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Distribution of pericellular matrix molecules in the temporomandibular joint and their chondroprotective effects against inflammation

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International Journal of Oral Science 2017年第1期9卷 43-52页

作者： Wern Cui Chu Shipin Zhang Timothy J Sng Yu Jie Ong Wen-Li Tan Vivien Y Ang Casper B Foldager Wei Seong Toh Faculty of Dentistry National University of SingaporeSingapore Orthopaedic Research Laboratory Aarhus University HospitalAarhusDenmark Tissue Engineering Program Life Sciences InstituteNational University of SingaporeSingapore

The objectives of this study were to （1） determine the distribution and synthesis of pericellular matrix （PCM） molecules （collagen VI, collagen IV and laminin） in rat temporomandibular joint （TMJ） and （2） investigate the effects of PCM molecules on chondrocytes against inflammation in osteoarthritis. Four zones （fibrous, proliferating, mature and hypertrophic） of condylar cartilage and three bands （anterior, intermediate and posterior） of disc were analysed by immunohistochemistry for the presence of PCM molecules in rat TMJs. Isolated chondrocytes were pre-treated with PCM molecules before being subjected to interleukin （IL）-II~ treatment to stimulate inflammation. The responses of the chondrocytes were analysed using gene expression, nitric oxide release and matrix metalloproteinase （MMP）-13 production measures. Histomorphometric analyses revealed that the highest areal deposition of collagen VI （67.4%）, collagen IV （45.7%） and laminin （52.4%） was in the proliferating zone of TMJ condylar cartilage. No significant difference in the distribution of PCM molecules was noted among the three bands of the TMJ disc. All three PCM molecules were expressed intracellularly by chondrocytes cultured in the monolayer. Among the PCM molecules, pre-treatment with collagen VI enhanced cellular proliferation, ameliorated IL-lp-induced MMP-3, MMP-9, MMP-13 and inducible nitric oxide synthase gene expression, and attenuated the downregulation of cartilage matrix genes, including collagen I, aggrecan and cartilage oligomeric matrix protein （COMP）. Concurrently, collagen VI pretreatment inhibited nitric oxide and MMP-13 production. Our study demonstrates for the first time the distribution and role of PCM molecules, particularly collagen VI, in the protection of chondrocytes against inflammation.

关键词： cartilage chondrocytes collagen iV collagen W inflammation laminin pericellular matrix temporomandibular joint

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Effects of in vitro cultivated Calculus Bovis compound on pulmonary lesions in rabbits with schistosomiasis

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World Journal of Gastroenterology 2010年第6期16卷 749-754页

作者： Li, Tao Yang, Zhen Cai, Hong-Jiao Song, Li-Wei Lu, Ke-Yu Zhou, Zheng Wu, Zai-De China Mei Tan Gen Hosp Dept Gen Surg & Oncol Beijing 100028 Peoples R China Huazhong Univ Sci & Technol Dept Gen Surg Tongji Hosp Tongji Med Coll Wuhan 430030 Hubei Province Peoples R China

AIM:To explore the interventional effects and mechanism of in vitro cultivated Calculus Bovis compound preparation(ICCBco) on pulmonary lesions in portal hypertensive rabbits with schistosomiasis. METHODS:The experimental group included 20 portal hypertensive rabbits with schistosomiasis treated by ICCBco.The control group included 20 portal hypertensive rabbits with schistosomiasis treated by praziquantel. The morphological changes of the pulmonary tissues were observed under light and electron microscopy.The expression of fibronectin(FN) and laminin(LN) in the lung tissues was analyzed by immunohistochemistry. RESULTS:Under light microscope,the alveolar exudation in the lung tissue was more frequently observed in the control group,while the alveolar space was fairly dry in the lung tissue of ICCBco group.Under electron microscope,more alveolar exudation in the lung tissue,and moremacrophages,alveolar angiotelectasis and the blurred three-tier structure of alveolar-capillary barrier could be seen in the control group.In ICCBco group,fibers within the alveolar interspace slightly increased in some lung regions,and the structure of typeⅠepithelium,basement membrane and endodermis was complete,and no obvious exudation from the alveolar space,and novascular congestion could be observed.There was a positive or strong positive expression of FN and LN in the lung tissue of the control group,while there was a negative or weak positive expression of FN and LN in ICCBco group. CONCLUSION:ICCBco can effectively prevent pulmonary complications in portal hypertensive rabbits with schistosomiasis by means of improving lung microcirculation and lowering the content of extracellular matrix.

