Basement membranes in the kidney of the dromedary camel (Camelus dromedarius): An immunohistochemical and ultrastructural study

Basement membranes consist of various proteins, the major ones being laminin and collagen type IV. Primary defects in these two proteins have been extensively associated with kidney pathologies. This study aimed to establish baseline information on the immunohistochemical distribution of laminin and collagen type IV, and to corelate these with the ultrastructure of basal laminae in the uriniferous tubules of the dromedary camel. Tissue samples were taken from the kidneys of eight adult female camels, and processed for immunohistochemical and ultrastructural investigations. Strong intensity of collagen type IV was observed within the basement membranes of Bowman’s capsule. The thickness of the basal lamina of the parietal layer of Bowman's capsule varied extensively depending on the region of the renal corpuscle; the thicker areas were always associated with cuboidal epithelial cells. The glomerular basement membrane revealed strong immunostaining of laminin, whereas the mesangial matrix was strongly immunoreactive to collagen type IV. Abundant amount of laminin was found in the basement membranes of proximal convoluted tubules, thin limbs of the loop of Henle, and collecting ducts. Dense immunostainings of laminin and collagen type IV were observed in the medullary regions of uriniferous tubule, in which numerous projections extended from the basal laminae into the subjacent connective tissue. Overall, the present study revealed marked variations in the distribution of the basement membrane markers laminin and collagen type IV in the uriniferous tubules of camel kidney. The results have also shown difference in the thickness of basal laminae. This variation in thickness, however, was unlikely to be influenced by the amount of laminin and collagen type IV.     

层粘连蛋白对腺样囊性癌细胞侵袭和转移的影响

目的 研究层粘连蛋白（ＬＮ）与人涎腺腺样囊性癌转移的关系。方法 应用免疫组织化学染色,Ｂｏｙｄｅｎ小室体外移动试验和体外粘附试验,对人涎腺腺样囊性癌细胞系Ａｃｃ－２和其克隆出的肺高转移细胞株Ａｃｃ－Ｍ进行比较研究。结果 发现ＬＮ和层粘连蛋白受体（ＬＮＲ）在Ａｃｃ－Ｍ表达明显高于Ａｃｃ－２。在加入外源性ＬＮ后,Ａｃｃ－２体外移动性、粘附率均明显提高,而Ａｃｃ－Ｍ无明显变化。结论 层粘连蛋白在Ａｃｃ转     

Structure and vascularization of the cruciate ligaments of the human knee joint

The structure and vascularization of the human anterior and posterior cruciate ligament were investigated by light microscopy, transmission electron microscopy, injection techniques and by immunohistochemistry. The major part of the anterior and posterior cruciate ligament is composed of bundles of type I collagen. Type III collagen-positive fibrils separate the bundles. The major cell type is the elongated fibroblast, lying solitarily between the parallel collagen fibrils. The histologic structure of the cruciate ligaments is not homogeneous. In both ligaments there is a zone where the tissue resembles fibrocartilage. In the anterior cruciate ligament the fibrocartilaginous zone is located 5–10 mm proximal of the tibial ligament insertion in the anterior portion of the ligament. In the posterior cruciate ligament the fibrocartilage is located in the central part of the middle third. Within those zones the cells are arranged in columns and the cell shape is round to ovoid. Transmission electron microscopy reveals typical features of chondrocytes. The chondrocytes are surrounded by a felt-like pericellular matrix, a high content of cellular organelles and short processes on the cell surface. The pericellular collagen is positive for type II collagen. The major blood supply of the cruciate ligaments arises from the middle geniculate artery. The distal part of both cruciate ligaments is vascularized by branches of the lateral and medial inferior geniculate artery. Both ligaments are surrounded by a synovial fold where the terminal branches of the middle and inferior arteries form a periligamentous network. From the synovial sheath blood vessels penetrate the ligament in a horizontal direction and anastomose with a longitudinally orientated intraligamentous vascular network. The density of blood vessels within the ligaments is not homogeneous. In the anterior cruciate ligament an avascular zone is located within the fibrocartilage of the anterior part where the ligament faces the anterior rim of the intercondylar fossa. The fibrocartilaginous zone of the middle third of the posterior cruciate ligament is also avascular. According to Pauwel’s theory of the ”causal histogenesis” (1960) the stimulus for the development of fibrocartilage within dense connective tissue is shearing and compressive stress. In the anterior cruciate ligament this biomechanical situation may occur when the ligament impinges on the anterior rim of the intercondylar fossa when the knee is fully extended. Compressive and shearing stress in the center of the middle third of the posterior cruciate ligament may result from twisting of the fiber bundles. Accepted: 12 March 1999     

