实时荧光定量聚合酶链式反应检测瘢痕疙瘩中核心蛋白多糖的表达

目的 探讨瘢痕疙瘩成纤维细胞中核心蛋白多糖的表达、含量,及其在瘢痕疙瘩形成中的作用和机制.方法对瘢痕疙瘩、正常瘢痕以及正常皮肤成纤维细胞进行体外培养,采用光镜、透射电镜观察成纤维细胞形态、活性及凋亡;应用实时荧光定量聚合酶链式反应(FQ-PCR)对核心蛋白多糖以及β1转化生长因子(TGF-β1)的mRNA表达进行检测、分析.结果 瘢痕疙瘩成纤维细胞形态不规则、排列紊乱,线粒体增多,粗面内质网扩张呈囊,细胞核常染色质丰富,表明其合成蛋白的功能活跃;瘢痕疙瘩成纤维细胞中核心蛋白多糖mRNA含量较正常瘢痕或正常皮肤成纤维细胞降低,而TGF-β1 mRNA表达则较正常皮肤及瘢痕组织成纤维细胞升高.结论 核心蛋白多糖在瘢痕疙瘩成纤维细胞内含量较正常皮肤明显减少,提示其对成纤维细胞增殖、合成的抑制作用随之减弱,同时使TGF-β1表达上调,导致成纤维细胞的大量增生、迁移,合成过量胶原.表明核心蛋白多糖是抑制瘢痕疙瘩形成的重要因子.     

内质网分子伴侣GRP94、GRP78和XBP1在病理性瘢痕中的表达初探

目的：检测内质网应激反应的关键分子葡萄糖调节蛋白94（GRP94）、葡萄糖调节蛋白78（GRP78）和X-盒结合蛋白1（XBP1）mRNA在病理性瘢痕的表达,探讨其基因表达及内质网应激反应与病理性瘢痕形成及转归的关系。方法：用RT-PCR法检测GRP94、GRP78和XBP1mRNA在：①瘢痕疙瘩（13例）、增生性瘢痕（17例）和正常皮肤（15例）组织以及体外培养的瘢痕疙瘩（5例）、增生性瘢痕（5例）和正常皮肤（6例）成纤维细胞中的表达;②体外培养瘢痕疙瘩和增生性瘢痕成纤维细胞（各5例）中经0.1mg/ml氢化可的松作用前后的表达。结果：①GRP94、GRP78和XBP1mRNA在瘢痕疙瘩、增生性瘢痕和正常皮肤组织及其成纤维细胞中的表达均无显著性差异（P〉0.05）;②0.1mg/ml氢化可的松作用后,GRP94mRNA在瘢痕疙瘩和增生性瘢痕来源的成纤维细胞中的表达量均显著下降,具有统计学意义（P〈0.01）;而GRP78和XBP1mRNA在瘢痕疙瘩和增生性瘢痕来源的成纤维细胞中的表达量改变均无显著性差异（P〉0.05）。结论：内质网分子伴侣GRP94、GRP78和XBP1在瘢痕疙瘩和增生性瘢痕组织及其成纤维细胞中基因转录与正常皮肤组织及其成纤维细胞中基因转录水平一致;GRP94mRNA的表达量显著下降可能是糖皮质激素治疗病理性瘢痕的机制之一。     

P57kip2和Maspin在病理性瘢痕组织中的表达

目的 研究P57kip2和Maspin在病理性瘢痕组织中的表达情况及相互关系,探讨它们在病理性瘢痕形成中的作用及机制.方法 应用免疫组化SP法结合计算机病理图像分析和逆转录聚合酶链反应( RT-PCR)检测正常皮肤、成熟瘢痕、增生性瘢痕和瘢痕疙瘩组织中P57kip2和Maspin的表达并对其表达进行统计学分析.结果 病理性瘢痕组织中P57kip2蛋白的表达定位于成纤维细胞的细胞核内,且P57kip2蛋白及mRNA的表达减少,与正常皮肤、成熟瘢痕对照组比较差异有统计学意义(P＜0.05).病理性瘢痕组织中Maspin蛋白的表达定位于成纤维细胞的细胞质和细胞核内,且Maspin蛋白及mRNA的表达减少,与正常皮肤、成熟瘢痕对照组比较差异有统计学意义(P＜0.05).P57kip2与Maspin蛋白表达存在明显正相关(P＜0.O1).结论P57kip2与Maspin在病理性瘢痕组织中均表达减少,是病理性瘢痕相关基因,两者存在明显正相关,是病理性瘢痕的形成机制之一.P57kip2和Maspin可能通过成纤维细胞在病理性瘢痕的形成过程中发挥着重要作用.     

