CD147抗体对破骨细胞中基质金属蛋白酶活性影响的体外研究

目的:建立人破骨细胞(Osteoclast,OCs)体外诱导分化模型,研究CD147单克隆抗体对OCs分化过程中基质金属蛋白酶-9(Matrix metalloproteinases 9,MMP-9)和基质金属蛋白酶-2(Matrix metalloproteinases 2,MMP-2)表达及活性的影响。方法:通过采集健康成年志愿者外周血所分离的单个核细胞贴壁培养,应用NF-κB配体激活因子(Receptor or Activator ofNF-KB Ligand,RANKL)与巨噬细胞集落刺激因子(Macrophage colony stimulating factor,M-CSF)诱导单个核细胞向OCs分化。本实验分为抗体组(RANKL+M-CSF+CD147单克隆抗体)与对照组(RANKL+M-CSF)。抗酒石酸酸性磷酸酶染色(Tartrate-resistant acid phosphatase,TRAP)与骨吸收实验检测鉴定OCs分化及活性情况,Real-Time PCR技术检测CD147、MMP-9和MMP-2 mRNA在破骨前体细胞(Osteoclast precursor cells,OPCs)中表达情况,明胶酶谱法检测细胞培养上清中MMP-9、MMP-2酶蛋白的活性变化情况。结果:①对照组经TRAP染色和骨吸收实验检测到破骨样细胞形成并具有骨吸收功能,而抗体组OCs分化及活性均受到抑制;②在24、48小时时相点上抗体组CD147、MMP-2及MMP-9 mRNA的相对表达量均低于相应的对照组(P<0.05),且MMP-2、MMP-9 mRNA的相对表达量与CD147 mRNA的表达量呈正相关。③明胶酶谱检测细胞培养上清,可见在24、48小时两时相点上抗体组OCs细胞中MMP-2、MMP-9酶原及活性酶的酶解量均较相应对照组明显降低(P<0.05)。结论:CD147单克隆抗体可以抑制OCs分化成熟过程中MMP-9及MMP-2的表达与活性;CD147对MMP-2、MMP-9活性的调节,可能是其对OCs活化调节的机制之一。     

CCR1参与肺癌分泌因子诱导破骨细胞生成

目的建立肺癌细胞与单核/巨噬细胞间接共培养模型,观察在肺癌细胞条件培养基作用下抑制前体破骨细胞(RAW264.7)的CCR1对破骨细胞生成的影响。方法收集肺癌细胞A549的条件培养基,将条件培养基以不同浓度加入RAW264.7培养基中共培养,在共培养体系中加或不加CCR1特异性拮抗剂BX471。细胞培养至第5天进行TRAP染色,提取不同条件作用后RAW264.7细胞的总RNA,采用Real-time PCR检测CCR1及破骨细胞标志基因cathepsin K、TRAP mRNA的表达,使用细胞免疫荧光和Western blot检测不同时期RAW264.7细胞中CCR1及Cathepsin K蛋白的表达量。结果加入A549细胞条件培养基的RAW264.7细胞TRAP阳性细胞显著增多,条件培养基明显上调RAW264.7的CCR1、Cathepsin K、TRAP的表达且呈浓度依赖,CCR1特异性拮抗剂BX471可抑制肺癌分泌因子对破骨细胞的活化。结论肺癌细胞分泌因子可促进破骨前体细胞向破骨细胞分化,CCR1参与了肺癌细胞分泌因子对破骨细胞的活化过程。     

CCN3在肺癌骨转移灶中高表达及其对破骨细胞的作用

背景与目的:肺癌易转移至骨并导致溶骨性破坏的具体机制目前还不清楚,本研究观察CCN3在肺癌骨转移灶中的表达水平以及在前体成骨细胞(MC3T3-E1)与前体破骨细胞(RAW264.7)共培养体系研究CCN3重组蛋白对前体破骨细胞分化的影响。方法:采用免疫组化检测9例肺癌骨转移转灶组织及相应癌旁组织中CCN3的表达;构建RAW264.7细胞与MC3T3-E1细胞共培养模型,向共培养体系中加或不加800 ng/mL的CCN3重组蛋白,采用Real-time PCR检测RAW264.7细胞的破骨细胞标志基因抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)、组织蛋白酶K(cathepsin K,CK)、降钙素受体(calcitonin receptor,CTR)mRNA的表达,并通过TRAP染色法对破骨细胞进行鉴定。结果:肺癌骨转移灶组织中的CCN3表达量明显高于癌旁组织;CCN3重组蛋白可显著促进共培养体系中的RAW264.7细胞的破骨细胞标志基因TRAP、CK、CTR的mRNA的表达量;TRAP染色可见对照组TRAP阳性细胞数[(6.2±1.48)个]明显低于加了CCN3重组蛋白的试验组[(21.4±3.04)个],差异具有统计学意义(P<0.05)。结论:CCN3在肺癌骨转移灶中高表达及其可诱导破骨细胞分化,CCN3可能在肺癌骨转移和溶骨性破坏中具有重要作用。     

脊髓损伤后星形胶质细胞的病理变化及相关治疗措施的进展

脊髓损伤(spinal cord injury,SCI)往往预后较差,致残率高。其病理过程尚不十分清楚,治疗也一直是困扰医学界的难题。脊髓损伤后星形胶质细胞(astrocyte,AS)的病理变化及其相关的治疗措施一直是研究的重点。笔者就近年来这一领域的研究进展综述如下。     

