二醛则均低于对照组[(11±5)nmol/L比(14±7)nmol/L,(11±5)nmol/L比(14±6)nmol/L,P＜0.05、＜0.01].术中低血压需持续应用血管活性药物支持者对照组多于试验组(37例比19例,P＜0.01),试验组总住院日、ICU住院日都短于对照组[(21±9)d比(23±9)d,(10±7)d比(13±9)d,均P＜0.05],行不停跳冠状动脉旁路移植术患者试验组术后cTn Ⅰ和LVEF与术前和对照组相比差异均有统计学意义(均P＜0.05).结论 老年重症患者术中应用依达拉奉可防止丙二醛升高、SOD下降,降低术中低血压发生率,减少老年重症患者总住院日与ICU住院日.特别是行不停跳冠状动脉旁路移植术患者术中应用依达拉奉,术后cTn Ⅰ与LVEF测量值试验组与术前和对照组相比都有明显改善.

Abstract：

Objective To observe the effects of intraoperative application of radical scavenger edaravone in severe elderly cases. Methods A total of 400 severe elderly patients scheduled for surgery were randomly assigned to receive edaravone 60 mg/40 ml ( Group Y) or an equal volume of normal saline (Group C). The arterial blood samples were harvested at immediately after pricking, 1 hour after the beginning of surgery and before saturation to determine the levels of superoxide dismutase (SOD) and malondialdehyde (MDA). The operative duration, fluid volume, blood loss, blood transfusion volume,urine output, intraoperative adverse events, mortality rate, total hospital stay, intensive care unit (ICU)stay, postoperative mechanical ventilation time and complications were recorded. Patients undergoing offpump coronary artery bypass graft (OPCABG) were evaluated for troponin Ⅰ (cTn Ⅰ ) and left ventricular ejection fraction (LVEF) before and after 24 hours of surgery. Results SOD was higher and MDA lower in Group Y than those in Group C at 1 hour intraoperation and before saturation[SOD: (87 ± 14)U/ml vs(78 ±14)U/ml, (83±13)U/mlvs(77±14)U/ml, P＜0.01, ＜0.05; MDA : (11 ±5)nmol/Lvs(14±7)nmol/L,( 11 ± 5 ) nmol/L vs ( 14 ± 6) nmol/L, P ＜ 0. 05, ＜ 0. 01]. There were more intraoperative hypotension cases requiring a continuous application of vasoactive drugs in Group C(37 cases vs 19 cases),total hospital stay[(21 ±9 )d vs (23±g)d, P＜0.05] and ICU stay[(10±7)dvs (13±9)d, P＜0. 05] were also longer. Postoperative cTn Ⅰ and LVEF of Group Y significantly improved in OPCABG cases ( all P ＜ 0. 05 ). Conclusion The intraoperative application of edaravone in severe elderly patients may prevent MDA increase and SOD decrease and reduce free radical damage. Especially in OPCABG patients,cTn Ⅰ and LVEF improve significantly.     