关键词： In vitro cultivated Calculus Bovis compound preparation Schistosomiasis Portal hypertension Lung lesion Fibronectin laminin Pulmonary microcirculation

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Comparative study on the stem cell phenotypes of C6 cells under different culture conditions

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Chinese Medical Journal 2011年第19期124卷 3118-3126页

作者： ZHANG Suo-jun YE Fei XIE Rui-fan HU Feng WANG Bao-feng WAN Feng GUO Dong-sheng LEI Ting Department of Neurosurgery and Neuro-Oncology Molecular Biology Laboratory Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei 430030 China

Background Glioma stem cell （GSC） hypothesis posits that a subpopulation of cells within gliomas have true clonogenic and tumorigenic potential. Significantly, a more controversial correlate to GSC is that cells in different culture conditions might display distinct stem cell properties. Considering these possibilities, we applied an approach comparing stem cell characteristics of C6 glioma cells under different culture conditions.Methods C6 cells were cultured under three different growth conditions, i.e., adherent growth in conventional 10% serum medium, non-adherent spheres growth in serum-free medium, as well as adherent growth on laminin-coated flask in serum-free medium. Growth characteristics were detected contrastively through neurosphere formation assay and cell cycle analysis. Markers were determined by immunofluorescence, relative-quantitative reverse transcription （RT）-PCR,Western blotting and flow cytometry. Side population cells were analyzed via flow cytometry. Tumor models were detected by magnetic resonance imaging and hematoxylin ＆ eosin staining. Data analyses were performed with SPSS software （17.0）.Results C6 cells （C6-Adh, C6-SC-Sph and C6-SC-Adh） showed distinctive growth patterns and proliferation capacity.Compared to suspending C6-SC-Sph, adherent C6-Adh and C6-SC-Adh displayed higher growth ratio. C6-SC-Sph and C6-SC-Adh showed enhanced capability of neurosphere formation and self-renewal. High side population ratio was detected in C6-SC-Sph and C6-SC-Adh. CD133 was not detected in all three kinds of cells. Conversely, Nestin and β-Ⅲ-tubulin were demonstrated positive, nonetheless with no statistical significance （P 〉0.05）. Interestingly, lower expression of glial fibrillary acidic protein was demonstrated in C6-SC-Sph and C6-SC-Adh. C6-Adh, C6-SC-Sph and C6-SC-Adh were all displayed in situ oncogenicity, while statistical difference of survival time was not confirmed.Conclusions C6 glioma cell line is endowed with some GSC phenotypes that can be moderately enriched in vitro when transfe

关键词： C6 glioma stem cell neurosphere CD133 side population laminin

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Expression of extracellular matrix proteins and vimentin in testes of azoospermic man： an immunohistochemical and morphometric study

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Asian Journal of Andrology 2002年第1期4卷 55-60页

