不同浓度牛和人血清培养软骨细胞中糖醛酸的测定

本实验用测定培养液中葡萄糖醛酸的方法,作为培养软骨细胞生长情况的指标,以观察软骨细胞的代谢情况,了解不同种类和不同含量血清对培养软骨细胞的影响.我们用传代兔软骨细胞于不同含量小牛血清和人血清的培养液中培养,定期镜下观察并收集培养液,测定其葡萄糖醛酸含量.结果表明5%至15%牛血清或人血清培养一代兔软骨细胞均可生长;测定培养液內葡萄糖醛酸量作为反映软骨细胞生长的检测指标是可行的。     

牛关节软骨培养稳态模型的实验研究方法

参照Ｈａｓｃａｌｌ实验方法建立了牛关节软骨体外培养，并通过组织学观察，３５Ｓ掺入率测定及已糖醛酸测定的方法证明在加入２０％小牛血清的ＤＭＥＭ培养液中，培养７ｄ后，软骨蛋白多糖的代谢达到了稳态，这为研究关节软骨蛋白多糖代谢及其影响因素提供了一个较好的实验模型。     

植骨术治疗颞颌关节强直的疗效观察

<正> 颞颌关节强直的手术治疗,其目的均是切开骨性愈着,保持切开间隙,防止再愈着而复发,达到恢复下颌运动功能,保持正常的咬(牙合)关系。临床上,根据关节的X线片检查,将强直分为下列三型:第Ⅰ型强直,骨性愈着的范围仅限于关节的颗状突。第Ⅱ型强直,骨性愈着由颗状突扩大到乙状切迹。第Ⅲ型强直,骨性愈着包括颗状突、乙状切迹和喙状突。按     

PREVENTIVE EFFECT OF BORON ON DAMAGE TO CARTILAGE OF RATS WITH INTAKE EXCESSIVE FLUORIDE

Althoughfluorideinthebodymayhelpthe growthofbonesandteethinitsrangeofsafety,ex cessiveintakeoffluoridefromtheenvironmentcan damagetissuesandorgansinhumanandanimal, thusinducingskeletalfluorosis(SF)[1,2].Control lingoffluoridesources(suchastodefluoridate…     

谷胱甘肽对Cu~(2+)-H_2O_2导致关节软骨蛋白聚糖降解的影响

应用小牛关节软骨短期器官温育的方法,研究了谷胱甘肽(GSH)对Cu~(2+)-H_2O_2导致关节软骨蛋白聚糖(PG)降解的影响。结果表明:GSH能非常显著地抑制Cu(2+)-H_2O_2对软骨PG的降解作用,并呈剂量依赖型。用Sepharosc 6B柱层析分析软骨PG降解产物,发现GSH还能消除Cu~(2+)-H_2O_2导致的PG链的断裂,保护PG结构的完整。GSH对Cu~(2+)-H_2O_2的作用机理可能一方面与Cu~(2+)形成了复合物,另一方面使H_2O_2还原为H_2O,而消除了两者对软骨PG的降解作用。     

EFFECT OF LOW SELENIUM ON CHONDROCYTE DIFFERENTIATION AND DIFFERENTIAL EXPRESSION OF COLLAGEN TYPES Ⅰ , Ⅱ AND X IN ARTICULAR CARTILAGE FROM MINI-PIGS