抑郁情绪障碍对围术期老年冠心病患者外周血单核细胞CD40表达及血浆白细胞介素-8的影响

目的 观察合并抑郁情绪障碍的老年冠心病(CHD)患者围术期外周血单核细胞表面CD40表达及血浆白细胞介素8(IL-8)浓度的变化,探讨抑郁情绪障碍和冠心病间的相关免疫学机制。方法 选择2009年12月至2010年12月河北医科大学第三医院高龄冠心病患者100例,冠状动脉粥样硬化性心脏病诊断明确,无认知障碍。所有患者均行单节段腰后路全椎板减压植骨融合内固定术。术前1d采用汉密尔顿抑郁和焦虑量表评定患者的抑郁和焦虑评分,并据此将患者分为3组：无抑郁组（A组,n =30）：抑郁评分≤20分,且焦虑评分≤6分;抑郁干预组（B组,n=35）：为抑郁评分≥30分,且焦虑评分≤6分,白术前1d至术后第7天进行心理辅助治疗;抑郁非干预组（C组,n=35）：抑郁评分≥30分,且焦虑评分≤6分。术前1d及术后第7天抽取外周静脉血测定单核细胞表面CD40表达及血浆IL-8浓度。并于术后第7天再次测定抑郁评分。结果 术后B组的抑郁评分(25.1±2.9)低于术前(33.2±1.4)和C组(34.2±0.8,P＜0.05);术前A组的CD40荧光强度(123±18)低于B、C组（197±23,204±26）,(P＜0.05),术后A组(132 ±20)低于B组(147±19),但B组低于C组(212±18,P＜0.05),且CD40荧光强度和抑郁总分呈显著性正相关(r=0.597,P＜0.01);术前A组的IL-8浓度(85 ng/L±16 ng/L)低于B、C组(151 ng/L±18 ng/L,164 ng/L±22 ng/L),(P＜0.01),B和C组差异无统计学意义,但术后B组( 158 ng/L±19 ng/L)低于C组(197 ng/L±24 ng/L,P＜0.05),且血浆IL-8浓度与抑郁总分呈显著性止相关(r=0.751,P＜0.01)。结论 抑郁情绪障碍可能会造成围术期老年冠心病患者血浆IL-8分泌增多及单核细胞CD40表达上调。     

山莨菪碱预处理对兔肺缺血再灌注后肺水肿的保护作用

目的探讨山莨菪碱预处理对兔肺缺血再灌注后肺水肿的保护作用及作用机制。方法建立在体兔肺缺血再灌注动物模型,10~12周龄健康家兔24只,雌雄不拘,体重2 200~2 600 g,随机分为3组,每组8只。山莨菪碱组于左肺缺血再灌注处理前静脉给予山莨菪碱3 mg/kg预处理,对照组进行左肺缺血再灌注处理,假手术组只开胸并套带不行缺血再灌注处理。各组左肺缺血1 h,再灌注3 h后结扎并切取左肺下叶,部分行组织学观察,部分通过测量肺湿干重的方法计算肺含水量,余肺组织按试剂盒说明书检测组织髓过氧化酶(MPO)含量。最后经兔股静脉注射2%伊文思蓝(1.5 mL/kg),然后取左肺上叶,参照Udaka的方法比较3组肺血管通透性。结果缺血再灌注处理后,对照组肺含水量、MPO含量及伊文思蓝含量分别为(6.59±0.53)g/g干重、(1.16±0.14)U/g.prot和(172.8±15.5)μg/g湿重,明显高于假手术组的(4.34±0.37)g/g干重、(0.53±0.09)U/g.prot和(103.3±11.2)μg/g湿重(P均<0.05),组织损伤严重并可见严重的肺水肿,山莨菪碱组上述指标水平分别为(5.62±0....     

