兔关节软骨细胞原代培养及生物学鉴定

目的探讨兔膝关节软骨细胞体外分离培养及鉴定方法,分析软骨细胞在体外不同阶段的生物学特性。方法取4周龄健康新西兰兔膝关节软骨,以机械分离和酶消化相结合的方法获取膝关节软骨细胞,经体外培养、传代。观察不同培养阶段细胞形态和生长,CCK-8法测定细胞增殖并绘制生长曲线,甲苯胺蓝和Ⅱ型胶原免疫荧光染色鉴定软骨细胞。结果兔软骨细胞经两步消化,可获取高纯度的软骨细胞,细胞活性率可达95%以上,甲苯胺蓝染色及免疫荧光呈阳性结果。原代细胞和第一代细胞呈多角形,传代4~5代后,细胞形态逐渐由多角形变为梭形,基质分泌氨基多糖和胶原染色变浅。结论成功建立了简单易行的软骨细胞分离、培养方法,通过传代可以获取大量实验细胞,但仅限于5代以内软骨细胞适用于组织工程研究。     

体外培养大鼠软骨细胞退变现象的连续观察

目的对体外培养大鼠软骨细胞退变现象进行连续观察，为进一步研究软骨退变机制实验及药物干预实验提供基础资料。方法进行大鼠软骨细胞取材并传代培养，对各代次细胞进行连续性观察。采用倒置相差显微镜、透射电镜观察细胞形态与微结构，免疫组化法检测增殖细胞核抗原（PCNA），阿力新蓝染色检测胞外基质硫酸糖氨多糖（GAG）含量和结构，反转录-聚合酶链式反应方法检测软骨细胞Ⅱ型胶原和Aggrecan表达，观察软骨细胞退变。结果第四代软骨细胞出现长梭形改变，电镜下超微结构出现核固缩或水肿、细胞器减少或变形等退变表现，PCNA表达百分率明显减少（P〈0．01），GAG含量、长链分子百分比显著减少（P〈0．01），Ⅱ型胶原及Aggrecan mRNA表达显著减少（P〈0．01）。结论体外培养软骨细胞第四代出现明显功能退变。     

兔外周血间充质干细胞的分离培养及诱导分化

目的 研究间充质干细胞(MSCs)在创伤修复中的作用并为MSCs作为组织工程种子细胞奠定实验基础.方法 抽取创伤后兔外周血,用密度梯度离心法分离纯化单个核细胞;α-MEM培养基培养,并测定细胞生长曲线;应用成骨和成软骨诱导剂定向诱导,检测分化能力.结果 原代培养的MSCs呈圆形、梭形、多角形,传代纯化后呈均一的长梭形.其生长曲线均呈典型的"S"形.外周血MSCs成骨诱导后7 d碱性磷酸酶染色阳性;21 d出现钙沉积,茜素红染色呈阳性.成软骨诱导2周后Ⅱ胶原免疫细胞化学染色阳性.结论 皮肤损伤刺激后可从外周血中获取MSCs,体外分离培养的MSCs具备向成骨及成软骨细胞分化特性;可用于组织工程种子细胞的研究.     

吖啶橙染色法用于湖北钉螺血淋巴细胞的观察

将湖北钉螺软体组织挤压后获取血淋巴液制成涂片,采用荧光染料吖啶橙染色,荧光显微镜下观察血淋巴细胞。结果可见12种形态的血淋巴细胞:红色多浆型无颗粒圆形淋巴细胞、红色多浆型有颗粒圆形淋巴细胞、红色少浆型无颗粒圆形淋巴细胞、红色少浆型有颗粒圆形淋巴细胞、绿色多浆型无颗粒圆形淋巴细胞、绿色多浆型有颗粒圆形淋巴细胞、绿色少浆型无颗粒圆形淋巴细胞、绿色少浆型有颗粒圆形淋巴细胞、多浆伸展型淋巴细胞、少浆伸展型淋巴细胞、红色胞质梭形淋巴细胞、绿色胞质梭形淋巴细胞,表明该方法可用于湖北钉螺血淋巴细胞的染色,为探讨贝类免疫防御、研究灭螺药物或各种生物与理化因素对湖北钉螺血淋巴细胞的影响提供技术支持。     

