不同时期增生性瘢痕组织内透明质酸含量与含水量的相关性*

目的：增生性瘢痕是由于成纤维细胞过度增殖，细胞外基质胶原纤维和蛋白多糖等过度合成及水的过量形成的，但有关增生性瘢痕的透明质酸量、水含量及它们之间关系的报道较少。观察不同时期瘢痕组织内透明质酸含量以及含水量，并分析其相关性。 方法：①试验对象：选择2004-06/2007-08入住本院要求手术的烧伤后增生性瘢痕患者18例。瘢痕形成时间6个月至2年以内者8例，男7例，女1例；瘢痕形成时间2年以上者10例，男8例，女2例(2~4年3例，4~10年5例，12年1例，19年1例)。纳入标准：患者无系统性疾病，瘢痕未进行过治疗，部位在四肢和背部。所有烧伤后增生性瘢痕组织均为手术时取得，另取其腹股沟处正常皮肤作为对照。患者对治疗及试验均知情同意，治疗方案经医院医学伦理委员会批准。②试验方法及评估：对正常皮肤及瘢痕组织进行病理检测，分别用苏木精－伊红染色、胶原纤维染色及李世荣等介绍的复合染色法染色并观察两者的形态。测量正常皮肤及瘢痕组织的含水量。ELISA法检测透明质酸含量，求出它们之间的相关系数以了解它们之间的关系。 结果：纳入烧伤后增生性瘢痕患者18例，均进入结果分析。①苏木精-伊红染色和胶原纤维染色显示，正常皮肤的真皮中，胶原纤维排列疏松，呈束状定向排列。增生性瘢痕组织中，胶原纤维增厚，排列不规则。复合染色法显示胶原纤维呈红色，酸性粘多糖呈蓝色，两种瘢痕均含大量酸性粘多糖。②正常皮肤含水量69.59%，2年以内的烧伤后增生性瘢痕为69.88%，2年以上为71.40%，3者之间相互比较，差异无显著性意义(P > 0.05)。③透明质酸在2年以上瘢痕组织中的含量为（3.36±0.08）mg/L，与正常皮肤中的含量（0.67±0.41）mg/L和2年内瘢痕组织含量（1.70±0.54）mg/L相比较，差异显著（P < 0.01，P < 0.05）。正常皮肤中的含量与2年内瘢痕组织之间相比较，差异有显著性意义(P < 0.05)。透明质酸含量与含水量之间的相关系数 γ=0.27，差异无显著性意义（P > 0.05）。 结论：随着瘢痕形成时间的延长，透明质酸含量增加，可能与瘢痕成熟有关；含水量在正常皮肤、2年以内及2年以上瘢痕组织间相似，透明质酸量的变化没有相应地导致含水量的变化。     

胎儿和成人皮肤相关基因的差异表达*

背景：胎儿皮肤创伤后无瘢痕愈合是胎儿发育过程特定阶段的特殊现象,其机制目前尚未明确,可能与调控基因的表达有关。目的：运用人类基因芯片技术,研究人胎儿及成人正常皮肤差异表达基因及其特征和可能的生物学意义。方法：选择人胎儿(孕20~24周)皮肤和成人(18~48岁)皮肤各10例,提取各标本的总DNA与RNA,制成荧光标记的cDNA探针,与含10 724个人类靶基因的芯片杂交,经扫描、生物信息学分析,比较两种组织基因的差异表达。结果与结论：基因芯片高通量地揭示了胎儿及成人正常皮肤基因信息的差异表达。与成人正常皮肤比较,胎儿皮肤基因显著差异表达83个,已明确功能基因26个,涉及多种信号传递及基因调控的改变。说明胎儿与成人正常皮肤差异表达基因的存在在一定程度上导致了胎儿创伤的无瘢痕愈合。     

