兔骨髓基质细胞诱导成骨细胞复合同种异体生物衍生骨实验研究

目的探讨体外培养的骨髓基质细胞与自体来源的生物衍生骨复合后的生长特性，为进一步研究骨髓基质细胞作为种子细胞，以及探索一种良好的支架材料提供实验依据。方法分离纯化兔骨髓基质细胞并诱导成成骨细胞后与同种异体来源的生物衍生骨复合后体外培养，并在相差显微镜和扫描电镜下观察其生长情况。结果骨髓基质细胞在生物衍生骨上贴附生长、增殖良好。结论骨髓基质细胞可作为骨组织工程的理想种子细胞，与生物衍生骨复合后可作为骨组织工程的载体。     

骨水泥强化椎弓根螺钉固定治疗老年退行性腰椎疾病的疗效观察

目的评价骨水泥强化椎弓根螺钉固定治疗老年退行性腰椎疾病的近期临床疗效。方法回顾性分析2011年6月~2013年5月采用聚甲基丙烯酸甲酯(PMMA)骨水泥强化椎弓根螺钉固定结合后路椎体间植入聚醚醚酮(PEEK)材质椎间融合器治疗老年退行性腰椎疾病30例。所有患者术前骨密度检测均符合骨质疏松诊断(超声骨密度值测定-2.5)。结果 30例患者均顺利完成手术,术中无神经及硬膜损伤,骨水泥无严重渗漏,术后复查X线、CT显示骨水泥分布均匀。随访10~21个月,平均(16±2.11)个月,神经受压症状均得到改善。VAS评分术前(7.01±1.44)、术后6个月随访为(3.00±0.57)、末次随访为(2.23±1.19);JOA评分术前为(9.98±5.64)、术后6个月随访为(17.99±1.41)、末次随访为(18.42±1.47);ODI评分术前为(0.64±0.24)、术后6个月为(0.27±0.07)、末次随访为(0.22±0.09)。三项评分术后6个月、末次随访分别与术前对比差异有统计学意义;术后6个月和末次随访对比差异无统计学意义。末次随访时复查X线或CT显示椎弓根螺钉无松动,椎间融合器无下沉,椎间融合满意,融合率为86.7%。结论使用骨水泥强化椎弓根螺钉能够提高螺钉对伴有骨质疏松的椎体的握持力,防止椎弓根螺钉松动,保证较高的椎间融合率,是治疗老年退行性腰椎疾病一种安全而有效的手术方式。     

股骨粗隆间骨折合并同侧股骨干骨折的手术治疗

目的探讨股骨粗隆间骨折合并同侧股骨干骨折的手术治疗方法及疗效。方法回顾分析我院自1998年1月至2005年12月收治股骨粗隆间骨折合并同侧股骨干骨折患者18例，全部病例均采用手术切开复位内固定，股骨粗隆间骨折应用动力髋内固定，股骨干骨折采用记忆合金环抱器内固定。结果术后随访2～8年，平均4．2年。全部病例均获骨性愈合，疗效满意。结论股骨粗隆间骨折合并同侧股骨干骨折在下肢骨折中相对较少，但处理较为复杂和困难，治疗有一定难度，使用动力髋与记忆合金环抱器同时进行内固定，方法简单，固定牢靠，患肢可早期进行功能锻炼，这种方法是治疗单侧肢体多处骨折较好的方法之一。     

细胞因子对肾小球系膜细胞合成PAF的影响

本文首先建立了血小板活化因子（ＰＡＦ）的生物活性检测法，并通过体外细胞培养技术，测定了肾小球系膜细胞在经ＬＰＳ、ＩＬ-１β、ＩＬ-６和ＴＮＦα的短暂刺激后，其培养上清中ＰＡＦ的活性。结果发现，它们均能诱导系膜细胞合成和分泌ＰＡＦ，ＩＬ-１β、ＩＬ-６和ＴＮＦα的诱生作用并呈一定的剂量依赖关系。实验提示上述因子在肾小球免疫病理损伤中，除直接作用外，也可通过诱生ＰＡＦ而间接发挥作用。应用ＰＡＦ拮抗剂来阻断这一途径，可能具有一定的临床应用价值。     

