胃腺癌癌巢和癌间质中巨噬细胞的分布及其与患者预后关系

目的了解胃腺癌组织中巨噬细胞（tumor associated macrophages,TAMs）浸润情况,探讨TAMs分布及其与预后的关系。方法采用免疫组织化学SP法,检测92例胃腺癌癌巢和癌间质中CD68^＋巨噬细胞数,结合随访资料分析TAMs与患者术后无复发生存率关系。结果胃腺癌间质及癌巢组织中均有大量TAMs浸润,且癌间质中TAMs表达高于癌巢（9．78个／HFP与4．00个／HFP,P=0．000）;Ⅰ＋Ⅱ期癌巢内TAMs高于Ⅲ＋Ⅳ期（4．73个／HFP与3．27个／HFP,P=0．001）,Ⅰ＋Ⅱ期癌巢TAMs／癌间质TAMs比值高于Ⅲ＋Ⅳ期（0．58个／HFP与0．29个／HFP,P=0．004）,而Ⅰ＋Ⅱ期癌间质TAMs低于Ⅲ＋Ⅳ期（8．85个／HFP与11．03个／HFP,P=0．000）。结论Cox风险比例回归模型显示,癌巢TAMs和癌巢／癌间质TAMs比值越大,患者无复发生存率越高,而癌间质TAMs越大患者无复发生存率越小。癌巢TAMs、癌间质TAMs及癌巢／癌间质TAMs比值可作为临床评估胃腺癌患者的预后指标。     

子宫颈鳞状细胞癌纤维结合素、层黏蛋白及Ⅳ型胶原的异常表达

目的 通过对纤维结合素（Fibronectin,FN）和层黏连蛋白（Laminin,LN）及Ⅳ型胶原蛋白的表达进行检测,探讨 基底膜重构与宫颈癌分化程度的关系。方法 应用双重免疫荧光染色技术,对正常人及不同分化程度的子宫颈鳞状细胞癌病人子宫颈石蜡包埋组织的FN、LN及Ⅳ型胶原和LN蛋白呈线状环绕于血管及上皮下基底膜内,FN呈细条索分布于基底膜的间质内。子宫颈癌组织中,癌细胞团周围基底人的Ⅳ型胶原的线状荧光减弱、断裂甚至缺失,尤其以低分化鳞状细胞癌明显,而N和LN蛋白表达增多,并与癌细胞分化程度负相关。一些癌细胞浆内可见FN和LN蛋白的阳性染。结论 子宫颈鳞状细胞癌时,基底膜变薄、断裂,Ⅳ型胶原降解。而癌细胞浆及邻近癌组织的间质内,FN及LN表达增多,并与细胞分化程度呈负相关。     

婴儿室间隔缺损合并肺动脉高压心房肌Ⅰ、Ⅲ型胶原的表达改变

[目的]探讨室间隔缺损合并肺动脉高压婴儿心房肌Ⅰ、Ⅲ胶原蛋白含量的变化,并分析心肌间质胶原成分含量及比值的改变与肺动脉压力的相关性.[方法]试验组为30例室间隔缺损合并肺动脉高压婴儿,体外循环建立时留取少量右心房肌标本,对照组为8例非心血管疾病及胶原疾病死亡的儿童,尸体解剖时留取右心房肌,并应用免疫组化及数字图像分析技术观测其心房肌Ⅰ、Ⅲ胶原的表达.[结果]正常心房肌Ⅰ型胶原为粗、细不等的条状纤维,彼此连接成网,Ⅲ型胶原呈散在的斑片状分布,而婴儿室间隔缺损合并肺动脉高压患儿Ⅰ型胶原呈现出大斑片状增加,Ⅲ胶原呈现出条索状增加.免疫组化图像分析显示：对照组Ⅰ型胶原含量明显高于Ⅲ型胶原,有显著差异（P〈0.01）,Ⅰ型/Ⅲ型比值为5.66±0.43,试验组中：Ⅰ型和Ⅲ型胶原含量均较对照组显著增加（P〈0.01）,Ⅰ型/Ⅲ型比值为3.04±2.06,Ⅲ型胶原蛋白增加幅度较大,Ⅰ型/Ⅲ型比值较前明显减少.线性回归模型拟合显示肺动脉压力与Ⅰ型/Ⅲ型呈显著相关,肺动脉压力=0.927-0.058（Ⅰ/Ⅲ）.[结论]室间隔缺损合并肺动脉高压婴儿心房肌中,Ⅰ型及Ⅲ型胶原含量均较正常心房肌中明显增加,且以Ⅲ型胶原含量增加明显,表明室间隔缺损合并肺动脉高压婴儿未成熟心肌组织亦存在心肌间质重构.室间隔缺损合并肺动脉压力患儿中,肺动脉压力与Ⅰ型及Ⅲ型胶原含量之比（Ⅰ/Ⅲ值）呈线性关系,即：肺动脉压力=0.927～0.058（Ⅰ/Ⅲ）,表明随着肺动脉压力的增高,心肌间质重构将逐步加重.     