作者： GulkKH ErdoT DepartmentofBiostatistics DepartmentofUrology

OBJECTIVE:To investigate the changes in the extracellular matrix protein expression and the morphology of seminiferous tubules in the testis of 88 azoospermic men.;METHODS:The patients were of the following categories: (1) 22 cases of Sertoli-cell-only syndrome, (2) 20 cases of spermatogenic arrest, and (3) 46 cases with hypospermatogenesis. Testicular sections were immunohistochemically stained for fibronectin, vimentin, laminin and collagen type IV. The seminiferous tubular diameter and the connective matrix zone (CMZ, the acellular zone between the basement membrane [BM] and the peritubular cells) thickness were measured. Seminiferous tubules were typed according to the thickness of the connective matrix in the lamina propria. The predominant tubule type and the Johnsen and Silber scores were determined.;RESULTS:The mean tubular diameter were 119 +/- 27, 117 +/- 20, and 140 +/- 38 microm for Groups 1, 2, and 3, respectively. Both the laminin and the type IV collagen were localized to the epithelial BM and peritubular cells. In most of the tubules, BM and peritubular cells were separated by a homogenous acellular layer, the CMZ, in which laminin, type IV collagen, fibronectin and vimentin were not present. It is perceived that the worse the testicular histology, the higher the thickness of the CMZ.;CONCLUSIONS:In testis with no or low sperm production, the diameter of the seminiferous tubules is decreased, the thickness of the seminiferous tubular wall is increased and a CMZ is formed between the peritubular cells and the BM. The thickness of CMZ is increasing with the advancement of testiclar deterioration. The most important morphologic predictive factor for spermiogenesis is the predominant

关键词： Azoospermia Collagen Extracellular matrix Fibronectins Immunohistochemistry laminin Male infertility Seminiferous tubules Testis Vimentin

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应用laminin促进周围神经再生的研究

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中华实验外科杂志 1999年第3期16卷 244-245页

作者：张键陈中伟张光健陈峥嵘上海医科大学附属中山医院骨科 200032

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... 详细信息

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。

关键词： laminin 周围神经损伤神经再生

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出生前后不同生长发育期狨猴(Callithrix jacchus)肝脏Ⅳ、Ⅵ型胶原,laminin和Nidogen的免疫荧光定位

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东北师大学报(自然科学版) 1993年第3期 93-98页

作者：郭玉莹郭焱高尚民李大宁徐大群吉林医学院吉林省火电公司职工医院德国柏林自由大学

应用免疫荧光定位法观察狨猴不同生长发育期肝脏内 IVVI 型胶原.laminin 和 Nidogen 的分布。在窦壁和肝小叶外的所有基板中,各时期均能见到大量的 IV 型胶原。VI 型胶原在早期胎儿窦壁显示微弱荧光,而后期荧光较强并持续到成年,在门静... 详细信息

应用免疫荧光定位法观察狨猴不同生长发育期肝脏内 IVVI 型胶原.laminin 和 Nidogen 的分布。在窦壁和肝小叶外的所有基板中,各时期均能见到大量的 IV 型胶原。VI 型胶原在早期胎儿窦壁显示微弱荧光,而后期荧光较强并持续到成年,在门静脉周它主要定位于小叶与结缔组织的交界处。laminin 数量很少,只是在出生前较明显。相反,Nidogen 出现于各期。因此,IV、VI 型胶原不适于做病理指标.而在成年肝脏消失的 laminin 的重新出现可做为病理指征。

关键词： Ⅳ、Ⅵ型胶原 laminin Nidogen 免疫荧光肝脏

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Non-invasive diagnostic screening of hepatic fibrosis in patients with chronic hepatitis B

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Chinese Medical Journal 2004年第7期117卷 1109-1112页

作者：陈永鹏冯筱榕戴琳章廉侯金林 Department of Infectious Diseases

Liver cirrhosis is usually associated with complications which can be effectively prevented. In the case of compensated cirrhosis, early diagnosis is essential. Liver biopsies are used as the primary method for the diagnosis of compensated cirrhosis. In clinical practice, the use of liver biopsies has several limitations including complications, which lead to a death rate of 0.015%, discomfort, and longer hospitalization. These limitations prevent the use of liver biopsies as a general screening procedure for cirrhosis.

