Analysis of Hurthle cell neoplasms of the thyroid by interphase fluorescence in situ hybridization.

Recent studies have indicated that numerical chromosomal abnormalities including changes in p53 and cyclin D1 may be involved in Hurthle cell tumorigenesis. We analyzed a series of Hurthle cell neoplasms of the thyroid to evaluate the diagnostic and prognostic utility of numerical anomalies by DNA fluorescent probes for cyclin D1 and p53 gene loci and chromosomes 5, 7, 11, 12, 17, and 22. Interphase fluorescence in situ hybridization (FISH) analysis was performed on paraffin-embedded tissue sections from 10 Hurthle cell adenomas, 19 Hurthle cell carcinomas, and 7 normal thyroid tissues used as controls. Directly labeled fluorescent DNA probes for the centromere region of chromosomes 7, 11, 12, and 17 and locus-specific probes for chromosomes 5 and 22, cyclin D1, and p53 were utilized for dual-probe hybridizations. Sixty percent (6 of 10) Hurthle cell adenomas and 63% (12 of 19) Hurthle cell carcinomas showed chromosome gains. Twenty percent (2 of 10) Hurthle cell adenomas and 26% (5 of 19) Hurthle cell carcinomas showed chromosome losses. Normal thyroid tissues used as controls showed no chromosomal abnormalities. Among Hurthle cell tumors with chromosomal abnormalities, adenomas averaged 2.7 gains and 0.3 losses per case, and carcinomas averaged 3.3 gains and 0.6 losses per case. The two adenomas with chromosome losses each showed loss of one chromosome, whereas the five carcinomas with losses averaged 1.8 losses per case. Chromosome 22 was the most common loss identified, occurring in three of the 11 patients who died of disease. These results indicate that chromosomal imbalances as gains are common in both benign and malignant Hurthle cell neoplasms, but Hurthle cell carcinomas tend to have more chromosome losses than adenomas. Among Hurthle cell carcinomas in this study, chromosome losses were identified only from patients who died of disease. The loss of chromosome 22 may have prognostic value in Hurthle cell carcinoma of the thyroid.     

Biosynthetic response and mechanical properties of articular cartilage after injurious compression.

Traumatic joint injury is known to produce osteoarthritic degeneration of articular cartilage. To study the effects of injurious compression on the degradation and repair of cartilage in vitro, we developed a model that allows strain and strain rate-controlled loading of cartilage explants. The influence of strain rate on both cartilage matrix biosynthesis and mechanical properties was assessed after single injurious compressions. Loading with a strain rate of 0.01 s(-1) to a final strain of 50% resulted in no measured effect on the cells or on the extracellular matrix, although peak stresses reached levels of about 12 MPa. However, compression with strain rates of 0.1 and 1 s(-1) caused peak stresses of approximately 18 and 24 MPa, respectively, and resulted in significant decreases in both proteoglycan and total protein biosynthesis. The mechanical properties of the explants (compressive and shear stiffness) were also reduced with increasing strain rate. Additionally, cell viability decreased with increasing strain rate, and the remaining viable cells lost their ability to exhibit an increase in biosynthesis in response to low-amplitude dynamic mechanical stimulation. This latter decrease in reparative response was most dramatic in the tissue compressed at the highest strain rates. We conclude that strain rate (like peak stress or strain) is an important parameter in defining mechanical injury, and that cartilage injuriously compressed at high strain rates can lose its characteristic anabolic response to low-amplitude cyclic mechanical loading.     

rhPTH(1-34)对成骨细胞增殖、Collagen I及Osterix mRNA表达的影响

目的 观察间歇给予重组人甲状旁腺激素(1-34)[rhPTH(1-34)]对体外成骨细胞增殖、I型胶原蛋白(CoHagen I)及Osterix mRNA表达的影响,初步探讨rhPTH(1-34)对体外成骨细胞的作用机制.方法 体外培养新生大鼠成骨细胞,间歇循环给予0、10-11、10-10、10-9、10-8、10-7 M rhPTH(1-34)干预,(24 h为一循环,前12h给药),共2次,用MTT法检测细胞的增殖;RT-PCR法半定量测定成骨细胞Collagen I、Ostefix mRNA的表达.结果 显示rhPTH(1-34)可明显促进成骨细胞的增殖(P<0.05),促进成骨细胞Collagen I和Ostefix mRNA表达(P<0.05),101-9 M增殖、表达最明显,呈剂量依赖关系.结论 rhPTH(1-34)可促进成骨细胞的增殖、分化,可能是通过Collagen I和Ostefix mRNA表达来调节.     