Objective According to the distribution of low selenium areas, low nutrition state of the residents and the affecting cartilage growth and articular cartilage of Kashin-Beck Disease(KBD),the chondrocyte differentia- tion and differential expression of collagen types Ⅰ , Ⅱ and Ⅹ in articular cartilage from Chinese mini-pigs treated with low selenium were investigated in order to gain insight into the effects of these conditions on chondrocyte differ- entiation in KBD cartilage. Mothods Eleven male juvenile mini-pigs, aged from 4 weeks to 6 weeks after birth, were divided into 3 groups. The Se content in the diet of the “low Se” group was 0. 035mg/kg diet, and 0. 175 mg/kg diet in the control. For Se-supplemented group 0. 390mg /kg diet was added. The content of Se in blood was assayed at the beginning and at the end of each experiment. Samples of articular cartilage were taken from the right femur condylus, and collagen types Ⅰ , Ⅱ and Ⅹ in articular cartilage were analyzed by immunohistochemistry and in situ hybridization. Results ①All cartilage samples from juvenile mini-pigs fed with low selenium diet revealed a re- duction in type Ⅹ collagen mRNA expression in the hypertrophic chondrocytes as shown by in situ hybridization, and reduced type Ⅹ collagen deposition in the lower hypertrophic zone as shown by immunohistochemistry. ②Addition of selenium to the diet restored the type Ⅹ collagen to normal level. ③Type Ⅱ collagen was evenly distributed over the entire articular cartilage in all experimental and control groups. Type Ⅱ collagen mRNA signals were most prominent in the upper articular layer as well as in the hypertrophic zone in all groups. Type Ⅱ collagen expression was restrict- ed to the zone of endochondral ossification in all experimental groups and the control. Conclusion Low selenium has an down-regulatory role on the synthesis and deposition of collagen type Ⅹ in hypertrophic chondrocytes in articular cartilage of mini-pigs. Supplement of the low Se diet with additional Se restored the signals of collagen type Ⅹ to nor- mal levels. These findings indicate that selenium deficiency may disturb chondrocyte differentiation to hypertrophic cells in the growth plate,and worthy to be investigated further.     

关节内注射川芎嗪对兔膝关节软骨退变的影响

目的观察川芎嗪关节腔内注射对骨关节炎过程中软骨退变的保护作用,探讨川芎嗪关节腔注射治疗骨关节炎的应用前景。方法健康成熟新西兰大白兔26只,设立正常组、模型组、对照组和实验组。模型组、对照组及实验组建立骨关节炎模型。对照组与实验组分别关节腔注射地塞米松0.3 mL和川芎嗪0.3 mL。每周一次。在模型建立6周和9周时各组分别处死一半动物,去除关节炎模型,取材进行大体标本病理学观察,并经光镜和透射电镜观察膝关节软骨的病理变化。结果在6和9周时,实验组的大体标本软骨损伤分级明显优于其他3组;Mankin软骨组织学评分与上述两组相比,有非常显著性差异(P<0.01);电镜超微结构观察其病理改变明显轻于对照组和模型组。对照组大体标本软骨损伤分级与模型组基本相同;Mankin软骨组织学评分与模型组比较无显著性差异(P>0.05);电镜超微结构观察对照组与模型组在细胞形态、细胞内细胞器改变等方面相似。结论关节腔内注射川芎嗪能减轻兔膝关节炎模型的关节软骨退变程度,并可促进软骨的自身修复;激素关节腔内注射对损伤软骨的修复作用不明显。     

Microstructure of hot-dip galvanized Zn-Al-Mg alloy coating

The microstructure of hot-dip galvanized Zn-11%Al-3%Mg-0.2%Si alloy coating was studied in this article. X-ray diffraction analysis revealed the coating is composed by Zn, Al and MgZn₂ phase. Optical microscope (OM) and scanning electron microscope (SEM) observations showed the coating is occupied by snowflake-like dendrite, double hexagonal organization and eutectic. The coating backbone was the dendrite considered to be a phase of hexagonal close-packed (HCP) structure judging form its morphology according to the crystal growth way in the coating. Transmission electron microscopy (TEM) and energy dispersive spectroscopy (EDS) researches on the dendrite suggested that an intermediate Zn-Al phase was formed at high temperature, then decomposed into a type of Zn-Al granular eutectoid after cooling down to room temperature, while the eutectoid Zn and Al had a certain crystallographic relationship. The coating solidification process and the grain crystal structure were discussed.     

STUDY ON THE EFFECT OF T-2 TOXIN AND SELENIUM ON CD44 EXPRESSION IN THE CULTURED HUMAN FETAL CHONDROCYTES IN VITRO

KBD (Kashin BeckDisease)isasevere ,en demicosteoarthrosisandthepathogenesisofitisstillapendentproblem[1] .WiththestudyofetiologyofKBD ,thehypothesisofselenium deficiencyinthesurroundingandfoodcontaminatedbyfungaltoxinisanoticeableone .The pathologicalfeaturesofKBDarethedegenerationandnecrosisofthearticu larcartilage ,disorderofmetabolismofcartilagematrix .Thestudyontheosteoarthrosissuggeststhatthereductionoftheaggregationandincreaseofdeg radationofaggrecanareinitialaffairsifcartilagehasdeg…     

