Objective: To investigate the relationship between the altered expression of the RNA helicase DDX3 and DDX5 in neuroepithelial cells with neural tube defect(NTD) induced by cyclophosphamide and the occurrence of NTD in order to provide experimental evidence for elucidating the molecular mechanism of the occurrence of NTD.Methods: Paraffin sections of the experimental group treated with cyclophosphamide and the control group treated with normal saline were chosen at different time points,and then immunofluorescence assay was performed to detect the expression of DDX3 and DDX5 in neuroepithelial cells with NTD induced by cyclophosphamide.Results: Compared with normal saline control group,integrated optical density of DDX3 and DDX5 positive cells in the neural tube neuroepithelial cells of the cyclophosphamide experimental group remained unchanged 4 hours after treatment,increased 8,12,24 hours after injection,but decreased 48 hours after injection.The difference between these groups was statistically significant.Conclusion: Abnormal expression of DDX3 and DDX5 in neuroepithelial cells of the neural tube of rat embryos induced by cyclophosphamide may be involved in the occurrence of NTD.

Hepatocellular carcinoma (HCC) is the fifth most common malignancy in men, and the seventh in women worldwide. Despite development in the therapy of HCC, the prognosis of HCC patients remains poor. Therefore, it is of great significance to explore the molecular mechanism underlying HCC progression, and investigate novel therapeutic strategies for the treatments of HCC. MicroRNAs (miRs) are known to be involved in the pathogenesis of HCC. The present study aimed to investigate the expression patterns and potential roles of miR‑296‑5p in HCC. Results revealed that miR‑296‑5p was frequently downregulated in HCC tissue samples and cell lines. Additionally, reduced miR‑296‑5p expression levels were correlated with tumor size, TNM stage and metastasis in HCC. Gain‑of‑function demonstrated that miR‑296‑5p inhibited HCC cell proliferation, migration and invasion in vitro. Furthermore, AKT2 was identified as a novel direct and functional target of miR‑296‑5p in HCC. These findings indicated that miR‑296‑5p/AKT2 axis serves important roles in HCC carcinogenesis and progression, and miR‑296‑5p/AKT2 based target therapy hampers HCC tumor growth and metastasis.

Terminal complement membrane attack complex (MAC) formation is induced initially by C5b, followed by the sequential condensation of the C6, C7, C8. Polymerization of C9 to the C5b-8 complex forms the C5b-9 (or MAC). The C5b-9 forms lytic or non lytic pores in the cell membrane destroys membrane integrity. The biological functionalities of MAC has been previously investigated by using either the mice deficient in C5 and C6, or MAC’s regulator CD59. However, there is no available C9 deficient mice ( mC9 −/− ) for directly dissecting the role of C5b-9 in the pathogenesis of human diseases. Further, since C5b-7 and C5b-8 complexes form non lytic pore, it may also plays biological functionality. To better understand the role of terminal complement cascades, here we report a successful generation of mC9 −/− . We demonstrated that lack of C9 attenuates anti-erythrocyte antibody-mediated hemolysis or LPS-induced acute shock. Further, the rescuing effect on the acute shock correlates with the less release of IL-1β in mC9 −/− , which is associated with suppression of MAC-mediated inflammasome activation in mC9 −/− . Taken together, these results not only confirm the critical role of C5b-9 in complement-mediated hemolysis and but also highlight the critical role of C5b-9 in inflammasome activation.

目的 探讨AKT-糖原合成激酶-3β(GSK-3β)信号通路对SOD1G93A突变N2a细胞的作用及机制.方法 选取小鼠成神经瘤细胞系N2a,分别转染pEGFP-WT-SOD1和pEGFP-G93A-SOD1质粒,应用Western blotting和免疫荧光染色方法检测AKT、GSK-3β和细胞周期蛋白D1(cyclin D1)在细胞模型中的表达变化.应用RT-PCR和Western blotting技术检测siRNA沉默AKT后GSK-3β和cyclin D1在SOD1突变N2a细胞中的表达变化,通过MTS方法,检测细胞增殖和存活的改变.结果 与pEGFP-WT-SOD1转染的N2a细胞比较,pEGFP-G93A-SOD1转染的N2a细胞中AKT及GSK-3β总蛋白在转染后24 h和48 h表达均无明显变化,p-AKT(Ser473)、p-GSK-3β(Ser9)和cyclin D1表达均升高.免疫荧光染色结果显示,转染后24 h和48 h,p-AKT(Ser473)、p-GSK-3β(Ser9)和cyclin D1在pEGFP-G93A-SOD1转染的N2a细胞中表达均升高.应用siRNA沉默AKT后与对照组相比,在转染后48 h和72h,AKT、p-AKT(Ser473)、GSK-3β、p-GSK-3β(Ser9)和cyclin D1蛋白均降低.MTS实验结果显示,在AKT沉默后72h、96 h、120 h,N2a细胞增殖和活力降低.结论SOD1G93A突变影响N2a细胞中AKT、GSK-3β翻译后磷酸化修饰及cyclin D1的表达,AKT可能通过调控GSK-3β和cyclin D1影响SOD1G93A突变N2a细胞的增殖和存活.