关键词： chronic hepatitis B liver cirrhosis hyaluronic acid collagen type Ⅲ laminin collagen type Ⅳ ultrasonography

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p63、Ki-67及laminin在泪腺上皮性肿瘤的表达及意义

p63、Ki-67及Laminin在泪腺上皮性肿瘤的表达及意义

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作者：杨娜河北医科大学

学位级别：硕士

目的:通过观察p63蛋白、细胞增殖核蛋白Ki-67及粘附因子层粘连蛋白(laminin,LN)在泪腺良、恶性肿瘤的表达情况,分析这些生物学指标与泪腺良、恶性肿瘤的临床和组织病理的关系,进一步探明它们在泪腺良、恶性肿瘤发生、发展中的作用,为在... 详细信息

目的:通过观察p63蛋白、细胞增殖核蛋白Ki-67及粘附因子层粘连蛋白(laminin,LN)在泪腺良、恶性肿瘤的表达情况,分析这些生物学指标与泪腺良、恶性肿瘤的临床和组织病理的关系,进一步探明它们在泪腺良、恶性肿瘤发生、发展中的作用,为在泪腺良、恶性肿瘤的鉴别诊断、治疗以及预后判断提供依据。方法:收集武警总医院眼眶病研究所于2003年1月至2006年1月手术切除并经病理组织学检查确诊的34例泪腺多形性腺瘤(pleomorphic adenoma,PA)、29例泪腺腺样囊性癌(adenoid cystic carcinoma,ACC)的石蜡标本,利用免疫组织化学染色SP法,观察p63、Ki-67及laminin在泪腺良、恶性肿瘤中的表达情况,并以8例正常泪腺组织作为对照,采用SPSS13.0统计软件包进行统计分析。结果:1 p63在正常泪腺及泪腺良、恶性肿瘤中的表达分别为31.13±9.67、34.65±24.95、20.20±18.53。经统计学分析,泪腺PA与正常泪腺和泪腺ACC组间均有显著性差异(P0.05)。2 Ki-67在正常泪腺及泪腺良、恶性肿瘤中的表达分别为0、6.65±4.78、37.35±23.11。Ki-67LI依次升高。经统计学分析,三组间有显著性差异(Plaminin在正常泪腺及泪腺良、恶性肿瘤中的阳性表达率分别为8例正常泪腺组织标本中有2例(25.00%)标本中有表达,34例泪腺PA标本中有11例( 32.35%)标本中有表达,29例泪腺ACC标本中有15例( 51.72%)标本中有表达。经统计学分析三组间有显著性差异(Plaminin表达的高低与泪腺ACC预后的关系。4.1 p63在术后复发组的表达为14.11±6.31,未复发组为21.65±8.93,统计学分析有显著性差异(Plaminin在术后复发组的阳性表达率为25.00%(3/12),未复发组阳性表达率为85.71%(12/14),统计学分析有显著性差异(Plaminin在泪腺上皮性肿瘤中的表达经统计学分析均无显著性差异(P>0.05)。结论:1 p63显示泪腺肌上皮细胞具有相对较高的敏感性和特异性,可作为肌上皮细胞的又一有效的标记物。p63蛋白表达的强度在良性肿瘤较强,恶性肿瘤最弱,提示p63蛋白表达可能与肿瘤的分化相关。p63在恶性肿瘤中的高表达可能能抑制肿瘤的复发和转移。2 Ki-67可能参与了泪腺肿瘤的发生、发展,可作为鉴别泪腺良、恶性疾病的一种辅助免疫标记物。也是判断泪腺肿瘤的增殖能力,反映其生物学行为的功能性指标。Ki-67在泪腺ACC组织中的表达与其临床预后有关,Ki-67的高表达预示着泪腺ACC患者的预后较差。3 LN可能参与了泪腺肿瘤的发生、发展,可作为鉴别泪腺良、恶性疾病的一种辅助免疫标记物。LN的表达类型可作为判定泪腺ACC的分化程度及患者预后的一个独立指标。4性别、年龄及肿瘤的大小对p63、Ki-67和LN在泪腺上皮性肿瘤中的表达无明显影响。

关键词：泪腺上皮性肿瘤 p63 Ki-67 laminin 免疫组织化学

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Improvement of human embryonic stem cell-derived retinal pigment epithelium cell adhesion, maturation, and function through coating with truncated recombinant human vitronectin

