严重烫伤后高迁移率族蛋白-1基因表达的改变及意义

目的　观察烫伤后肝、肺组织高迁移率族蛋白-1（HMG-1）mRNA表达的变化规律及其与器官功能损害的关系。　方法　采用大鼠35%Ⅲ度烫伤模型,动物随机分为正常对照组、烫伤组和重组杀菌/通透性增加蛋白(rBPI21)治疗组,留取组织和血标本分别检测组织内毒素含量、HMG-1mRNA表达及器官功能指标。　结果　严重烫伤后早期肝、肺组织HMG-1基因表达改变不明显（正常对照组肝组织0.202±0.097;肺组织0.263±0.091）,伤后24h则明显增多(肝组织0.487±0.189;肺组织0.513±0.069,P＜0.05,0.01),且一直持续至伤后72h(肝组织0.687±0.142;肺组织0.520±0.076,P＜0.01)。给予rBPI21治疗可有效抑制肝和肺中内毒素水平的升高,并显著抑制肝、肺组织HMG-1mRNA水平(P＜0.01)。相关分析结果显示,肝组织HMG-1mRNA表达与血清谷丙转氨酶、谷草转氨酶水平、肺组织HMG-1mRNA表达与髓过氧化物酶活性呈显著正相关（P＜0.05,0.01）。　结论　严重烫伤后肝、肺组织HMG-1表达显著增多,且持续时间较长,局部组织HMG-1诱生与烧伤后内毒素介导的重要器官功能损害关系密切。     

严重烫伤后高迁移率族蛋白-1基因表达的改变及意义

目的:观察烫伤后肝、肺组织高迁移率族蛋白-1(HMG-1)mRNA表达的变化规律及其与器官功能损害的关系.方法:采用大鼠35%Ⅲ°烫伤模型,动物随机分为正常对照组(n=7)、烫伤组(n=24)和重组杀菌/通透性增加蛋白(rBPI21)治疗组(n=12),留取组织和血标本分别检测组织内毒素含量、HMG-1 mRNA表达及器官功能指标.结果:严重烫伤后早期肝、肺组织HMG-1基因表达改变不明显,伤后24小时则明显增多(P＜0.05-0.01),且一直持续至伤后72小时(P＜0.01).给予rBPI21治疗可有效防治肠源性内毒素移位的发生,并显著抑制肝、肺组织HMG-1 mRNA水平.相关分析结果显示,肝组织HMG-1 mRNA表达与血清谷丙转氨酶、谷草转氨酶水平、肺组织HMG-1 mRNA表达与髓过氧化物酶活性呈显著正相关(P＜0.05-0.01).结论:严重烫伤后肝、肺组织HMG-1表达显著增多,且持续时间较长,局部组织HMG-1诱生与烧伤后内毒素介导器官功能损害关系密切.     

高迁移率族蛋白2在结直肠癌组织中的表达及临床意义

目的:探讨结直肠癌癌组织中高迁移率族蛋白2(HMGB2)的表达及其临床意义。方法:收集2012年1月—2014年1月茂名市人民医院胃肠外科收治的82例结直肠癌患者术后组织标本及临床资料,用免疫组化方法检测标本中HMGB2表达,分析HMGB2表达与结直肠癌患者临床病理因素的关系及无瘤生存率和总生存率的关系,并分析影响结直肠癌患者无瘤生存率和总生存率的危险因素。结果:HMGB2高表达者比例为60.9%(50/82),HMGB2低表达者比例为39.1%(32/82)。HMGB2高表达与结直肠癌患者的组织分化程度低、临床分期、T分期、N分期和M分期有关(均P0.05)。HMGB2高表达患者5年无瘤生存率与总生存率均明显低于HMGB2低表达患者(27.7%vs. 38.7%,P0.05;32.1%vs. 41.5%,P0.05)。多因素分析及结果显示,HMGB2与M分期均为结直肠癌患者无瘤生存率(HR=1.771,95%CI=1.146～4.923,P=0.027;HR=3.874,95%CI=1.886～7.824,P=0.002)与总生存率(HR=1.823,95%CI=1.035～3.198,P=0.005;HR=3.865,95%CI=1.770～8.278,P=0.001)的独立危险因素。结论:HMGB2高表达与结直肠癌患者恶性临床病理特征密切相关,且是影响结直肠癌患者预后的独立危险因素。     

