Ⅵ型胶原放免检测方法及其初步应用

从人胎盘中经胃蛋白酶消化，氯化钠分级沉淀分离，ＥＤ－５２纤维素，ＳｅｐｎａｃｒｙｌＳ－２００柱层析纯化制备Ⅵ型胶原。用酸性介质致敏新西兰兔产生的抗体效价为１：２０００，天与Ⅰ，Ⅱ，Ⅳ型胶原等无交叉。     

大鼠肺间质纤维化支气管肺泡灌洗液透明质酸含量的变化

经单次气管内灌注博莱霉素Ａ５复制大鼠肺间质纤维化模型，在不同时间收集大鼠气管肺泡灌洗液。用放射免疫分析测定气管肺泡灌洗液中的透明质酸。结果发现，经博莱霉素处理后气管肺泡灌洗液中的透明质酸于３天和７天明显升高。透明质酸的升高与粒细胞数和蛋白质浓度的改变呈平行关系，提示透明质酸的升高与博莱霉素所致的急性肺泡炎有关。     

血清透明质酸放射免疫分析的临床应用

透明质酸(Hyaluronic acid, HA)是一种氨基多糖。1985年Kujuwa等首先报告人血清HA在肝病时升高,以后陆续有所报道。我国于1989年由张鲁榕等率先建立血清HA放射免疫分析法,并证实本法能灵敏准确地反映肝脏的病变程度。近年来国内不少学者对HA进行了大量研究,并发现HA浓度增高与多种疾病有关。     

烧伤病人血清透明质酸水平的变化

本文用放射免疫分析测定２３例烧伤病人的血清透明质酸水平。烧伤病人的血清透明质酸水平明显增高，并与烧伤的程度和疤痕增生程度有联系。血清透明质酸测定可作为烧伤病人的监测指标之一。Ｐ＜０．０１３．烧伤深度与ＨＡ水平本组２３例中，以单纯浅二度７例与深二度或三度为主的９例比较，ＨＡ水平见表３。     

血清透明质酸放射免疫分析在血液病诊断中的应用

本文用放射吏疫分析法测定50例正常人及71例血液病患者血中HA含量，藉以探讨HA在血液病患者中的临床意义。     

酶联免疫法（ELISA）在检测肝纤维化指标中的应用

目的 探讨酶联免疫法（ELISA）在检测血清透明质酸（HA）、Ⅲ型前胶原（PcⅢ）、Ⅳ型胶原（CIV）、层粘蛋白（LN）中的应用。方法 利用ELISA与放射免疫分析法（RIA）分别检测116例慢性肝炎组、215例正常组和36例肝硬化组的血清HA、PCⅢ、CIV、LN，比较二者的优缺点。结果 两种方法结果无差异（p〉0．05）。结论 ELISA法准确、简便和安全，可代替RIA法。     

透明质酸、甘胆酸和前白蛋白联合测定诊断肝硬化的应用

为探讨透明质酸(HA)、甘胆酸(CG)和前白蛋白(PA)在肝实质损伤中的临床意义,采用放射免疫法和免疫比浊法测定了85例肝病患者和20名正常人血清HA、CG和PA浓度,并分析和比较三者联合测定在肝病的诊断、疗效及预后判断中的临床应用.现报道如下.     

透明质酸及其相关蛋白与大鼠肝纤维化的关系

目的:探讨血清透明质酸(HA)及其相关蛋白(SHAP-HA)水平与肝纤维化间的关系.方法:建立实验性肝纤维化动物模型,检测血清HA、SHAP-HA、谷-丙转氨酶(ALT)、谷-草转氨酶(AST)、白蛋白(ALB),观察大鼠肝组织病理变化.结果:随着肝纤维化程度的加重,血清HA、SHAP-HA表达量均增加,与ALT、AST呈正相关,差异具有统计学意义.结论:实验性肝纤维化过程中HA、SHAP-HA表达水平不仅与肝组织的炎症坏死程度有关,而且与纤维化过程关系密切,HA、SHAP-HA可作为肝纤维化评估指标之一.     

人瘦素放免分析法的建立及初步临床应用

瘦素是由肥胖基因编码, 由脂肪细胞合成和分泌的一种亲水性激素蛋白.本文用高纯的重组人瘦素抗原免疫家兔, 得到高效价的兔抗人瘦素多克隆抗体, 氯胺-T法制备125I-瘦素标记物, 建立人血清瘦素RIA.本方法特异性强, 灵敏度较高, 精密度较好, 其测定值与Linco公司的放免药盒有很好的一致性.用本法建立的正常参考值(BMI 18-25): 男性(34例)为2.15±1.46ng/mL,女性(36例)为7.86±3.60ng/mL.将112名健康者的检测结果分成男、女两组, 血清瘦素值与BMI之间有明显的正相关(r=0.72,P＜0.001).瘦素水平与年龄之间无明显相关性(P＞0.05).女性的促黄体激素、促卵泡激素、泌乳素、孕酮和雌二醇与瘦素无明显相关性(P＞0.05).而对于男性, 睾酮与瘦素水平成明显的负相关(r=0.792,P＜0.01).     