鹿瓜多肽联合hBMP-2基因对软骨细胞增殖和Ⅱ型胶原分泌的影响

目的 观察鹿瓜多肽联合hBMP-2基因对软骨细胞的增殖和软骨细胞分泌Ⅱ型胶原是否有协同作用.方法 分离培养原代兔软骨细胞,用第3代细胞为模型,实验分为未转染组、腺病毒-绿色荧光蛋白(Ad-GFP)组、Ad-hBMP2-GFP组、鹿瓜多肽+Ad-hBMP2-GFP组,培养48 h后.RT-PCR检测hBMP-2mRNA的表达,流式细胞仪检测软骨细胞的周期.培养48、72 h后.MTT法检测软骨细胞的增殖,Western印迹检测hBMP-2蛋白和Ⅱ型胶原的表达.结果 培养48 h后,Ad-hBMP2-GFP组和鹿瓜多肽+Ad-hBMP2-GFP组hBMP-2mRNA呈阳性,而未转染组、Ad-GFP组表达呈阴性;与未转染组、Ad-GFP组、Ad-hBMP2-GFP组比较,鹿瓜多肽+hBMP-2基因组能使软骨细胞S-G2/M期细胞比例增加,软骨细胞数量增加(P<0.05),软骨细胞hBMP-2蛋白表达和Ⅱ型胶原分泌增加(P<0.01);培养48与72 h比较,鹿瓜多肽联合hBMP-2基因对软骨细胞增殖和Ⅱ型胶原分泌无显著性差异(P>0.05).结论 鹿瓜多肽能促进hBMP-2基因在软骨细胞中合成hBMP-2蛋白,鹿瓜多肽联合hBMP-2基因对软骨细胞增殖和分泌Ⅱ型胶原的功能有协同作用.     

人胎肝干细胞体外分离、培养及鉴定

研究人胎肝干细胞体外分离纯化方法，并对其进行初步鉴定及生物学特性分析。采用胶原酶消化、重力沉降及密度梯度离心方法分离人胎肝干细胞，通过免疫细胞化学方法对其进行初步鉴定，以及应用流式细胞仪等对其生长状况进行评估。所获得的原代细胞共传8代，细胞呈小梭形或三角形，体积较小，胞核较大，而胞浆较少；第4代细胞中进入生长期的细胞约占88．2％；免疫细胞化学染色显示细胞胞质中ALB、CK19染色阳性。人胎肝中存在具有肝干细胞特征的细胞，它们可能会成为细胞移植及肝移植良好的细胞资源。     

Ad-hBMP-2对促进软骨细胞分泌Ⅱ型胶原影响的实验研究

目的观察腺病毒-人骨形态发生蛋白-2-绿色荧光蛋白(Ad-hBMP-2-GFP)转染兔软骨细胞后,hBMP-2蛋白的表达和其对软骨细胞分泌Ⅱ型胶原功能的影响。方法包装hBMP-2-GFP腺病毒载体,测定病毒滴度。分离培养并鉴定原代兔软骨细胞,通过基因转染技术,转染Ad-hBMP-2-GFP至兔第三代软骨细胞,RT-PCR检测hBMP-2mRNA的表达,细胞免疫荧光法和Western印迹检测软骨细胞中hBMP-2蛋白的表达和Ⅱ型胶原的变化情况,用未转染组和空病毒组(Ad-GFP)做对照。结果转染后,hBMP-2的mRNA在48h有明显阳性条带表达,对照组呈阴性,hBMP-2蛋白在48、72h的表达量随时间而逐渐增强(P0.05),同时,软骨细胞分泌Ⅱ型胶原的量也逐渐增加(P0.05)。结论Ad-hBMP-2-GFP能成功转染兔软骨细胞,并能促进软骨细胞分泌Ⅱ型胶原,为软骨组织工程提供功能良好的种子细胞,且为hBMP-2基因治疗骨性关节炎提供实验依据。     

成年C57BL／6小鼠空肠Cajal间质细胞的分离、培养及鉴定

目的尝试优化体外培养成年小鼠小肠Cajal间质细胞的操作方法,为深入研究该细胞的生理功能提供一定细胞模型。方法6～8周龄C57BL／6小鼠禁食24h,无菌条件下截取3～5cm中段空肠,分离平滑肌肌条并剪至小块后使用Ⅱ型胶原酶消化20min,离心后进行组织块原代培养,观察不同时期细胞形态。使用c—Kit免疫荧光染色鉴定细胞表型是否为Cajal间质细胞。Fluo-3AM染色细胞内钙,观察细胞能否自发产生钙离子振荡。结果分离所得组织不含空肠上皮和固有层组织,仅为小肠肌层。联合使用胶原酶消化和组织块原代培养能够较方便地培养出原代细胞,该细胞呈梭形,且具有细胞突起,彼此交织成网状。该细胞免疫荧光染色呈c—Kit阳性,平滑肌标志物SMA阴性。钙成像分析显示其能够产生钙离子振荡。结论联合使用酶消化和组织块培养能够较方便地培养出成年小鼠的空肠Cajal间质细胞,该方法为后续探讨Cajal间质细胞的生理功能及机制提供了一定的细胞模型。     