肾纤维化及γ-干扰素的治疗

肾纤维化是各种肾脏病终末期的基本病理变化。肾纤维化的特点是Ⅰ、Ⅲ和Ⅳ型胶原、纤维连接蛋白、各种蛋白聚糖等在肾组织基底膜和肾间质不恰当的积聚和分布，导致正常组织结构破坏，肾功能丧失。因此，肾纤维化已成为近年来肾脏病学领域的研究热点之一。本文拟对近年来肾纤维化研究的重要成果和γ-干扰素在其中作用的研究作一系统介绍。     

Ⅰ、Ⅲ、Ⅳ、Ⅴ型胶原和层连蛋白在人肝硬化和肝细胞癌中的分布

正常肝脏肝窦壁Ⅳ型胶原和层连蛋白染色阳性,说明存在基底膜结构,是为功能性基底膜.肝硬化时肝窦毛细血管化,使功能性基底膜破坏,肝硬化时各种细胞外基质成分显著增多,以Ⅰ型胶原为最多.由于各成分增多程度不一致,使细胞外基质的构成发生了改变.原发性肝细胞性肝癌组织中细胞外基质的分布与含量与癌组织的分化程度密切相关.分化程度高,细胞外基质沿癌组织小梁间的血窦壁排列成连续或断续线状;分化程度低,癌组织的细胞外基质仅见于血管壁.     

Ⅰ、Ⅲ、Ⅳ、Ⅴ型胶原和层连蛋白在人肝硬化和肝细胞癌中的分布

正常肝脏肝窦壁Ⅳ型胶原和层连蛋白染色阳性,说明存在基底膜结构,是为功能性基底膜。肝硬化时肝窦毛细血管化,使功能性基底膜破坏。肝硬化时各种细胞外基质成分显著增多,以Ⅰ型胶原为最多。由于各成分增多程度不一致,使细胞外基质的构成发生了改变,原发性肝细胞性肝癌组织中细胞外基质的分布与含量与癌组织的分化程度密切相关,分化程度高,细胞外基质沿癌组织小梁间的血窦壁排列成连续或断续线状;分化程度低,癌组织的细胞外基质仅见于血管壁。     

胶原和层连蛋白在人肝硬化和肝细胞癌中的分布

正常肝脏肝窦壁Ⅳ型胶原和层连蛋白染色阳性,说明存在基底膜结构,是为功能性基底膜.肝硬化时肝窦毛细血管化,使功能性基底膜破坏.肝硬化时各种细胞外基质成分显着增多,以Ⅰ型胶原为最多.由于各成分增多程度不一致,使细胞外基质的构成发生了改变.原发性肝细胞性肝癌组织中细胞外基质的分布与含量与癌组织的分化程度密切相关.分化程度高,细咆外基质措癌组织小梁间的血窦壁排列成连续或断续线状;分化程度低,癌组织的细胞外基质仅见于血管壁.     

