1000/基于PC机上颅脑CT二维图像的三维重建

目的: 在普通个人计算机上利用颅脑CT二维图像实现对颅骨的三维重建。方法: 利用正常人颅脑CT扫描数据集，采集二维图像数据，利用Amira软件对二维图像数据进行颅骨的图像分割、提取其轮廓线、三维可视化重建。结果：利用颅脑CT二维图像数据在普通计算机上得到了颅骨精细的三维模型。此模型能够多个视角显示和观察，并能单独显示某一分离的结构，同时可对其进行编辑和精确距离测量。结论：在普通计算机上应用软件进行三维重建可以获得颅骨精细的三维模型，解决了传统上只能依赖螺旋CT机进行三维重建的难题，为科研、教学及建立颅脑模型数据库提供方法和积累了资料，为三维重建技术在普通计算机上的广泛应用提供重要理论依据。     

TOLL样受体4拮抗剂干预周围神经损伤后的瓦勒变性

BACKGROUND: The mechanism underlying Wallerian degeneration following peripheral nerve injury is complex. Immune regulation on Wallerian degeneration is beneficial for early repair of perpheral nerve injury. OBJECTIVE: To investigate the effects of Toll-like receptor 4 (TLR4) antagonist on Wallerian degeneration and axonal regeneration after early peripheral nerve injury in rats. METHODS: Fifty male Wistar rats were recruited and randomly divided into treatment group (n=20), model group (n=20) and sham group (n=10). The right sciatic nerves of rats in treatment and model groups were cut and sutured end-to-end, while the sciatic nerves of rats in sham group were only exposed. In the treatment group rats were intravenously injected with 0.15 mg/kg TAK-242 via tail vein 1 hour preoperatively and 7 days postoperatively, and the rats in the other two groups were given intravenous injection of the same volume of normal saline. The sciatic nerves were removed at 24 hours, 3, 4 and 7 days after surgery. RESULTS AND CONCLUSION: Real-time PCR indicated that the mRNA expressions of interleukin-1β and monocyte chemoattractant-1 were significantly increased in the model group compared with the sham group at 24 hours after surgery (both P < 0.001), while the expressions were significantly decreased after TAK-242 injection (both P < 0.001). Immunofluorescence showed that compared with the model group, down-regulated expression of CD68+ and iba1+ cells appeared in the treatment group at 3 days after surgery (P < 0.01, P < 0.05). Luxol fast blue staining revealed that demyelination at the sciatic nerve stump appeared in both model and treatment groups at postoperative 7 days, but myelin debris clearance in the treatment group was significantly reduced compared with the model group (P < 0.05). Hematoxylin-eosin staining showed that a lot of inflammatory cells, Schwann cells and regenerated nerve fibers at the sciatic nerve stump were found in the model group, while there were few inflammatory cells, Schwann cells and regenerated nerve fibers in the treatment group at 7 days after surgery. Immunohistochemistry found that the expression of growth-associated protein-43 in the treatment group was significantly lower than that in the model group at 4 days postoperatively (P < 0.05). Besides, compared with the model group, a significantly decreased sciatic functional index was found in the treatment group at 20, 30 and 40 days after surgery (P < 0.05). These results show that TLR4 antagonists delay early nerve regeneration in rats after sciatic nerve injury probably by inhibiting the TLR4 signaling pathway. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

人参皂苷对体外培养低氧海马神经元的保护作用

目的 观察人参皂苷（Rg2）对大鼠低氧海马神经元的保护作用及其机制。方法 取新生Wistar大鼠海马神经元体外培养14d，随机分为对照组、5μmol/L尼莫地平组（尼莫地平组）、Rg2 0．025mmol/L组（Rg2 1组）、Rg2 0．050mmol/L组（Rg2 2组）。将相应药物加入到培养液中孵育4h后，连续充以体积分数0．95N2＋体积分数0．05CO2混合气体，建立急性低氧细胞模型。台盼蓝染色计数存活细胞，吉姆萨染色观察细胞形态，荧光分光光度计法测细胞内Ca^2＋浓度。结果 尼莫地平组、Rg2两个剂量组的细胞死亡率均明显低于对照组，以Rg2 2组最低，差异均有显著性（X^2=3．37，P〈0．05）。海马神经元细胞内Ca^2＋浓度在尼莫地平组、Rg2两个剂量组均明显低于对照组，以Rg2 2组最低，差异均有显著性（F=466．58，q=6．76～48．82，P〈0．01）。结论 Rg2可显著降低体外培养大鼠低氧海马神经元的细胞死亡率，其机制可能是通过减少Ca^2＋内流而发挥作用。     

静脉曲张过程中血管内皮细胞NO合成酶表达变化

目的 探讨下肢静脉曲张过程中管壁血管内皮的变化及一氧化氮（NO）合成酶的改变与静脉曲张发生的关系。方法 选用17例曲张大隐静脉的内皮细胞作为观察组,选择3例正常静脉的内皮细胞作为对照组,应用苏木精-伊红染色观察内皮的一般形态学变化,采用免疫组织化学染色观察内皮细胞NO合成酶的表达情况。结果 正常静脉管壁厚度均匀一致,内弹力膜完整紧密,无断裂,内皮细胞分布均匀;而曲张静脉的管壁厚度不一,内膜不完整,内皮细胞大部分脱落。与正常静脉相比,代偿期曲张静脉内皮细胞NO合成酶表达增高（F=260.83,q=4.96,P〈0.05）,而失代偿期曲张静脉内皮细胞NO合成酶的表达则明显降低（q=11.83,P〈0.05）。结论 曲张静脉内皮组织损伤随病变进程逐渐加剧;曲张静脉内皮细胞NO合成酶的表达从最初的增高到逐渐减低,可作为曲张静脉由代偿到失代偿的标志。     