Blood and lymph supply of the posterior cruciate ligament: a cadaver study

Injection techniques, immunohistochemical (antibodies against laminin), and histochemical (5′-nucleotidase activity) methods were employed to describe the vascular pattern of the human posterior cruciate ligament (PCL); in parallel we used conventional light microscopy and immunohistochemical techniques to visualize the histological structure of the PCL. The blood supply of the PCL mainly arises from the middle geniculate artery. The ligament is covered by a synovial fold where the terminal branches of the middle geniculate artery form a periligamentous network. From the synovial sheath, the blood vessels penetrate the ligament in a horizontal direction and anastomose with a longitudinally orientated intraligamentous network. Within the ligament the blood vessels are located in the loose connective tissue that is sited between longitudinal fibre bundles. Histologically the longitudinal fibre bundles of the ligament consist of dense connective tissue. Lymphatics accompany most of the smaller blood vessels, showing similar regional distribution. Compared to the surrounding synovial layer, the amount of vessels in the substance of the ligament is lower. The distribution of blood vessels within the PCL is not homogenous: we detected three avascular areas within the ligament. Both fibrocartilaginous entheses of the PCL are devoid of blood vessels, and a third avascular zone is located in the central part of the middle third. The histological structure of this zone varies from the rest of the PCL which consists of the characteristic dense connective tissue. In the central part of the PCL the tissue resembles fibrocartilage: the cell shape is round, the pericellular matrix of those cells is rich in acid glycosaminoglycans and the immunohistochemical demonstration of type II collagen is positive. The occurrence of an avascular zone within the central part of the middle third of the PCL where the tissue consists of fibrocartilage has not been described in the literature. Received: 10 March 1998 Accepted: 8 July 1998     

应用Laminin促进周围神经再生的研究

目的探讨局部应用Laminin（LN）对周围神经再生的影响。方法将34只SD大鼠分为4组,1～3组每组10只,手术切除双侧10mm坐骨神经并用硅胶管套接,左侧套管内注入LN0．6μg,右侧注入生理盐水作对照。术后1、3、4个月分别进行电生理和组织学检查。另4只鼠于术后3个月行辣根过氧化物酶逆行示踪检查。结果LN治疗组术后1、3、4个月时的神经肌肉动作电位、运动神经传导速度、再生有髓神经纤维数量及髓鞘厚度均明显优于对照组。相应节段脊髓前角和背根神经节标记神经元明显多于对照组。结论LN可促进周围神经损伤后的再生。     

血清HA、PLD、PCⅢ、LN、Ⅳ—C联合检测对慢性肝病的诊断价值探讨

目的:寻找肝纤维化等慢性肝病敏感可靠的血清学诊断指标。方法:对196例各型肝病联合检测血清透明质酸(HA)、脯氨酸肽酶(PLD)、Ⅲ型前胶原(PCⅢ)、层粘蛋白(LN)、Ⅳ型胶原(Ⅳ-C),并与10例肝组织学和30例健康献血员做对照研究。结果:五项指标分别在急性肝炎、慢性肝炎和肝硬变中依次升高,差异显著(P<0.05或P<0.01),其改变情况与轻、中、重型纤维化程度呈同步性,有良好的正相关。结论:五项指标联合检测可极大地增加慢性肝病临床诊断的准确性与可靠性,动态观察其变化可掌握病情演变。     

