应用蛋白质组学和组织芯片研究annexin A2在胃癌中的表达

目的: 研究胃癌中膜联蛋白A2(ANXA2)的差异表达及其与胃癌临床病理参数的关系。方法: 激光捕获显微切割技术(LCM)纯化15例胃腺癌细胞(GAC)和其癌旁正常胃黏膜上皮细胞(NGEC),采用¹⁸O/¹⁶O分别标记2种细胞样本酶切后的多肽混合物。纳升级反相液相色谱串联质谱(Nano-RPLC-MS/MS)定量鉴定GAC和NGEC的差异表达蛋白质。Western blotting验证差异蛋白ANXA2的表达。免疫组化方法检测组织芯片中75对胃癌组织和癌旁组织中ANXA2的表达,分析ANXA2表达与胃癌临床病理参数的关系。结果: 共筛选出78个差异表达蛋白质,其中ANXA2蛋白的表达水平在GAC中较NGEC中明显上调(2.32∶ 1),Western blotting检测ANXA2蛋白表达在GAC组升高(P<0.01);免疫组化显示:ANXA2的表达与胃癌的浸润深度、淋巴结转移、分化程度、TNM分期和肿瘤大小相关(P<0.01),与年龄、性别和远处转移无关(P>0.05)。结论: ANXA2蛋白的上调可能在胃癌的生物学行为中发挥重要作用。     

膝关节前交叉韧带后外束股骨止点位置的解剖

目的 通过对膝关节前交叉韧带后外束股骨止点的解剖测量,找到确定前交叉韧带后外束股骨止点的简单可行的方法,为双束重建前交叉韧带手术中的骨道定位提供理论依据。方法 解剖20例新鲜膝关节标本（25~45岁）。在屈膝90°位,测量前交叉韧带后外束股骨止点中心点距股骨髁间窝外侧壁前方、后方和下方软骨缘的距离,再对测量数据进行评估和对比。结果前交叉韧带后外束股骨止点中心点距离股骨前方软骨缘(8.74±1.39)mm,距离后方软骨缘(8.69±1.57)mm（P =0.926）。后外束止点中心点距离股骨下方软骨缘(5.06±0.77)mm。结论膝关节屈膝90°位时,前交叉韧带后外束的股骨止点中心点位于股骨髁间窝外侧壁,距离下方软骨缘5mm,距离前方和后方软骨缘的距离相等。在前交叉韧带双束重建的手术中,应用本研究的结果能够简单、快捷地确定前交叉韧带后外束股骨骨道位置。     

Ndrg2对成年大鼠皮层机械性损伤后神经发生的影响

目的:探讨Ndrg2对成年大鼠皮层机械损伤后神经发生的影响及可能作用机制。方法:选取成年雄性Sprague-Dawley大鼠24只,利用立体定位法对大鼠皮层进行针刺机械损伤。随后分为4组(n=6):对照组(注射生理盐水)、病毒对照组(注射空载体病毒)、上调Ndrg2组(注射AAV-Ndrg2病毒)及抑制剂组(注射AAV-Ndrg2病毒和-分泌酶抑制剂DAPT)。术后7 d免疫荧光检测损伤区周边Ndrg2及Nestin表达变化。结果:在对照组和病毒对照组,针道周边的Ndrg2+细胞明显增多,并表达神经干细胞标记物Nestin;与病毒对照组相比,上调Ndrg2组的Ndrg2+及Ndrg2+/Nestin~+细胞进一步增多(P0.01);注射Notch信号抑制剂DAPT后,与上调Ndrg2组相比,针道周边Nestin~+细胞数目明显减少(P0.01),但Ndrg2+细胞数目无明显改变(P0.05)。结论:大鼠皮层机械损伤后,Ndrg2可参与损伤区的神经发生过程;Notch信号通路可能在Ndrg2促神经发生中发挥关键作用。     

狂犬病高发地区犬只感染情况调查分析

目的 分析自然状况下犬只感染狂犬病毒状况及其与人间狂犬病的关系.方法 2005年10月到2006年12月间,在狂犬病疫情最严重的贵州、广西和湖南3省区的狂犬病高、中、低发病地区分别选择1-2个市作为标本采样点,用直接免疫荧光法和RT-PCR两种方法 检测标本.结果 在3省区的15个市共收集犬脑标本2887份,检测阳性标本66份,感染率为2.3%,不同地区犬感染率与当地人狂犬病发病率不呈线性相关.结论 犬只感染狂犬病毒普遍存在,这是我国狂犬病高发区疫情严重的一个重要因素.     