微纤维蛋白1在病理性瘢痕中的表达及意义

目的 检测微纤维蛋白1(FBN1)在瘢痕疙瘩和增生性瘢痕的表达情况,研究其与TGF-β1的相关性,探讨病理性瘢痕发生的分子机理.方法 用RT-PCR法检测瘢痕疙瘩、增生性瘢痕和正常皮肤三种组织中FBNI和TGF-β1mRNA的表达量并进行相关性分析;用免疫组化的方法对FBN1蛋白在人类皮肤组织的表达进行定位及半定量研究.结果 FBN1 mRNA在瘢痕疙瘩(0.802±0.116)高于正常皮肤(0.252±0.067)218.25%(P<0.01),增生性瘢痕(0.628±0.144)较正常皮肤增高149.21%(P>0.05).TGF-β1在瘢痕疙瘩较正常皮肤和增生性瘢痕分别增高200.27%和92.81%(均为P<0.01);FBNI和TGF-β1 mRNA在上述标本中的表达量呈正性相关(r=0.820,P<0.01).FBN1蛋白主要在人类皮肤组织的表皮、毛囊和汗腺的基底膜以及真皮层的血管内皮细胞、部分成纤维细胞和细胞外基质中表达阳性.在真皮层,FBN1蛋白在瘢痕疙瘩(0.117±0.042)表达量较正常皮肤(0.185±0.043)和增生性瘢痕(0.181±0.048)分别减少36.76%和35.36%(均为P<0.01).结论 FBN1在瘢痕疙瘩表达异常,它是瘢痕疙瘩相关基因(瘢痕相关基因),可能通过参与TGF-β1信号通路的调节影响病理性瘢痕的发生.     

病理性瘢痕中VEGF与突变型P53基因表达的关系

目的 研究病理性瘢痕中血管内皮生长因子(VEGF)与突变型抑癌基因P53的表达情况及其相互关系,探讨它们在病理性瘢痕形成中的作用及机制.方法 应用免疫组化SP法检测正常皮肤、成熟瘢痕、增生性瘢痕和瘢痕疙瘩组织中VEGF和突变型P53基因的表达及其相关性.结果 病理性瘢痕组织中VEGF的表达增高,与正常皮肤、成熟瘢痕对照组比较,差异有统计学意义(P＜0.005).增生性瘢痕与瘢痕疙瘩之间VEGF蛋白的表达,差异无统计学意义(P＞0.05).病理性瘢痕组织中突变型P53的表达增高,与正常皮肤、成熟瘢痕对照组比较差异有统计学意义(P＜0.005).增生性瘢痕与瘢痕疙瘩之间突变型P53蛋白的表达,差异无统计学意义(P＞0.05).VEGF与突变型P53蛋白表达,存在明显正相关(P＜0.01).结论 VEGF与突变型P53在病理性瘢痕组织中均表达增高,与病理性瘢痕的形成密切相关,可能对病理性瘢痕的形成起着重要作用,两者之间存在正相关.     

病理性瘢痕成纤维细胞中Cx43及Cx43 mRNA的表达

目的 探讨连接蛋白43(Cx43)在病理性瘢痕中的调控作用.方法 选择瘢痕疙瘩20例、增生性瘢痕23例、普通瘢痕18例、正常皮肤12例四组标本,应用免疫组织化学与原位杂交技术,从蛋白及核酸水平检测Cx43及Cx43 mRNA在四组标本成纤维细胞中的表达.结果 Cx43及Cx43 mRNA在增生性瘢痕和搬痕疙瘩成纤维细胞中的表达明显少于普通瘢痕和正常皮肤(P＜0.05).结论 成纤维细胞Cx43表达下调可能造成病理性瘢痕组织的成纤维细胞间缝隙连接细胞间的通讯异常,从而导致病理性瘢痕的发生,且Cx43表达下调可能是由于Cx43基因转录受到抑制造成的.     