人Ⅳ型胶原放射免疫分析方法的建立

本文将人胎盘用胃蛋白酶消化、氯化钠分级沉淀及DE-52、CM-52层析提取、纯化N M胶原（ⅣC）。提取物紫外最大吸收峰在230nm,富含羟脯氨酸、羟赖氨酸,两个α链分子量分别为95000和70000,与Sigma产品特性相似。免疫新西兰兔产生的抗血清效价为1:512（间接血凝法）,与其它间质成份无或弱交叉反应。建立的放射免疫分析方法,灵敏度为8μg/L,批内及批间C.V.分别为7.3％和11.2％,测得正常献血员血清ⅣC浓度为49.77±15.00μg/L,慢性肝炎患者显著升高,并与PANASSAY ⅣC EIA法检测结果相关显著,提示本方法灵敏可靠,有推广应用价值。     

透明质酸凝胶在成肌细胞体外培养中的应用

目的探讨透明质酸凝胶与成肌细胞的生物相容性,为成肌细胞组织工程研究提供新型载体。方法将体外培养的成肌细胞接种于含透明质酸凝胶的培养液中常规培养,通过倒置显微镜和扫描电镜观察细胞形态,描绘细胞生长曲线,并进行免疫细胞组化鉴定。结果透明质酸凝胶组与对照组成肌细胞体外培养生长曲线相似,均成desmin阳性,透明质酸凝胶组亲水性和吸附力优于对照组。扫描电镜下透明质酸呈三维框架结构,有利于成肌细胞附着。结论透明质酸凝胶是成肌细胞良好的载体材料。     

慢性无症状HBV感染者血清CG和HA RIA的临床意义

目的:探讨慢性无症状HBV感染者血清甘胆酸(CG)和透明质酸(HA)测定的临床意义.方法:对35例慢性无症状HBV感染者和100例正常对照组用放射免疫分析检测血清CG和HA的含量.结果:慢性无症状HBV感染者组血清CG的水平显著高于正常对照组(P＜0.01),慢性无症状HBV感染者组HA的水平亦显著高于正常对照组(P＜0.05).结论:慢性无症状HBV感染者血清CG和HA的测定能反映慢性无症状HBV感染者有一定程度的肝损情况.     

慢性肝病患者TPO水平与肝纤维化指标关系的探讨

目的：探讨慢性肝病患者血清中血小板生成素（TPO）水平与肝纤维化指标的关系。方法：ELISA测定正常人群及肝病患者的TPO水平,肝纤维化指标用放射免疫分析。结果：血小板生成素水平在慢性肝炎[（120.41±99.73）pg/ml]轻、中、重组,肝硬化[（125.84±100.40）pg/ml]与正常人群[（144.18±48.01）pg/ml]中比较均无显著性差异（P〉0.05）;肝硬化组（18例）的ⅣC与TPO水平有显著相关性（r=0.517,P〈0.05）,其余各组HA、LN、PCⅢ、ⅣC与TPO均无相关性（P〉0.05）。结论：慢性肝病患者血清TPO水平与疾病的严重程度无关,而肝硬化患者TPO水平与肝纤维化程度有一定的关系。     

Prognostic Factors after Intra-Articular Hyaluronic Acid Injection in Ankle Osteoarthritis

Purpose

The goal of this study was to identify baseline prognostic factors of outcome in ankle osteoarthritis patients after intra-articular hyaluronic acid injection.

Materials and Methods

Patients with ankle osteoarthritis who received hyaluronic acid injection therapy were retrospectively reviewed. Each patient received weekly intra-articular hyaluronic acid injections (2 mL) for 3 weeks. Six predictors including gender, age, symptom duration, radiographic osteoarthritis stage, radiographic subchondral cyst, and fracture history were evaluated. Visual analogue scale (VAS) and patient satisfaction were evaluated as outcome measures. These predictors and outcome measurements were included in a logistic regression model for statistical analysis.

Results

Total of 40 consecutive patients (21 male, 19 female) were included in this study. Mean age was 60.6. Average follow up period was 13 months. The mean VAS recorded 3, 6, and 12 months after the first injection was 3.6 (SD 2.54, p<0.001), 4.33 (SD 2.9, p<0.001), and 5.3 (SD 2.7, p=0.0071), respectively, when compared to baseline VAS. Early stage disease was identified as an independent predictor associated with ''positive VAS outcome'' at 3 and 6 months. Early stage disease and duration of pain less than 1 year were independent predictors associated with higher satisfaction.