五加补骨方及阿仑膦酸钠对模拟失重大鼠骨丢失干预比较

目的:比较五加补骨方及阿仑膦酸钠对3周尾吊模拟失重大鼠前后肢骨胳及肌肉丢失的干预作用.方法:自2009年3月至5月,6周龄雄性Wistar大鼠40只,按体重用随机区组法分为:五加补骨方组(HUC)、阿仑膦酸钠组(HUA)、空白组(CON)、模型组(HU),每组10只.动物实验周期为4周,HUC组全程给予五加补骨方(含刺五加、熟地黄、怀牛膝、牡蛎等)水煎剂,按体重10 ml/kg剂量每日灌胃1次,药液浓度0.704 g/ml;HUA组全程给定量阿仑膦酸钠片溶解混悬剂灌胃,按体重10 ml/kg剂量每周灌胃1次,药液浓度0.9 mg/ml;CON组和HU组均以上述方法全程给予蒸馏水.自第2周始,HU、HUC、HUA组均以尾部悬吊模拟失重3周.第4周末处死大鼠,分别测量血清钙、磷含量及碱性磷酸酶活性(ALP),肱骨和股骨骨密度(BMD)、肱骨和胫骨生物力学性能(biomechanical property)以及肱二头肌和腓肠肌重量指数.结果:与CON组比较,HU组血清Ca显著升高(P＜0.05),后肢BMD、力学性能及肌肉指数均显著降低(P＜0.01);与CON组比较,HUA组血清Ca显著升高(P＜0.05).HUC组血清ALP显著高于其他3组(P＜0.01).与HU组比较,HUC组及HUA组股骨BMD均显著升高,胫骨最大载荷、最大桡度及弹性载荷均有升高趋势;与HU组相比,HUC组及HUA组腓肠肌萎缩分别缓解12.5%和50%(P＞0.05),肱骨BMD均无显著变化,而HUA组肱骨最大桡度(P＜0.01)及弹性桡度(P＜0.05)均较HU组有显著降低.结论:中药复方和阿仑膦酸钠均可有效抑制模拟失重造成的后肢骨及肌肉丢失,改善其力学结构.中药复方在缓解上肢力学性能改变方面表现出一定优势.在治疗航天失重骨丢失类疾病上,五加补骨方与阿仑膦酸钠效果相当.     

介入栓塞术联合术后注射无水乙醇治疗颌面部动静脉畸形

目的：探讨以数字减影成像显影下血管介入栓塞技术联合术后注射无水乙醇治疗颌面部动静脉畸形的疗效。方法：对6例颌面部动静脉畸形患者行血管介入造影检查,并超选择性插管栓塞供血动脉及病灶,注入聚乙烯醇颗粒、无水乙醇等栓塞剂栓塞畸形血管团,术后根据恢复情况定期在病灶血管腔内注射无水乙醇。结果：6例颌面部动静脉畸形患者均栓塞成功,达到临床治疗目的,无严重并发症发生。结论：血管介入造影下超选择性插管栓塞联合术后无水乙醇是治疗颌面部动静脉畸形的一种有效、安全的方法。     

新辅助治疗在直肠癌脉管癌栓中的临床意义

目的 研究新辅助治疗后直肠癌脉管癌栓在中低位直肠癌组织中的分布规律,探讨脉管癌栓对直肠癌预后评价的意义.方法 按照入选标准收集2002年8月至2005年8月北京大学临床肿瘤学院连续收治的接受根治性切除的中低位直肠癌患者297例,根据是否接受术前辅助治疗将患者分成新辅助治疗组和对照组,观察两组患者术后病理标本中的脉管癌栓并根据术后随访资料研究脉管癌栓与预后的关系.采用x2检验分析其相关性,Kaplan-Meier生存法分析无病生存率和总生存率.结果 脉管癌栓总体阳性率为23.9%(71/297),新辅助治疗组阳性率为21.5%(31/144),对照组阳性率为26.1%(40/153),两组比较,差异无统计学意义(x2=0.872,P＞0.05).新辅助治疗组和对照组的脉管癌栓均与病理T、N分期及组织学分化程度有关(x2=13.490,27.401,7.323;16.188,21.623,16.534,P<0.05).新辅助治疗组脉管癌栓与局部复发无关(x2=0.000,P＞0.05),对照组的脉管癌栓与局部复发有关(x2=4.010,P<0.05).两组的脉管癌栓均与远处转移有关(x2=4.950,14.332,P<0.05).脉管癌栓阳性者比阴性者的无病生存率和总体生率低,分别为46.4%(26/56)和75.1%(148/197)、56.7%(34/60)和79.4%(166/209),两者比较,差异有统计学意义(x2=16.720,12.660,P<0.05).结论 新辅助治疗并未使脉管癌栓减少,但脉管癌栓在生物学行为上已经发生了变化,且脉管癌栓阳性的患者有可能从新辅助治疗中获益.     