层黏连蛋白和Ⅳ型胶原在前列腺不同疾病中的表达及意义

目的 探讨层黏连蛋白(LM)和Ⅳ型胶原在前列腺不同疾病中的表达及意义。方法将100例诊断明确的前列腺标本分为5组，即正常组、良性增生组(BPH)、高级别上皮内瘤组(HPIN)、偶发癌组和前列腺癌组，每组20例。应用SP法检测LM和Ⅳ型胶原在5组标本中的表达。结果 LM和Ⅳ型胶原的表达基本一致，在正常组、BPH及大部分HPIN组织中呈棕黄色连续环线状包绕前列腺腺泡及导管外周，且分布均匀；但在部分HPIN组织中排列不规则、分布不均匀或紊乱；在偶发癌病灶区有片状缺失、间断或分布杂乱；在前列腺癌组织中均呈大片状或多灶性缺失，或杂乱排列。结论 LM和Ⅳ型胶原在前列腺不同疾病中的表达有差异，偶发癌和前列腺癌的基底膜损伤明显。提示LM和Ⅳ型胶原的表达不仅有助于预测HPIN的癌变潜能，也对前列腺良、恶性疾病的鉴别有帮助。     

利用组织芯片技术观察p53及其同系物p63在恶性肿瘤中的表达

目的 探讨p53和p63在部分肿瘤发生中的作用，为其人群大规模筛查、早期诊断及预后评估提供依据。方法 选取病理科档案材料中食管鳞癌、胃腺癌、大肠腺癌、膀胱移行细胞癌、肺癌、肝细胞性癌、乳腺浸润导管癌、肾透明细胞癌、卵巢腺样囊腺癌和胰腺癌石蜡包埋组织各6例及相应正常组织各3例。肿瘤标本先观察苏木精—伊红(HE)切片以确定肿瘤部位，采用组织芯片仪及手工法制作180点列阵组织芯片，进行p53和p63抗体免疫组化染色。光镜观察p53、p63阳性细胞在各组的分布，并进行对比分析。结果 p53表达类型：(1)正常组织p53阳性细胞数很少；(2)3例食管癌、3例胃癌、2例大肠癌、1例肺癌、1例乳腺癌、2例卵巢癌的绝大多数肿瘤细胞呈强阳性棱染色反应，癌旁组织细胞和间质细胞阴性，1例膀胱癌中有少量阳性肿瘤细胞散在分布于阴性肿瘤细胞中。p63表达类型：(1)正常食道复层扁平上皮基底层细胞呈棱染色，p63阳性细胞散在分布于棘细胞层；(2)6例食道癌、6例膀胱癌和4例肺鳞癌均为强阳性反应，大多数肿瘤细胞呈强棱染色，癌旁组织细胞和间质细胞p63阴性；两例肺腺癌p63阴性；(3)6例胃癌中的4例、6例大肠癌中的3例、6例乳腺浸润导管癌中的2例、6例肝细胞性癌中的1例可见较强的肿瘤细胞胞质染色。其中3例食管癌、3例胃癌、1例大肠癌、1例肺癌、1例乳腺癌的肿瘤细胞同时呈p53和p63阳性反应。结论 p53在正常组织中表达量很少，在多种肿瘤细胞中表达量升高；p63阳性细胞主要位于正常上皮组织的增殖区，而在多种癌细胞中表达增强，两者在肿瘤细胞中不同的分布情况提示其在肿瘤的发生发展中作用途径不同。     

