旋转铰链型与单纯铰链型膝关节肿瘤假体置换的临床比较研究

目的 研究膝关节肿瘤保肢手术两种假体置换的术后效果及其比较.方法 随访我院1999年6月2006年3月治疗的72例膝关节肿瘤患者，失访9例，余63例中40例采用单纯铰链型假体置换(A组)，23例采用旋转铰链型假体置换（B组）63例随访时间9月~8年平均3年9个月，按MSTS膝关节评分标准、ambulation scores、关节屈伸度及感染、复发、手术时间进行评估。结果：A组：手术时间2.6625±0.3362h，复发5例，转移死亡3例，感染2例； B组：手术时间2.5722±0.2878h，复发3例，转移死亡2例，感染2例 。B组1例术后发生旋转轴聚乙烯轴套断裂关节脱位，再次手术更换轴套后未再出现并发症，两组无假体松动病例。两组手术时间无统计学意义；去除复发及转移死亡病例后,剩50例, A组32例， MSTS评分平均23.0/30分。 B组18例，MSTS评分平均23.7/30分。两组MSTS评分p>0.05，无统计学意义；ambulation scores各分项总分均p<0.05，有统计学意义．两组关节伸直时为0°或接近0°,屈曲度A组119.3750±17.49°，B组125.0000±15.34°，无统计学意义（p>0.05）。结论： 1. 按ambulation scores标准，在步行舒适度和快速步行方面，旋转型铰链假体置换组与单纯铰链型假体置换比较有统计学意义；2.而MSTS标准则未能体现出两组术后功能方面的统计学意义，因此ambulation scores较MSTS评分更灵敏，更适合对膝关节保肢术后功能评价；3.两组在手术时间、术后临床屈伸功能及并发症方面，如复发、感染，未见显著差别，单纯铰链型不会出现脱位，而可旋转铰链型假体有脱位的可能。     

B cells as a target of immune modulation

B cells have recently been identified as an integral component of the immune system; they play a part in autoimmunity through antigen presentation, antibody secretion, and complement activation. Animal models of multiple sclerosis (MS) suggest that myelin destruction is partly mediated through B cell activation (and plasmablasts). MS patients with evidence of B cell involvement, as compared to those without, tend to have a worse prognosis. Finally, the significant decrease in new gadolinium-enhancing lesions, new T2 lesions, and relapses in MS patients treated with rituximab (a monoclonal antibody against CD20 on B cells) leads us to the conclusion that B cells play an important role in MS and that immune modulation of these cells may ameliorate the disease. This article will explore the role of B cells in MS and the rationale for the development of B cell–targeted therapeutics. MS is an immune-mediated disease that affects over 2 million people worldwide and is the number one cause of disability in young patients. Most therapeutic targets have focused on T cells; however, recently, the focus has shifted to the role of B cells in the pathogenesis of MS and the potential of B cells as a therapeutic target.     

Involvement of ADAM10 in axonal outgrowth and myelination of the peripheral nerve

The disintegrin and metalloproteinase 10 (ADAM10) is a membrane‐anchored metalloproteinase with both proteolytic and disintegrin characteristics. Here, we investigate the expression, regulation, and functional role of ADAM10 in axonal outgrowth and myelination of the peripheral nerve. Expression pattern analysis of 11 ADAM family members in co‐cultures of rat dorsal root ganglia (DRG) neurons and Schwann cells (SCs) demonstrated the most pronounced mRNA expression for ADAM10. In further studies, ADAM10 was found to be consistently upregulated in DRG‐SC co‐cultures before the induction of myelination. Neurons as well as SCs widely expressed ADAM10 at the protein level. In neurons, the expression of ADAM10 was exclusively limited to the axons before the induction of myelination. Inhibition of ADAM10 activity by the hydroxamate‐based inhibitors GI254023X and GW280264X resulted in a significant decrease in the mean axonal length. These data suggest that ADAM10 represents a prerequisite for myelination, although its activity is not required during the process of myelination itself as demonstrated by expression analysis of myelin protein zero (P0) and Sudan black staining. Hence, during the process of myelin formation, ADAM10 is highly upregulated and appears to be critically involved in axonal outgrowth that is a requirement for myelination in the peripheral nerve. © 2009 Wiley‐Liss, Inc.     