高迁移率族蛋白B1联合肠型脂肪酸结合蛋白对新生儿坏死性小肠结肠炎的诊断价值

目的探究联合检测高迁移率族蛋白B1（HMGB1）、肠型脂肪酸结合蛋白（I-FABP）诊断新生儿坏死性小肠结肠炎（NEC）的价值。 方法选择2016年7月至2018年7月西北妇女儿童医院收治的NEC新生儿119例（NEC组）以及同期非NEC患儿30例（对照组）。运用酶联免疫吸附法（ELISA）检测患儿粪便样本中HMGB1蛋白以及血清中I-FABP蛋白的表达水平,ROC曲线分析单项检测与联合检测对NEC的诊断效能。 结果Bell Ⅲ期NEC患儿的HMGB1、I-FABP蛋白表达水平均显著高于Ⅰ、Ⅱ期患儿（P<0.05）,随着病情加重,蛋白水平呈逐渐上升趋势。NEC组患儿HMGB1、I-FABP蛋白水平显著高于对照组,差异有统计学意义（P<0.05）。联合应用两项指标诊断NEC的敏感度为89.60%,特异度为86.50%,ROC曲线下面积为0.985（P<0.01）,诊断效能明显高于单项检测（P<0.05）。 结论HMGB1、I-FABP联合检测诊断NEC患儿敏感度及特异度高,动态测定HMGB1、I-FABP指标水平,有助于疑似NEC新生儿的早期筛查、治疗以及病程进展的判断。     

高迁移率族蛋白B1诱导小鼠腹腔巨噬细胞凋亡的受体机制

目的 了解高迁移率族蛋白B1(HMGB1)对小鼠腹腔巨噬细胞凋亡的影响及其受体机制.方法 分离培养小鼠腹腔巨噬细胞,在巨噬细胞中加入不同的刺激物,分为HMGB1组:加入10 μg/ml的HMGB1;HMGB1+抗晚期糖基化终末产物受体(RAGE)组:先加入RAGE多克隆抗体5μg/ml孵育2 h后,再加入HMGB1;HMGB1+重组鼠(rm)RAGE/Fc组:将10 μg/ml的HMGB1与10μg/ml的rmRAGE/Fc混合作用2 h后,再加入巨噬细胞;对照组:加入磷酸盐缓冲液.采用流式细胞仪检测细胞表面RAGE的表达强度.激光共聚焦显微镜观察细胞凋亡情况,流式细胞仪检测细胞凋亡率.结果 HMGB1组RAGE阳性细胞率(54±12)%明显高于对照组[(13±5)%,P＜0.01],其荧光强度(126±10)也显著高于对照组(34±8,P＜0.01).HMGB1+rmRAGE/Fc组、HMGB1+抗RAGE组凋亡细胞明显多于对照组,而HMGB1组晚期凋亡及坏死细胞明显多于其他3组.HMGB1组细胞凋亡率(39.5±2.3)%高于HMGB1+rmRAGE/Fc组[(17.3±3.6)%]、HMGB1+抗RAGE组[(14.8±4.8)%]及对照组[(5.4±2.3)%,P＜0.01].结论 HMGB1可诱导RAGE表达上调,RAGE是HMGB1诱导巨噬细胞凋亡的主要受体之一.     

高迁移率族蛋白1在内毒素性急性肺损伤大鼠肺血管重构中的作用

目的 评价高迁移率族蛋白1 (HMGB1)在内毒素性急性肺损伤大鼠肺血管重构中的作用.方法 健康清洁级雄性Wistar大鼠30只,体重220 ～ 250 g,采用随机数字表法,将大鼠随机分为3组(n=10)∶对照组(C组)、内毒素性急性肺损伤模型组(M组)和HMGB1抗体组(H组).C组尾静脉输注生理盐水5 ml/1.5 h;M组输注生理盐水3 ml/1.0h,再输注LPS 2 ml(1 mg/kg)/0.5 h;H组于注射LPS后12、24和36 h时尾静脉注射HMGB1抗体2 mg/kg.于注射LPS后72 h时处死取肺,光镜下观察肺组织病理学结果,采用图像分析软件测量并计算肺小动脉中膜/血管面积百分比,免疫组化法检测肺血管增殖细胞核抗原(PCNA)表达,Western bolt法检测HMGB1表达.结果 与C组比较,M组和H组肺小动脉中膜/血管面积百分比、PCNA和HMGB1表达水平升高(P＜0.05),肺组织急性炎症细胞增多,血管壁明显增厚;与M组比较,H组肺小动脉中膜/血管面积百分比、PCNA和HMGB1表达水平降低(P＜0.05),肺组织急性炎症和血管壁增厚明显减轻.结论 HMGB1可能是诱发内毒素性急性肺损伤大鼠肺血管重构的重要因素.     