胶原-聚羟基乙酸支架与骨髓间质干细胞构建的组织工程肌腱

背景：肌腱组织工程中应用的支架材料大致分为合成高分子材料和天然细胞外基质材料两类。 目的：探讨用胶原一聚羟基乙酸复合材料作为支架材料，以家兔骨髓间质干细胞为种子细胞预构组织工程化肌腱重建兔跟腱的可能性。 设计、时间及地点：单一样本观察，于2004—06／2005-06在中南大学湘雅医学院动物学部及湘雅医院中心实验室完成。 材料：健康家兔6只，体质量1．0～1．5kg，雌雄不限；体质量250g左右大白鼠1只用于Ⅰ型胶原溶液的制备。 方法：以鼠尾胶原-聚羟基乙酸为支架，将分离、培养的自体骨髓间质干细胞作为种子细胞，在体外混合培养后回植至兔跟腱缺损处，设为实验组。对照组以不含种子细胞的胶原聚羟基乙酸支架修复肌腱缺损。 主要观察指标：①骨髓问充质干细胞原代接种后5d半量换液时在倒置显微下观察，并以CD44原位杂交检测该细胞。②组织工程化肌腱的体外构建结果。③于术后12周取移植处新生组织进行大体、组织细胞学观察，并做Ⅰ型和Ⅲ型胶原免疫组织化学染色。 结果：原代细胞以CD44原位杂交显色，胞浆呈棕黄色，胞核经苏木精复染呈蓝色。提示为骨髓间充质干细胞。术后12周实验组新生组织与受体肌腱组织完好连接，呈条索状，与周围其他组织无粘连；组织学切片结果类似于正常肌腱组织；免疫组织化学染色示Ⅰ型胶原阳性，Ⅲ型胶原阴性。而对照组新生组织较细小、与周围组织有粘连；组织学切片示胶原纤维呈松散网丝状，细胞排列紊乱；免疫组织化学染色示Ⅰ型、Ⅲ型胶原阳性。 结论：以胶原聚羟基乙酸为支架，以自体骨髓间质干细胞作为种子细胞可望构建组织工程化肌腱。     

肠镜检查并发肠穿孔42例报告

随着大肠内镜 (纤维 /电子 )检查日趋普及 ,并发肠穿孔者仍时有发生 ,内镜医师应需警惕。为此 ,笔者截至 1995年调查收集全国 2 6所医院 ( 11个省市自治区 )在 382 86例次肠镜检查中并发肠穿孔 4 2例 ,发生率为 0 .11% ,其中死亡 3例。现就本组资料分析报告如下。1　一般资料4     

Changes in distribution of connective tissue components of human placentae with maturation

The distributional changes of type IV collagen and laminin with normal maturation of human placentae were examined in relation to those of fibronectin by the histochemical methods including immunofluorescence staining. In the early chorionic villi, these components were detected along the trophoblastic basement membrane, around the fetal blood vessels, and in the villous stroma. Laminin was detected also in the pericellular matrices of nonvillous cytotrophoblasts where type IV collagen was rarely detected. In the late and term placentae, laminin and type IV collagen were detected along the trophoblastic basement membrane, while this structure was virtually not stained for fibronectin. These observations suggest that type IV collagen and laminin are the constituents of the trophoblastic basement membrane throughout the maturation period of the placentae, while fibronectin is a transient constituent.     

Role of basement membrane collagen and elastin in the autofluorescence spectra of the colon.

BACKGROUND: Autofluoresence can be used to detect neoplasia in the colon. Two known fluorophores, collagen and elastin, are probably partly responsible for colonic emission spectra. Their contribution to colonic autofluorescence was investigated. METHODS: Autofluorescence spectra of normal, dysplastic, and malignant colonic tissue were studied by using excitation wavelengths from 280 nm to 350 nm. The wavelengths of peak emission and their widths at half maximum intensity were measured. Similar measurements were performed on collagen types I, III, IV, V, IX, and elastin. Colonic spectra were compared to those of collagen and elastin. Spectral differences between collagen types IV (basement membrane) I, III, V, and IX were studied. RESULTS: Four major emission peaks were noted whose wavelength of peak emission and full widths at half maximum intensity were independent of tissue histology. The emission spectra of type IV collagen differed markedly from that of nonbasement membrane collagens and elastin. CONCLUSIONS: Type IV (basement membrane) collagen is most likely responsible for the emission peak at 365 nm. The spectra of basement membrane collagen and not other types of collagen should be used in studies of epithelial tissue spectra. Elastin did not appear to be responsible for any of the four autofluorescence peaks observed in colonic tissue.     

Changes in the collagen of synovial membrane in rheumatoid arthritis and effect of D-penicillamine.

