兔骨髓间充质干细胞/壳聚糖-胶原支架复合体构建软骨组织工程支架*

背景：软骨组织工程的核心是利用少量的细胞经体外培养、扩增后,在一定环境下附着在三维多孔支架上,再将细胞/支架复合体移植到体内形成新的组织。 目的：拟构建兔骨髓间充质干细胞/壳聚糖-胶原支架复合体,探讨该支架作为软骨组织工程支架的可行性。 设计、时间及地点：细胞-支架学体外观察,于2008-03/2009-02在武警医学院生物化学教研室完成。 材料：日本大耳白兔6只用于分离培养骨髓间充质干细胞。医用壳聚糖粉末为山东奥康生物科技有限公司产品。Ⅰ型胶原为Sigma公司产品。 方法：取3%的壳聚糖和2%的胶原混合的醋酸溶液,倒入72孔板内,-20 ℃预冷冻10 h,冻干机冷冻抽干24 h制作成壳聚糖-胶原支架。取第3代兔骨髓间充质干细胞,以1×109 L-1密度接种于支架内,构建细胞/支架复合体。 主要观察指标：傅里叶红外光谱、扫描电镜、液体置换法测定支架的理化性质,观察三维培养后细胞在支架上的生长情况。 结果：壳聚糖-胶原支架孔径为160~380 μm,平均为270 μm,孔相通性好,支架孔隙率为(86.00±5.12)%。傅里叶红外光谱仪测定数据表明复合支架组成物有典型的壳聚糖、胶原峰,未发现有聚乙二醇峰。细胞/支架共培养24,48,72 h后,骨髓间充质干细胞可渗入支架多孔结构内,并黏附在支架上成簇生长,部分细胞已与支架融合。 结论：壳聚糖-胶原支架基本符合软骨组织工程支架要求,能够作为种子细胞的承载体。 关键词：壳聚糖-胶原支架;骨髓间充质干细胞;冻干法;软骨组织工程     

福州地区老年男性雌激素受体α基因多态性与骨密度关系：150例数据分析

背景：关于男性骨质疏松症候选基因的研究,不同国家和地区的学者针对不同的种族和人群,得出的结论并不一致。 目的：分析福州地区汉族老年男性雌激素受体α基因XbaⅠ及PvuⅡ多态性分布,进一步研究其与骨密度的关系。 设计、时间及地点：骨密度及基因型相关性分析,于2007-02/2008-09在福建省中医药研究院门诊部及中医药管理局经络三级实验室完成。 对象：福州地区60岁以上汉族男性150例,年龄(68.92±5.33)岁,体质量(66.47±9.08) kg,体质量指数(24.23±3.12) kg/m2。 方法：双能X射线骨密度仪检测受试者正位L2~4、左侧股骨颈、大转子和Ward’s区骨密度,应用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)检测雌激素受体α基因XbaI及PvuⅡ多态性。 主要观察指标：骨密度;雌激素受体α基因型;年龄;身高;体质量。 结果：150例受试对象中,雌激素受体α XbaI基因型分别为XX型10例(占6.7%)、Xx型56例(占37.3%),xx型84例(占56.0%),X等位基因频率为25.3% 、x等位基因频率为74.7% ;PvuⅡ基因型分别为PP型18例(占12.0%)、Pp型78例(占52.0%),pp型54例(占36.0%),P等位基因频率为38.0% 、p等位基因频率为62.0% ;基因型分布均符合Hardy-Weinberg定律。分析基因型与骨密度的关系显示：与XX基因型相比,xx基因型人群在Ward’s区、大转子具有较高骨密度值,差异具有显著性 (P 分别为0.0192及0.087);xx基因型人群在大转子比Xx基因型具有较高骨密度值 (P < 0.05);PvuⅡ多态性各基因型间骨密度值均无差异。 结论：雌激素受体α基因XbaⅠ多态性与福州地区汉族老年男性骨密度相关,x基因是骨密度保护因素。     

Axon terminals expressing vesicular glutamate transporter VGLUT1 or VGLUT2 within the trigeminal motor nucleus of the rat: Origins and distribution patterns