独活寄生汤抑制白细胞介素-1β诱导的软骨细胞凋亡的作用机制

目的探讨独活寄生汤抑制白细胞介素(IL)-1β诱导的软骨细胞凋亡的作用机制。方法从8只SPF级SD大鼠膝关节分离获取软骨细胞并培养,采用Ⅱ型胶原免疫组化染色鉴定。噻唑蓝(MTT)检测不同浓度独活寄生汤对IL-1β诱导的软骨细胞凋亡的影响,从而优化独活寄生汤最佳干预浓度。将第2代软骨细胞随机分为空白组、模型组、独活寄生汤组,空白组以含10%胎牛血清(FBS)的DMEM培养液培养,模型组以10 ng/ml IL-1β和10%FBS的DMEM培养液培养,独活寄生汤组采用300μg/ml独活寄生汤、10 ng/ml IL-1β和10%FBS的DMEM培养液培养。干预48 h,逆转录PCR检测各组软骨细胞中miR-98的表达;Western印迹法检测各组软骨细胞中B细胞淋巴瘤(Bcl)-2、Bcl-2相关X蛋白(Bax)、Caspase-3、Caspase-9蛋白表达。结果Ⅱ型胶原免疫组化染色后阳性组软骨细胞胞质为棕黄色。100、200、300、400μg/ml独活寄生汤均能抑制IL-1β诱导的软骨细胞凋亡,以300μg/ml独活寄生汤最为明显(P0.01)。与模型组比较,独活寄生汤组软骨细胞中miR-98表达明显下降(P0.05)。与模型组比较,独活寄生汤组软骨细胞中的Bcl-2蛋白表达显著升高,而Bax、Caspase-3、Caspase-9蛋白表达明显下降(P0.05)。结论独活寄生汤能够抑制IL-1β诱导的软骨细胞凋亡,可能是通过下调miR-98起作用的。     

体外蓝氏贾第鞭毛虫滋养体的形态学观察

目的观察体外培养的蓝氏贾第鞭毛虫滋养体超微结构,探究贾第虫滋养体的核仁。方法用改良TYI-S-33培养基培养蓝氏贾第鞭毛虫滋养体,姬姆萨(Giemsa)染色和铁苏木素染色后光镜观察虫体的形态结构;以透射电镜观察虫体的超微结构;以核仁特异性抗体作用虫体产生间接免疫荧光,用激光共聚焦显微镜检测核仁。结果光学显微镜下观察姬姆萨染色后的贾第虫胞质为淡监色,可见1对细胞核,核内有分散的染色体,数目为10条;4对鞭毛、1对半月形的中体、轴柱。铁苏木素染色可见1对椭圆形的泡状细胞核内各有1个细小的核仁。透射电镜观察虫体外观颜面状,呈倒置的梨形:质膜下可见单层排列、大小均匀的小囊泡;可见虫体的细胞器:游离核糖体颗粒、内质网;细胞骨架:腹吸盘、鞭毛、基体、中体的微管结构;核结构:核膜、核孔、核仁、染色体。激光共聚焦显微镜观察发现分裂间期虫体细胞核内有绿色荧光并聚集为核仁,在分裂期虫体内绿色荧光呈弥散分布。结论光镜和电镜观察结果显示体外蓝氏贾第鞭毛虫滋养体的形态结构复杂,有典型的核仁、染色体及复杂的细胞骨架结构。     

Cartilage tissue engineering using differentiated and purified induced pluripotent stem cells