椎板间隙后路显微内镜治疗腰椎间盘突出症724例

目的　报道经后路椎板间隙显微内镜 (MED Ⅱ手术 )治疗腰椎间盘突出症的手术技术、临床效果及适应证选择。方法　局麻或持硬膜外麻醉下 ,床旁C 臂机定位 ,沿导针依次扩张 ,置放手术通道和内镜 ,电视监控下扩大后路椎板间隙 ,咬除局部增生内聚小关节及增厚黄韧带 ,扩大及清理神经根通道 ,摘除突出椎间盘髓核组织。结果　MED手术共 72 4患者 ,86 7腰椎间盘 ,平均住院时间 16 .8d。平均手术出血量 4 5ml。平均手术时间 5 8分钟。疗效优良率达 93.2 %。结论　MED手术比常规手术创伤小 ,恢复快 ,基本保持了脊柱后柱完整 ,对脊柱稳定性影响小 ,手术视野清晰 ,手术安全 ,疗效可靠。随临床经验积累和手术技巧提高 ,MED -Ⅱ手术适应证不断扩大     

CTGF在软骨中的研究进展

<正>结缔组织生长因子(CTGF)是CCN家族成员之一。CCN(CTGF、Cef10/cyr61和Nov的缩写)家族包括半光氨酸富集蛋白(cysteine-rich61,CYR61/CCN1)、CTGF(CCN2)、肾母细胞瘤过度表达蛋白(ne-     

成人腰椎峡部裂型滑脱症的微创治疗

Objective To investigate the clinical experiences and technical skills of adult isthmic spondylolisthesis. Methods Twenty-one patients with adult isthmic spondylolisthesis underwent minimally invasive surgery. There were 12 men and 9 women, with the mean age of 51.7 years. Isthmic spondylolisthe-sis occurred at the L4.5 in 7 patients, and at the L5S1 in 14 patients. According to Meyerding classification, 13 cases were of Grade Ⅰ, 7 of Grade Ⅱ, 1 of Grade Ⅲ. Under general anesthesia, guiding by fluoroscopy, the surgeries performed minimally invasive microendoscopic(METRx) techniques for posterior decompression, interbody cage fusion and novel Sextant-R percutaneous pedicle screw techniques for deformity reduction and fixation. Results Seventeen cases were followed up 1 year postoperatively. The average low back pain VAS reduced from preoperative 6.0±2.6 to postoperative 2.9±2.5. The average leg pain VAS decreased from preoperative 6.7±3.3 to postoperative 2.8±1.6. The average ODI decreased from preoperative 44.3% to post-operative 27.1%. The Nakai good and excellent rate was 90%. The mean operative time 170 min, blood loss 160 ml, and postoperative stay in bed 7.5 d. The sagittal spondylolisthesis rate significantly decreased from preoperative 35.5%±2.5% to postoperative 8.3%±7.5%. The lordotic angle from preoperative 11.5°±1.7° in-creased to postoperative 16.8°±9.5°; the intervertebral disc height from preoperative (5.4±2.5) mm increased to postoperative (9.1±3.0) mm. According to Lenke judgement for fusion, complete fusion rate was 76%, in-complete fusion rate was 12%, nonfusion rate was 12%. Conclusion The minimally invasive microendo-scopic (METRx) assisted with a novel Sextant-R percutaneous pedicle screw systems for deformity reduction and fixation to treat adult isthmic spondylolisthesis, is not only a minimally invasive and safe surgical tech-nique, also an effective treatment for deformity reduction and fixation.     

Ad-huVEGF121基因转染对兔股骨头Ⅰ型胶原表达及ALP活性的影响

目的观察VEGF在坏死股骨头内诱导成骨的作用.方法重组腺病毒Ad-hVEGF121注射入坏死的股骨头,应用酶学和免疫组织化学技术检测血清碱性磷酸酶(ALP)的活性和Ⅰ型胶原(collagen Ⅰ)的表达.结果基因转染组ALP活性和collagen Ⅰ表达在各个时相点显著高于空病毒对照组.结论VEGF具有在坏死股骨头内诱导成骨的功能.     

肝移植术后并发脊柱结核1例

1临床资料 患者,男性,37岁,因"肝移植术后5年,胸背痛3月,双下肢麻木,乏力40 d"于2006年3月13日入骨科.2001年7月13日在西南医院行了"肝移植术",术后予强的松冲击治疗5 d,长期服用环孢素A至今.     

Treatment of extremely-lateral lumbar disc herniation with METRx through intertransversal route

To study the feasibility and effects of disc resection in patients with extremely-lateral disc herrniation (ELDH) with microendoscopic tubular retractor (METRx) through intertransversal route. Methods: In the period from March 2000 to December 2002, 9 patients with ELDH underwent disc resection with METRx system through intratransversal route and the clinical outcome was evaluated with Nakai standard. Results: The operation time ranged from 60 to 120 min with an average of 75 min; the blood loss ranged from 50 to 120 ml with an average of 60 ml; and the hospitalization time ranged from 5 to 19 d with an average of 13 d. No wound infection, neurological damages, pseudomeningocele or other complications occurred. The rate of excellent and good outcome was 88.9%. Conchusion: Disc removal in patients with ELDH can be done with METRx system and satisfactory restdts are obtained.