MBD1甲基化调控胰腺癌BxPC-3细胞株mdr1转录及其表达的研究

目的 观察MBD1通过对mdr1转录区域结合位点甲基化影响,在转录水平间接调控胰腺癌mdr1基因表达.方法 应用RNA干扰技术对人胰腺癌细胞株BxPC-3中MBD1基因进行抑制;应用定量荧光聚合酶链式反应(FQ-PCR)技术、MSP(甲基化专用聚合酶链式反应)方法分别检测转染前后细胞株中的mdr1 mRNA的表达差异和转录区域结合位点甲基化变化.结果 成功构建并转染MBDI siRNAs至人原位胰腺腺癌细胞BxPC-3(BxPC-3)细胞中,证实MBDI mRNA水平明显下调(下调幅度为93.19%).MBD1下调后,mdr1 DNA转录区域(-110GC和-50GC)甲基化分别上调(上调幅度分别为181.98%和409.57%);mdr1 mRNA表达明显下调(下调幅度为97.79%).结论 MBD1可通过对mdr1转录区域结合位点甲基化的影响,在转录水平间接调控胰腺癌mdr1基因表达.     

白花丹素体外诱导前列腺癌PC-3细胞凋亡的研究

目的 探讨白花丹素对前列腺癌PC-3细胞增殖、凋亡的作用及其和RelA(p65)表达的关系.方法 应用不同浓度梯度的白花丹素(1、5、1O、15、20 μmol/L)共同培养PC-3细胞24、48 h,噻唑蓝(MTT)比色法检测PC-3细胞增殖活力,双染流式细胞仪检测凋亡细胞,透射电镜观察超微病理变化,计算药物半数抑制浓度(IC50).逆转录-聚合酶链反应(RT-PCR)法扩增检测RelA(p65).结果 24 h组在10～20 μmol/L,48 h组在5～20 μmol/L时均出现生长抑制,IC50分别为12.88、3.71 μmol/L.双染流式细胞仪检测显示PC-3随作用浓度的上升凋亡率增加并呈现浓度依赖关系.透射电镜观察作用后PC-3呈现典型凋亡表现.RT-PCR结果提示其细胞凋亡率同Rel A(p65)表达呈负相关.结论 白花丹素体外实验可能通过抑制Rel A(p65)表达诱导PC-3的凋亡.     

大鼠异体皮瓣移植后不用免疫抑制剂情况下移植物坏死的主要原因

目的 探讨大鼠异体皮瓣移植后不用免疫抑制剂情况下移植物坏死的主要原因.方法 实验分为3组,每组受者均为8只,以股动、静脉为蒂进行下腹部皮瓣移植.A组:以BrownNorway(BN)大鼠为供者,Lewis(LEW)大鼠为受者,移植术后2周内使用免疫抑制剂西罗莫司4mg·kg-1·d-1腹腔注射;B组:以BN大鼠为供者,LEW大鼠为受者,移植后不使用任何药物;C组:为LEW大鼠自体移植对照组,术后不使用任何药物.术后观察各组移植物的大体变化,并取移植物进行病理学检查,比较各组的病理学变化.结果 A组自术后17 d起,陆续出现皮瓣红斑,表皮脱落,真皮及皮下组织则最后坏死;B组受者排斥反应进展迅速,表皮不出现脱落现象,皮瓣直接完全发黑、坏死.C组移植物长期存活,未见排斥反应征象.A、B两组从出现排斥反应迹象到组织完全坏死的时间分别为(16.7±3.0)d和(5.4±1.1)d.病理检查显示:A组表皮组织首先被排斥,真皮和皮下组织由于大量淋巴细胞浸润而坏死.B组则为移植皮瓣真皮下血管网内皮细胞坏死,血管栓塞,皮肤内细胞缺血而坏死.结论 微血管栓塞是大鼠异体皮瓣移植后不应用免疫抑制剂情况下移植物坏死的主要原因.     

术中联合125Ⅰ粒子植入治疗中晚期食管鳞状细胞癌前瞻性研究

目的 探讨术中联合125Ⅰ粒子植入治疗胸段中晚期食管鳞状细胞癌(ESCC)的安全性及疗效.方法 前瞻性队列研究,入组时间为2000年1月至2004年8月.298例Ⅱ～Ⅲ期胸段ESCC患者随机分为术中联合125Ⅰ粒子植入组(A组,150例)及单纯手术组(B组,148例).A组术中直视下植入125Ⅰ粒子,术后通过CT和胸部X线片行粒子验证和质量评估.所有患者根据术中情况行食管癌根治术、姑息减瘤术或食管胃转流术.临床观察患者术后并发症,CT监测肿瘤影像学和局部复发情况,按WHO相关肿瘤评定标准评价患者近期疗效,随访术后1、3、5和7年生存率.结果 随访截至2008年8月31日,中位随访42个月(95%CI:37～55个月).A组术后粒子验证无移位、脱落,质量评估满意.A组及B组局部复发率分别为14.9%、38.7%,差异有统计学意义(P<0.05).术后3个月,A组有效率78.8%,与B组30.3%比较差异有统计学意义(P<0.05).两组并发症差异无统计学意义(P>0.05).A组及B组的3、5和7年生存率分别为64.0%比52.0%、42.7%比34.5%、25.1%比12.6%,差异有统计学意义(P<0.05).结论 术中联合125Ⅰ粒子植入治疗中晚期ESCC是简单、安全、有效的方法,可降低局部复发率、延长患者生存期.