联合检测血清透明质酸、Ⅳ型胶原及层黏蛋白对肝纤维化的诊断价值

目的 :探讨血清透明质酸 (HA)、Ⅳ型胶原 (CIV)及层黏蛋白 (LN)联合检测对肝纤维化的诊断价值。方法 :采用酶联免疫法对健康人组 2 2例、急性肝炎组 10例、慢性肝炎组 10例、肝硬化组 2 5例及慢性重症肝炎组 10例 ,分别检测血清HA ,CIV及LN的水平 ;对肝硬化组三项指标分别检测和联合检测所得的敏感性、特异性及准确性进行比较。结果 :血清中HA ,CIV及LN的水平以肝硬化组和慢性重症肝炎组最高 ,均明显高于其它组 (P <0 0 1)。肝硬化组联合检测的敏感性高于单项检测的敏感性 ,但其特异性降低。结论 :联合检测HA ,CIV及LN可提高诊断肝纤维化敏感性 ,但特异性降低 ;其中HA +CIV的敏感性和特异性较高 ,为最优联合     

多种血管新生指标在肝细胞肝癌中的表达及敏感性比较

目的　探讨血管新生指标CD34、CD31、vWF、Ⅳ型胶原纤维及层粘连蛋白在肝细胞肝癌(HCC)中的表达及意义 ,同时比较上述几种血管新生因子与增殖细胞核抗原 (PCNA)、病理指标及预后的相关性 ,以便筛选出有效的临床预后指标。方法　采用免疫组化方法 ,对 5 3例肝细胞肝癌的标本进行CD31、CD34、vWF、Ⅳ型胶原纤维及层粘连蛋白的染色、计数 ,并用检测数据与患者的临床资料进行统计分析。结果　统计染色的血管面积后发现 ,CD34与多种临床病理指标无相关性 ;CD31与肝内门静脉浸润相关 ;vWF与肿瘤的TNM分期及肝内门静脉浸润呈正相关 ;CollⅣ与肝内门静脉浸润呈正相关、与术后生存期呈负相关 ;Lam与肝硬化及术中出血量呈负相关、与术后生存期呈正相关。PCNA与肿瘤TNM分期有关。结论　在HCC中 ,CollⅣ、vWF、及CD31为肝细胞肝癌的有效血管新生及预后指标 ;Lam则与肝硬化及术中出血相关 ;PCNA指数肿瘤分期有关 ;CD34不能用作血管新生或预后指标     

乳腺癌层粘连蛋白的免疫组织化学研究

本研究应用层粘连蛋白（ＬＮ）抗体，对２７例乳腺癌、５例乳腺良性病变进行了免疫组织化学研究。结果发现乳腺良性病变时，ＬＮ以均匀完整的线型阳性反应显示在乳腺导管及小叶基膜处。乳腺癌时，ＬＮ的染色表现为癌细胞胞浆内弥漫状阳性反应。２７例乳腺癌中，癌细胞ＬＮ染色强度不同，伴有转移的乳腺癌ＬＮ染色多数较强，强度在＋＋～的为７／１２（５８．３％）；不伴有转移的病例，ＬＮ染色多数较浅或为阴性，强度在＋＋～＋＋＋的病例为３／１５（２０％）。说明癌细胞内源性ＬＮ染色强弱与肿瘤转移密切相关，内源性ＬＮ的合成可能改变肿瘤细胞的浸润活性。     

Adhesion in skeletal muscle during regeneration.

Adhesion molecules were studied in regenerating skeletal muscle immunohistochemically and ultrastructurally after a standardized trauma. In normal muscle, extracellular matrix (ECM) protein tenascin was restricted to myotendinous junctions (MTJ), while the integrin beta 1-subunit was present also on the sarcolemma. After injury, tenascin increased on the outer surface of regenerating myofibers, where cellular fibronectin also accumulated. Later, tenascin concentrated at the tips of regenerating myofibers, where new MTJs were formed. The beta 1-subunit disappeared on necrotized myofibers and reappeared on regenerating fibers in a thicker layer. The regenerating myofibers were invested by a basal lamina, except for the growth cones at the distal ends, which were laminin-negative until the formation of MTJs occurred. These results indicate that regenerating muscle cells are attached to the ECM in a way that allows both growth of the muscle cells across the scar and their use before the regeneration is completed.     

