大骨节病儿童及成人关节软骨中甲状旁腺激素相关肽的表达

目的 探索甲状旁腺激素相关肽(PTHrP)在大骨节病儿童及成人关节软骨中的表达及分布。方法 应用免疫组化方法比较正常儿童和成人与大骨节病儿童和成人关节软骨中PTHrP的表达。结果 ①大骨节病儿童关节软骨表层、中层、深层PTHrP阳性细胞表达率分别为30％、24％和31％，中层、深层表达率比正常儿童明显增高(P＜0．01)；②大骨节病成人关节软骨表、中、深层PTHrP阳性细胞表达率分别为54％、46％和16％，各层细胞阳性表达率均明显高于正常组(P＜0．01)。②大骨节病儿童与成人比较各层软骨细胞PTHrP的阳性表达差异均有显著意义(P＜0．01)，表现为表、中层成人高于儿童，深层儿童高于成人。结论 大骨节病患者关节软骨中PTHrP阳性表达率高于正常对照。     

过量氟对大鼠肾肝心损害的形态学研究

目的 探讨过量氟对实验大鼠肾、心、肝脏的损害作用。方法 用光镜观察摄入不同过量氟后3个月、5个月大鼠的肾、心、肝脏的病理形态学变化,用免疫组织化学方法观察大鼠肾组织内增殖细胞核抗原（PCNA）的表达。结果 不同剂量过量氟组大鼠肾、心、肝脏的实质细胞肿胀和单个细胞坏死。肾脏损害病变严重而复杂,特别是5个月高氟组肾组织内出现多种灶状病变、肾小管阻塞和扩张;间质灶状纤维化、小管萎缩、上皮细胞增生伴单个核炎细胞浸润;肾内细胞PCNA高表达;灶状坏死伴钙化;肉芽肿和感染灶等。结论 过量氟可损害多种内脏器官,以肾脏损害最重,其病变具有多样性特征。     

雪腐镰刀菌烯醇对培养软骨细胞形态和细胞凋亡的影响

目的：研究雪腐镛刀菌烯醇（ＮＩＶ）对培养软骨细胞形态和细胞凋亡的影响。方法：分别用细胞爬片的电镜观察和姬母萨染色光学显微镜观察,以及体外细胞培养再建软骨组织切片ＨＥ染色观察等手段,以图发现其对软骨细胞的损伤。结果：ＮＩＶ低浓度组只引起细胞超微结构轻微改变,ＮＩＶ中、高浓度组引起细胞明显改变,且有凋亡小体的存在,结论：设定ＮＩＶ之低浓度可能是引起软骨细胞病理损伤的最小浓度;ＮＩＶ中、高浓度引起软骨细胞过度凋亡等明显病理损伤。     

低硒、低碘及联合对亲代和子代大鼠骨、软骨生长的影响

Objective To study the effects of selenium deficiency,iodine deficiency and combined selenium and iodine deficiency on bone and cartilage growth in the parental and the first filial generation rats. Methods Forty-eight weanling healthy SD rats were randomly divided into selenium deficieney, iodine deficiency, combined selenium and iodine deficiency and control groups according to their body mass. These rats were fed with selenium deficiency, iodine deficiency, combined selenium and iodine deficiency, and normal fodder, respectively. The parental rats (about 3 months old) were mated in each group 8 weeks after the beginning of the experiment. Right tibias and left knee joints were collected when the parental generation rats were about 6 months and the first filial generation rats were about 3 months old. Tibial length, mid-shaft tibial diameter, and articular cartilage diameters of the right tibias were measured by vernier caliper. Left knee joints were embedded and cut into sections and the thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes in growth plate cartilage were observed under the light microscope. Results The selenium deficiency had significant effect on serum selenium level of the parental and the first filial generation rats(F value were 239.56,232.68, P< 0.01), and also on serum T4 level of the first filial generation rats(F value were 6.95, P < 0.05). The iodine deficiency had significant effect on serum T3 and T4 level in the two generations rats(F value were 14.11,14.05,30.29,34.53, P < 0.01 ). There were interactions between selenium deficiency and iodine deficiency on serum T4 level in the first filial generation rats (F= 5.99, P< 0.05). The serum selenium of selenium deficiency group[ (30.28 ± 6.34), (43.95 ± 9.75)μg/L],combined selenium and iodine deficiency group[ (30.33 ± 5.18), (35.40 ± 3.16)μg/L] were significantly lower than iodine deficiency group[(345.83 ± 29.55), (245.24 ± 9.95)μg/L] and the controls[ (358.64 ± 30.50), (236.50 ±9.75) μg/L] in the two generations. The serum T3 of combined selenium and iodine deficiency group [(0.55 ± 0.05 ),(0.88 ± 0.14)nmol/L] were significantly lower than the controls[(0.75 ± 0.08), (1.26 ± 0.26)nmol/L] in the two generations. The serum T4 of iodine deficiency [ (24.11 ± 2.29), (42.10 ± 8.92) nmol/L ] and combined selenium and iodine deficiency group[ (20.66 ± 1.93), (26.55 ± 5.98)nmol/L] were significantly lower than the controls[ (36.15 ±2.74), (52.79 ± 8.84)nmol/L] and selenium deficiency group[ (28.12 ± 3.33), (52.02 ± ll.99)nmol/L] in the two generations. The selenium deficiency and iodine deficiency had significant effect on tibial length, thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes in first filial generation rats(F values were 24.31,6.98,40.76,56.15,25.24,82.82, 10.07,5.57, P <0.05 or <0.01). There were interactions between selenium deficiency and iodine deficiency on tibial length, thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes (F values were 5.68,24.86,41.82,9.12, P <0.05 or <0.01 ). The tibial length of the selenium deficiency group[ (33.17 ± 0.34)mm] and combined selenium and iodine deficiency group[ (31.30 ± 0.87)mm] were significantly lower than the controls[ (34.12 ± 0.32)mm, P< 0.05]. Thickness of the growth plate cartilage [ (1.60 ± 0.18)mm ], layers of proliferative chondrocyte (8.54 ± 0.81), and hypertrophic chondrocyte (4.95 ± 0.37)of the combined selenium and iodine deficiency group were significantly decreased when compared to the selenium deficiency group[ (3.03 ± 0.10)mm, 14.68 ± 0.84,6.60 ± 0.31], iodine deficiency group[ (2.90 ± 0.09)mm, 13.75 ±0.33,6.61 ± 0.84 ] and the controls [ (3.19 ± 0.09) mm, 14.94 ± 0.36, 6.64 ± 0.26, P <0.05]. Thickness of the growth plate cartilage, layers of proliferative chondrocyte of the iodine deficiency group were lower than the controls(P<0.05). Conclusions Selenium deficiency impair tibial growth in first filial generation rats, iodine deficiency retarded the chondroncyte proliferation and decreases the thickness of growth plate cartilage in first filial generation rats, and combined selenium and iodine deficiency significantly impair the growth of bone and cartilage in first filial generation rats.     