增生性瘢痕中血管内皮细胞生物学功能的研究

目的 探讨增生性瘢痕中血管内皮细胞的生物学功能及其与瘢痕形成的关系。方法 取人增生性瘢痕组织和正常皮肤组织，进行组织学观察。分离和纯化2种标本中的血管内皮细胞，应用酶联免疫吸附测定法分别检测单个血管内皮细胞中转化生长因子β1，（TGF-β1）、成纤维细胞生长因子2（FGF2）、血小板源性生长因子（PDGF）、内皮素1（ET-1）和血管内皮生长因子（VEGF）的水平。结果光学显微镜下可见正常皮肤微血管数目较少；增生性瘢痕微血管数目增多，血管狭长扭曲甚至闭塞。透射电镜可见增生性瘢痕中毛细血管管腔狭窄，有内皮细胞脱落。增生性瘢痕中血管内皮细胞分泌TGF-β1、PDGF、ET-1、VEGF、FGF2的水平分别为（60±8）、（30±4）、（0．12±0．03）、（52±5）、（18．1±1．2）μg／个细胞，明显低于正常皮肤（P〈0．05）。结论 增生性瘢痕中血管内皮细胞生物学功能减退，可能与瘢痕中胶原的大量产生和缺氧有关。     

脂肪来源间充质干细胞对大鼠肝移植的免疫调节作用

目的探讨大鼠脂肪来源间充质干细胞(MSC)对大鼠肝移植术后急性排斥反应的作用。方法分离、培养SD大鼠MSC,体外混合淋巴细胞培养(MLC)体系中,研究MSC对Wistar大鼠T细胞增殖的抑制作用。以SD与Wistar大鼠为供受体建立肝移植模型。随机分为MSC处理组与空白对照组,术后第7天检测肝功能、血清白细胞介素(IL)-2和白细胞介素(IL)-10水平、肝组织病理形态及肝细胞凋亡。结果体外MLC中,Wistar大鼠T细胞增殖明显受抑,抑制率为48.44%。实验组血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆红素(TBIL)、IL-2、IL-10分别为(134.2±45.0)、(162.5±30.5)U/L、(30.6±5.4)μmol/L、(187.35±18.26)、(193.95±37.62)μg/L;对照组上述指标分别为(355.6±54.3)、(296.4±71.2)U/L、(145.7±28.6)μmoL/L、(295.73±57.15)、(75.12±11.23)μg/L,两组差异有统计学意义(P<0.05);病检提示实验组排斥反应较对照组明显减轻;脱氧脲核苷酸缺口末端标记(TUNEL)检测提示实验组肝细胞凋亡程度明显低于对照组(P<0.05)。结论供体来源MSC能明显抑制MLC体系中受体源T细胞的增殖,并能显著减轻大鼠肝移植术后急性排斥反应。     

Determination of serotonin and 5-hydroxyindoleacetic acid in guinea pig and human prostate using HPLC

The finding of significant numbers of endocrine-paracrine (EP) cells in the prostate glands of guinea pigs and man suggests that these cells may be important in the regulation or modulation of prostatic function. Serotonin is a biogenic amine common to most prostatic EP cells. In order to extend current knowledge relating to these cells, an assay was developed using high-performance liquid chromatography to quantitate serotonin and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) in guinea pig and human prostatic tissue extracts. Levels of serotonin and 5-HIAA in the guinea pig whole-gland preparation were 105.4 +/- 70.6 ng/g and 48.4 +/- 95.7 ng/g, respectively. Normal human prostatic tissue contained 1423.9 +/- 750.8 ng/g serotonin and 66.7 +/- 92.8 ng/g 5-HIAA. Recoveries ranged between 60 and 100%. The detection limits were 24 pg/injection for serotonin and 12 pg/injection for 5-HIAA. This assay provides an expeditious, specific and highly sensitive method for the simultaneous determination of monoamines in guinea pig and human prostatic tissue.     

Bacterial ghosts (BGs)—Advanced antigen and drug delivery system

Bacterial ghosts (BGs) are empty bacterial envelopes of Gram-negative bacteria produced by controlled expression of cloned gene E, forming a lysis tunnel structure within the envelope of the living bacteria. BGs are devoid of cytoplasmic content and possess all bacterial bio-adhesive surface properties in their original state while not posing any infectious threat. BGs are ideally suited as an advanced drug delivery system (ADDS) for toxic substances in tumor therapy. The inner space of BGs can be loaded with either single components or combinations of peptides, drugs or DNA which provides an opportunity to design new types of (polyvalent) drug delivery vehicles. Uptake of BGs loaded with Doxorubicin (Dox) by CaCo2 cells led to effective Dox release from endo-lysosomal compartments and accumulation in the nucleus. Viability and proliferative capacity of the cells were significantly decreased (2–3 orders of magnitude) after internalization of Dox loaded BGs as compared to cells incubated with free Dox. The same effect was observed with leukemia cells. Melanoma cells also revealed a high capability to internalize BGs. These results indicate that BGs are able to target a range of types of cancer. BGs have also been investigated as DNA delivery vectors. Studies show DNA loaded BGs are efficiently phagocytosed and internalized by both professional APCs and tumor cells with up to 82% of cells expressing the plasmid-encoded reporter gene. Our studies with BGs as an ADDS system contribute (i) to optimize drug delivery for the treatment of cancer; (ii) define specific conditions for selection and preparation of BG formulations; (iii) and provide a background for the clinical application of BGs in cancer therapy.     