脑脊液中TGF-β1、PICP、PⅢNP、HA、LN在蛛网膜下腔出血后脑积水发生中的意义

目的探讨蛛网膜下腔出血后转化生长因子β1(Transforming growth factor-β1,TGF-β1)、Ⅰ型前胶原前肽(procollagenⅠCpropeptide,PICP)、Ⅲ型前胶原前肽(procollagenⅢN-propeptide,PⅢNP)、透明质酸(hyaluronic acid,HA)及层黏蛋白(laminin,LN)在脑积水形成的过程中的作用。方法收集我院颅脑损伤及自发性颅内出血(波及到蛛网膜下腔及脑室系统)的患者,入院时不同时间点检查患者脑脊液中TGF-β1、PICP、PⅢNP、HA、LN的水平,同时随访上述患者在半年内发生脑积水的情况,比较出现与未出现脑积水患者脑脊液中TGF-β1、PICP、PⅢNP、HA、LN水平的差异。分析患者脑脊液中TGF-β1分别与PICP、PⅢNP、HA、LN表达的相关性及其时间-浓度关系。结果收集病例83例,随访半年,53例未出现脑积水(A组),30例出现脑积水(B组),结果显示,出现脑积水的患者在脑脊液各个时间点的TGF-β1、PICP、HA、LN水平均显著高于未出现脑积水的患者。83例患者脑脊液中TGF-β1与PICP、PⅢNP、HA、LN的表达呈正相关,相关系数分别为0.8248、0.7951、0.9078、0.7572。脑脊液中TGF-β1在7天时达到高峰期,之后逐渐下降,而PICP、PⅢNP、HA、LN在脑脊液中的表达是在TGF-β1达到高峰后快速上升,后缓慢下降。结论脑脊液中TGF-β1、PICP、PⅢNP、HA、LN与脑脊液系统出血后慢性脑积水的发生过程中发挥着重要作用。TGF-β1在脑脊液系统出血后的表达可能促进了PICP、PⅢNP、HA、LN的表达。     

肝纤维化血清学指标水平与肝癌的相关性研究

目的 研究血清Ⅲ型前胶原、Ⅳ型胶原、层粘连蛋白和透明质酸在肝癌诊断中的价值.方法 采用放射免疫法测定52例原发性肝癌、34例转移性肝癌、34例肝外恶性肿瘤患者及100例健康体检人员血清Ⅲ型前胶原、Ⅳ型胶原、层粘连蛋白和透明质酸水平,并进行分析比较.结果 治疗前和治疗1、3个月后,原发性肝癌组Ⅲ型前胶原、Ⅳ型胶原、层粘连蛋白、血清透明质酸水平(计量单位为μg/L)分别为(176±43)、(166±38)、(129±38);(139±34)、(108±31)、(85±32);(123±23)、(100±23)、(93±18);(347±136)、(299±120)、(266±113)μg/L,转移性肝癌组分别为(97±25)、(93±22)、(79±16);(96±31)、(80±28)、(77±19);(93±24)、(80±21)、(62±15);(254±108)、(199±98)、(132±93)μg/L.原发性肝癌组血清透明质酸和Ⅳ型胶原水平在治疗前与对照组[(67±11)、(55±15)μg/L]比较差异均有统计学意义(均P＜0.01),Ⅲ型前胶原和层粘连蛋白水平明显高于对照组[(55±11)、(54±13)μg/L,P＜0.05)].转移性肝癌组及肝外恶性肿瘤组治疗前透明质酸[(149±68)μg/L]与对照组比较,差异均有统计学意义(均P＜0.05).原发性肝癌组治疗前后血清Ⅲ型前胶原、Ⅳ型胶原、层粘连蛋白和透明质酸水平均明显高于同一时间转移性肝癌组的水平(均P＜0.05).原发性肝癌组各项指标治疗前和治疗1、3个月后之间相比均差异有统计学意义(均P＜0.05).结论 血清Ⅲ型前胶原、Ⅳ型胶原、层粘连蛋白和透明质酸水平与原发性肝癌具有相关性,可作为反映原发性肝癌严重程度和预后的指标.