自噬相关蛋白PTEN、MAP1LC3在舌癌中的表达与意义

目的 探讨自噬相关蛋白人第10号染色体缺失的磷酸酶及张力蛋白同源基因(phosphatase and ten-sinhomology deleted on chromosome ten,PTEN)、微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,MAP1LC3)在舌鳞状细胞癌(tongue squamous cell carcinoma,TSCC)、癌前病变及癌旁组织中的表达及相关性,探讨其与TSCC发生、发展及转移的关系.方法 采用免疫组化SP法检测46例TSCC手术标本及13例癌前病变、13例癌旁组织标本中PTEN、MAP1LC3的蛋白表达,分析其与临床病理特征间的关系.结果 PTEN(X2=9.905,P=0.007)、MAP1LC3(X2=9.577,P=0.008)在TSCC组织、癌前病变、癌旁组织中的蛋白表达差异有统计学意义.在TSCC中,PTEN蛋白的阳性表达率(34.78%)明显低于癌旁组织(69.23%,X2=4.926,P=0.026)及癌前病变组织(76.92%,X2=7.302,P=0.007);MAP1LC3蛋白的阳性表达率(28.26%)明显低于癌旁组织(61.53%,X2=4.896,P=0.027)及癌前病变组织(69.23%,X2=7.275,P=0.007),差异均有统计学意义.PTEN与MAP1LC3蛋白表达与患者年龄、性别、吸烟、饮酒、细胞分化无关(P>0.05).颈淋巴结从无转移到转移以及临床分期越晚,PTEN蛋白及MAP1LC3蛋白的阳性表达率降低,差异有统计学意义(P<0.05).PTEN和MAP1LC3两种蛋白在复发组中阳性表达率皆较低,复发组与非复发组PTEN的蛋白表达差异有统计学意义(X2=4.269,P=0.039),而复发组与非复发组MAP1LC3的蛋白表达差异尚无统计学意义(X2=0.343,P=0.453).PTEN与MAP1LC3蛋白阳性表达呈正相关.结论 自噬相关蛋白PTEN与MAP1LC3的低表达与TSCC的临床病理特征相关,对TSCC的诊断与预后有一定参考价值.     

Biomimetic Approach to Perforation Repair Using Dental Pulp Stem Cells and Dentin Matrix Protein 1

Introduction

Dentin regeneration could be an ideal treatment option to restore tissue function. This study was conducted to evaluate the ability of dental pulp stem cells (DPSCs) and dentin matrix protein 1 (DMP1) impregnated within a collagen scaffold to regenerate dentin.

Methods

Simulated perforations were created in 18 dentin wafers made from freshly extracted human molars. Six groups were established. They were (1) empty wafers, (2) mineral trioxide aggregate, (3) collagen scaffold, (4) scaffold with DMP1, (5) scaffold with DPSCs, and (6) scaffold with DPSCs and DMP1. One sample was placed subcutaneously in each mouse with three mice in each group. After 12 weeks, the samples were subjected to radiographic, histological, and immunohistochemical evaluations.

Results

DPSCs impregnated within a collagen scaffold differentiated into odontoblast-like cells forming a highly cellular, vascular, and mineralized matrix in the presence of DMP1.

Conclusions

A triad consisting of DPSCs, DMP1, and a collagen scaffold promotes dentin regeneration in a simulated perforation repair model.     