人类不同年龄段腰椎间盘组织细胞凋亡及其调控蛋白Bcl-2和Bax的表达

目的 探讨细胞凋亡及其调控蛋白在人类不同年龄段椎间盘组织退变中的作用及其可能存在的基因调控机制。方法 对0～58岁不同年龄段正常人腰椎间盘髓核细胞的形态结构进行苏木精-伊红染色光镜观察、TUNEL细胞凋亡检测、Bcl -2和Bax蛋白免疫组织化学染色。结果从胚胎后期开始,椎间盘髓核细胞数量随年龄增长而逐渐减少,到老年阶段髓核细胞的数量已经很少（H=95.123,P〈0.05）。从胚胎后期到成年,TUNEL阳性细胞率随年龄增长而逐渐降低,并降到整个生命过程中的最低点;继之,TUNEL阳性细胞率又逐年升高（F=205.391,P〈0.05）。自胚胎后期开始,Bcl-2蛋白就开始有较高水平的表达,但呈现逐年下降的趋势（F=81.385,P〈0.05）。Bax蛋白在胚胎后期即呈现较高水平表达,随年龄增长其表达水平逐渐降低（F=102.134,P〈0.05）;到成年后,Bax蛋白表达水平又有所升高,但不明显。结论 在整个生命过程中,随年龄增长大量腰椎间盘髓核细胞发生凋亡,细胞数量明显减少。Bcl-2蛋白可能参与了椎间盘细胞凋亡的调节,但表达水平较低,不能阻止细胞凋亡的发生。Bax蛋白的表达可能参与了整个生命过程中椎间盘髓核细胞的凋亡。     

调控Tim-3通路在免疫性肝损伤大鼠中对Th17细胞的影响

目的 建立免疫性肝损伤模型,探讨Tim-3通路在阻断或激活时对Th17细胞的影响。方法 大鼠尾静脉注射刀豆蛋白A (ConA) 8周,建立免疫性肝损伤模型。无菌取脾脏制备脾淋巴细胞,取外周血分离血清后于-20℃保存,取肝脏组织放入4%的多聚甲醛中固定备用。在96孔板上将脾淋巴细胞平均分成5组,即阻断实验组、阻断对照组、激活实验组、激活对照组和ConA对照组。用Tim-3的单克隆抗体即Anti-Tim-3来阻断Tim-3通路;用Tim-3的重组配体galectin-9来激活Tim-3通路,37℃、5% CO₂培养箱中培养72 h,收集细胞上清液于-20℃保存。生化分析法测血清中ALT、AST和ALB的表达;HE染色观察肝脏组织的病理变化;免疫组化法测肝脏组织中IL-17A和ROR-γt蛋白的表达;ELISA法测细胞上清液中IL-17A及IL-6的表达;实时定量PCR测各组脾淋巴细胞ROR-γt mRNA的表达。结果 与对照组相比,模型组HE染色可见明显的炎性细胞浸润、肝脏组织损伤及较多的假小叶,免疫组化可见IL-17A和ROR-γt蛋白的表达明显升高;与阻断对照组相比,阻断实验组中IL-17A和IL-6的水平升高(P<0.05);与激活对照组相比,激活实验组中IL-17A和IL-6的水平降低(P<0.05);实时定量PCR显示与阻断对照组相比,阻断实验组中ROR-γt mRNA的表达明显增加(P <0.05)。结论 免疫性肝损伤的发病与Th17细胞密切相关,免疫调控Tim-3通路可通过影响Th17细胞效应,进而参与免疫性肝损伤的发病机制。     

甲壳质抗特应性皮炎的实验研究

目的:探讨甲壳质对卵清蛋白(OVA)诱导的小鼠特应性皮炎(AD)的作用。方法:28只BALB/c小鼠随机分成3组:对照组(N组)(8只)、模型组(M组)(10只)、甲壳质组(E组)(10只)。通过小鼠腹腔注射及背部皮肤外敷OVA建立AD动物模型(模型组),甲壳质组在建模过程中同时给予3 mg/d甲壳质灌胃4周。建模成功后处死小鼠,取背部皮肤,进行石蜡切片HE和甲苯胺蓝染色,显微镜下观察表皮和真皮厚度的变化以及细胞浸润情况;ELISA法检测血清中总Ig E、总Ig G2a和OVA-specific Ig E的水平;取脾细胞进行体外培养,ELISA法测定培养液中细胞因子的含量。结果:与模型组小鼠比较,甲壳质组小鼠皮肤激发处炎症症状较轻,皮肤表皮层与真皮层增厚程度降低,真皮内炎症细胞浸润总量(P0.05)、嗜酸性粒细胞数量及肥大细胞数量(P0.01)明显减少。甲壳质组小鼠血清中总Ig E和OVA-specific Ig E的水平都明显降低(P0.05~0.001),Ig G2a水平显著升高(P0.001)。甲壳质组小鼠脾细胞体外培养产生的IL-12及IFN-γ水平明显高于模型组,而IL-4水平明显低于模型组(P0.05)。结论:甲壳质能有效减轻OVA诱导的小鼠AD症状,降低小鼠血清中Ig E水平,甲壳质的抗过敏作用可能与其诱导AD小鼠脾细胞产生Th1型细胞因子(IL-12、IFN-γ)有关。