谷氨酸脱羧酶65片段与IL－10双基因真核表达载体的构建与鉴定

 目的 优化以往构建的以预防 1 型糖尿病为目的的全长谷氨酸脱羧酶（GAD）65 DNA疫苗，尝试构建GAD 65片段与IL-10双基因真核表达载体。方法 从GAD65质粒中扩增出GAD190-315片段和GAD490-570片段的cDNA，以overlap PCR法将之分别与hIL-2信号肽cDNA拼接，得到SGAD190-315、SGAD490-570融合基因。以p43.2-mIL-10质粒为模板，用PCR方法扩增出IL-10基因。将SGAD190-315、SGAD490-570融合基因分别与IL-10基因依次克隆入双启动子真核表达载体pBudCE4.1中，构建出2种双基因重组真核表达载体pBud-SGAD190-315/IL-10和pBud-SGAD490-570/IL-10。2种重组真核表达载体经测序鉴定正确后，用脂质体介导的方法转染COS-7细胞，转染后48和72 h以蛋白质印迹法检测细胞裂解产物及上清液中SGAD190-315和SGAD490-570融合基因的表达，ELISA方法检测细胞上清液中IL-10的表达。结果　核酸序列测定表明克隆的SGAD190-315、SGAD490-570融合基因和IL-10基因序列与报告序列一致。蛋白质印迹法和ELISA方法均检测到SGAD190-315/IL-10和SGAD490-570/IL-10重组真核表达载体在COS-7细胞中的表达。结论　成功构建了2种GAD65片段与IL-10双基因真核表达载体，为1型糖尿病的基因疫苗预防研究提供了实验基础。     

Immunohistochemical detection of XIAP and p63 in adenomatous hyperplasia, atypical adenomatous hyperplasia, bronchioloalveolar carcinoma and well-differentiated adenocarcinoma.

The critical distinction of bronchioloalveolar carcinoma (BAC), well-differentiated adenocarcinoma (WDAC) of lung, adenomatous hyperplasia (AH) and atypical adenomatous hyperplasia (AAH), is based on morphological criteria alone, and is therefore potentially subjective. We examined expression of two markers, X-linked inhibitor of apoptosis protein (XIAP), the most potent of the inhibitor of apoptosis protein (IAP) family, and p63, a marker of bronchial reserve cells (BRC) and squamous cells, in these entities. H&E slides of 37 tissue blocks from 27 patients were reviewed and classified as AH (n=7), AAH (n=8), BAC (n=9) and WDAC (n=13). Immunostaining was performed on 4 mum sections with monoclonal anti-XIAP and monoclonal anti-p63. Granular or heterogeneous cytoplasmic staining for XIAP and nuclear staining for p63 were considered positive. Neither XIAP nor p63 were detected in normal lung alveolar cells. All seven AHs were negative for XIAP and negative or focally positive for p63. All eight AAHs were positive for XIAP and displayed p63 positivity in scattered cells. All BACs displayed XIAP positivity, which ranged from focal/weak to diffuse/strong. p63 was negative in seven and focally positive in two of nine BACs. Twelve of 13 WDACs showed XIAP positivity in a similar pattern to BAC; all were negative for p63. One aberrant case diagnosed on H & E as WDAC was negative for XIAP but strongly positive for p63. Significant XIAP expression appears to be useful for distinguishing AAH from AH. Commonality of XIAP staining in AAH, BAC and WDAC supports the possibility that AAH may be a pre-malignant lesion. The rarity of p63 expression confirms previous reports and supports a nonbronchial histogenesis of these entities. In contrast, diffuse p63 staining may facilitate the identification of rare cases that may have been misclassified as alveolar in origin based on morphology but may be of BRC origin.     

