全反式维甲酸对TGF-β1刺激的肝星状细胞COL1α2、MMP-2、TIMP-1以及信号通路的影响

目的观察全反式维甲酸对大鼠肝星状细胞(HSC-T6)增殖,以及经转化生长因子-β1(TGF-β1)刺激后Ⅰ型胶原α1链蛋白基因(COL1α2)、基质金属蛋白酶2(MMP-2)、金属蛋白酶组织抑制物1(TIMP-1)及Smad 2/3的表达变化,探讨其对肝纤维化的作用及其分子机制。方法不同浓度的全反式维甲酸(0.1、1、10μmol/L)分别作用HSC-T612、24、48h,采用噻唑蓝(MTT)比色实验检测HSC-T6增殖活性;另外,经TGF-β1(5ng/mL)刺激后的HSC-T6用不同浓度全反式维甲酸干预24h后,利用逆转录酶-聚合酶链反应(RT-PCR)技术测定COL1α2、MMP-2及TIMP-1mRNA的表达,细胞免疫化学染色法检测smad2/3蛋白的表达。结果全反式维甲酸能够抑制HSC-T6的增殖,而且具有浓度依赖性(P<0.05);受外源TGF-β1 5ng/mL刺激后,HSC-T6中COL1α2、MMP-2、TIMP-1 mRNA及Smad2/3蛋白表达较对照组明显增强(P<0.05),而全反式维甲酸干预能够有效降低HSC-T6受TGF-β1刺激后COL1α2、MMP-2、TIMP-1mRNA及Smad2/3蛋白的表达(P<0.05)。结论全反式维甲酸能够抑制HSC-T6的增殖,下调HSC-T6受TGF-β1刺激后COL1α2、MMP-2、TIMP-1mRNA的表达,并且可能通过抑制HSC-T6中TGF-β1的下游信号蛋白Smad 2/3的表达,影响TGF-β1/Smad信号通路,发挥其抗肝纤维化作用。     

太白楤木对肝星状细胞凋亡及相关基因表达的影响

目的研究太白楤木对肝星状细胞(HSC)凋亡及相关基因表达的影响,以探讨其抗肝纤维化作用的机制。方法 30只雄性SD大鼠随机平均分为阴性对照组、秋水仙碱组及太白楤木组,分别给予生理盐水、秋水仙碱及太白楤木灌胃,3d后制备含药血清。体外培养大鼠肝星状细胞株HSC-T6,应用上述各组含药血清进行干预,48h后采用流式细胞仪检测细胞凋亡率;采用RT-PCR法检测HSC中Bcl-2、Bcl-xl、Bax及Bcl-xs的mRNA表达水平;采用Western blot法检测HSC中Bcl-2、Bax及α-SMA的蛋白表达水平。结果 1与阴性对照组相比,秋水仙碱组和太白楤木组HSC凋亡率明显增加(P<0.05);与秋水仙碱组相比,太白楤木组凋亡率更高(P<0.05)。2秋水仙碱及太白楤木均可抑制HSC中Bcl-2的表达(P<0.05),并促进Bax的表达(P<0.05),其中太白楤木的作用更加明显(P<0.05)。3秋水仙碱及太白楤木对HSC中Bcl-xl及Bcl-xs的表达无明显影响。4秋水仙碱及太白楤木均可抑制HSC中α-SMA的表达(P<0.05),且太白楤木的作用更加明显(P<0.05)。结论太白楤木可促进活化的HSC凋亡,其促HSC凋亡的机制可能与下调Bcl-2表达、上调Bax表达有关。     