Conclusion

While hyaluronic acid injection for ankle osteoarthritis is a safe and effective treatment, careful selection of patients should be made according to the above prognostic predictors.     

A Molded Hyaluronic Acid Gel as a Micro-Template for Blood Capillaries

Abstract

Fabrication of blood capillaries in tissue-engineered tissue is necessary for creating thick three-dimensional (3D) tissue with a high cellular density. For inducing blood capillaries in the tissue in vitro, a molded hyaluronic acid (HA) capillary-shaped gel was made as a template for blood capillaries by photolithography and power free pumping techniques. The fabricated HA capillary-shaped gel was sandwiched between two cell sheets consisting of neonatal normal human dermal fibroblasts (NHDFs), human umbilical vein endothelial cells (HUVECs), or co-cultured NHDFs and HUVECs, and eventually covered with the cells. Although a slight degradation of the HA gel was observed in the sandwiched tissue with HUVEC or NHDF cell sheets, significant degradation of the HA gel was observed in the sandwiched tissue with co-cultured cell sheets. Moreover, by continuing to culture the co-cultured tissue with HA gel, a tube formation was observed at the HA gel site. A sandwiched HA capillary-shaped gel with two cell sheets has a potential for creating blood capillaries in vitro and fabricating vascularized artificial organs.     

肝硬化患者血浆Hcy检测的临床意义

目的：检测肝硬化患者血浆同型半胱氨酸（Hcy）的含量，探讨血浆Hcy水平与肝硬化的相关关系，评估血浆Hcy水平用于肝硬化诊断的价值。方法：采用酶联免疫法对64例肝硬化患者、42例非肝硬化患者及60例健康者血浆Hcy含量进行检测。结果：肝硬化患者血浆Hcy水平显著高于非肝硬化患者与健康志愿者，且肝硬化患者血浆Hcy水平与血浆透明质酸、层黏蛋白等肝硬化相关指标水平具有良好的相关性（r分别为0．87、0．88．P〈0.01），但与胆固醇、三酰甘油、高密度脂蛋白无相关性（均P〉0．05）。结论：肝硬化患者血浆Hcy含量显著升高，检测血浆Hcy含量有助于肝硬化的诊断，而且血浆Hcy水平与肝硬化的发生发展密切相关。     

Electrospun Biomimic Nanofibrous Scaffolds of Silk Fibroin/Hyaluronic Acid for Tissue Engineering

Abstract

This study aimed to fabricate nanofibrous scaffolds which could biomimic the natural extracellular matrix from aqueous solutions of silk fibroin and hyaluronic acid blends (SF/HA) by means of electrospinning. Scanning electronic microscopy results indicated that electrospun SF/HA nanofibers were ribbon-shaped and their average width obviously decreased with the increase of HA content. However, there is no fiber observed when the volume of HA further increased to 50% of overall volume. After being treated with 75% ethanol vapor for 24 h, the fibers still remained their fibrous morphologies and presented good capability of water-resistance. Fourier transform infrared attenuated total reflectance spectroscopy, ¹³C-CP-MAS nuclear magnetic resonance and differential scanning calorimetry results revealed that HA did not induce SF conformation from random coil to β-sheet. SF conformation converted from random coil to β-sheet after being treated with 75% ethanol vapor. Cell viability studies demonstrated that SF/HA nanofibrous scaffolds significantly promoted cell proliferation. Electrospun SF/HA nanofibers may provide an ideal biomimic tissue-engineering scaffold or vehicle for water-soluble drugs.     

Hyaluronic Acid Based Azo Polymer: Synthesis,Characterization, and Photo-Responsiveness

Hyaluronic acid based azo polymer (HA-Azo-COOH) is synthesized by post-azo coupling reaction. Controllable grafting percent of azobenzene pendant is achieved by adjusting the feed ratio. In selective solvents (dimethyl sulfoxide/water), uniform colloidal aggregates of the prepared polymer can be obtained by self-assembly process. The self-assembled colloids can be elongated along the polarization direction of the irradiation laser light. The relationship between the deformation of colloids and the irradiation time is explored.     

Collective Dynamics and Transient Behavior of Partially Hydrophobic Hyaluronic Acid Chains

The unique performance of a partially hydrophobic derivative of hyaluronic acid, HYADD4, bearing 2–3 hexadecylic side chains every 100 repeating units, enables hydrogels based on this polymer to be suitably used as a viscosupplement to replace synovial fluid in stressed or injured articular joints. This contribution addresses: i) as to whether there is a self‐assembly process driving the HYADD4 hydrogel stabilization and ii) what the characteristics of the long‐range dynamics governing the macroscopic viscoelastic behavior are, and to what extent these features are different from those present in the parent polymer (i.e., hyaluronic acid). Mesoscale dynamic and static properties are interpreted in the light of “sticky reptation theory” and evaluated in combination with small‐angle neutron scattering results.