透明质酸合成酶1在脑出血后脑组织透明质酸含量及相对分子质量变化过程中的作用

目的观察透明质酸合成酶1(HAS1)在脑出血后脑内透明质酸(HA)含量和相对分子质量变化过程中的作用。方法采用自体血注射法制作HAS1基因敲除小鼠和野生型C57BL/6J小鼠脑出血模型。分别分为出血后24 h组、出血后72 h组和对照组(不注自体血,余同),每组均n=6。提取出血侧半球脑组织的HA,经凝胶层析分离后用ELISA方法检测HA的总量和相对分子质量分布。Real time PCR方法观察HAS1、HAS2、HAS3和透明质酸分解酶(Hyal)mRNA表达水平的变化。通过对野生型C57BL/6J小鼠和HAS1基因敲除小鼠的比较,评价HAS1在此过程中的作用。结果脑出血后,①野生型C57BL/6J小鼠脑HA含量基本不变,仅在出血后72 h组呈轻微下降,但HA相对分子质量分布有显著变化,出血后24 h组呈现一过性的大分子HA的比例增加。②HAS1基因敲除小鼠脑内HA总量比野生型C57BL/6J小鼠减少1/4。出血后72 h组的HA总量明显下降,而且出血后24 h组的大分子HA比例不升反降,至出血后72 h后也未见恢复。③出血后,基因表达均随时间而增加,增幅大小顺序为HAS2>HAS3>HAS1,Hyal1>Hyal3>Hyal2。野生型C57BL/6J和HAS1基因敲除小鼠间只有HAS3的表达有差异。结论 HAS1基因敲除导致脑内HA总量减少1/4,脑出血后24 h HAS3基因表达代偿性增加,导致脑出血后脑内大分子HA含量下降。     

干细胞在皮肤组织工程中的应用

目前还没有一种理想的组织工程皮肤能满足临床严重皮肤缺损患者的需求,其重要原因之一是无合适的种子细胞来源。干细胞在皮肤组织工程中的应用已取得了很大进展,但仍存在一些问题需要解决。基于此,文章重点介绍了胚胎干细胞、表皮干细胞和间充质干细胞等有代表性的干细胞的分离、培养以及在皮肤组织工程中的应用现状,并提出了存在的问题和下一步的研究方向,以期研制出更理想的组织工程皮肤替代物。     

人工皮肤替代物的材料种类及其特征

目的：评价组织工程人工皮肤替代物各种生物材料的性能和应用,寻找适合人体的替代物。方法：以“组织工程,人工皮肤,支架材料”为中文关键词,“tissue engineering, Artificial skin,intravascular stent” 为英文关键词,采用计算机检索1993-01/2009-10相关文章。纳入与有关生物材料与人工皮肤修复相关的文章;排除重复研究或Meta分析类文章。以26篇文献为主重点进行了讨论组织工程人工皮肤替代物及其性能。结果：组织工程人工皮肤是应用组织工程技术将体外培养的上皮细胞和成纤维细胞扩增后,接种于具有良好生物相容性的材料上,经体外培养,形成含有与正常皮肤相似的表皮和真皮结构的皮肤替代物。然后将其移植于皮肤创面处,以实现创伤的修复和重建。将两种或两种以上的材料复合在一起,或对生物材料表面进行各种各样的修饰,促进细胞与材料之间的黏附、提高细胞的生物活性、维持生物功能成为目前组织工程生物材料研究的热点。结论：目前还没有一种人工材料能完全符合组织工程的要求。进一步提高支架材料的微观渗透性和生物活性,促进毛细血管的长入;制备结构仿生支架材料及高活性复合支架材料是今后的研究方向。关键词：组织工程皮肤材料;人工皮肤;支架材料;皮肤替代物;生物相容性doi:10.3969/j.issn.1673-8225.2010.29.037     

合成及天然支架材料在皮肤组织工程中的应用

目前用作皮肤组织工程支架的材料有很多种，大致可以分为合成支架材料和天然支架材料两大类。文章在广泛查阅国内外有关皮肤组织工程方面相关文献的基础上进行总结，发现天然支架材料可以更好地为细胞的黏附、增殖提供三维支架空间，具有更大的潜力应用于皮肤组织工程。     