The pattern and frequency of t(14;18) translocation and immunophenotype in Asian follicular lymphoma

AIMS: Follicular lymphoma is frequently associated with t(14;18)(q32;q21) translocation. This study was undertaken to determine the pattern of Bcl-2, CD10 and Bcl-6 expression in relation to t(14;18) translocation in follicular lymphoma from a cohort of a multi-ethnic Asian population. METHODS AND RESULTS: Sixty-two cases of follicular lymphoma were retrieved for immunohistochemistry, and t(14;18) translocation analysis by polymerase chain reaction and fluorescent in-situ hybridization techniques. Bcl-2 expression was present in 74% of the cases. CD10 expression was also relatively low (61%), with decreasing frequency of expression in high-grade tumours. Bcl-6 protein was expressed in most of the tumours (88%) regardless of the tumour grade. The t(14;18) translocation was detected in 46 cases (74%) with an extremely high rate of t(14;18) translocation in ethnic Indian cases (100%). CONCLUSION: The frequency of t(14;18) translocation in this series of follicular lymphomas was higher when compared with previous Asian reports, but in accordance with European and North American findings. CD10 expression is strongly associated with a t(14;18) translocation event, but the overall CD10 expression was relatively low, possibly due to the high proportion of high-grade tumours in the series. t(14;18) translocation was not associated with Bcl-2 or Bcl-6 expression.     

术前颈椎过伸功能与颈椎后路单开门椎管扩大成形术后前凸角度丢失的关系

目的 探讨术前颈椎过伸功能与颈椎后路单开门椎管扩大成形术后前凸角度丢失的关系。方法 回顾性分析首都医科大学大兴教学医院骨科2017年1月-2018年12月58例行颈椎后路单开门椎管扩大成形术患者临床资料,其中男45例、女13例,年龄49~85岁(平均64.8岁)。术前测量患者中立侧位X线片上的T₁倾斜角、矢状面垂直轴(SVA),以及中立侧位、过伸位X线片的C₂~C₇ Cobb角。随访12~24个月,术后再次测量中立侧位X线片上的C₂~C₇ Cobb角。术前颈椎过伸功能测量值为术前过伸位X线片C₂~C₇ Cobb角度减去术前中立侧位X线片C₂~C₇ Cobb角。前凸角度丢失量为术前中立侧位片C₂~C₇ Cobb角减去末次随访时中立侧位片C₂~C₇ Cobb角。依据58例患者术前颈椎过伸功能均值(8.7°)分为两组,≥8.7°为A组,＜8.7°为 B 组。比较两组患者术前及术后影像及临床资料,同时对58例患者的影像学资料与临床资料进行相关性分析。结果 A组25例患者年龄54~83岁,B组33例患者年龄49~85岁,两组患者术前年龄、性别、疾病种类差异均无统计学意义(P值均>0.05)。术前A组颈椎过伸功能(14.09°±4.75°)大于B组(4.62°±2.54°),A组T₁倾斜角(17.00°±3.40°)小于B组(29.68°±6.34°),颈椎前凸角度丢失[1.10(-0.85,4.00)]小于B组[8.60 (7.70,12.40)],差异均有统计学意义(P值均＜0.01)。颈椎过伸功能与前凸角度丢失之间呈负相关(r=-0.965, P＜0.01),T₁倾斜角与前凸角度丢失之间呈正相关(r=0.954, P＜0.01),颈椎过伸功能与T₁倾斜角呈负相关(r=-0.900, P＜0.01);SVA与T₁倾斜角、颈椎过伸功能、术后前凸角度丢失均无相关性(r=-0.065、0.216、-0.202, P＞0.05)。术后JOA评分改善率与过伸角度变化、SVA及T₁倾斜角均无相关性(r=0.201、-0.034、-0.213, P值均＞0.05)。A组术后JOA改善率为69%±23%,B 组术后JOA改善率为62%±23%,两组差异无统计学意义(t=1.147, P＞0.05)。术后Odom's分级评价A组优良率为88.0%(22/25),B组优良率为63.6%(21/33),差异有统计学意义(χ² =4.403, P<0.05)。结论 对于后路单开门椎管扩大成形术患者,颈椎过伸功能与前凸角度丢失存在相关性,术前过伸功能越低,术后越易发生前凸角度丢失,可作为术前预判术后颈椎曲度变化的参数之一。