Tenascin-C在瘢痕疙瘩和增生性瘢痕中的基因表达研究

目的 探讨Tenascin-C基因在瘢痕疙瘩和增生性瘢痕中的表达。方法 取正常成人皮肤组织RNA，构建正义、反义Tenascin-C(Tn-C)mRNA探针，运用原位杂交技术，观测10例瘢痕疙瘩、10例增生性瘢痕和5例正常成人皮肤组织中Tn-C mRNA的表达。结果 Tn-C mRNA在正常皮肤表皮中无表达，真皮中表达稀少，局限于乳头真皮层的成纤维细胞和皮肤附属器；10例瘢痕疙瘩表皮均有表达，真皮分布较广，如成纤维细胞、血管内皮和皮肤附属器；Tn-C mRNA在3例增生性瘢痕表皮表达，7例无表达，真皮中表达与瘢痕疙瘩相同但较弱，比正常皮肤增多，但差异无显著性。结论 Tenascin-C mRNA在瘢痕疙瘩表皮和真皮中有高表达。     

Bcl—2和Fas基因在瘢痕成纤维细胞中的表达

目的研究凋亡基因Fas/Apo-1和抑凋亡相关基因Bcl-2在瘢痕形成机制中的作用.方法采用免疫组织化学方法对10例正常皮肤、10例增生性瘢痕及10例瘢痕疙瘩成纤维细胞Fas/Apo-1和Bcl-2蛋白的表达进行检测.Fas蛋白稀释为1∶50;Bcl-2蛋白稀释为1∶40,阴性对照以PBS代替一抗.随机选择五个高倍视野,计算其细胞平均阳性率.结果Bcl-2蛋白在正常皮肤、增生性瘢痕及瘢痕疙瘩中阳性率分别为6.78%、38.6%和83.2%.瘢痕疙瘩、增生性瘢痕的表达阳性率高于正常皮肤,有非常显著性差异(P＜0.01),而瘢痕疙瘩的表达阳性率明显高于增生性瘢痕(P＜0.01).Fas/Apo-1蛋白在正常皮肤、增生性瘢痕及瘢痕疙瘩中阳性率分别为78.4%、80.4%和84.4%,三组间无显著性差异(P＞0.05).结论病理性瘢痕的形成与Bcl-2基因的过度表达有关,而对Fas基因介导的凋亡不敏感或凋亡受阻,亦是导致瘢痕过度增生原因之一.     

E2F1蛋白在病理性瘢痕组织中的表达

目的增生性瘢痕和瘢痕疙瘩是临床上常见的病理性瘢痕,是创伤后过度愈合反应的结果,以成纤维细胞的异常增殖及合成分泌大量细胞外基质为特征,其形成机理尚不清楚,研究表明基因失调是其中的关键.E2F基因是细胞周期G1向S期过渡的重要调控因子,在调节细胞周期进程和调节细胞增殖过程中起着关键作用.本实验的目的是检测E2F1基因在病理性瘢痕组织中的表达,以正常皮肤组织做对照,初步探讨E2F1在病理性瘢痕形成中的生物学作用.方法利用免疫组化ABC法检测正常皮肤、成熟瘢痕、增生性瘢痕和瘢痕疙瘩组织中E2F1蛋白的表达,并进行统计学分析.结果增生性瘢痕和瘢痕疙瘩组织中E2F1蛋白表达两组间无明显差异,与正常皮肤、成熟瘢痕对照组比较均有显著性差异(P＜0.01).结论 E2F1蛋白表达在病理性瘢痕组织中增高,促进瘢痕组织中细胞的增生,对病理性瘢痕的形成可能起着重要作用.     