细胞分化、凋亡及调节因子与大骨节病软骨细胞坏死发生的关系

大骨节病(KBD)是一种以侵犯青少年发育中透明软骨(骺软骨、骺板软骨和关节软骨)为主的地方性畸形性骨关节病,以15岁以下儿童发病为主,病区呈灶状分布,关节损害从四肢向脊柱发展.病理学研究认为软骨细胞坏死是大骨节病的主要病变,分布在软骨深层,近邻软骨内的血管渠,相当于肥大软骨细胞层,但也有在增殖层与肥大软骨细胞上层的移行带间.     

雪腐镰刀菌烯醇和硒对培养软骨组织CD44表达的影响

目的 研究大骨节病(KBD)有关病因因素对靶组织细胞的损伤和保护作用;探索引起软骨细胞变性坏死的机制。方法 采用细胞培养法于体外再建软骨组织模型,并加入KBD可疑致病因子雪腐镰刀菌烯醇(NIV)和保护因子硒,检测软骨细胞膜上透明质酸受体CD44和细胞培养液中可溶性CD44(SoCD44)。结果 软骨细胞膜上CD44的表达随着,NIV浓度的增加而减少,加硒后有增加趋势;细胞培养液中SoCD44浓度随NIV浓度升高逐渐降低,但高浓度组出现了增高,加硒后趋势不变;除对照组与加硒对照组外,组间差异有统计学意义(P<0.05)。结论 NIV能干扰软骨细胞表面粘附分子CD44表达,进而引起软骨细胞外基质代谢紊乱;补硒能够拮抗NIV对软骨细胞的损伤,但作用有限。     

大骨节病与骨关节病软骨组织死亡相关因子表达的比较

目的观察大骨节病软骨组织中程序化细胞死亡分子5（programmed cell death 5, PDCD5）和早期生长反应蛋白1（early
growth response protein-1, EGR-1）表达的变化及其在大骨节病软骨损伤中的作用。方法收集来自大骨节病患者关节软骨（KBD
组）10例,同时收集15例骨关节炎病人关节软骨（OA组）作为疾病对照,收集6例正常软骨作为健康对照（正常组）。采用免疫组化
染色法检测3组关节软骨组织中死亡受体调节因子PDCD5和EGR-1表达变化,并在显微镜下计数和分析3组关节软骨不同分层
间阳性表达率的显著性差异。结果（1）KBD软骨中层PDCD5阳性细胞表达率（41.35±2.97）%显著高于OA组（26.48±2.04）%和
正常组（19.02±1.88）%（P=0.001和P=0.000）,KBD软骨深层显著高于正常组和OA组（P=0.000和P=0.029）,OA组也高于正常组
（P=0.038）,而3组间的表层软骨细胞PDCD5阳性率无差异（P>0.05）;（2）在KBD软骨表层,EGR-1表达显著高于OA软骨和正常
软骨表层（P=0.000和P=0.000）,3组软骨的阳性表达率分别为（27.94±3.09）%、（3.20±1.49）%和（12.66±1.06）%,KBD软骨中层
EGR-1阳性细胞表达率显著低于OA组软骨（P=0.002）,而高于正常软骨（P=0.017）,KBD软骨和OA软骨深层阳性率均明显高于
正常组（P=0.000和P=0.001）,而KBD组和OA组的平均阳性率无统计学差异（P=0.187）;（3）KBD组与正常组的PDCD5和EGR-1
分别在3个软骨细胞层的表达均无相关性,而PDCD5和EGR-1在OA关节软骨表层呈强正相关。结论KBD软骨深层PDCD5显
著上调,而软骨表层和深层EGR-1显著高表达,提示这两种重要的细胞死亡相关因子在大骨节病软骨破坏过程中发挥重要作用。
     