Little is known about the significance of the two types of glutamatergic neurons (those expressing vesicular glutamate transporter VGLUT1 or VGLUT2) in the control of jaw movements. We thus examined the origin and distribution of axon terminals with VGLUT1 or VGLUT2 immunoreactivity within the trigeminal motor nucleus (Vm) in the rat. The Vm was divided into the dorsolateral division (Vm.dl; jaw‐closing motoneuron pool) and the ventromedial division (Vm.vm; jaw‐opening motoneuron pool). VGLUT1‐immunopositive terminals were seen within the Vm.dl only, whereas VGLUT2‐immunopositive ones were distributed to both the Vm.dl and the Vm.vm. Transection of the motor root eliminated almost all VGLUT1‐immunopositive axons in the Vm.dl, with no changes of VGLUT2 immunoreactivity in the two divisions, indicating that the VGLUT1‐ and VGLUT2‐immunopositive axons came from primary afferents in the mesencephalic trigeminal nucleus and premotor neurons for the Vm, respectively. In situ hybridization histochemistry revealed that VGLUT2 neurons were much more numerous than VGLUT1 neurons in the regions corresponding to the reported premotoneuron pool for the Vm. The results of immunofluorescence labeling combined with anterograde tract tracing further indicated that premotor neurons with VGLUT2 in the trigeminal sensory nuclei, the supratrigeminal region, and the reticular region ventral to the Vm sent axon terminals contacting trigeminal motoneurons and that some of the VGLUT1‐expressing premotor neurons in the reticular region ventral to the Vm sent axon terminals to jaw‐closing motoneurons. The present results suggested that the roles played by glutamatergic neurons in controlling jaw movements might be different between VGLUT1‐ and VGLUT2‐expressing neurons. J. Comp. Neurol. 512:595–612, 2009. © 2008 Wiley‐Liss, Inc.     

胰肾联合移植后应用连续性肾脏替代治疗1例

目的：探讨连续性肾脏替代治疗在胰肾联合移植后急性肾功能衰竭及多器官功能衰竭中应用的疗效。 方法：1例胰肾联合移植患者在术后出现移植肾急性肾功能衰竭、移植胰急性胰腺炎、肺感染、十二指肠空肠吻合口出血、腹膜炎等并发症,在积极的抗排斥、抗感染、抑制胰酶、止血、营养支持治疗的同时进行床边连续性肾脏替代治疗治疗,应用百特BM25机器进行连续性静脉-静脉血液滤过治疗,共治疗22 d。 结果：连续性肾脏替代治疗治疗期间生命体征、血流动力学稳定。并迅速控制了肺水肿,维持了水电解质酸碱平衡稳定。经过22 d共计180 h的连续性肾脏替代治疗治疗,患者移植胰肾功能逐渐恢复,患者住院40 d痊愈出院。 结论：连续性肾脏替代治疗在胰肾联合移植后急性肾功能衰竭及多器官功能衰竭的治疗中发挥了重要作用,是胰肾联合移植后良好的肾脏支持方式。     

腘窝淋巴结免疫细胞多种表面分子在PLNA评价致敏性中的综合分析

目的利用腘窝淋巴结实验(popliteal lymph node as-say,PLNA)分析腘窝淋巴结免疫细胞多个表面分子在PLNA评价致敏性中的综合变化,旨在提高PLNA的可靠性和灵敏性。方法选用♀BALB/c小鼠,选取具有致敏潜能物质氯化汞(HgCl2)、链脲佐菌素(STZ)、2,4,6-三硝基苯磺酸(TN-BS)和D-青霉胺(D-pen)为供试品,后肢足趾部注射免疫动物1次,5 d后处死动物,取注射侧腘窝淋巴结制备细胞悬液,流式细胞术检测腘窝淋巴结淋巴细胞、效应T细胞、抗原提呈细胞及其MHCⅡ、协同刺激和早期活化等十余种表面分子的变化,探讨这些表面分子在供试品致敏后的综合改变。结果 HgCl2和TNBS引起腘窝免疫细胞多种表面分子明显变化,D-pen和STZ所致的相应变化不明显。结论腘窝淋巴结免疫细胞表面分子变化的综合分析可以判断供试品的致敏潜能;不同供试品引起表面分子变化的种类和强弱有所不同,在致敏性评价中应综合考察各细胞表面分子的变化。     

