SOX9基因在软骨发育过程中的表达谱分析及其质粒转染鼠骨髓基质细胞的实验研究

目的 检测并分析SOX9基因在软骨发育过程中的表达规律;构建其质粒并转染至培养的骨髓源基质细胞(BMSCs)中,通过细胞培养观察SOX9基因对BMSCs生长特性的影响,为可能的骨关节炎(OA)基因治疗提供理论基础.方法 用基因芯片技术建立妊娠胎鼠肢芽软骨发育过程的基因表达谱,分析SOX9基因在软骨发育过程中的表达规律;用噻唑蓝(MTT)法、免疫组织化学、苏木素-伊红(HE)染色、反转录-聚合酶链反应(RT-PCR)及酶联免疫吸附试验(ELISA)法检测SOX9基因转染MSCs的效果及产物的表达.结果 SOX9基因在软骨发育过程中的软骨形成关键期表达显著上调;pDC316-SOX-9质粒转染骨髓基质细胞后可促进骨髓基质细胞的增殖,细胞有向软骨细胞分化趋势.结论 SOX9能够促进软骨形成,SOX9基因质粒转染的MSCs可望在软骨组织工程临床治疗中得到更广泛的运用.     

趋化性细胞因子及其受体与脊髓损伤的研究

继发性脊髓损伤是伤后几小时至几天发生一系列损伤激活的自身破坏过程，结果是最初病灶周围原来完整的组织也发生自身破坏性病变，神经功能损害进一步发展且常常较原发性损伤更为严重和广泛。脊髓继发性损伤的病理包括微循环改变、缺血缺氧、再灌注损伤、组织水肿、细胞凋亡及坏死，以及     

无症状HBV携带者的HA,PCⅢ检测及其动态变化

检测了１５７例无症状ＨＢＶ携带（ＡＳＣ）者的血清透明质酸（ＨＡ）、Ⅲ型前胶原（ＰＣⅢ）含量，结果显示，其与病理理论的符合率〉７０％。动态观察结果提示，ＨＡ、ＰＣⅢ可作为观察ＡＳＣ患者病情变化的一项较敏感指标。     

盐酸小檗碱片含量测定方法的改进

盐酸小檗碱片是用于治疗肠道感染的抗菌药，其含量测定方法，2000年版《中国药典》采用的是反滴定法，2005年版《中国药典》采用的是高效液相色谱法法，液相法与滴定法相比较，方法快速、简便、重复性好。2005年版《中国药典》测定方法中流动相的组成是：磷酸盐缓冲液-乙腈（60：40），其磷酸盐比例高，对色谱柱的损害较大，乙腈对人体的危害较大。鉴于此，笔者改变流动相，用甲醇和水作为流动相，并与标准方法相比较，结果较为满意。     

同种异体骨板覆盖预防椎板切除术后硬膜粘连的研究

目的探讨同种异体骨板覆盖在预防椎板切除术后硬膜粘连方面的作用。方法a)实验研究12只成年雄性公绵羊行L3~4和L4~5全椎板切除后,在其中一处用“H”型同种异体冻干辐照骨板覆盖,另一处作自身对照。于术后4、8、12、16、20、24周取材,行大体、X线平片和光镜观察;并对4、24周取材行CT扫描。b)临床应用21例节段性腰椎间盘突出或腰椎管狭窄患者行全椎板切除,“H”型同种异体冻干辐照骨板覆盖。术后随访0.5~1.5a,观察临床症状及CT、MRI表现。结果a)实验研究与对照节段相比重建节段硬膜粘连程度明显减轻(P<0.01),椎管内容积无明显改变,硬膜囊形状保持良好,无明显压迫;b)临床应用18例患者疼痛基本或完全消失,恢复原来的生活和工作,另3例较术前有明显好转;CT、MRI显示椎管扩大,骨板无倾斜、移位,脊髓无压迫;均未见排异反应;术后1.5a,椎管内容物形态良好,骨板两侧已与相邻骨组织融合,密度相同。结论同种异体冻干辐照骨板具有良好的生物相容性、生物力学性能和诱导成骨作用,是一种良好的硬膜外覆盖材料,能有效阻止硬膜外粘连,防止术后再狭窄,可用于节段性椎板覆盖成形术。     

Early total hip arthroplasty for severe displaced acetabular fractures

Objective ： To investigate the effect of early total hip arthroplasty for severe displaced acetabular fractures. Methods： Total hip arthroplasty was performed on 17 cases of severe fracture of the acetabulum from 1997 to 2003. The mean follow-up was 2.1 years（ 1-6 years） and the average period from fracture to operation was 8 days （5-21 day）. The average age of the patients was 53 years （ 26-69 years）. Restdts： At the final follow-up the Harris hip score averaged 82（69-100） points and 15 cases have got a good outcome. There was one case of heterotopic bone formation. There were no radiographic evidences of lateloosening of the prosthesis. One patient had severe central displacement of the cup. Conclusions： In patients with severe displaced acetabular fractures, particularly in elderly patients, early total hip arthroplasty is probably an alternative efficient way to achieve a painless and stable hip.     

