免疫抑制的

Neural cell adhesion molecule L1, discovered in the 80s, is one of the matters that can promote nerve regeneration. It can mediate the interaction of cell-to-cell and cell-to-matrix, take great efforts on neurite outgrowth and fasciculation, migration of neural cells, formation of the myelin, structure of neural trace, transmembrane signal transduction, inhibiting apoptosis of neural cells, even on immune system and formation of the tumor. Li is discovered on the mouse's Cerebellar membrane primarily. Thereafter, it also is found in post- mitotic neurons, pre- and non-myelinating Schwann cells and so on.

神经细胞粘附分子L1（L1）是80年代发现的具有促进神经再生作用的物质，它能介导神经细胞与神经细胞之间的粘附，在神经轴突的生长、聚集，神经细胞的转移，纤维髓鞘的形成，神经通路的构建及信号的转导，抑制神经细胞的凋亡甚至在免疫系统、肿瘤的发生等方面均具有非常重要的作用。L1最初是从小鼠的小脑膜中发现，后来发现在有丝分裂后的神经元，髓鞘形成前及非形成髓鞘的雪旺细胞等组织中同样存在。

Glioma is still one of refractory disease in the neurosurgical field; the development of new primary and adjuvant treatment is vital. Recently, the gene therapy of glioma is developed rapidly and there are many methods about the gene therapy that include: suicide gene therapy, immunologic gene therapy, drug resistangce gene therapy, angiostatin gene therapy and so on. The sucide gene therapy is the most potential approach of antitumer, these nonmammalian genes encode enzyme that convert nontoxic prodrugs into highly toxic metablites. Cells transfected with suicide genes are targeted for specific negative selection, witch can be induced by administrtion of the corresponding produg. Among the enzyme/produg combinations, two of the best characterized system are herpes simplex virus thymidine kinase /ganciclovir and Escherichia coli cytosine deaminase /5-flourocytosine (5-FC). The formor can convert the antiviral nucleoside analogs acyclovir , ganciclovir to their nucleoside monophosphate derivatives, the monophosphate forms are subsequently phosphorylated by endogenus cellular kinases to triphosphates, these molecules are potent inhibitors of DNA synthesis.

近年来脑胶质瘤的基因治疗发展迅速，应运而生的方法有自杀基因、免疫基因、多药耐药基因以及抗血管生成基因等，其中自杀基因被认为是最有前景的基因治疗方法，它又称病毒介导的酶／药物前体疗法，是利用转基因技术将哺乳动物细胞中所不含有的自杀基因转入到哺乳动物肿瘤细胞中，该基因表达的产物可将无毒的药物前体转化为毒性药物，从而选择性杀伤该肿瘤细胞，常用的自杀基因有单纯疱疹病毒-胸苷激酶基因和大肠杆菌胞嘧啶脱氨酶基因，前者催化无毒性抗病毒核苷类似物如丙氧鸟苷、无环鸟苷等成为单磷酸核苷衍生物，然后在内源性细胞激酶作用下转化为具有明显毒性的三磷酸核苷，作为DNA合成链的终止剂和DNA合成酶的抑制剂，干扰细胞DNA的合成；后者编码的胞嘧啶脱氨酶可催化5-氟胞嘧啶（5-FC）脱氨成为5-氟尿嘧啶（5-FU），然后代谢为有毒性的5-氟尿嘧啶-5′三磷酸（5-FUTP）和5-氟-2′脱氧尿嘧啶-5′磷酸（5-FdUTP），5-FUTP通过与UTP竞争性结合而抑制mRNA和tRNA的合成，5-FdUTP则作用于胸苷合成酶，导致TMP衰竭而阻止DNA的合成，最终诱导肿瘤细胞凋亡。

Light microscope and transmission electron microscopy showed that SMMC-7721 cells induced by SAHA had undergone the restorational alteration in morphology and ultrastructure, which were different from those of nontreated cells but were similar to those of normal cells, and the changes were as follows: the cells turned to be flat and spread; the nucleo-cytoplasmic ratio lessened and nuclear shape became rather regular; the number of nucleolus reduced and its volume lessened; euchromatin increased while heterochromatin decreased in nucleus; in the cytoplasm, mitochondria grew in number with relatively consistent structure and well-developed mitochondria cristae; Golgi complex turned to be well-developed and typical; rough endoplasmic reticulum increased. Immunocytochemistry assay showed that the expression of AFP and PCNA were declined significantly. FCM analysis showed SAHA could arrest SMMC-7721 cells in G0/G1 phase, with an accumulation of the cells in G0/G1 phase while a decrease of cells in S phase. Semi-quantitative RT-PCR detection revealed that the expression of p21WAFl mRNA was upregulated remarkably in the cells treated with SAHA.