目的 研究大鼠骨髓间充质干细胞(MSCs)体外培养后移植治疗帕金森病(PD)大鼠的作用及机制.方法 采用全骨髓贴壁培养法分离、体外培养大鼠骨髓MSCs,采用免疫细胞化学法鉴定第3代MSCs,将第3代MSCs作为移植实验组,磷酸盐缓冲液(PBS)作为对照组,分别注射到6-OHDA制备的PD大鼠模型损毁侧纹状体内,观察移植术后PD大鼠不同时间段的行为变化,并检测PD大鼠黑质酪氨酸羟化酶(TH)的表达变化.结果 骨髓MSCs定向移植术后,PD大鼠旋转行为从移植前的(11.08±2.47) r/min,降到移植后4周(3.75±0.96) r/min,移植后1周,2周,4周分别较移植前和对照组行为明显改善(P<0.05),注射PBS后,大鼠在1周,2周,4周旋转行为无明显改善,MSCs移植术后2周黑质TH阳性细胞表达较PBS对照组明显增多(P<0.05).结论 骨髓间充质干细胞移植后可能影响黑质多巴胺功能的神经元的改变,有助于改善PD大鼠的旋转行为.

传统的组织胚胎学实验课以观察玻璃切片为主,玻璃切片在实验教学使用中存在易褪色、不易永久保存和使用时受时间和空间限制等缺陷,利用全自动显微镜扫描系统将玻璃切片制作为数字切片,结合显微数码互动系统和校园网络系统应用于组织胚胎学实验教学,不仅能有效解决优质典型玻璃切片片源不足带来的教学问题,也有利于教师的备课讲解、学生的复习与讨论以及师生间的互动,推动了组织胚胎学实验教学和考试模式的改革.该文主要介绍组织胚胎学数字切片的特点、应用及发展前景.

The set up of Interactive digital microscopy laboratory is the first leap of a modern mode of teaching microscopic morphology,which has caused the tremendous changes in teaching experimental histology and embryology.Compared with the traditional teaching mode,it has multiple functions mode flexible options for application of experimental teaching of Histology and Embryology,Histology and Embryology,experimental teaching methods to teaching various forms,in improving the quality of teaching has a unique advantage.

实验课是医学免疫学的重要部分,为了提高生物技术专业实验课的教学质量,适应社会需求,我们结合近几年来的实践教学经验,对免疫学实验课程体系进行了尝试性的改革,取得了较好的教学效果。

Objective:To define the influence of cyclophosphamide on proliferation and apoptosis of neuroepithelial cells in developing rats,and to further explore the mechanism causing the neural tube defects,which may provide a theoretical basis on the prevention of human neural tube defects(NTDs).Methods:The pregnant rats were randomly divided into an experimental group and a control group.The expression of PCNA and Caspase-3 in the development of neural tube defects was detected using immunohistochemical labeling technology.Results:1.The immunohistochemistry results of PCNA showed that there were no significant changes in proliferation index in the experimental group compared with the control group at 4 and 8 hours after administration;At 12,24 and 48 hours,proliferation index in the experimental group was significantly decreased compared with that in the control group;2.The immunohistochemistry results of Caspase-3 showed that there were no significant changes about apoptosis index in the experimental group compared with the coutrol group at 4 and 8 hours after administration;At 12,24 and 48 hours,the number of Caspase-3 positive cells was increased,and the apoptosis index in the experimental group was increased compared with that in the control group with a significant difference.Conclusion:Cyclophosphamide can increase the expression of Caspase-3,decrease the expression of PCNA,inhibite the proliferation of the neuroepithelial cells,and induce their apoptosis,leading to imbalance in the ratio of proliferation and apoptosis imbalance,which might be one of the important causes of NTDs.