胃肠道肿瘤p16、cyclinD1基因表达的研究

[目的]探讨p16、cyclinD1在胃肠道肿瘤发生、分化中的变化规律。[方法]病人430例,用原位杂交技术观察不同病变状态胃肠道粘膜（正常粘膜、慢性炎症、癌前病变及肿瘤）中p16、cyclinD1的基因表达。[结果]正常胃、结肠粘膜p16阳性表达率为90.9％、86％,cyclinD1均为阴性,萎缩性胃炎、不典型增生、胃癌中p16阳性率分别为78.6％、37.5％、26.25％,cyclinD1阳性率依次为21％、58％、73.75％（P<0.05）;p16与cyclinD1有交叉共存现象,胃癌30％,结肠癌26.7％;同时,有反向表达现象,胃癌占70％,结肠癌为73.8％;p16阳性表达率与cyclinD1的阳性表达率呈显著负相关。[结论]p16的缺失、cyclinD1的激活,可单独或协同促进胃肠肿瘤的发生、发展;两者的反向表达趋势提示两者可能存在相互抑制机制。     

HBV携带者中抗—TB治疗所致肝损伤的临床分析

近年来,结核杆菌(TB)感染者有复发倾向,每年全世界约有1千万新发病人,约有3百万人死于结核病,是单因素所致的感染性疾病中病死率最高的疾病。HBV感染是人类最常见的病毒感染,全世界约有3亿,我国HBV感染率为10％～15％,因此有相当一部分人有HBV、TB混合感染。在长达6～8月抗TB治疗过程中,其主要副作用为肝损害。     

髋臼缺损重建及骨水泥和非骨水泥全髋关节置换术临床观察

目的：寻找全髋置换术髋臼缺损处理的方法。并对骨水泥和非骨水泥全髋关节置换术临床观察比较。方法：采用切下的股骨头,保留软骨修成帽状,软骨面位于腹腔侧,然后将臼和骨水泥与重髋臼相嵌。同时组同济髋晨骨水泥,而骨水泥固定股骨干假体,另一组采用骨不泥全髋关节置换。结果：进行临床和放射学随访。随访时间为１￣３．５年。发现非骨水泥组１６例优良率为９４％,采用骨水泥置换组１６例优良率为９５．６％。随访期间两者均未     

Induction of chondrogenesis of adipose-derived stem cells by novel recombinant TGF-β3 fusion protein

A new type of TGF-β3 fusion protein with targeted therapy function was constructed,and its feasibility and target specificity of inducing chondrogenesis were investigated by transfecting LAP-MMP-mTGF-β3 gene into adipose-derived stem cells (ADSCs).The recombinant pIRESEGFP-MMP was constructed by inserting the sense and antisense DNA of encoding the amino acid of the synthetic MMP enzyme cutting site into the eukaryotic expression vector pIRES-EGFP.LAP and mTGF-β3 fragments were obtained by using RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively,and the recombinant plasmid of pIRES-EGFPLAP-MMP-mTGF-β3 was constructed,which was transferred to ADSCs.The ADSCs were cultured and divided in three groups:experimental group (MMP group),negative control group (no MMP) and non-transfection group.The morphological changes were observed microscopically,and the expression of proteoglycan and type Ⅱ collagen (ColⅡ) was detected by using Alcian blue staining and immunohistochemistry staining at 7th,14th and 21st day after culture.The recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3 was correctly constructed by methods of enzyme cutting and sequencing analysis.The mTGF-β3 fusion protein was successfully expressed after transfection,and in the presence of the MMP,active protein mTGF-β3 was generated,which significantly promoted differentiation of ADSCs into chondrocytes and the expression of cartilage matrix.The novel fusion protein LAP-MMP-mTGF-β3 can targetedly induce differentiation of ADSCs into chondrocytes,which would open up prospects for target therapy of cartilage damage repair in future.