5-氟尿嘧啶-N-琥珀酰壳聚糖纳米粒在荷瘤小鼠体内组织分布分析方法

目的:建立5-氟尿嘧啶-N-琥珀酰壳聚糖纳米粒(5-FU-Suc-Chi/NPs)在荷瘤小鼠血浆和组织中高效液相色谱(HPLC)分析方法。方法:生物样品经甲醇沉淀蛋白质并提取药物,采用HPLC法测定药物含量。色谱柱:Hypersil ODS C18(4.6mm×200mm,5μm);流动相:水-甲醇(99∶1)以磷酸调pH值6.2;流速:0.8mL.min-1;紫外检测波长:266nm;柱温:室温;进样量:20μL;内标:利巴韦林。结果:该方法线性范围为0.1~10mg.L-1,线性关系良好(r2=0.993 2~0.997 9),方法回收率、日内和日间精密度的RSD均小于15%。结论:该分析方法灵敏性、准确性较高,适用于5-FU-Suc-Chi/NPs荷瘤小鼠肿瘤靶向性及药动学研究。     

Assessment of the Chinese-version SF-36 in the Chinese opiate addicts

This study was designed to assess the reliability and validity of the Chinese version of the Medical Outcomes Study Short Form 36 (SF-36) assessed with opiate addicts in China. The sample was composed of 442 questionnaires from three regions in China, in 2004. The reliability of Chinese-version SF-36 was obtained using Cronbach's α and split-half coefficients and the validity was calculated using correlation analysis and confirmatory factor analysis method. The results of reliability and validity provide the first evidence that Chinese-version SF-36 is validated for Chinese addicts. The study's limitations are noted.     

Antiproliferative and antimetastatic effects of emodin on human pancreatic cancer

Emodin (1, 3, 8-trihydroxy-6-methylanthraquinone) is an active constituent isolated from the root of Rheum palmatum L and is the main effective component of some Chinese herbs and plants. Pharmacological studies have demonstrated that emodin exhibits anti-cancer effects on several human cancers. However, the molecular mechanisms of emodin-mediated tumor regression have not been fully defined. This study was performed to investigate the antiproliferative and antimetastatic effects of emodin on pancreatic cancer in vitro and in vivo. Our results showed that emodin induced a higher percentage of growth inhibition and apoptosis in the pancreatic cancer cell line SW1990 compared to that of control, and emodin suppressed the migration and invasion of SW1990 cells in a dose-dependent manner. To investigate the possible mechanisms involved in these events, we performed electrophoretic mobility shift assay (EMSA) and Western blot analysis, and found that emodin significantly down-regulated NF-κB DNA-binding activity, survivin and MMP-9 in SW1990 cells. Moreover, the expression of cleaved caspase-3 was up-regulated in SW1990 cells after treatment with emodin. In addition, a metastatic model simulating human pancreatic cancer was established by orthotopic implantation of histologically intact human tumor tissue into the pancreatic wall of nude mice. Oral administration of emodin significantly decreased tumor weight and metastasis compared to control. Furthermore, the expression of NF-κB, survivin and MMP-9 were also suppressed in tumor tissues after treatment with emodin. Collectively, our results indicated that emodin exerts antiproliferative and antimetastatic activity on pancreatic cancer both in?vitro and in?vivo, which may be related to down-regulation of NF-κB and its regulated molecules such as survivin and MMP-9 proteins. Consequently, these results provide important insights into emodin as an anti-invasive agent for the therapy of human pancreatic cancer.     

新型医用钛合金生物相容性评价

目的:评价新型医用钛合金Ti12.5Zr2.5Nb2.5Ta (TZNT)的生物相容性,并和3 种标准医用钛材料Ti6Al4V、Ti6Al7Nb和TA2进行对比.方法:通过细胞毒性试验观察细胞在TZNT、Ti6Al4V和Ti6Al7Nb合金离子析出培养液中的生长情况,计算在不同时间组的相对细胞增殖率,评价其毒性等级;通过皮下埋植试验观察4 种材料植入白鼠体内4 周和8 周的组织变化情况,计算炎细胞密度和纤维膜厚度的变化情况,评价其引起的组织反应程度.结果:TZNT、Ti6Al4V和Ti6Al7Nb在3 个时间组(2、 4、 6 d)的相对细胞增殖率均不低于100%,细胞毒性等级评定为0 级.和Ti6Al4V和Ti6Al7Nb相比,TZNT和TA2植入白鼠体内4 周和8 周后,无论是炎细胞密度还是植入物周围的纤维膜厚度均有所降低,引起的组织反应程度较低.结论:TZNT具有良好的生物相容性,是一种理想的生物医用钛合金.