Objective To establish a human expression vector and observe its expression in HepG2 cells.Methods Total RNA was extracted from peripheral blood monocytes (PBMCs) of human treated with LPS (5g/ml),S_1 and S_2 of caspase-1 gene were obtained through RT-PCR.The two DNA fragments were spliced through overlapping PCR (SOE) to form the full length caspase-1 gene.The purified product digested by EcoR I and BamH I was cloned into pIRES_2-EGFP to construct the pIRES_2-EGFP-caspase-1 expression vector which was verified by PCR screening,restriction enzyme assay (EcoR I and BamH I) and DNA sequencing.Then the purified pIRES_2-EGFP-caspase-1 plasmid was transfected into HepG2 hepatoma cells through a jetPEI mediated method.The level of expression of caspase-1 was analyzed by RT-PCR 48h after transfection and compared with cells with pIRES_2-EGFP.Results A 365bp and a 883bp DNA segments were amplified respectively from total RNA isolated from PBMCs treated with LPS which was compatible to the sequence expected.The purified PCR products then spliced by SOE-PCR which produced a long DNA sequence of 1234bp as expected.SOE product and pIRES_2-EGFP digested with EcoR I and BamH I respectively were ligated together by T_4 liagase,in the following colony PCR it was showed that we got several positive clones. Assessed by restriction enzyme assay,it indicated that the positive clones gave rise to a 1234bp DNA fragment which was finally proved to be identical to the full length human caspase-1 reported by GeneBank.The recombinant vector were then transfected into HepG2 hepatoma cells by jetPEI,48h after transfection,RT-PCR indicated that the expression level of caspase-1 was significantly increased compared with cells transfected with pIRES_2-EGFP.Conclusion We successfully constructed a human caspase-1 expression vector,the recombinant vector could express exogenous gene effectively in human hepatoma cells.

目的 探讨移位锁骨骨折采用切开复位克氏针内固定治疗的方法,总结其临床经验.方法 自1998年3月-2005年6月,对68例移住锁骨骨折采用切开复位克氏针内固定治疗,随访61例,占89.9%,随访时间为6-12个月,平均8个月.结果 随访的61例中全部骨折愈合.锁骨及肩关节功能恢复情况:优59例,良2例,差0例.结论 克氏针内固定治疗移位锁骨骨折创伤小、操作简便、取针方便、手术费用低、愈合率高,是行之有效的治疗手段,值得在临床工作中推广应用.

根据我院的实际情况,结合青年实验技术人员的特点,对目前医学院校青年实验技术人员的现状进行了分析,同时根据医学院校发展建设的趋势,对如何培养和提高青年实验技术人员的整体素质提出了相应的措施。

Objective To investigate the method of induced differentiation of mouse mesenchymal stem cells into cadiocytes in vitro.Methods The mononuclear cells were isolated from tibias and fibulas of 2 weeks old mice and the mesenchymal stem cells of bone marrow were obtained with the combination of gradient centrifugation and cellular attaching culture method.Cells of the 2nd passage were divided into 4 groups at 48h,cells of group A were treated with simple upper liquid from cardiac tissue culture.10μmol/L 5-aza was added to group B.In group C,cells were given associated treatment with the two factors.Group D was given no inductor for control.Then the expression of α-actin from the 4 groups was detected by immunocytochemistry method.Results Cells treated with cardiac tissue upper liquid(group A)for 3 weeks showed a positive expression of α-actin.With treatment of 10μmol/L 5-aza(group B)at 3 weeks,cells were strongly positive stained.In group C,cells took on intensively positive staining at 2 weeks after co-treated with 10μmol/L 5-aza and cardiac tissue culture upper liquid and there was no positive cell all the time in group D.Comparison between every two groups among A,B,C and D showed statistical difference(P0.05).Conclusion Co-treatment of cardiac tissue culture upper liquid and 10μmol/L 5-aza may accelerate the differentiation from mesenchymal stem cells into cadiocytes in vitro.

卡介苗及卡介苗制剂用于支气管哮喘防治的研究已有十余年的历史.医学界普遍认为卡介苗及卡介苗制剂对支气管哮喘的防治有一定疗效.引起了制剂对支气管哮喘的防治研究热潮.但卡介苗及卡介苗制剂对支气管哮喘的防治到底有无价值,本文就此方面的有关研究进行简述.