高迁移率族蛋白HMGA1在肝细胞癌及胆管细胞癌中的表达差异及临床意义

目的：探讨高迁移率族蛋白A1（HMGA1）在肝细胞癌（HCC）、肝内胆管细胞癌（IHC）中的表达差异和临床意义。 方法：采用免疫组化SABC法对60例HCC、42例胆管细胞癌及10例正常肝脏组织标本中HMGA1的表达情况进行检测,并分析其与临床病理特点之间的关系。 结果：HMG组A1在10例正常肝脏组织中无一例表达,在HCC组中表达率为3.3％（2/60）,在IHC组中表达率为66.7％（28/42）。IHC组与HCC组中HMGA1表达差异有统计学意义（P = 0.000）;IHC组HMGA1与p53的表达呈正相关（r = 0.673,P＜0.05）;1年内死亡患者的HMGA1表达水平（80.0％）较1年内存活患者的（33.3％）高,差异有统计学意义（P 〈 0.05）。 结论：HMGA1可作为鉴别肝细胞癌与肝内胆管细胞癌的一个潜在指标。HMGA1在肝内胆管细胞癌的生成中可能发挥了重要作用,HMGA1在肝内胆管细胞癌的高表达提示预后不良。     

高迁移率族蛋白B1及其生物学效应研究进展

高迁移率族蛋白B1 （ high mobility group protein, HMGB1 ）是一种高度保守的核蛋白，广泛分布于哺乳动物细胞。随着其晚期促炎作用的发现，HMGBl成为近年来危重医学研究的热点之一。本文就其家族分类、基因和蛋白结构、细胞生物学效应方究进展进行综述。     

高迁移率族蛋白B1与肝损伤研究进展

1973年,Goodwin等[1]首先从细胞核中发现了高迁移率族蛋白B1(high mobility group box 1,HMGB1).HMGB1作为一种与DNA结合的非组蛋白,长期以来,人们一直关注其在维持核稳定方面的功能,包括稳定核小体结构、调节基因转录、参与DNA复制与修复.     

高迁移率族蛋白1和急性肺损伤

背景 高迁移率族蛋白1(high mobility group box protein 1,HMGB1)是一种非组蛋白染色体结合蛋白,进化上高度保守.HMGB1广泛存在于细胞核内,在核内作为DNA分子伴侣,被释放至细胞外将发挥促炎作用.急性肺损伤(acute lung injury,ALI)和急性呼吸窘迫综合征(acute respiratory distress syndrome,ARDS)是全身性炎症的肺部表现,特征为血管通透性增高致组织水肿、器官功能障碍.越来越多的研究证实细胞外的HMGB1能够通过促进NF-κB核转位和显著增加促炎因子释放引起ALI和致死性的炎症反应.目的 阐述HMGB1和ALI之间的关系以及HMGB1在治疗ALI中的研究现况,为临床治疗ALI提供新的靶点.内容 重点回顾有关HMGB1在ALI发病中的作用、HMGB1抑制剂在改善ALI中作用的研究.趋向 HMGB1抑制剂可以改善ALI结局,在治疗ALI中具有广阔前景.     

Organ changes and bacterial translocation in a rat model of chronic rejection after small bowel transplantation

Rejection after small bowel transplantation (SBTx) may allow bacterial translocation damaging the liver and lungs. This study investigated these issues in a rat model of chronic rejection. MATERIALS AND METHODS: Orthotopic SBTx was performed in syngeneic (SYN) (ACI-ACI, n=8) and allogeneic (ALLO) (ACI-Lewis, n=8) rat strain combinations. Cyclosporine was given to ALLO rats for 28 days. Animals were sacrified between 55 and 65 days. Lymph nodes and venous samples were cultured; Escherichia coli DNA was assessed by polymerase chain reaction. We measured intestine, liver, spleen, and lung protein and DNA contents. Chronic rejection was histologically confirmed. RESULTS: Two of eight and four of eight rats died in the first week after SYN and ALLO SBTx, respectively. There were no differences in organ weights or DNA and protein contents compared with the controls. Gram-negative enteric bacteria were found in two of four ALLO and two of six SYN rats (ns), and aerobic gram-positive were found in two of four and two of six (ns), respectively. Anaerobic growth occurred in mesenteric lymph nodes in only one ALLO rat. E. coli DNA was negative in all animals. Lungs were severely emphysematous in ALLO rats with no histologic changes observed in the other phagocytic organs. Mild rejection was found in the intestine of ALLO rats. CONCLUSIONS: Bowel lesions in ALLO rats might be consistent with chronic rejection and lung lesions could be related to bacterial translocation after SBTx. However, contrary to our expectations, no significant bacterial translocation was demonstrated in either group at the end of the experiments.     

A rat small bowel transplant model of chronic rejection: histopathologic characteristics.