The development of regenerative therapies for cartilage injury has been greatly aided by recent advances in stem cell biology. Induced pluripotent stem cells (iPSCs) have the potential to provide an abundant cell source for tissue engineering, as well as generating patient-matched in vitro models to study genetic and environmental factors in cartilage repair and osteoarthritis. However, both cell therapy and modeling approaches require a purified and uniformly differentiated cell population to predictably recapitulate the physiological characteristics of cartilage. Here, iPSCs derived from adult mouse fibroblasts were chondrogenically differentiated and purified by type II collagen (Col2)-driven green fluorescent protein (GFP) expression. Col2 and aggrecan gene expression levels were significantly up-regulated in GFP+ cells compared with GFP− cells and decreased with monolayer expansion. An in vitro cartilage defect model was used to demonstrate integrative repair by GFP+ cells seeded in agarose, supporting their potential use in cartilage therapies. In chondrogenic pellet culture, cells synthesized cartilage-specific matrix as indicated by high levels of glycosaminoglycans and type II collagen and low levels of type I and type X collagen. The feasibility of cell expansion after initial differentiation was illustrated by homogenous matrix deposition in pellets from twice-passaged GFP+ cells. Finally, atomic force microscopy analysis showed increased microscale elastic moduli associated with collagen alignment at the periphery of pellets, mimicking zonal variation in native cartilage. This study demonstrates the potential use of iPSCs for cartilage defect repair and for creating tissue models of cartilage that can be matched to specific genetic backgrounds.     

体外重建组织工程关节软骨的实验研究

目的　用胶原蛋白和人血纤维蛋白混合物为载体在体外进行软骨细胞三维立体培养 ,构建人工软骨组织。方法　取 2周龄的新生兔关节软骨 ,经消化 ,将获得的软骨细胞与牛 型胶原、人血冻干纤维蛋白原、凝血酶按一定比例混合 ,制成软骨培养物并在体外培养。培养第 3周时 ,取材进行 HE、甲苯胺蓝染色和透射电镜检查。结果　体外培养 3周 ,培养物内细胞均存活 ,形成软骨陷窝 ,同源性细胞簇出现 ,并分泌软骨基质。透射电镜下可见丰富的粗面内质网和线粒体 ,及少量的高尔基复合体。结论　用胶原蛋白和人血纤维蛋白为载体支架体外培养软骨细胞 ,可构建较大的组织工程软骨     

Osteoarthritis-like changes and decreased mechanical function of articular cartilage in the joints of mice with the chondrodysplasia gene (cho)

OBJECTIVE: To investigate whether heterozygosity for a loss-of-function mutation in the gene encoding the alpha1 chain of type XI collagen (Col11a1) in mice (chondrodysplasia, cho) causes osteoarthritis (OA), and to understand the biochemical and biomechanical effects of this mutation on articular cartilage in knee and temporomandibular (TM) joints. METHODS: Articular cartilage from the knee and TM joints of mice heterozygous for cho (cho/+) and their wild-type littermates (+/+) was examined. The morphologic properties of cartilage were evaluated, and collagen fibrils were examined by transmission electron microscopy. Immunohistochemical staining was performed to examine the protein expression levels of matrix metalloproteinase 3 (MMP-3) and MMP-13 in knee joints. In 6-month-old animals, fixed-charge density was determined using a semiquantitative histochemical method, and tensile stiffness was determined using an osmotic loading technique. RESULTS: The diameter of collagen fibrils in articular cartilage of knee joints from heterozygous cho/+ mice was increased relative to that in control cartilage, and histologic analysis showed OA-like degenerative changes in knee and TM joints, starting at age 3 months. The changes became more severe with aging. At 3 months, protein expression for MMP-3 was increased in knee joints from cho/+ mice. At 6 months, protein expression for MMP-13 was higher in knee joints from cho/+ mice than in joints from their wild-type littermates, and negative fixed-charge density was significantly decreased. Moreover, tensile stiffness in articular cartilage of knee joints from cho/+ mice was moderately reduced and was inversely correlated with the increase in articular cartilage degeneration. CONCLUSION: Heterozygosity for a loss-of-function mutation in Col11a1 results in the development of OA in the knee and TM joints of cho/+ mice. Morphologic and biochemical evidence of OA appears to precede significant mechanical changes, suggesting that the cho mutation leads to OA through a mechanism that does not initially involve mechanical factors.     

Localization of myosin-V in the centrosome

The perinuclear localization of myosin-V was investigated in a variety of cultured mammalian cells and in primary cultures of rat hippocampus. In all cells investigated, myosin-V immunoreactivity was associated with the centrosome. In interphase cells, myosin-V was found in pericentriolar material, and in both mother and daughter centrioles. These results were obtained by using two different fixation protocols with three different affinity-purified antibodies that recognized a single band in Western blots. During cell division, myosin-V staining was intense throughout the cytoplasm and was concentrated in a trail between migrating centrioles and in the mitotic spindle poles and spindle fibers. The centrosome targeting site was determined to reside within the globular tail domain, because centrosome association also was observed in living cells transfected with DNA encoding the tail domain fused with a green fluorescent protein tag, but not in cells transfected with the vector encoding green fluorescent protein by itself.     