宫内发育迟缓儿的分娩方式与围产儿转归

对１５７例足月体重≤２５００ｇ宫内发育迟缓儿的分娩方式与围产儿Ｌ转归进行了临床分析。结果表明，新生儿出生体重愈低，围产儿死亡率、新生儿窒息率和新生儿并发症的发生率愈高。剖宫产组围产儿死亡率不仅没有下降，反而增加了新生儿窒息率和并发症的发生率。胎吸和产钳助娩组新生儿并发症的发生率明显升高。臀位助娩和臀位牵引的新生儿窒息率、并发症发生率及围产儿死亡率显著高于自然分娩组，故对宫内发育迟缓儿应加强围产保健和产时监护，尽量使其自然分娩；臀位儿以剖宫产为宜。     

雌激素受体在肝海绵状血管瘤中的表达及意义

采用免疫组化Ｓ Ｐ法检测３０ 例肝海绵状血管瘤（Ｃａｖｅｒｎｏｕｓ ｈｅｍ ａｎｊｉｏｍ ａｏｆｌｉｖｅｒ,ＣＨＬ）组织中雌激素受体（Ｅｓｔｒｏｓｅｎ ｒｅｃｅｐｔｏｒ, ＥＲ）的表达,并以ＣＨＬ瘤旁正常肝组织作对照。结果ＥＲ 阳性表达于ＣＨＬ 血窦内皮细胞胞浆中,阳性率为４３３％ ;ＣＨＬ瘤旁正常肝组织中未见表达（ Ｐ＜ ０．０５）。提示肝海绵状血管瘤的生长与雌激素有关。雌激素可能通过其受体作用促进ＣＨＬ生长     

胎儿和幼儿跖趾骨血液供应的观察

采用微血管造影等方法,观察30例胎、幼儿跖趾骨的血供。结果表明:胎儿期跖趾骨骨干和两端软骨逐渐增大,出现滋养动脉和软骨管,幼儿期两端软骨逐渐变小,软骨管也逐渐消失。提示软骨管的出现与软骨块的大小有关,软骨管是胎儿软骨内的临时血管系统。跖趾骨血供特点:①跖趾骨血供的不均称性分布,即骺端软骨血供丰富,这与两端生长能力不同相适应;②远趾只骺端有软骨管分布,这与只骺端有生长能力相适应,说明跖趾骨血供与其生长发育有密切关系。     

蛋白聚糖与软骨结构、功能及骨关节病的关系

蛋白聚糖是一类由核心蛋白与1条或多条共价连接的氨基聚糖所组成的糖复合物,是细胞膜、基底膜、特别是细胞外基质的重要组成成份,与组织细胞的结构和功能息息相关。随着科学研究的发展,人们逐渐认识到蛋白聚糖代谢与骨软骨发育和退变、神经组织的发育和退变、心血管疾病和肿瘤的发生发展关系密切。本文仅对蛋白聚糖代谢与软骨结构和功能及骨关节病发病机制的研究进行探讨。     

βTCP/CPP/PLLA软骨组织工程支架制备及性能

以提纯的PLLA为基体材料,以自制CPP纤维（直径10－20μm）及β-磷酸三钙（-βTCP,粒径：30－60μm）为增强材料,以二氯甲烷（分析纯）为溶剂,以碳酸氢氨NH4HCO3（颗粒尺寸：200－500μm）为致孔剂,采用溶媒浇铸/粒子滤取技术与气体发泡相结合的方法制备了-βTCP/CPP/PLLA软骨组织工程支架复合材料.测试了β-TCP/CPP/PLLA软骨组织工程支架复合材料的物理、力学性能,并借助扫描电镜对其微结构进行了观察.结果表明,支架复合材料的初始物理、力学性能和微结构满足软骨组织工程对其支架材料的要求.     