大骨节病病区和非病区胎儿手指骨软骨的病理形态学对比观察

大骨节病的基本病理变化是软骨的变性,坏死,以及继发的修复性变化,导致骨骼的软骨内化骨障碍和随后的骨关节病。根据临床和X 线观察,以手指骨出现变化最早。关于最早的发病年龄,国内报告了出生三个月零七天的     

松质骨骨基质明胶复合软骨细胞构建组织工程软骨

目的 观察软骨细胞在同种异体松质骨骨基质明胶(BMG)上的生长、增殖和分化,探讨用松质骨BMG与软骨细胞体外构建组织工程软骨的效果.方法 分离1月龄兔关节软骨细胞,扩增后种植在松质骨BMG上体外构建组织工程软骨.于1、2、4、6周取材进行组织学、Ⅱ型胶原免疫组织化学及透射电镜观察.结果 软骨细胞在BMG上生长良好,分泌蛋白聚糖和胶原.随培养时间延长,细胞增殖,松质骨BMG空隙内细胞数量增多,软骨陷窝形成,基质分泌增强.培养6周时软骨细胞在BMG表面及孔隙内形成软骨样组织,免疫组织化学染色显示细胞周围基质富含Ⅱ型胶原,分布均匀:透射电镜显示软骨细胞超微结构正常,细胞周围有大量基质产生.结论 同种异体松质骨BMG可以促进软骨细胞的生长增殖,维持软骨细胞的分化表型;在体外成功构建了组织工程软骨,它是一种较好的软骨组织工程支架材料.

Abstract：

Objective To evaluate the use of cancellous bone matrix gelatin (BMG) combined with chondrocytes in constructing tissue-engineered cartilage by observing the growth, proliferation and differentiation of chondrocytes on allogeneic cancellous BMG. Methods The articular chondrocytes isolated from a 1-month-old rabbit were multiplied to a monolayer and seeded onto cancellous BMG to construct tissue-engineered cartilage in vitro during a period of 6 weeks. Samples were taken from the construct after 1, 2,4, and 6 weeks of culture and evaluated by histology, immunohistochemistry and transmission electron microscopy (TEM). Results The chondrocytes excreted matrix proteoglycan and collagen on cancellous BMG. With the prolongation of the culture time, the cells proliferated in the construct and the cells in the lacunae increased. Numerous chondrocytes were present the central region of the cancellous BMG and surrounded by extracellular matrix. By 6 weeks of culture, the BMG was covered with 15-20 layers of chondrocytes and cartilaginous tissue occurred in the pores throughout the cancellous BMG. Immunohistochemical staining showed rich and evenly distributed type Ⅱ collagen around the chondrocytes, and TEM revealed an ultrastructure of the chondrocyte similar to that of native chondroctyes, with abundant extracellular matrix produced around the cells. Conclusion Tissue-engineered cartilage can be constructed in vitro using allogeneic cancellous BMG combined with chondrocytes. Allogeneic cancellous BMG serves as a good scaffold material for tissue-engineered cartilage to promote the growth and proliferation of the seeded chondrocytes and allows maintenance of the differentiation phenotype of the cells.     

儿童与成人大骨节病关节软骨X型胶原表型和bFGF的表达

目的探讨儿童与成人大骨节病关节软骨中X型胶原表型和bFGF表达的分布特点。方法用单克隆和多克隆免疫组化法检测大骨节病与对照组关节软骨X型胶原表型和bFGF表达。结果(1)儿童大骨节病关节软骨深层X型胶原表型表达率(35.0%)比儿童对照组(63.0%)明显减少(P<0.01);(2)儿童大骨节病关节软骨中层X型胶原表型表达(0.0%)较成人大骨节病组(34.0%)减弱(P<0.01);(3)儿童大骨节病软骨表层bFGF表达率(6.0%)低于对照组(21.0%)(P<0.01),而中、深层表达(分别为12.0%、37.0%)与对照组(无表达)相比明显增多(P<0.01)。成人大骨节病关节软骨bFGF表达阳性率(分别为47.0%、37.0%、11.0%)与对照组(分别为19.0%、15.0%、1.0%)比较明显增多;(4)成人大骨节病关节软骨bFGF阳性表达在表、中层表达明显高于儿童组(P<0.01),深层儿童大骨节病组高于成人组(P<0.01)。结论X型胶原表型和bFGF表达在儿童与成人大骨节病关节软骨的分布不同。