透明质酸、透明质酸酶与垂体腺瘤侵袭性的关系研究

目的研究透明质酸、透明质酸酶在垂体腺瘤侵袭性生物学行为中的意义。方法应用放射免疫法测定瘤组织中透明质酸,用酶联吸附测定法测定瘤组织中透明质酸酶活性及应用逆转录聚合酶链式反应(RT-PCR)测定瘤细胞中透明质酸酶基因表达。结果侵袭性垂体腺瘤患者瘤组织中透明质酸及透明质酸酶显著增高,透明质酸酶基因PH-20高表达,与非侵袭性垂体腺瘤组及正常脑组织组的差异有显著统计学意义(P<0.05)。结论透明质酸、透明质酸酶在垂体腺瘤侵袭性生物学行为中发挥作用,测定透明质酸和透明质酸酶对侵袭性垂体腺瘤患者诊断及预后判断有参考价值。     

含有生肌液的组织工程皮肤修复兔皮肤缺损

背景：在皮肤开放创面上进行缺损修复与重建是组织修复领域的研究焦点。构建含有中药的组织工程皮肤修复创面缺损的研究并不多见。 目的：将含有生肌液的皮肤支架与自体皮肤细胞复合，构建含药组织工程皮肤，修复兔皮肤缺损。 设计：自体对照动物实验。 单位：实验于2005-02/08在天津市天津医院骨科研究所细胞工程室完成。 材料：同窝大耳白兔8只，体质量1.6~1.8 kg。自行提取生肌液(1 500 g/L)，成分为当归、生地、龟板、象皮、血余、生石膏、炉甘石；明胶-壳聚糖支架材为料天津大学提供，分别修剪成1.3 cm直径的圆片，60Co照射灭菌；DR无细胞真皮基质由江苏省启动市医疗用品研究所提供。 方法：分离培养兔皮肤细胞，获取第3代角质形成细胞与成纤维细胞。延兔脊柱两侧制作直径1.5 cm的圆形全层皮肤缺损创面4个。分为4组，阴性对照组（明胶-壳聚糖支架贴附于创面后滴加生理盐水），人工真皮组（DR无细胞真皮基质贴附于创面后接种细胞悬液），非含药组织工程皮肤组（明胶-壳聚糖支架贴附于创面后接种细胞悬液），含药组织工程皮肤组（生肌液-明胶-壳聚糖支架贴附于创面后接种细胞悬液）。 主要观察指标：①术后13 d测量剩余创面面积。②术后7 d、10 d取材行组织学与免疫组织化学染色检测。 结果：①术后13 d各组创面面积均有不同程度的缩小，剩余面积阴性对照组>人工真皮组>非含药组织工程皮肤组>含药组织工程皮肤组，非含药组织工程皮肤组、含药组织工程皮肤组与阴性对照组比较差异有显著性意义（P < 0.05)。②术后10 d，阴性对照组、人工真皮组、非含药组织工程皮肤组均有炎性肉芽组织增生，无表皮组织形成；含药组织工程皮肤组形成接近正常的上皮组织及幼稚的真皮组织。非含药组织工程皮肤组与人工真皮组BrdU标记弱阳性，含药组织工程皮肤组BrdU标记强阳性。 结论：实验采用的方法使种子细胞在含有生肌液的明胶-壳聚糖支架上增殖，在体修复皮肤损伤，能够有效扩展皮肤缺损处的覆盖面积，缩短皮肤缺损的愈合时间。     