双功能域重组FN多肽抑制癌细胞浸润能力的研究

在制备了两个Cell Ⅰ-Hep Ⅱ 双结构域重组FN多肽(CH50和CH56)的基础上,研究其抑制肿瘤细胞浸润能力的作用。两个多肽的结构差异是CH50中删除了Cell I和HepⅡ之间的Ⅲ-11和ED-A结构顺序。CH50(ED_(50)为30.2 nmol/L)结合细胞的能力略高于CH56(ED_(50)为45.4 nmol/L)。两种多肽均可显著抑制黑色素瘤B16/F1细胞结合层粘素的能力,抑制作用相同。在体内肿瘤浸润抑制试验中,两种多肽均可显著抑制癌细胞浸润能力,使肺转移结节数降低80％左右。结果提示:Ⅲ-11和ED-A结构顺序对Cell Ⅰ-Hep Ⅱ 双结构域多肽结合细胞的能力有一定的影响,但删除Ⅲ-11和ED-A不是重组多肽抑制肿瘤转移的决定因素,Cell I和Hep Ⅱ 这两个结构域单独连接在一起是其抑制肿瘤细胞转移的结构基础。     

Schistosoma mansoni infection in a recently exposed community in Senegal: lack of correlation between liver morphology in ultrasound and connective tissue metabolites in serum

Summary Four hundred and seventy villagers of Ndombo, a village with recently established intensive transmission of Schistosoma mansoni in the Senegal River Basin, were enrolled in a study with the intention to assess hepatosplenic morbidity. All patients were examined parasitologically and by ultrasound. Hepatic fibrosis serum markers were determined in 153 adult patients (aminoterminal propeptide of procollagen type III, hyaluronan and laminin). By ultrasound, about 60% of the patients showed early stages of hepatic involvement, 3% of the patients unequivocally showed severe hepatosplenic pathology (grade 3 according to the Managil classification), whereas in another study performed in the same village 3 years earlier, no patients with severe hepatosplenic pathology had been found. No correlation between the aminoterminal propeptide of procollagen type III, hyaluronan or laminin and the ultrasound findings could be established. These hepatic fibrosis serum markers do not seem to be a sensitive method to detect early hepatic fibrosis in schistosomiasis.     

Clinical and molecular study in congenital muscular dystrophy with partial laminin alpha 2 (LAMA2) deficiency

Complete laminin alpha2 (LAMA2) deficiency causes approximately half of congenital muscular dystrophy (CMD) cases. Many loss-of-function mutations have been reported in these severe, neonatal-onset patients, but only single missense mutations have been found in milder CMD with partial laminin alpha2 deficiency. Here, we studied nine patients diagnosed with CMD who showed abnormal white-matter signal at brain MRI and partial deficiency of laminin alpha2 on immunofluorescence of muscle biopsy. We screened the entire 9.5 kb laminin alpha2 mRNA from patient muscle biopsy by direct capillary automated sequencing, single strand conformational polymorphism (SSCP), or denaturing high performance liquid chromatography (DHPLC) of overlapping RT-PCR products followed by direct sequencing of heteroduplexes. We identified laminin alpha2 sequence changes in six of nine CMD patients. Each of the gene changes identified, except one, was novel, including three missense changes and two splice-site mutations. The finding of partial laminin alpha2 deficiency by immunostaining is not specific for laminin alpha2 gene mutation carriers, with only two patients (22%) showing clear causative mutations, and an additional three patients (33%) showing possible mutations. The clinical presentation and disease progression was homogeneous in the laminin alpha2-mutation positive and negative CMD patients.     

Integrin α6 expression in human prostate carcinoma cells is associated with a migratory and invasive phenotypein vitro andin vivo

Cell adhesion and migration are important features in tumor invasion, being mediated in part by integrins (extracellular matrix receptors). Integrins are significantly decreased in human prostate cancer. An exception is 6 integrin (laminin receptor) which persists during prostate tumor progression. We have selected high (DU-H) and low (DU-L) expressors of 6 integrin from a human prostate tumor cell line, DU145, to assess experimentally the importance of 6 integrin in tumor invasion. DU-H cells exhibited a four-fold increased expression of 6 integrin on the surface compared to DU-L cells. Both cell types contained similar amounts of 3 and 5 integrin. The DU-H cells contained 6 subunits complexed with both the 1 and 4 subunits whereas DU-L cells contained 6 complexed only with 4. DU-H cells were three times more mobile on laminin as compared to DU-L, but adhered similarly on laminin. Adhesion and migration were inhibited with anti-6 antibody. Each subline was injected intraperitoneally into SCID mice to test its invasive potential. Results showed greater invasion of DU-H compared to DU-L cells, with increased expression of a6 integrin on the tumor at the areas of invasion. These data suggest that 6 integrin expression is advantageous for prostate tumor cell invasion.     