BACKGROUND: The major impediment to long-term success in solid organ transplantation is the development of chronic rejection (CR). The vascular lesion of CR, transplant vascular sclerosis (TVS) is characterized by neointimal smooth muscle cell proliferation, and is driven by both immune- and nonimmune-mediated mechanisms. Although the features of chronic heart and kidney allograft rejection have been well characterized, the more immunogenic small bowel allograft has not received similar study. METHODS: F344 small bowel (SB) was transplanted heterotopically into Lewis recipients that were treated with low-dose Cyclosporine A for 15 days. Lewis recipients of F344 or Lewis SB grafts without immunosuppression, served as controls. Grafts were assessed histologically when recipients showed clinical signs of rejection or at predetermined time points. The immunological components involved in the chronic rejection process were evaluated by immunohistochemical staining. RESULTS: All SB allografts (100%) developed histologic evidence of CR Cyclosporine A. TVS was seen in 36 of the 46 (78%) of these allografts. The median time to develop TVS was 45 days. Immunohistochemical staining of chronically rejected grafts showed infiltration predominantly by CD4+ cells and macrophages, uniform up-regulation of class II MHC molecule expression, moderate to intense ICAM-1 staining in grafts harvested at postoperative day 45, and uniform neointimal cell staining for smooth muscle cell alpha-actin in the TVS lesions. CONCLUSIONS: This F344 to Lewis SB transplant model is a useful model that reproduces significant features of CR. The highly immunogenic nature of the SB allografts allows this model to serve as a stringent test for protocols designed to prevent CR.     

Experimental assessment of small intestinal submucosa as a small bowel graft in a rat model

Background/Purpose:

Small intestinal submucosa (SIS) is an extracellular matrix used in tissue engineering. The purpose of this study is to evaluate the feasibility of using SIS as a scafford for small bowel regeneration in a rat model.

Methods:

A 2-cm length tubular SIS graft from donor Sprague Dawley rats was interposed with bilateral anastomosis in the median tract of an isolated ileal loop of Lewis rats used to construct an ileostomy. The grafts were harvested and analyzed at each of the time-points ranging from 2 weeks to 24 weeks after operation using histology and immunohistochemistry.

Results:

Macroscopic examination found no adhesion in the surrounding area of neointestine by 24 weeks, and no stenosis was visible. The shrinkage of neointestine was indicated from 20% to 40%. Histologic and immunohistochemical evaluation showed that SIS grafts were colonized by numerous inflammation cells by 2 weeks. Neovascularization was evident, but the luminal surface was not epithelized. By 4 weeks, transitional mucosal epithelial layer began to line the luminal surface of the graft, and nearly 70% luminal surface of the graft had been covered by mucosal epithelium at 8 weeks. By 12 weeks, the luminal surface was covered completely by a mucosal layer with distinct bundles of smooth muscle cells in the neointestine. At 24 weeks, the neointestine wall showed 3 layers of mucosa, smooth muscle, and serosa.

Conclusions:

The preliminary study suggested that SIS allow rapid regeneration of mucosa and smooth muscle and might be a viable material for the creation of neointestine.     

Influence of pneumoperitoneum on small bowel anastomoses: a histological analysis in the rat model.

Laparoscopic techniques are increasingly applied for the treatment of diverse gastrointestinal diseases. With regard to reports of a pronounced decrease of intra-abdominal blood flow with increasing intra-abdominal pressure, the present study investigates the impact of pressure and gas type on ischemia in small bowel anastomoses in the rat model. Laparotomy and ileoileal anastomosis were performed in 39 male Sprague-Dawley rats. A CO2 or helium pneumoperitoneum of 3 mm Hg or of 6 mm Hg was maintained before and after anastomoses. Rats in the control group received no pneumoperitoneum. Animals were sacrificed after 5 d, and the anastomotic region was explanted for subsequent histopathological examinations. In hematoxylin and eosin (HE)-stained sections, the Chiu score, villi configuration, and number of goblet cells were analyzed. Proliferation (Ki67) and expression of a matrix metalloproteinase (MMP-8) were examined by immunohistochemistry. Mucosal damage according to the scoring system by Chiu, the number of goblet cells, the villus length, the proliferation (Ki67), and the submucosal expression of MMP-8 was similar in all groups. Our results suggest that within a certain range of pressures and time, laparoscopic assisted surgery using CO2 pneumoperitoneum can be performed safely. Helium gas offers no advantages over CO2.     

大鼠肝小肠联合移植模型的建立

本文报告一种封闭群大鼠进行的肝小肠联合移植模型。术中供体肝与小肠整块游离和灌注，分开切取。供肝原位、供肠异位移植于受体大鼠。主要血管用Ｋａｍａｄａ袖套法吻合，正式实验２１次，３天以上存活率为４３％。结果表明：减少手术时间和简化操作技术是提高成功率的关键因素和基本原则。