树突状细胞Wnt/β-catenin信号通路对哮喘小鼠气道变态反应性炎症的调控作用

目的探讨Wnt/β-catenin信号通路对哮喘小鼠气道变态反应性炎症的调控作用。方法通过小鼠骨髓细胞诱导分化建立树突状细胞(DCs)体系,并用氯化锂(LiCl)和PKF118-310干预细胞,光镜下观察细胞形态;同种异体混合淋巴细胞反应检测DCs刺激淋巴细胞的增殖能力;Western blot方法检测DCs中GSK-3β和β-catenin蛋白的表达水平。选用BALB/c雌性小鼠为研究对象,使用OVA联合氢氧化铝构建哮喘模型,并用LiCl和PKF干预小鼠,干预结束后收集标本,观察支气管肺泡灌洗液(BALF)中细胞总数和嗜酸性粒细胞百分比;HE染色观察小鼠肺部变态反应性炎症情况;ELISA检测BALF、脾细胞培养上清液中干扰素-γ(IFN-γ)和白介素-4(IL-4)的含量及血清总IgE抗体水平;Western blot检测肺脏组织中Wnt/β-catenin信号通路的重要组分GSK-3β和β-catenin蛋白的表达变化。结果(1)LiCl组DCs对同种异体T细胞的刺激和促增殖能力明显弱于PKF组DCs(P<0.05);(2)LiCl组DCs中GSK-3β的蛋白表达水平明显低于PKF组,而其β-catenin的蛋白表达水平显著高于后者(P<0.05);(3)动物实验中,与哮喘组相比,LiCl组BALF中白细胞总数和嗜酸性粒细胞百分比均较低,而PKF组与此相反(P<0.05);(4)肺组织病理表明,LiCl组肺部炎症程度最轻,PKF组最重(P<0.05);(5)LiCl组小鼠BALF及脾脏中IL-4水平最低,IFN-γ水平最高;PKF组与此相反,哮喘组介于两者之间(P<0.05);(6)各组小鼠血清中总IgE含量,与正常对照组比较,实验组IgE显著升高(P<0.05);LiCl组总IgE水平明显低于哮喘组和PKF组(P<0.05);(7)LiCl组肺组织中GSK-3β的蛋白表达水平明显低于各组(P<0.05),而其β-catenin的蛋白表达水平显著高于各组(P<0.05),PKF组β-catenin的蛋白表达水平明显低于各组(P<0.05),哮喘组和PKF组之间GSK-3β的蛋白表达水平未见明显差异(P>0.05)。结论 LiCl和PKF118-310可通过调控哮喘小鼠肺组织中Wnt/β-catenin信号通路重要组分GSK-3β和β-catenin的蛋白表达而改变哮喘严重程度,为哮喘的治疗提供了新方向。     

人类宫颈癌中白细胞介素-6与髓样细胞白血病-1表达的相关性及临床意义

目的研究白细胞介素-6(IL-6)、髓样细胞白血病-1(MCL-1)蛋白及MCL-1mRNA在宫颈癌、癌前病变及正常宫颈组织中的表达情况及其相关性。方法采用免疫组化法检测30例宫颈鳞癌、10例宫颈上皮内瘤变(CIN)及10例正常宫颈组织中IL-6和MCL-1蛋白表达情况,采用逆转录PCR法检测组织中MCL-1 mRNA的表达情况。结果IL-6和MCL-1蛋白在宫颈癌组织中的表达高于正常宫颈组织及CIN组织(P<0.01),且IL-6和MCL-1蛋白的表达呈正相关性(r=0.566,P<0.01);MCL-1蛋白与MCL-1mRNA表达呈正相关性(r=0.772,P<0.01)。结论IL-6和MCL-1的表达在宫颈癌的发生、发展过程中起着共同调节作用,这一发现有助于对早期癌前病变的筛查,并为宫颈癌的治疗提供了一个细胞因子靶点。     

Study of the mechanism on the apoptosis induced in Human leukemia cell line K562 by the combination of indole-3-acetic acid and horseradish peroxidase

Objective To investigate the mechanisms of apoptosis induced in Human leukemia cell line K562 by the combination of indole-3-acetic acid and horseradish peroxidase. Methods Human leukemia cell line K562 were exposed to indole-3-acetic acid （IAA） at 20, 40, 60, 80 or 100 mol/L and horseradish peroxidase（HRP） at 1.2 g/mL for varying times. MTT assay was applied to detect the cell proliferation. Flow cytometry was performed to detect the arrest of cell cycle. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay was used to measure apoptosis. 2, 7-dichlorofluorescin diacetate （DCFH-DA） uptake was measured to determine free radical by confocal microscope. Content of malondiadehyde （MDA） and activity of superoxide dismutase （SOD） were measured by biochemical methods. Results IAA/HRP initiated growth inhibition of K562 cells in a dose- and time-dependent manner. Flow cytometry revealed that cell cycle arrested at G1/G0 after 24 hours treatment. After 72 hours treatment, apoptotic rate of 100 mol/L IAA group increased to 43.9 %, which was 5 times that of control（P〈0.01）. Content of MDA and activity of SOD increased respectively in treatments compared with control. Meanwhile, IAA/HRP stimulated the formation of free radical, which was increased by IAA concentration-dependently. Conclusion The combination of IAAand HRP can inhibit the growth of Human leukemia cell line K562 in vitro by inducing apoptosis which is associated with the increase of free radical. The combination of IAA and HRP might be a promising chemopreventive and chemotherapeutic agent against human leukemia.     