Cell surface laminin-like substances and laminin-related carbohydrates of rat ascites hepatoma AH7974 and its variants with different lung-colonizing potential

Rat ascites hepatoma AH7974 cells strongly expressed antilaminin antibody-reactive substances (laminin-like substances) andGriffonia simplicifolia isolectin B4 (GS)-reactive carbohydrate (alpha)-d-galactose; alpha-Gal) on their cell surface. The alpha-Gal expression was not apparently influenced by the pretreatment of cells with methanol. The cell membrane laminin-like substances had approximate molecular weights of 150, 62 and 56 kDa in denaturating reducing conditions, of which the 62 and 56 kDa bands were stained with GS. The cell membrane molecules bearing alpha-Gal were 62 and 56 kDa and were stained with antilaminin antibody. Therefore, the major molecules bearing alpha-Gal residues of AH7974 cell membrane are considered to be laminin-like substances. To determine the role of the substances in metastasis, we selected four cell lines (74AD, 74AD-f, 74FL, 74FL-a) from AH7974 in culture. 74AD and 74FL-a are adherent lines and 74AD-f and 74FL are floating lines. All of these cell lines strongly expressed laminin-like substances, but a marked difference was found in expression of alpha-Gal, which was most strongly expressed by 74FL, followed by 74AD, and rarely by 74AD-f and 74FL-a; the staining intensity was positively correlated with their experimental lung-colonizing potential. Cell membrane laminin-like substances were 200, 97, 62, 56 and 46 kDa and among them 62 and 56 kDa molecules were glycosylated with alpha-Gal. The pretreatment of 74FL cells with antilaminin antibody or with human type A serum (containing natural antibody to alpha-Gal epitope) depressed remarkably the lung-colonizing potential of the cells. These results suggest that the expression of 62 and 56 kDa laminin-like substances with alpha-Gal residues on tumor cell surfaces is one of the determinants associated with lung-colonizing potential of these cells.     

The expanding phenotype of laminin alpha2 chain (merosin) abnormalities: case series and review

Initial reports of patients with laminin alpha2 chain (merosin) deficiency had a relatively homogeneous phenotype, with classical congenital muscular dystrophy (CMD) characterised by severe muscle weakness, inability to achieve independent ambulation, markedly raised creatine kinase, and characteristic white matter hypodensity on cerebral magnetic resonance imaging. We report a series of five patients with laminin alpha2 deficiency, only one of whom has this severe classical CMD phenotype, and review published reports to characterise the expanded phenotype of laminin alpha2 deficiency, as illustrated by this case series. While classical congenital muscular dystrophy with white matter abnormality is the commonest phenotype associated with laminin alpha2 deficiency, 12% of reported cases have later onset, slowly progressive weakness more accurately designated limb-girdle muscular dystrophy. In addition, the following clinical features are reported with increased frequency: mental retardation (~6%), seizures (~8%), subclinical cardiac involvement (3-35%), and neuronal migration defects (4%). At least 25% of patients achieve independent ambulation. Notably, three patients with laminin alpha2 deficiency were asymptomatic, 10 patients had normal MRI (four with LAMA2 mutations reported), and between 10-20% of cases had maximum recorded creatine kinase of less than 1000 U/l. LAMA2 mutations have been identified in 25% of cases. Sixty eight percent of these have the classical congenital muscular dystrophy, but this figure is likely to be affected by ascertainment bias. We conclude that all dystrophic muscle biopsies, regardless of clinical phenotype, should be studied with antibodies to laminin alpha2. In addition, the use of multiple antibodies to different regions of laminin alpha2 may increase the diagnostic yield and provide some correlation with severity of clinical phenotype.