Regulation of matrix metallo-proteinase expression by extracellular matrix components in cultured hepatic stellate cells

Hepatic stellate cells (HSC) changed their morphology and function including production of matrix metalloproteinases (MMPs) in response to extracellular matrix (ECM) component used as a substratum in culture. We examined in this study the regulatory role of ECM component on expression of MMPs and tissue inhibitor of metalloproteinase (TIMP) in rat HSCs cultured on polystyrene, type I collagen-coated surface, type I collagen gel, or Matrigel, respectively. When cultured on type I collagen gel, HSCs showed the asteroid cell shape and MMP-1 activity, as detected by in situ zymography. Expression of MMP-1 protein and mRNA were examined by using immunofluorescence staining and RT-PCR analysis in HSCs cultured on type I collagen gel. Active form of MMP-2 was detected by gelatin zymography in the conditioned medium of HSCs cultured on type I collagen gel, whereas it was not detected when HSCs were cultured on polystyrene, type I collagen-coated surface, or Matrigel. Increased MMP-2 mRNA was detected by RT-PCR in HSCs cultured on type I collagen gel. Increased MT1-MMP proteins were shown to localize on the cell membrane by using immunofluorescence staining in HSCs cultured on type I collagen gel. Elevated expression of membrane-type matrix metallproteinase-1 (MT1-MMP) mRNA and tissue inhibitor of metalloproteinase-2 (TIMP-2) mRNA was detected by RT-PCR in HSCs cultured on type I collagen-coated surface or type I collagen gel. These results indicate that expression of MMPs and TIMP-2 is regulated by ECM components in cultured HSCs, suggesting an important role of HSCs in the remodeling of liver tissue.     

Development and characteristics of pannus-like soft tissue in osteoarthritic articular surface in rat osteoarthritis model

OBJECTIVES: Pannus is invasive granulation tissue found on the articular cartilage having rheumatoid arthritis (RA). However, pannus-like tissue has also been found in osteoarthritis (OA). Our previous study showed that pannus-like tissue in OA (OA pannus) was frequently found in human OA samples. The purpose of the study is to investigate the development and the characteristics of OA pannus in a rat OA model. DESIGN: Ligaments of the knee joint were transected in Wister rats to induce OA. The knee joints were removed at weeks 1, 2, 4 and 6, and subjected to histological study. Samples were stained with hematoxylin and eosin (HE), Safranin-O and immuno-stained for vimentin, CD34, type II collagen and MMP-3. The whole knee joint of OA rats was implanted in SCID mice and kept for a further 3 weeks. Then the histological findings were evaluated in HE sections. RESULT: OA pannus appeared at week 2 and extend over the articular surface. OA pannus cells were positive for vimentin and/or CD34. At week 6, a part of articular surface was restored with matrix. OA pannus cells expressed MMP-3 as well as type II collagen. Histological study of rat OA knees implanted in SCID mice showed that OA pannus cells filled the joint space and invaded articular cartilage. CONCLUSIONS: The presence of OA pannus was found in a rat OA model and its features were similar to those in human OA. OA pannus had both catabolic and reparative features, and the latter feature were speculated to be dominant in the later phase of the disease under a certain environmental condition.     

Characterization of human synovial fluid cells of 26 patients with osteoarthritis knee for cartilage repair therapy

Aim: To investigate the possibility of chondrogenic differentiation and cartilage repair of synovial fluid cells of osteoarthritis (OA) knee. Methods: Synovial fluids from 26 patients with OA knee were aspirated from each knee joint and cultured in vitro. The morphology of cultured synovial fluid cells, cell proliferation rate, the phenotype, and chondrogenic differentiation were analyzed in in vitro. Also, human autologous synovial fluid cells were transplanted to OA cartilage, and the cells were traced in ex vivo. Results: In 19 of 26 materials, the cells proliferated satisfactorily. The cell proliferation in six materials was very slow and one material contaminated. Culture‐expanded synovial fluid cells had a fibroblastic morphology and the phenotype was negative for CD10, CD14, and CD45, and positive for CD13, CD44, and CD105. Pellet culture of synovial fluid cells showed chondrogenic differentiation. In the ex vivo study, autologous transplanted synovial fluid cells were observed in repaired or enhanced regenerative cartilage areas and showed a tendency to infiltrate the original degenerative cartilage of OA. Conclusions: This study proved the possibility of chondrogenic differentiation of synovial fluid cells of OA knee joints despite the pathologic environment within a diseased joint. Synovial fluid cells were actually heterogeneous cells but they showed chondrogenic differentiation, similar to that of bone marrow‐derived mesenchymal progenitor cells (BMMPCs). The Ex vivo study suggested that synovial fluid cells had a tendency to adhere to OA degenerative cartilage in humans.     