hTERT启动子调控hNIS基因介导肺癌细胞碘摄取和131I治疗的实验研究

目的 构建含人端粒酶反转录酶(hTERT)核心启动子调控的人钠/碘同向转运体(hNIS)基因重组腺病毒,并靶向转染至肺癌A549细胞中特异性表达.探讨hTERT启动子调控的hNIS基因介导放射性碘治疗肿瘤的可能性.方法 应用AdEasy系统构建重组腺病毒Ad-hTERT-hNIS,同时构建巨细胞病毒(CMV)启动子调控的hNIS重组腺病毒Ad-CMV.hNIS作为阳性对照,不含hNIS的重组腺病毒Ad-CMV作为阴性对照.应用反转录.聚合酶链反应(RT-PCR)方法验证hTERT在转染肿瘤细胞中的转录活性,摄碘实验检测表达的hNIS蛋白功能,细胞克隆形成实验评价131I对转染肿瘤细胞的毒性作用.结果 成功构建重组腺病毒Ad-hTERT-hNIS、Ad-CMV-hNIS及Ad-CMV,并经PCR验证正确.RT-PCR证实hNIS cDNA能从Ad-hTERT-hNIS转染的细胞中扩增出来.Ad-hTERT-hNIS和Ad-CMV-hNIS转染的肺癌A549细胞摄碘能力比阴性对照组Ad-CMV转染的细胞分别提高了23和31倍,且摄碘能力可以被NaClO4抑制.Ad-hTERT-hNIS和Ad-CMV-hNIS转染的肺癌A549细胞均可被131I杀死,2组细胞成活率分别为(31.2±1.45)%和(23.6±4.08)%,而阴性对照组和未转染病毒组分别为(89.0 ±2.99)%和(91.2 ±4.63)%.结论 hTERT启动子调控的hNIS重组腺病毒转染肿瘤细胞后,应用131I治疗有望成为一种新的基因靶向治疗手段.     

脑源性神经生长因子和神经干细胞联合移植对大脑中动脉阻塞大鼠神经功能恢复的作用

目的 将脑源性神经生长因子(BDNF)和神经干细胞(NSCs)单独及联合移植应用于大脑中动脉阻塞(MCAo)模型大鼠,观察BDNF和NSCs移植对大鼠缺血性脑卒中神经功能恢复的作用及BDNF对内源性和外源性NSCs增殖、迁移及分化的影响.方法 体外分离、培养新生大鼠海马NSCs,BrdU标记.实验动物随机分为A组(MCAo组);B组(MCAo+BDNF组);C组(MCAo+NSCs组);D组(MCAo+BDNF+NSCs组),每组16只,移植后进行神经功能损害评分(NSS),用免疫组织化学行BrdU、nestin、BrdU/NSE检测,分析结果.结果 移植后的2、4周神经功能评分分别为:A组5.3±0.5、5.3±0.5;B组4.0±0.8、3.8±0.5;C组3.5±0.6、3.5±0.6;D组2.0 ±0.8、1.8±1.0,D组显著好于其他3组(P<0.05),B组与c组显著好于A组(P<0.05).nes.tin阳性细胞数:A组1.24±1.13,B组2.59±1.44(P<0.05),BrdU阳性细胞数:A组0.52±0.68,B组1.65±1.10(P<0.05).BrdU阳性细胞数:C组6.08±1.52,D组10.26±1.96(P<0.05),BrdU/NSE双阳性细胞数:C组1.74±1.04,D组3.58±1.20(P<0.05).结论 BDNF和NSCs移植单独及联合应用对MCAo大鼠的神经功能恢复均有作用,两者联合具有协同作用.BDNF对内源性NSCs的激活、增殖有促进作用,对外源性NSCs的增殖、迁移及分化有促进作用.     

复合干细胞生物陶瓷在裸鼠异位成骨中的评价

目的 研究h-BMP-2基因转染骨髓间充质干细胞(BMSCs)在复合煅烧骨、β-TCP或直接植入裸鼠股部后的成骨能力。方法 通过影像学、组织学和形态计量学等方法,观察未经诱导、OS液诱导和h-BMP-2基因转染BMSCs在复合煅烧骨,或多孔β-TCP后植入裸鼠皮下,或直接制成细胞悬液注入,在4、8、12周诱导成骨和材料降解情况。结果 在裸鼠皮下,单纯生物陶瓷不能诱导成骨,而复合了未诱导、OS液诱导和h-BMP-2基因转染BMSCs的生物陶瓷均能成骨,成骨量为h-BMP-2基因转染组>OS液诱导组>未经诱导组(P<0.05),B-TCP可随骨长入而降解;注入裸鼠肌肉的OS液诱导的和h-BMP-2转染的BMSCs均能诱导成骨,而未经诱导MSCs则不能成骨。结论 复合人BMP基因转染BMSCs的β-TCP是一种理想的骨修复材料。