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。     

精神分裂症子女个性行为特征的对照研究

目的 探索材神分裂症子女的个性和行为特征。方法 分别用艾森克个性问卷(EPQ)和Achenbach儿童行为量表(CBCL)，对研究组53例精神分裂症子女，对照组50例正常儿童评定。结果 两组儿童EPQ各项因子分无显著差异，在CBCL分析中，研究组退缩、社交问题和内向因子分显著高于对照组。研究组男童焦虑抑郁，女童躯体主诉分值也显著高于对照组。结论 精神分裂症子女个性无异常。存在有内向，退缩．社交困难等行为问题．应对其早期干预。     

先天性软骨发育不全成纤维细胞生长因子受体3基因1138位核苷酸点突变的检测

目的 验证成纤维细胞生长因子受体3(fibroblast growth factor receptor 3，FGFR3)跨膜区1138位核苷酸为先天性软骨发育不全突变热点及用变性梯度电泳方法筛查的效果。方法 对17例临床诊断为先天性软骨发育不全患者进行基因组DNA聚合酶链反应－限制性酶切片段长度多态性（polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP)分析，并用变性梯度凝胶电泳（denaturing gradient gel electrophoresis,DGGE)进行其它突变位点的筛查。结果 17例患者中14例RFLP检测显示能被Sfc I酶切，提示存在1138位核苷酸G→A的转换，且均为杂合子。17例用Msp I酶切均阴性，提示无G→C的颠换。DGGE方法的筛查中，酶切阳性的14例标本均显示存在杂合型突变。3例PCR－RFLP检测阴性的标本未显示突变位点，提示在该扩增区域确实不存在突变位点。结论 FGFR3跨膜区1138核苷酸G→A的突变是软骨发育不全的主要发病机理，DGGE可作为筛查某一区域基因突变的敏感而可靠的检测手段，尤其是对杂合子的检测。     

人类ZAKβ与YWHAZ蛋白相互作用的初步研究

目的研究ZAKβ与YWHAZ蛋白之间的相互作用。方法以ZAKβ为诱饵蛋白,利用酵母双杂交筛选人类肝脏cDNA文库,经GST Pull-down、免疫共沉淀和细胞共定位实验分别验证ZAKβ与猎物蛋白在体外和细胞内的相互作用,构建ZAKβ的截短体以确定相互作用的区域,用磷酸化抗体检测ZAKβ在相互作用中的磷酸化作用,并通过流式细胞仪检测这对蛋白相互作用对细胞周期的影响。结果利用酵母双杂交筛选到一个能与ZAKβ相互作用的蛋白YWHAZ,并在体外和细胞内都验证了这二者的相互作用,发现这两个蛋白都能共定位于293T细胞的胞质中,确定了ZAKβ的N端1~250氨基酸为与YWHAZ蛋白相互作用的区域。体外实验发现了这二者的相互作用涉及到磷酸化反应,发现这对蛋白的相互作用能提高293T细胞的G2期水平。结论首次发现并证实了ZAKβ和YWHAZ之间的相互作用,发现该相互作用涉及到磷酸化过程并能够对细胞周期产生影响。     

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。     

Photodynamic therapy induced Fas-mediated apoptosis in human carcinoma cells

Photodynamic therapy (PDT) is a clinical approach that utilizes light-activated drugs for the treatment of a variety of pathologic conditions. Human poorly (CNE2) and moderately differentiated (TW0-1) human nasopharyngeal carcinoma (NPC) cells undergo rapid apoptosis when treated with PDT sensitized with Hypocrellin A (HA) and Hypocrellin B (HB). It has been shown that these compounds have a strong photodynamic effect on tumors and viruses. The initiating events of PDT sensitized HA and HB-induced apoptosis are poorly defined. In the current study, we sought to determine whether Fas/FasL upregulation and involvement of mitochondrial events are an early event in HA and HB-treated PDT induced apoptosis. Loss of mitochondrial transmembrane potential, release of cytochrome c, involvement of caspases-8 and -3 and the status caspase-3 specific substrate PARP, were evaluated in PDT treated tumor cells. Photoactivation of HA and HB enhanced both CD95/CD95L expression and induced CD95-signaling dependent cell death in all tumor cell lines studied. CD95/ CD95L expression appeared within 2 h following light activation and appeared to be a primary event in PDT induced apoptosis. Furthermore, these results indicate that release of mitochondrial cytochrome c into the cytoplasm is a secondary event following the activation of initiator caspase-8 preceding caspase-3 activation, cleavage of PARP and DNA fragmentation. Cytochrome c appeared in the cytosol within 2-3 h post PDT. Cleavage of PARP was observed at 3-4 h following PDT and caspase-3 specific inhibitor DEVD-CHO and broad-spectrum caspases inhibitor z-VAD-fmk blocked caspase-3 activation and PARP cleavage suggesting that caspase-3 plays an important role in HA and HB-induced apoptosis.