血管内皮祖细胞在组织工程皮肤构建中的意义

背景：血管内皮祖细胞可加速缺血部位的血管化。目的：综合分析血管内皮祖细胞加速组织工程皮肤血管化的方法和途径。方法：计算机检索中国期刊全文数据及PubMed数据库1999-09/2009-09相关文章,检索词为“血管内皮祖细胞,组织工程皮肤,血管新生,endothelial progenitor cell(EPC),tissue engineered skin,vascularization ”。纳入与血管内皮祖细胞与组织工程皮肤研究现状与发展密切相关文章,包括：①血管内皮祖细胞的生物学特性、动员方法及促进血管新生的可能机制。②组织工程皮肤的应用现状。③组织工程皮肤中血管内皮祖细胞的研究现状。结果与结论：血管内皮祖细胞是指特异性归巢于血管新生组织,并能分化增生为成熟内皮细胞的一群祖细胞。近年来大量的动物实验证实,通过移植体外培养的血管内皮祖细胞到肢体缺血部位,可以加速缺血部位的血管化。在心肌梗死方面已经进行了自体血管内皮祖细胞治疗心肌梗死的临床试验,使其临床应用逐渐成为可能。而目前构建的组织工程皮肤中缺少血管成分,严重影响人工皮肤中活细胞的存活及其功能实现,如何构建出含有血管的人工皮肤及加速人工真皮替代物血管化引起了广大的关注。应用血管内皮祖细胞促进组织工程皮肤血管化进而提高其成活率有重要的临床意义。     

颌面部间隙感染的观察与护理

口腔颌面部间隙感染,是颌面及其内周围软组织的炎症,当炎症弥漫时称蜂窝组织炎,局限时称间隙脓肿.颌面部各间隙是潜在的,其中充满脂肪或疏松的结缔组织.一旦感染,发生炎性反应时,间隙才存在,分为牙源性、腺源性、血源性,以牙源性较为常见.     

Hyaluronic acid is increased in the skin and urine in patients with amyotrophic lateral sclerosis

We performed morphological studies of skin and measured glycosaminoglycans in the urine from patients with sporadic amyotrophic lateral sclerosis (ALS) and control subjects. The wide spaces separating collagen bundles reacted strongly with alcian blue stain in ALS patients and stained more markedly as ALS progressed. Staining with alcian blue was virtually eliminated by Streptomyces hyaluronidase. The urinary excretion of hyaluronic acid (HA) (mg/day) was significantly increased (P < 0.01) in ALS patients compared with that of control subjects, and there was a significant positive correlation between the excreted amount of HA and the duration of illness in advanced ALS patients with a duration of more than 2 years from clinical onset (r = 0.72,P < 0.02). We suggest that sporadic ALS includes a metabolic disorder of HA in which an accumulation of HA in the skin is linked to an increased urinary excretion of HA.     

Amyotrophic lateral sclerosis: unusually low content of collagen in skin

The skin tissues from patients with amyotrophic lateral sclerosis (ALS) and controls were studied by electron microscopy, and their amino acid compositions were examined. On electron microscopy, the most conspicuous findings in ALS were (1) a marked increase in amorphous material separating collagen bundles, and (2) the smaller diameter of collagen fibrils. These were more marked with longer duration of illness. Amino acid analysis showed that the levels of hydroxyproline, hydroxylysine, and glycine were significantly low (P < 0.001, < 0.01, andP < 0.001, respectively) in ALS patients as compared with those of controls, and there was a significant negative correlation between the level of hydroxyproline and duration of illness in ALS patients (r = − 0.88,P < 0.01). In addition, the collagen content per dry weight (mg) of the tissues in ALS was significantly smaller (P < 0.001) than in controls. These results indicate that the metabolism of skin collagen might be affected in the disease process of ALS.     

透明质酸在脑出血和腔隙性脑梗塞中病理作用的探讨

本文对５７例急性脑出血和脑腔梗患者及３５例对照组血清透明质酸（ＨＡ）含量进行了测定，并与相应的临床指标（病程、血压及神经功能缺损评分等）、血液生化指标（血糖、血脂、纤维蛋白原及血液流变学等）和影像学指标进行了相关分析。结果表明：脑出血和脑腔梗组的ＨＡ含量明显高于对照组，腔梗组明显高于出血组（Ｐ＜０．０１）。脑出血组ＨＡ含量与纤维蛋白原含量呈明显负相关，与其它指标无相关性。而脑腔梗组ＨＡ含量与纤维蛋白原及其它指标均无相关性。     