RhoA-ROCK通路在血管紧张素Ⅱ刺激人胚肺成纤维细胞表达结缔组织生长因子中的作用

目的 探讨RhoA-ROCK通路在血管紧张素Ⅱ(AngⅡ)刺激人胚肺成纤维细胞(HFL-1)表达结缔组织生长因子(CTGF)中的作用.方法 培养HFL-1,设置无刺激对照组、AngⅡ组(10-7 mol/L的AngⅡ刺激)、Irbesartan+AngⅡ组(以10-6 mol/L的AT-1受体拮抗剂Irbesartan预处理1 h后再予10-7 mol/L的AngⅡ刺激)和ROCK抑制剂Y27632+AngⅡ组(10-6 mol/L的Y27632预处理1 h后再予10-7 mol/L的AngⅡ刺激).利用Western印迹和QuantiGene多基因定量方法检测RhoA-ROCK激活及下游因子CTGF表达情况.结果 观察Irbesartan对AngⅡ诱导CTGF蛋白表达的实验,结果:AngⅡ组CTGF蛋白表达较对照组增强(0.89±0.05比0.48±0.10,P＜0.01),Irbesartan+AngⅡ组(0.72±0.05)较AngⅡ组减弱(P＜0.05).AngⅡ组RhoA蛋白表达较对照组增强(3.40±0.46比1.77±0.37,P＜0.01),Irbesartan+AngⅡ组(2.27±0.45)较AngⅡ组减弱(P＜0.05).重新培养细胞留取标本观察Y27632对AngⅡ诱导CTGF蛋白表达的影响,结果:AngⅡ组CTGF蛋白表达较对照组增强(0.62±0.15比0.16±0.05,P＜0.01),Y27632+AngⅡ组(0.17±0.04)较AngⅡ组减弱(P＜0.01).从基因水平重复上述实验,结果:AngⅡ组CTGF mRNA表达较对照组增强(1.16±0.06比1.00±0.01,P＜0.01),Irbesartan+AngⅡ组(0.99±0.07)较AngⅡ组减弱(P＜0.01),Y27632+AngⅡ组(1.04±0.08)较AngⅡ组减弱(P＜0.05);AngⅡ组RhoA mRNA表达较对照组增强(1.21±0.07比1.00±0.06,P＜0.01),Irbesartan+AngⅡ组(1.00±0.12)较AngⅡ组减弱(P＜0.05),Y27632+AngⅡ组(1.10±0.05)与AngⅡ组比较差异无统计学意义(P＞0.05).结论 AngⅡ通过AT-1受体诱导HFL-1细胞CTGF蛋白和mRNA表达,RhoA-Rock通路参与其中.

Abstract：

Objective To explore the production of connective tissue growth factor(CTGF)by Angiotensin Ⅱ(Ang Ⅱ)in human embryonic lung fibroblast via the RhoA-ROCK pathway. Methods Human embryonic lung fibroblast(HFL-1)was divided into 4 groups:(1)control group:no stimulation;(2)AngⅡgroup:stimulation of AngⅡ(10-7 mol/L);(3)Irbesartan plus Ang Ⅱ group:stimulation by Ang Ⅱ(10-7 mol/L)with AT-1 receptor antagonist irbesartan(10-6 mol/L)pre-treatment;(4)Irbesartan plus Ang Ⅱ group:stimulation by Ang Ⅱ(10-7 mol/L)with ROCK inhibitor Y27632(10-6 mol/L)pretreatment.Then the products of protein and RNA were collected.Western blot and QuantiGene were used to detect the activation of RhoA-Rock pathway and CTGF.Results Exploring the affect of irbesartan on Ang Ⅱ through the Western blot analysis of CTGF and RhoA protein expression:the CTGF level was up-regulated by AngⅡ(0.89±0.05 vs control 0.48 ±0.10,P ＜0.01).Such an effect was markedly blocked by a pretreatment of irbesartan(0.72 ± 0.05,P＜0.05).After the use of Ang Ⅱ,the expression of RhoA protein was significantly enhanced(3.40 ± 0.46 vs control 1.77 ± 0.37,P＜0.01)and blunted by a pretreatment of irbesartan(2.27 ± 0.45,P＜0.05).The Western blot analysis of CTGF protein expression showed that Ang Ⅱ caused a robust increase in CTGF(0.62 ± 0.15 vs control 0.16 ± 0.05,P＜0.01).Such an effect was markedly blocked by a pretreatment of Y27632(0.17 ± 0.04,P＜0.01).The result was similar at the gene level.Ang Ⅱ significantly increased the expression of CTGF mRNA(1.16 ± 0.06 vs control 1.00 ± 0.01,P＜0.01).And it was markedly blocked by a pretreatment of irbesartan(0.99 ±0.07,P＜0.01)or Y27632(1.04 ±0.08,P＜0.05).AngⅡ significantly increased the expression of RhoA mRNA(1.21 ± 0.07 vs control 1.00 ± 0.06,P＜0.01).And it was markedly blocked by a pretreatment of irbesartan(1.00 ± 0.12,P＜0.05)but not Y27632(1.10 ± 0.05,P＞0.05).Conclusion Ang Ⅱ activates HFL-1 to produce CTGF through the AT-1 receptor.And the RhoA-Rock pathway is involved.