结果：倒置显微镜和透射电镜观察显示，经SAHA处理的细胞增殖速度显著减慢，细胞体积增大，细胞核较小，形状较为规则，核仁数量减少、体积变小，核内常染色质增多而异染色质减少，核质比例减小，细胞质内线粒体数量增多、线粒体嵴发达，高尔基体较为典型，粗糙型内质网增多，呈现出与正常上皮细胞相似的形态变化；MTT比色法测定结果显示不同浓度(2.5、5.0、7.5、10.0uM)SAHA对SMMC-7721细胞的增殖均有抑制作用，并有明显的剂量依赖和时间依赖关系；免疫细胞化学检测显示SAHA能显著降低PCNA和AFP在SMMC-7721细胞中的表达；流式细胞仪检测结果显示，SMMC-7721细胞经SAHA处理后，G0/G1期细胞明显增加，S期细胞则明显减少，细胞被阻滞于G0/G1期；RT-PCR检测结果表明，SAHA作用12h后SMMC-7721细胞中p21WAF1 mRNA的表达即有增加，24h后更为明显。

Results:Comparing to the control group, the proliferation of overexpressed p16 QBC939 cells was inhibited and p16 overexpress induced G1 phase blockade.

结果：与对照组相比，p16高表达的人胆管癌细胞QBC939的增殖受到明显抑制；流式细胞光度术表明p16的高表达导致了较为明显的G1期阻滞作用，但对细胞周期的其他时相并无影响；Western免疫印迹分析结果表明癌基因c-myc的蛋白水平表达下降。

He experiment results indicate that (1) The extract of Rubia Cordifolia L has the effects of anti-proliferation and inducing APO;(2) Whenadministrating with 5-fu, the extract of Rubia Cordifolia L obviously enhanced the effects of anti-proliferation and inducing APO of 5-fu;(3) The basement of molecular biology of the extract of Rubia Cordifolia L for inducing APO of MGC-803 cells is correlated with inhibiting the expression of bcl-2 of anti-apoptosis gene.

UNEL法与PCNA的免疫细胞化学法的结果：0.2、0.4、0.8、1.0mg/ml浓度的茜草提取物与2、4、8、16μg/ml浓度的5-Fu，分别诱导细胞凋亡和抑制细胞增殖，表现出浓度、时间依赖性关系，除个别浓度与个别时间之间无显著性差异（P＞0.05）外，不同浓度与不同作用时间之间均有显著性差异（P＜0.05或P＜0.01），并且两种药物联合使用时，MGC-803细胞的APO%显著提高，降低PCNA的阳性表达率（P＜0.05或P＜0.01）。

This preliminary case control study reconfirms that TWHf is a very potent herbal remedy to suppress mononuclear expressions of many inflammatory cytokines and pro-inflammatory molecules.

尽管只有少数的研究对象，本篇研究提供了在免疫致病机转上的可能证据来支持雷公藤对於僵直性脊椎炎患者周边血液单核球细胞内的许多发炎性细胞激素及其它和发炎反应有关的分子的基因表现具有强烈的抑制作用的说法。

The inhibition of SPG-Rg3 on the growth of MCF-7 cells was detected by MTT assay,IC50 was calculated to obtain the effective concentration; flow cytometry was used to observe the cell cycle of MCF-7 cells; AO/EB fluorescence double-dye technology was performed to observe the apoptosis of cells in morphology,immunocellularchemistry and RT-PCR were utilized to investigate the apoptosis of MCF-7 cells and its relationship with caspase-8 from protein level and molecule level.