Abstract：

Objective To investigate the significance of serum fibrosis markers in liver fibrosis in liver neoplasms. Methods Radioimmunoassay was used to measure the level of serum procollagen (PC) Ⅲ, collagen type Ⅳ(Ⅳ-C), laminin LN and hyaluronic acid(HA) in 120 patients with liver neoplasms and 100 healthy people.Results Before therapy, the level of Ⅳ-C in the hepatocellular carcinoma group showed significant difference compared with those in control group (P＜0. 01 ). The levels of PCⅢ and LN in the hepatocellular carcinoma group were higher than those in control group ( P ＜ 0.05 ). Before therapy the levels of HA in metastatic cancer of liver group and extrahepatic malignant tumor group were higher than those in control group ( P ＜ 0. 05 ). Before and after the therapy the levels of hepatocellular carcinoma group serum PC Ⅲ, Ⅳ-C, LN and HA were significantly higher than those in the metastatic cancer of liver group ( P ＜ 0. 05). Before and after the therapy the levels of hepatocellula carcinoma group serum PC Ⅲ, Ⅳ-C, LN and HA were significantly difference ( P ＜ 0.05 ). Conclusions The levels of serum PC Ⅲ, Ⅳ-C, LN and HA are significantly related with the hepatocellular carcinoma. These markers can reflect the fibrosis degree of chronic hepatocellular carcinoma and will be an indicator of prognosis.     

Serum surrogate markers of liver fibrosis in primary biliary cirrhosis

Background

Hyaluronan, leptin, laminin and collagen IV have been used extensively for the assessment of liver fibrosis. The aim of this study was to assay these markers in the peripheral and hepatic vein blood of primary biliary cirrhosis (PBC) patients and to study their ability to discriminate early from advanced disease.

Methods

Sera from 62 PBC patients were compared to 60 controls, 44 chronic Hepatitis C, 38 hepatocellular carcinoma and 34 viral cirrhosis patients. Serum from the hepatic vein of 15 cirrhotic PBC patients and 17 patients with viral cirrhosis was also assayed.

Results

All disease groups had significantly increased levels of hyaluronan and collagen IV, compared to controls, while laminin was significantly increased only in viral cirrhosis. Hyaluronan levels were statistically different between early (54.5 ng/ml; 95%CI 27.3-426.9) and late PBC (154.5 ng/ml; 95%CI 55.3-764.4, p < 0.05). The area under the curve (AUC) for the identification of late PBC was 0.74 for hyaluronan, 0.63 for leptin, 0.59 for laminin and 0.70 for collagen IV. Hyaluronan had high sensitivity and NPV in identifying late stages of PBC (96% and 90%, respectively). Short term UDCA had no effect on these markers.

Conclusion

No single measurement can differentiate between advanced and early fibrosis in PBC. However serum hyaluronan is a promising single serum marker for longitudinal studies in PBC.     

Sequential neuroradiological and neurophysiological studies in a Japanese girl with merosin-deficient congenital muscular dystrophy

We describe the early manifestation and sequential assessment of the central and peripheral nervous system in a Japanese girl with merosin-deficient congenital muscular dystrophy. She showed severe hypotonia (‘‘floppy infant”) and suffered mild respiratory failure postnatally. Serum creatine kinase was elevated to 11,487 IU/L. The muscle biopsy showed dystrophic changes with negative expression of merosin (laminin α2), thereby confirming merosin-deficient congenital muscular dystrophy. Her motor milestones were severely delayed, but she could sit without support at the age of 3 years. After 3 years, her motor ability deteriorated and by the age of 5 years, she could not sit and control her neck. Magnetic resonance imaging (MRI) at 2 months of age revealed patterns that were appropriate for her age. At 1 year of age, the T2 weighted images showed diffuse high signal intensities throughout the centrum semiovale, and periventricular and subcortical white matter of the frontal and occipital lobes, while the U fibers, the corpus callosum and the internal capsule were spared. At the age of 7 years, these white matter abnormalities decreased. MR spectroscopy (MRS) revealed normal values of N-acetylaspartate (NAA)/creatine (Cr) and choline (Cho)/Cr metabolite ratios as well as slightly increased myoinositol (mI)/Cr metabolite ratios. Neurophysiological motor nerve conduction velocity (MCV) and compound muscle action potential (CMAP) of the median nerve were in the normal range at the age of 2 months. After the child reached 1 year of age, the MCV and CMAP lagged behind those of healthy controlled children. The sensory nerve conduction velocity of the median nerve demonstrated a mild delay at the age of 15 months. It improved to normal range at the age of 6 years but decreased at 7 years of age. These sequential findings suggest not only that muscular degeneration and dysmyelination had occurred but also that various other factors, including demyelination and the vasogenic system, may influence the pathology of MDC1A.