结缔组织生长因子及I型和Ⅲ型胶原在脊柱关节病骶髂关节组织中的表达

目的 了解脊柱关节病(SpA)患者骶髂关节中结缔组织生长因子(CTGF)、I型胶原、Ⅲ型胶原的表达情况,探讨CTGF在spA关节软骨纤维化、骨化、关节强直中的作用.方法 30例spA患者(17例双侧影像学骶髂关节炎≥Ⅱ级,13例影像学骶髂关节炎Ⅰ级)均接受CT引导下骶髂关节穿刺活检术,取得骶髂关节组织.组织标本均行苏木素一伊红(HE)染色确认存在骶髂关节炎后,通过免疫组织化学染色方法,标记CTGF、Ⅰ型胶原及Ⅲ型胶原的表达情况.统计学方法采用单因素方差分析和t检验.结果 30例SpA患者骶髂关节组织中CTGF主要在血管翳炎症细胞及骨髓细胞的胞质中高度表达,阳性细胞数明显多于正常组织对照组[(57.9±42.4)腐倍视野和(2.7±2.5),高倍视野);Ⅰ型胶原及Ⅲ型胶原明显沉积于骨、部分软骨及韧带,平均吸光度均明显高于对照组(分别为0.298±0.080和0.044±0.024;28.254±41.165和0.105±0.054).结论 SpA骶髂关节中存在CTGF的高表达,Ⅰ型胶原及Ⅲ型胶原的沉积增多,提示CTGF参与了SpA骶髂关节局部胶原沉积、软骨纤维化变性过程,可能在SpA关节软骨纤维化、关节强直中起重要的作用.

Abstract：

Objective To investigate the expression of connective tissue growth factor(CTGF),coll agen I and collagen Ⅲ in sacroiliac joint(SIJ)of patients with spondyloarthropathy(SpA).Methods Thirty patients with SpA,including 17 patients with grade Ⅱ saeroiliitis and 13 patients with grade Ⅰ sacroiliitis,were performed on CT guided needie biopsy of SIJ.After sacroiliitis were confirmed by staining with hematoxylin and eosin in sacroiliac joint tissue sample,immunohistochemical assay was performed to determine the expression of CTGF,collagen Ⅰ and collagen Ⅲ in sacroiliac ioint tissue.Univariate Chi-square test was used for data comparison between multiple groups and t-test was used for two group data comparison.Results Contrast to healthy controls,CTGF were found upexpressed on the cytoplasm of inflammatory cells in pannus and bone marrow of sacroiliac tissue samples of patients with SpA,while collagen I and collagen Ⅲ were found up-expressed in bone,cartilage and ligament tissue[(57.9±42.4)/HP vs(2.7±2.5)/HP P<0.05,0.298±0.080 vs 0.044±0.024 and 28.254±41.165 vs 0.105±0.054.P<0.05 respectively].Conclusion CTGF,collagen Ⅰ and collagen Ⅲ are up-expressed in SIJ of SpA patients.CTGF may play an important role in articular cartilage fibrosis and ossification of SpA.     

Early myocardial infarction. A feasible histologic diagnostic procedure

The Barbeito-López trichrome stain (BLTS) was compared with hematoxylin and eosin (HE), the basic fuchsin picric acid (BFP) and nitro-blue tetrazolium (NBT) dye tests. Two types of hearts were studied: myocardial necrosis induced by isoproterenol (ISP) in rats and human hearts with early coagulation necrosis (Early CN). The BLTS was much more sensitive than HE for diagnosis of Early CN in rats. By 24 hours, both BLTS and HE showed established coagulation necrosis (Established CN) although BLTS did so more clearly. In human material BLTS was a useful indicator of Early CN. It showed patchy areas of coagulation necrosis in the cytoplasm of damaged myocardial cells which were not visualized with HE. Normal myocardium remained green or pale blue. Early CN appears as patchy yellow areas while the remainder of the cytoplasm stains green. Established CN appears "golden yellow" or "orange yellow".