采用MTT法观察SPG-Rg3对MCF-7细胞生长的抑制作用，并计算出IC50，依据IC50值确定SPG-Rg3的有效浓度；流式细胞术检测SPG-Rg3作用后MCF-7细胞周期的变化；AO/EB双染从形态学上观察SPG-Rg3对MCF-7细胞凋亡的作用；免疫细胞化学染色和RT-PCR技术分别从蛋白水平和分子水平上检测SPG-Rg3对MCF-7细胞的诱导凋亡作用及其与caspase-8基因的关系。

Mechanical strains also regulated the protein and mRNA expression of several differentiation markers, as well as the activation of extracellular signal-regulated kinases, p38 MAP kinase and protein kinase B in a frequency-dependent manner. Furthermore, the inhibition of p38 pathway could block the strain-frequency induced the phenotype modulation of VSMCs, neither ERKs nor Akt. Frequency of mechanical strain, not conditioned medium, regulated the phenotype of VSMCs in a frequency-dependent manner. Rho-GDI alpha was suppressed by the mechanical strain at 1Hz.

采用免疫细胞化学法检测VSMCs形态和排列的变化；RT-PCR和Western blotting检测表型标志分子α-肌动蛋白、肌球蛋白重链(SM1/2)、肌动蛋白相关蛋白SM22α和调宁蛋白(h1-calponin)的mRNA和蛋白水平的变化；抑制剂或RNA干扰阻断可能的信号调节分子的活性或表达，包括p38、细胞外信号调节激酶1/(2extracellular signal-regulated kinases， ERK1/2)、蛋白激酶B和Rho-鸟苷酸解离抑制因子(Rho-guanine nucleotide dissociation inhibitor， Rho-GDI alpha)，研究了不同频率张应变对VSMCs表型转化的影响及其调节机制。

Objective:The aim of the experiment is to search for the regulation that the hepatic stem cells abnormally differentiate into cancer cells. In the experiment,the variating regulation of PCNA,c-kit and c-myc in the formation and development of HCC will be interpreted with immunohistochemistry and RT-PCR. So,the initial mechanism that hepatic stem cells abnormally differentiate into cancer cells from the levels of cell,protein and gene will be clarified. The last,the inhibition to HCC with ulinastain will be studied,from which,a new way of clinical prevention and treatment for HCC will expect to be explored.

二、目的本实验旨在探讨以卵圆细胞为代表的肝干细胞在HCC发生、发展过程中在肝组织内分布规律，并通过免疫组化、RT-PCR等技术阐明PCNA、c-kit、c-myc在HCC发生、发展过程中的动态演变规律，从细胞、蛋白、基因三个水平初步研究卵圆细胞向癌细胞异常分化的机制，同时观察乌司他丁对HCC发生、发展的抑制作用，为肝癌的临床预防和治疗探索新的途径。

San-Huang, so called Mesima in Japan and Korea, has been considered to be a traditional Chinese and Japanese medicine that was reported to be used as a stypti, both internally and externally, for treating uterine bleeding, and for blood in the urine. The modern researches proved that it is also an effective emmenogogue for amenorrhea and for treating leukorrhea. Extracts from hot water have been experimented to have antitumor and inhibit Sacroma 180 cancer cell abilities. The powdered fruiting body is taken in liquid or in pill form, and preparations can also be applied externally for pains in the abdominal region.

在传统的药用真菌中，桑黄在明代李时珍的「本草纲目」即有记载，性寒，味微苦，能利五脏，宣肠气，排毒气，压丹石，人热发，止血等，在中国及日本有长久的使用历史，现代化科学研究证实桑黄的热水抽出物具有抑制老鼠Sacroma 180肿瘤细胞及一般抗肿瘤的功能，治妇女月经不顺，提高免疫机能及强化T细胞能力，对癌症肿瘤切除后化疗的患者具有相乘的功效且长期服用并不会有任何副作用及毒性产生，这些证据显示「桑黄」确实具有开发潜力。

According to the clear water experiment, aeration performance of the new equipment is good with high total oxygen transfer coefficient and oxygen utilization ratio.

曝气设备的动力效率在叶轮转速为120rpm～150rpm时取得最大值，此时氧利用率和充氧能力也具有较高值。

The environmental stability of that world - including its crushing pressures and icy darkness - means that some of its most famous inhabitants have survived for eons as evolutionary throwbacks, their bodies undergoing little change.

稳定的海底环境─包括能把人压扁的压力和冰冷的黑暗─意谓海底某些最知名的栖居生物已以演化返祖的样态活了万世，形体几无变化。