1. Normal synovial membrane contains approximately equal proportions of two genetically distinct forms of collagen, types I and III. The proportion of these two collagens is unchanged in rheumatoid synovium but in addition a small amount of basement membrane collagen is present. Tissue culture of rheumatoid synovium confirms the synthesis of both type I and III collagens. 2. In young normal synovium both type I and type III collagens are stabilized by a reducible keto cross-link, which is replaced in adult tissue by an as yet unknown non-reducible cross-link. During the proliferation of the collagen in adult rheumatoid synovium a high proportion of the keto cross-link is present. This cross-link is not susceptible to cleavage by D-penicillamine, nor does the drug have any effect on the rate of synthesis in vitro. The mode of action of D-penicillamine in rheumatoid arthritis does not appear to involve a direct effect on the synovial membrane collagen.     

Rheumatic fever-associated Streptococcus pyogenes isolates aggregate collagen

Acute rheumatic fever is a serious autoimmune sequel of Streptococcus pyogenes infection. This study shows that serotype M3 and M18 S. pyogenes isolated during outbreaks of rheumatic fever have the unique capability to bind and aggregate human basement membrane collagen type IV. M3 protein is identified as collagen-binding factor of M3 streptococci, whereas M18 isolates bind collagen through a hyaluronic acid capsule, revealing a novel function for M3 protein and capsule. Following in vivo mouse passage, conversion of a nonencapsulated and collagen-binding negative M1 S. pyogenes into an encapsulated, collagen-binding strain further supports the crucial role of capsule in mediating collagen binding. Collagen binding represents a novel colonization mechanism, as it is demonstrated that S. pyogenes bind to collagen matrix in vitro and in vivo. Moreover, immunization of mice with purified recombinant M3 protein led to the generation of anti-collagen type IV antibodies. Finally, sera from acute rheumatic fever patients had significantly increased titers of anti-collagen type IV antibodies as compared with healthy controls. These findings may suggest a link between the potential of rheumatogenic S. pyogenes isolates to bind collagen, and the presence of collagen-reactive autoantibodies in the serum of rheumatic fever patients, which may form a basis for post-streptococcal rheumatic disease. These anti-collagen antibodies may form a basis for poststreptococcal rheumatic disease.     

Type IV and type "A-B" collagens do not elicit platelet aggregation or the serotonin release reaction.

Human collagens were isolated from kidney, lung, skin, aorta, cartilage, and placenta. Five different types were obtained, including two new molecular species, one characteristic of basement membranes, or type IV collagen, and the other the recently described "A-B" collagen derived from fetal membranes. All the collagens were purified and separated by combination of heat-gelation fractionation and salt fractionation. In neutral solution at 37 degrees neither type IV nor type "A-B" collagen elicited platelet aggregation or 14C-serotonin release. Preincubation of platelets with both types IV and "A-B" collagen did not inhibit aggregation upon subsequent addition of collagen types I, II, or III.     

Extracellular matrix gene expression increases preferentially in rat lipocytes and sinusoidal endothelial cells during hepatic fibrosis in vivo.

Whether parenchymal or nonparenchymal liver cells play a predominant role in the pathophysiology of hepatic fibrosis has not been firmly established in vivo. We have addressed this question by quantitating the relative abundance of specific mRNAs for collagen types I, III, and IV, and laminin in purified populations of hepatocytes, sinusoidal endothelial cells, and lipocytes from normal and fibrotic rat liver. In normal liver, type I collagen gene expression was minimal in all cell types; mRNA for types III and IV collagen were apparent in endothelial cells and lipocytes, but not in hepatocytes. Laminin mRNA was present in all cell types. Induction of fibrogenesis by either bile duct ligation or carbon tetrachloride administration was associated with a substantial increase in mRNA for types I and III collagen in nonparenchymal cells. Lipocytes from fibrotic animals exhibited a greater than 30-fold increase in type I collagen mRNA relative to normal lipocytes, and greater than 40-fold relative to hepatocytes. Type III collagen mRNA reached 5 times that in normal lipocytes and greater than 120 times that in hepatocytes. Endothelial cells exhibited an isolated increase in type I collagen mRNA, reaching five times that in normal liver. Type IV collagen and laminin gene expression were not significantly increased in nonparenchymal cells during fibrogenesis; in fact, mRNA for type IV collagen and laminin decreased by up to 50% in endothelial cells. Despite the pronounced changes that occurred in matrix gene expression in nonparenchymal cells during fibrogenesis, no change was noted in hepatocytes. We conclude that nonparenchymal liver cells, particularly lipocytes, are important effectors of hepatic fibrosis in vivo.     