颞颌关节上腔灌洗术治疗不可复性关节盘前移位的临床研究

目的：评价颞下颌关节穿刺灌洗术治疗不可复性关节盘前移位的临床疗效。方法：关节上腔灌洗术治疗30例因不可复性关节盘前移位而引起张口受限的患者。结果：张口受限病程在6个月之内，关节冲洗疗效显著。结论：关节上腔灌洗术所需器械价廉易得，操作简单，疗效显著，在我国目前关节镜的普及率较低的情况下，具有较好的临床应用价值。     

Dual release of dexamethasone and TGF-β3 from polymeric microspheres for stem cell matrix accumulation in a rat disc degeneration model

Low back pain is frequently caused by nucleus pulposus (NP) degeneration. Tissue engineering is a powerful therapeutic strategy which could restore the normal biomechanical motion of the human spine. Previously we reported that a new nanostructured three-dimensional poly(lactide-co-glycolide) (PLGA) microsphere, which is loaded with dexamethasone and growth factor embedded heparin/poly(l-lysine) nanoparticles via a layer-by-layer system, was an effective cell carrier in vitro for NP tissue engineering. This study aimed to investigate whether the implantation of adipose-derived stem cell (ADSC)-seeded PLGA microspheres into the rat intervertebral disc could regenerate the degenerated disc. Changes in disc height by plain radiograph, T2-weighted signal intensity in magnetic resonance imaging (MRI), histology, immunohistochemistry and matrix-associated gene expression were evaluated in normal controls (NCs) (without operations), a degeneration control (DC) group (with needle puncture, injected only with Dulbecco’s modified Eagle’s medium), a PLGA microspheres (PMs) treatment group (with needle puncture, PLGA microspheres only injection), and PLGA microspheres loaded with ADSCs treatment (PMA) group (with needle puncture, PLGA microspheres loaded with ADSC injection) for a 24-week period. The results showed that at 24 weeks post-transplantation, the PM and PMA groups regained disc height values of ～63% and 76% and MRI signal intensities of ～47% and 76%, respectively, compared to the NC group. Biochemistry, immunohistochemistry and gene expression analysis also indicated the restoration of proteoglycan accumulation in the discs of the PM and PMA groups. However, there was almost no restoration of proteoglycan accumulation in the discs of the DC group compared with the PM and PMA groups. Taken together, these data suggest that ADSC-seeded PLGA microspheres could partly regenerate the degenerated disc in vivo after implantation into the rat degenerative intervertebral disc.     

Hepcidin对小鼠脑组织中铁沉积及运动行为能力的影响

目的：观察hepcidin对小鼠脑组织铁沉积及运动行为能力的影响,并探讨其作用机制。方法：将雌雄各半的昆明小鼠随机分为7组,分别为空白对照组、铁负荷组（2mg·ml^-1、1mg·ml^-1、0.5mg·ml^-1）、hepcidin 5μg＋铁负荷组（2mg·ml^-1、1mg·ml^-1、0.5mg·ml^-1）,以不同浓度的右旋糖酐铁隔日大腿肌肉注射建立铁过载模型。8周后,血细胞计数仪检测血液中血红蛋白含量,全自动生化分析仪检测血清铁含量,试剂盒检测血清中转铁蛋白（Tf）和总铁结合力（TIBC）;原子吸收光谱法检测测定脑组织中铁的含量;ROS试剂盒检测脑组织中活性氧簇（Reactive oxygen species,ROS）含量;Morris水迷宫实验观察小鼠运动行为的改变。结果：与空白对照组比较,铁负荷组小鼠血清铁及血红蛋白含量明显增加,转铁蛋白（Tf）和总铁结合力（TIBC）明显减少,小脑、中脑和纹状体中铁含量和ROS明显升高,且呈剂量依赖性;Morris水迷宫实验结果显示,铁负荷组小鼠与空白对照组比较,平均逃逸时间明显增加,而靶象限活动时间明显下降（p〈0.05）;而hepcidin组均出现相反方向变化：hepcidin组小鼠平均逃逸时间明显下降,而靶象限活动时间明显增加。结论：铁过负荷对小鼠小脑、中脑和纹状体造成明显毒性损害,并引起脑的氧化应激,认知行为和空间记忆力下降;而hepcidin可抑制铁负荷引起的脑铁沉积和运动行为能力变化。