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International Journal of Ophthalmology(English edition) 2021年第8期14卷 1160-1167页

作者： Xin-Yue Zhu Yu-Hong Chen Ting Zhang Su-Jun Liu Xin-Yue Bai Xian-Yu Huang Mei Jiang Xiao-Dong Sun Department of Ophthalmology Shanghai General HospitalShanghai Jiao Tong University School of MedicineShanghai 200080China National Clinical Research Center for Eye Diseases Shanghai 200080China Shanghai Key Laboratory of Fundus Diseases Shanghai 200080China Shanghai Engineering Center for Visual Science and Photomedicine Shanghai 200080China

AIM:To explore an xeno-free and defined coating substrate suitable for the culture of H9 human embryonic stem cell-derived retinal pigment epithelial(hES-RPE)cells in vitro,and compare the behaviors and functions of h ESRPE cells on two culture substrates,laminin521(LN-521)and truncated recombinant human vitronectin(VTN-N).METHODS:hES-RPE cells were used in the experiment.The abilities of LN-521 and VTN-N at different concentrations to adhere to hES-RPE cells were compared with a high-content imaging system.Quantitative real-time polymerase chain reaction was used to evaluate RPE-specific gene expression levels midway(day 10)and at the end(day 20)of the time course.Cell polarity was observed by immunofluorescent staining for apical and basal markers of the RPE.The phagocytic ability of hES-RPE cells was identified by flow cytometry and immunofluorescence.RESULTS:The cell adhesion assay showed that the ability of LN-521 to adhere to hES-RPE cells was dosedependent.With increasing coating concentration,an increasing number of cells attached to the surface of LN-521-coated wells.In contrast,VTN-N presented a strong adhesive ability even at a low concentration.The optimal concentration of LN-521 and VTN-N required to coat and adhesion to hES-RPE cells were 2 and 0.25μg/cm^(2),respectively.Furthermore,both LN-521 and VTN-N could facilitate adoption of the desired cobblestone cellular morphology with tight junction and showed polarity by the hES-RPE cells.However,hES-RPE cells cultivated in VTN-N had a greater phagocytic ability,and it took less time for these hES-RPE cells to mature.CONCLUSION:VTN-N is a more suitable coating substrate for cultivating hES-RPE cells.

关键词： retinal pigment epithelial stem cell surface coating laminin vitronectin cell adhesion phagocytosis

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胃肠道类癌中Ki-67、PCNA、laminin的表达及意义

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临床与实验病理学杂志 2004年第6期20卷 712-714页

作者：李淑霞王立峰戚基萍哈尔滨医科大学附属第一医院病理科

目的　研究胃肠道类癌Ki 6 7、PCNA、laminin(LN)的表达与侵袭性和增殖性的关系。方法　应用免疫组织化学S P法检测 36例胃肠道类癌组织中 3种指标的表达。结果　 36例胃肠道类癌组织中Ki 6 7、PCNA、LN阳性表达率分别为5 0 0 %、5 5 ... 详细信息

目的　研究胃肠道类癌Ki 6 7、PCNA、laminin(LN)的表达与侵袭性和增殖性的关系。方法　应用免疫组织化学S P法检测 36例胃肠道类癌组织中 3种指标的表达。结果　 36例胃肠道类癌组织中Ki 6 7、PCNA、LN阳性表达率分别为5 0 0 %、5 5 5 %、5 5 5 % ;随肿瘤分化程度降低、浸润深度加深、淋巴结的转移 ,各指标阳性表达率逐渐升高。结论　Ki 6 7、PC NA可作为评估胃肠道类癌预后的指标。Ki 6 7比PCNA在评价胃肠道类癌增殖性方面更准确 ;PCNA免疫染色反应性较敏感 ,其类癌诊断的阳性标准应适当提高。LN的表达可预测胃肠道类癌组织的分化。

关键词：胃肠肿瘤类癌 Ki-67 PCNA laminin 侵袭增殖免疫组织化学

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