平阳霉素治疗口腔颌面部静脉畸形的临床观察

目的观察平阳霉素治疗口腔颌面部静脉畸形的临床效果。方法160例确诊的颌面部静脉畸形患者,随机分为2组,每组80例。分别于瘤腔注射平阳霉素 地塞米松和鱼肝油酸钠。结果注射平阳霉素 地塞米松组总有效率为92.5%,治愈率为80%;注射鱼肝油酸钠组总有效率为62.5%,治愈率为42.5%。两组比较有显著性差异(P<0.01)。结论平阳霉素 地塞米松局部注射是治疗颌面部静脉畸形的一种安全有效的方法。     

miRNA-21通过TPM1参与高糖诱导的人主动脉平滑肌细胞表型转化与增殖

目的探讨miRNA-21在高糖诱导的人主动脉平滑肌细胞表型转化与增殖中的作用,以及对靶基因原肌球蛋白1(tropomyosin 1,TPM1)表达的影响。方法体外培养人主动脉平滑肌原代细胞,利用脂质体转染miRNA-21mimic和inhibitor至细胞中,BrdU法检测高糖刺激miRNA-21过表达和缺失情况下人主动脉平滑肌细胞的增殖情况,荧光定量PCR检测转染前后高糖刺激时细胞miRNA-21和TPM1的表达情况,Western blot检测人主动脉平滑肌细胞中TPM1、骨桥蛋白(osteopontin,OPN)、平滑肌蛋白22α(smooth muscle 22alpha,SM22α)的表达情况。结果 10、20、25mmol/L D-葡萄糖刺激时人主动脉平滑肌细胞较正常对照组(0.613±0.022)显著增殖(0.725±0.026、0.913±0.024、1.513±0.082),差异有统计学意义(P<0.01);10、20、25mmol/L D-葡萄糖刺激时miRNA-21表达较对照组升高1.1、2.0、3.1倍(P<0.01);正常组细胞转染miRNA-21mimic后细胞增殖能力增强1.6倍,转染miRNA-21inhibitor后高糖组细胞增殖能力减弱(P<0.01);转染miRNA-21mimic后TPM1表达减弱,SM22α表达下调,OPN表达增加(P<0.01),转染miRNA-21inhibitor后TPM1表达上调,SM22α表达上调,OPN表达减少(P<0.01)。结论 miRNA-21可下调细胞TPM1的表达,从而促进人主动脉平滑肌细胞表型转化与增殖。     

Metrigel-VEGF-SW982细胞支架复合物的构建及在滑膜肉瘤小鼠模型上的应用

目的建立Metrigel-VEGF-SW982细胞支架复合物并利用复合物制作移植瘤动物模型,为其他软组织肿瘤动物模型的建立提供思路。方法培养扩增SW982细胞,消化收集细胞,用Metrigel对SW982细胞进行重悬,同时加入VEGF,应用Transwell制作Metrigel-VEGF-SW982细胞支架复合物,并进行培养,HE染色观察细胞支架复合物形态结构。选取10只4周龄雌性SCID小鼠,随机分成支架组和对照组。支架组小鼠腋下移植细胞支架复合物,对照组注射移植细胞培养基悬液。8周后,比较两组造模成功率,取材HE染色观察肿瘤的组织形态特点。结果利用Metrigel对SW982细胞进行立体式培养,能够很好的模拟细胞在体的生长状态。采用细胞支架复合物构建滑膜肉瘤动物模型,成瘤率较对照组明显提高,且肿瘤体积更大,细胞密度更高,肿瘤内部有新生血管。结论利用MetrigelVEGF-SW982细胞支架复合物构建滑膜肉瘤动物模型,能够极大地提高模型成瘤率,为滑膜肉瘤的研究提供便利。     

Radiosensitivity of β-elemene on rabbit VX2 renal transplant carcinoma model

Objective To investigate the effects of low dosage ofβ-elemene on the radiosensitivity of rabbit VX2 renal transplant carcinoma model. Methods We took the rabbit VX2 renal transplant carcinoma as the model. Experimental rabbits were divided into three groups： the control group, the radiation group, and the radiation ＋β-elemene （radiosensitivity） group. The change of tumor was observed by Spiral CT and B ultrasound to compare its regrowth period. The tumor was measured by light microscopy and electron microscopy. Results The tumor in radiosensitivity group was restrained obviously and the sensitization enhancement ratio （SER） of β-elemene was 1.89. Different apoptosis was observed under transmission electron microscopy. Conclusion Low dosage β-elemene can enhance the radiosensitivity of rabbit VX2 renal transplant carcinoma model and induce the apoptosis of tumor cells, but the mechanism needs further study. It promotes apoptosis in mechanisms in vitro.     

Study on Distributed Motion of Self-Reconfigurable Robot Based on Local Rules

The eigenvector of a module with six adjacent module's state was constructed according to self-reconfigurable robot M-Cubes and the configuration of system was expressed with the eigenvectors of all modules.According to the configuration and motion characteristics of the modules,a 3-dimension motion rule set was provided.The rule sets of each module was run according to eigenvector of the module after the motion direction of system decided and motion rules were selected.At last,the rapid and effective motion and metamorphosis were realized in system.The rule sets are operated on three systems and the distributed motion of system is fully realized.The result of simulation shows that the 3-dimension motion rule sets has perfect applicability and extensibility.The motion steps and communication load of the modules increase with the module number in linear.