应用激光捕获显微切割技术制备肺癌蛋白质组学研究样品

目的探讨应用激光捕获显微切割(laser capture microdissection,LCM)技术获取高同质性肺癌组织细胞及正常对照肺泡上皮细胞的方法及可行性。方法将新鲜肺癌组织标本及其配对正常组织常规制备8μm冰冻切片,采用改良的HE方法染色;应用LCM技术大宗切割肺鳞癌、腺癌及正常对照肺泡上皮细胞;选择性获取同质肺癌细胞和配对正常细胞。结果通过LCM技术可以准确、快速地分离癌细胞、间质及其他坏死组织,得到同质性不低于95%的肿瘤细胞和正常细胞。按照Duration 15.5 s的激光脉冲频率和平均8 000 shots条件,既可保证每个帽子的细胞收集数量,也能够有效地缩短实验时间,提高实验可控性,从而使操作过程标准化。结论应用LCM技术可以成功获取同质肺癌细胞和正常细胞。其所需标本量少、组织同质性高、操作过程稳定、可控性好,是肺癌分子生物学及蛋白质组学研究有价值的样品制备方法。     

CHONDROCYTE APOPTOSIS IN ARTICULAR CARTILAGE WITH KASHIN-BECK DISEASE

Kashin BeckDisease(KBD)isachronic,en demicosteoarthritisaffectingover0.81millionspa tientsand101millionsofpeopleatriskinChina[1]. Thebasicpathologicalfeaturesofthediseaseis chondrocytesdegenerationandnecrosisinthedeep zoneofarticularcartilageandgrowthpl…     

海藻酸盐小球中培养的软骨细胞的生长

目的为海藻酸盐水凝胶作为软骨细胞三维培养环境提供实验依据。方法将酶消化法获取的P1代软骨细胞包裹在20 g/L海藻酸钙凝胶小球中持续培养4周,在培养的不同时间点取适量的小球行倒置相差显微镜观察,HE、番红"O"染色及Ⅱ型胶原免疫组化染色;并通过MTT法检测细胞活性及增殖;二甲基亚甲兰法进行氨基葡聚糖(GAG)含量测定。结果在海藻酸钙小球内培养的软骨细胞能够保持球形状态,且随时间的延长,细胞增殖逐渐聚集成簇;细胞外基质沉积逐渐增加,GAG含量增多。Ⅱ型胶原免疫组化染色阳性,鉴定证实为具有正常表型的软骨细胞。结论软骨细胞在海藻酸盐凝胶小球中培养有效维持了细胞的分化状态,细胞增殖并具有良好的分泌蛋白功能。海藻酸钙凝胶适合于软骨细胞的生长。     

软骨细胞-骨基质明胶复合物修复兔关节软骨缺损的初步观察

目的　初步观察骨基质明胶 (bonematrixgelatin ,BMG)复合软骨细胞移植修复关节软骨缺损的可行性。方法　BMG复合软骨细胞体外培养 ,并将培养复合物植入同种异体兔膝关节软骨缺损处 ,术后行大体及组织学观察。结果　BMG复合软骨细胞体外培养后能形成软骨组织 ,此“工程软骨”植入兔膝关节软骨缺损处能继续生长发育。结论　BMG复合软骨细胞体外培养及体内移植均能形成软骨组织 ,有可能修复关节软骨缺损     

镓盐对骨质疏松骨细胞超微结构的影响

目的探讨镓盐对骨质疏松骨细胞超微结构的影响。方法用8月龄雌性SD大鼠造模,骨质疏松组开腹切除双侧卵巢后缝合,对照组开腹切一小块脂肪后缝合。12周后,取胫骨上段骨组织,观察模型复制情况。嗣后随机分为3组:对照组与骨质疏松组正常饲养12周;硝酸镓治疗组给予硝酸镓治疗(1 mg/kg),3次/周,腹腔注射。应用脱钙骨切片观察光镜下骨形态;制作透射电镜样本,在透射电镜下观察骨细胞形态及骨细胞内部结构情况。结果对照组及硝酸镓治疗组骨细胞结构正常,细胞核染色质分布均匀,核为卵圆形;骨质疏松组部分骨细胞核呈固缩状,形态变长。对照组成骨细胞中细胞器较少,有胶原分泌;骨质疏松组成骨细胞中有大量内质网存在,有大量胶原分泌,硝酸镓治疗组成骨细胞可见有较多胶原分泌,细胞中存在大量内质网及线粒体。结论镓盐对骨质疏松引起的骨细胞超微结构的损伤有改善作用。     

组织工程化软骨的构建及应用

"组织工程"(tissue engineering)是上世纪80年代正式提出来的新概念。它是生物医学和材料科学多学科交叉融合的新兴分支学科。近20年来,该学科有了很大的发展,其研究范围也更加广泛。本文结合我们近年的研究成果,着重论述组织工程化软骨的构建及其在软骨损伤和修复研究中的应用。