不同浓度铜孵育对肝豆状核变性离体培养细胞内铜含量的影响

本实验研究了不同浓度铜孵育对肝豆状核变性患者、杂合子及对照组离体培养皮肤成纤维细胞内铜含量的影响,发现经20μg/ml铜孵育12或24小时后,患者细胞浆内的铜/蛋白比值显著高于后两组(P<0.01),且与后两组完全无重叠,提示该方法可用于早期、临床表现不典型患者的诊断,并有可能成为症状前患者及产前诊断的有效方法。     

Retinoic acid induced myelomeningocele in fetal rats: characterization by histopathological analysis and magnetic resonance imaging

The prevention of human neural tube defects by folic acid administration and the potential for fetal surgical intervention for myelomeningocele (MMC) have renewed interest in the molecular pathways and pathophysiology of spina bifida. Animal models for assessment of the early developmental biology and pathophysiology of this lesion are needed. The goal of this study was to develop and characterize a non-surgical rat model of MMC. Time-dated Sprague-Dawley rats were gavage fed different doses of retinoic acid (RA) dissolved in olive oil at E10 (maternal n = 55, fetal n = 505). Control animals received olive oil alone (maternal n = 20, fetal n = 265) or were untreated (maternal n = 5, fetal n = 63). Fetuses were analyzed by detailed histopathology and MRI. Overall, isolated MMC occurred in 60.7% (307/505) of RA-exposed fetuses and no controls. Histopathology confirmed the entire spectrum of severity observed in human MMC, ranging from exposure of the cord with intact neural elements to complete cord destruction. MRI of the brain of MMC fetuses confirmed structural changes similar to humans with Arnold-Chiari malformation, including downward displacement of the cerebellum to just above the foramen magnum and compression of the developing medulla into a small posterior fossa. In conclusion, the RA-induced rat model of MMC is developmentally and anatomically analogous to human MMC. This relatively efficient and cost-effective model of MMC should facilitate investigation of the developmental biology and pathophysiology of MMC, and may be useful for the evaluation of further strategies for prenatal treatment.     

Mechanisms of docosahexaenoic acid accretion in the fetal brain

Docosahexaenoic acid (DHA, 22:6 n-3) is the major polyunsaturated fatty acid (PUFA) in the adult mammalian brain. DHA is an essential fatty acid (FA) since it, or its short chain precursor, α-linolenic acid (LnA, 18:3 n-3), have to be obtained in the diet. Moreover, dietary n-3 FA deficiency is associated with biochemical changes in the brain and with disturbances in vision and other neurological parameters. Under normal nutritional conditions, fetal brain DHA accumulation is substantial, with a “DHA accretion spurt” being demonstrated in the last period of gestation. This accumulation is supported by the maternal supply of DHA or LnA, but selectivity of DHA accumulation is probably a placental function whose mechanism is lately being clarified. The fetal gastrointestinal (G-I) tract may be instrumental in supplying DHA to the fetal brain under certain conditions, such as following intra-amniotic administration of ethyl-docosahexaenoate (Et-DHA). In this pathway, DHA is supplied independently of the maternal metabolism, and the fetal liver is apparently involved. The fetal G-I tract may be advantageous for DHA supply in cases of maternal–placental insufficiency resulting in intrauterine growth retardation. The fetal brain itself is capable of metabolizing LnA to DHA, without the participation of the fetal liver, thus contributing to the accumulation of its own DHA during one of the most crucial periods of its development. J. Neurosci. Res. 52:129–136, 1998. © 1998 Wiley-Liss, Inc.     

Hyaluronic acid through a new injectable nerve guide delivery system enhances peripheral nerve regeneration in the rat

The use of non-neural conduits to bridge gaps in peripheral nerves has been noted in the literature for many years. A logical extension of this concept is the introduction of neurotrophic or growth promoting factors into the lumen. We present here an injectable nerve guide that allows percutaneous access to the microenvironment of the regenerating peripheral nerve within the guide's lumen. Hyaluronic acid, a compound associated with decreased scarring and improved fibrin matrix formation, is added sequentially to the regenerating peripheral rat sciatic nerve via this injectable nerve guide. Assessment of nerve regeneration and reinnervation shows better conduction velocity, higher axon counts, and a trend toward earlier myelination with hyaluronic acid compared with saline. This work not only implies hyaluronic acid's role as an agent that aids nerve growth but also describes a new tool that allows percutaneous access to the milieu of a regenerating nerve. © 1995 Wiley-Liss, Inc.     