Collagen type IV and procollagen type III during granulation tissue formation: a serological, biochemical, immunohistochemical and morphometrical study on the viscose cellulose sponge rat model

The serum concentrations of collagen type IV,7S, collagen type IV,nc1, and aminoterminal type III procollagen peptide immunoreactive components were measured by means of specific radioimmunoassays during development of granulation tissue in rats. The results were compared with tissue deposition of basement membranes and interstitial collagens in the granulation as measured morphometrically. A parallel sequential pattern in tissue deposition of collagen types III and IV, and serum increase of collagen types III- and IV-related fragments, was observed. Serum collagen type IV was less sensitive as a marker for development of granulation tissue than the serum procollagen type III N-peptide. This was in accordance with a low collagen type IV/interstitial collagen ratio in the granulation tissue. However, a cross-sectional study showed that serum collagens types IV,7S and IV,nc1 may be useful as early quantitative indicators of granulation tissue formation. Simultaneously, measurement of collagen type IV- and procollagen type III N-peptide-related antigens in serum provides a differentiated reflection of the dynamic matrix processes in developing granulation tissue.     

Amyloid P-component is a constituent of normal human glomerular basement membrane

Glomerular and other vascular basement membranes were found to contain an antigen that was immunochemically indistinguishable from serum amyloid P-component. There was no immunological cross-reactivity between antisera to serum amyloid P-component and to collagen types I, III, IV, or V. The amyloid P-component antigen was confined to the endothelial aspect, the lamina rara interna, of glomerular basement membrane. It could not be eluted by high-ionic-strength saline, EDTA, dithiothreitol, or either polar or nonpolar detergents, but was released into solution when isolated glomerular basement membrane was digested by highly purified bacterial collagenase. Most of these P- component molecules and their constituent polypeptide chains were of higher molecular weight and lower isoelectric point than serum amyloid P-component. These findings indicate that, as well as being a normal plasma protein and a universal constituent of amyloid deposits, P- component is also a normal matrix glycoprotein of basement membrane in which it is covalently linked to collagen and/or other matrix proteins. This may be relevant both to the pathogenesis of amyloidosis and to other aspects of physiology and pathology of basement membranes.     

Colorectal cancer metastatic phenotype stimulates production by fibroblasts of N-terminal peptide of type III collagen: clinical implications for prognosis

In this study we assessed whether the serum levels of the N-terminal peptide of type III collagen (PIIIP), an index of type III collagen synthesis, are influenced by colorectal cancer stage, and whether "in vitro" fibroblast growth and PIIIP production could be altered by tumor tissues obtained from metastatic and nonmetastatic colorectal cancer. 208 colorectal cancer patients (115 colon and 93 rectum) were studied; 54 were stage I, 62 stage II, 37 stage III and 55 stage IV. PIIIP serum levels were significantly higher in stage IV as compared to all other patient groups. The 5-year survival of stage I, stage II, stage III and stage IV patients were 87%, 88%, 32% and 20%, respectively. In the subgroup of stage I and stage II patients considered together, PIIIP (> 0.5 U/ml), but not CEA (> 5 microg/l) serum levels, were predictive for survival. Fibroblast growth was significantly inhibited, while PIIIP production was significantly enhanced, when these cells were conditioned with colorectal cancer homogenates obtained from patients with distant metastases, than from those without distant metastases. In conclusion, colorectal tumors, when metastatic, stimulate fibroblasts' PIIIP synthesis and the serum levels of this peptide might predict patients' outcome after radical surgery.