Body composition assessment and methodology in nonathletic and athletic adolescent and adult males and females

The purpose of this review is to outline methodology for assessing body composition utilizing anthropometric and densitometric techniques. The objective of body composition assessment is to measure body fat and lean body mass. The quantity of these components varies due to growth, physical activity, dietary regimens, and aging. Anthropometric techniques incorporate selected skinfolds, circumferences, skeletal widths, or other variables to estimate body composition within k2.0-4.0%. These techniques are adequate for field testing of groups or individuals, but are population specific. Densitometry measures body volume irrespective of physique, sex, or age. This laboratory technique estimates body composition within 1.0-2.0%, is more difficult to administer, but is not population specific. Some limitation exists with any present technique due to biological variability and incomplete research of reference body composition in children, females, and the aged. J Orthop Sports Phys Ther 1984;5(6):336-347.     

Postoperative nausea and vomiting and opioid-induced nausea and vomiting: guidelines for prevention and treatment

Postoperative nausea and vomiting (PONV) causes patient discomfort, lowers patient satisfaction, and increases care requirements. Opioid-induced nausea and vomiting (OINV) may also occur if opioids are used to treat postoperative pain. These guidelines aim to provide recommendations for the prevention and treatment of both problems. A working group was established in accordance with the charter of the Sociedad Espa?ola de Anestesiología y Reanimación. The group undertook the critical appraisal of articles relevant to the management of PONV and OINV in adults and children early and late in the perioperative period. Discussions led to recommendations, summarized as follows: 1) Risk for PONV should be assessed in all patients undergoing surgery; 2 easy-to-use scales are useful for risk assessment: the Apfel scale for adults and the Eberhart scale for children. 2) Measures to reduce baseline risk should be used for adults at moderate or high risk and all children. 3) Pharmacologic prophylaxis with 1 drug is useful for patients at low risk (Apfel or Eberhart 1) who are to receive general anesthesia; patients with higher levels of risk should receive prophylaxis with 2 or more drugs and baseline risk should be reduced (multimodal approach). 4) Dexamethasone, droperidol, and ondansetron (or other setrons) have similar levels of efficacy; drug choice should be made based on individual patient factors. 5) The drug prescribed for treating PONV should preferably be different from the one used for prophylaxis; ondansetron is the most effective drug for treating PONV. 6) Risk for PONV should be assessed before discharge after outpatient surgery or on the ward for hospitalized patients; there is no evidence that late preventive strategies are effective. 7) The drug of choice for preventing OINV is droperidol.     

Perioperative and long-term morbidity and mortality after above-knee and below-knee amputations in diabetics and nondiabetics

We performed a retrospective review of a vascular surgery quality assurance database to evaluate the perioperative and long-term morbidity and mortality of above-knee amputations (AKA, n = 234) and below-knee amputations (BKA, n = 720) and to examine the effect of diabetes mellitus (DM) (181 of AKA and 606 of BKA patients). All patients in the database who had AKA or BKA from 1990 to May 2001 were included in the study. Perioperative 30-day cardiac morbidity and mortality and 3-yr and 10-yr mortality after AKA or BKA were assessed. The effect of DM on 30-day cardiac outcome was assessed by multivariate logistic regression and the effect on long-term survival was assessed by Cox regression analysis. The perioperative cardiac event rate (cardiac death or nonfatal myocardial infarction) was at least 6.8% after AKA and at most 3.6% after BKA. Median survival was significantly less after AKA (20 mo) than BKA (52 mo) (P < 0.001). DM was not a significant predictor of perioperative 30-day mortality (odds ratio, 0.76 [0.39-1.49]; P = 0.43) or 3-yr survival (Hazard ratio, 1.03 [0.86-1.24]; P = 0.72) but predicted 10-yr mortality (Hazard ratio, 1.34 [1.04-1.73]; P = 0.026). Significant predictors of the 30-day perioperative mortality were the site of amputation (odds ratio, 4.35 [2.56-7.14]; P < 0.001) and history of renal insufficiency (odds ratio, 2.15 [1.13-4.08]; P = 0.019). AKA should be triaged as a high-risk surgery while BKA is an intermediate-risk surgery. Long-term survival after AKA or BKA is poor, regardless of the presence of DM.