Serotonin and amino acid content in platelets of autistic children

The platelet levels of serotonin and the amino acids aspartic acid, glutamine, glutamic acid and gamma-aminobutyric acid were measured in 18 drug-free autistic (DSM-III criteria) and 14 age-matched healthy children. Serotonin was significantly increased while the amino acids aspartic acid, glutamine, glutamic acid and gamma-aminobutyric acid were significantly decreased in comparison with the controls. It is suggested that the decline of the amino acids in platelets from autistic children represents a biochemical marker related to infantile autism.     

Intraparenchymal fetal striatal transplants and recovery in kainic acid lesioned rats

Striatal kainic acid (KA) lesions induce behavioral and biochemical deficits which resemble symptoms encountered in patients suffering from Huntington's disease. In rats with KA lesions, fetal striatal transplants have shown to reverse the pervasive nocturnal hyperactivity induced by the lesion. In the present study 4.6 mm3 of fetal striatal tissue were delivered bilaterally into the anterodorsal portion of the lesioned caudate nucleus. Care was taken to deliver the transplant within the host parenchyma and away from the lateral ventricles. Locomotor behavior analyzed using the Digiscan animal activity monitors before and after the transplants demonstrated a reversal of the hyperactivity following transplants in 70% of lesioned animals. Microinjections of horseradish peroxidase delivered into the globus pallidus and substantia nigra of a small group of functionally recovered transplanted animals, did not reveal evidence for reinnervation between host nigra or pallidum and the transplant at 10 weeks post-transplantation. Other laboratories have reported anatomical connections by 6 months post-transplantation. Ventricular/brain ratios demonstrated that intraparenchymal transplants significantly reduced the ventricular dilation following KA lesion. These results suggest that functional recovery can be obtained when the transplant is immersed into the host's striatal parenchyma regardless of the existence of long-range anatomical connections.     

Replenishment of docosahexaenoic acid in n-3 fatty acid-deficient fetal rats by intraamniotic ethyl-docosahexaenoate administration

A procedure for intraamniotic ethyl-docosahexaenoate (Et-DHA) administration was used to restore the docosahexaenoic acid (DHA; 22:6 n-3) levels in n-3-deficient fetal rats. The state of deficiency, characterized by a 34% and 60% decrease in DHA content of fetal brain and liver, respectively, was attained by feeding the pregnant dams from day 8 and up to 20 days gestation, with an n-3 linolenic acid-deprived diet. After a single intraamniotic administration of Et-DHA on day 18 or 19, a rapid increase in both fetal brain and liver DHA was achieved. This increase was accompanied by a decrease in the docosapentaenoic acid (DPA; 22:5 n-6) level. After 48 hr following Et-DHA administration, the major phospholipids (PLs) phosphatidylserine (PS), phosphatidylethanolamine (PE), and phosphatidylcholine (PC), together accounting for more than 90% of total lipid phosphorus in sunflower oil (SFO)-treated animals, regained the DHA content to levels similar to control animals in both fetal brain and liver tissues. Unlike brain, however, most of the DHA content in liver PLs was restored by 24 hr, suggesting that the fetal liver may have a higher metabolic turnover. The DHA/DPA ratio was used to assess the degree of DHA correction. Fetal brain PS, PC, and PE ratios following Et-DHA administration grew steadily over a period of 48 hr but reached only approximately 60% of the control levels. Liver PS regained a value similar to the control, while those of PC and PE were 33% and 46% lower than the controls, respectively. Alterations in the PL polar head-group composition were observed following the dietary manipulations and Et-DHA administration. Although the intraamniotic injection is an invasive approach, the ability to rapidly enhance DHA acylation during intrauterine life may hold potential clinical value whenever an indication for DHA deficiency exists. J. Neurosci. Res. 48:264–272, 1997. © 1997 Wiley-Liss, Inc.