细胞外的

Compared with classical cytotoxic T lymphocytes , NK cells are superior in the following aspects: NK cells are among the first cell types to be activated after intracellular pathogen stimuli, the activity of NK cells peaked between 1 to 3 days, retained during the first 7-10 days, and the T cells response emerged only after 7 days when the NK cell responses begin to decline; though comprising of only 10％-15％ of peripheral lymphocytes, NK cells mobilize most of the cell populations in a strong and high level response to different invasions, and act as key factors at early defense before a specific immune response could mount, while only certain clones were involved in T cell response; NK cells kill virus-infected or malignant transformed cells without pre-sensitization and without restriction by major histocompatibility antigen, which is usually a mechanism of immune escape to T cell recognition by virus-infected or tumor cells, thus NK cell in complementary with T cells play crucial roles in tumor and virus eradication.

NK细胞以其强大的细胞毒活性为特征，与细胞毒性T细胞相比，NK细胞具有如下特点：NK细胞虽然只占外周血淋巴细胞的10-15％，但是对刺激性因素产生应答的过程十分迅速，一次可以调动大多数细胞共同参与，而不是几个克隆的活化，免疫应答的强度高；病毒感染或恶性转化细胞的一个主要特征是细胞表达的MHCⅠ类分子下降或消失，并借助这一机制逃避特异性T细胞应答的识别，NK细胞介导的杀伤活性无需抗原刺激，不受MHCⅠ分子表达的限制，从而与T细胞应答互为补充发挥免疫防御的功能；NK细胞的免疫活性可以被多种细胞因子上调，其中IFNα，IL-2，IL-12，IL-15和IL-18具备更强的作用；NK细胞具有强大的细胞因子分泌功能，对于启动T细胞特异性应答必不可少。

The mechanism probably is that Pishang can inhibit the growth, movement and adheresion to extracellular matrix and endothelial cell of liver tumor cells, up-regulate the expression of nm23, down-regulate the expression of CD44, MMP-2, and inhibit the tumor neovascular by supressing the vascular endothelial cell, down-regulating the expression of VEGF, damaging the primitive mesenchymal cells and inhibiting neovascular gemmation.

砒霜具有明显的抗肝癌细胞作用，并有一定的抗侵袭与转移作用，其机制可能与其能抑制肝癌细胞增殖、迁移、阻止肝癌细胞与细胞外基质和内皮细胞粘附、上调肿瘤转移抑制基因nm23表达、下调肿瘤侵袭分子CD44、MMP-2表达以及通过抑制内皮细胞增殖、下调VEGF表达、损伤间充质细胞、阻止新生血管芽生等途径从而抑制肿瘤新生血管形成有关。

Results:①In normal extracellular solution, the non-selective cholinergic agonist acetylcholine and the muscarinic agonist muscarine both caused the increace of [Ca2+]i in separately 21 out of 25 and 14 out of 18 VHCs I, while only 3 out of 20 and 2 out of 16 VHCs I had a weak increase of [Ca2+]i if investigated in calcium free extracellular solution; nicotine could increase the [Ca2+]i in 7 out of 32 VHCsⅠonly at enough high concentration(up to 10 mmol/L) in normal extracellular solution, this [Ca~(2+)]_i increase by nicotine at high concentration could not be investigated if at calcium free solution.

结果①非选择性胆碱能受体激动剂乙酰胆碱、M型胆碱能受体激动剂毒蕈碱在正常细胞外液中均可引起大部分(21/25， 14/18)单离VHCⅠ[Ca~(2+)]_i的升高，但无钙外液中ACh、muscarine仅使少部分(3/20，2/16)VHCⅠ[Ca~(2+)]_i升高且作用减弱；正常外液中，N型受体激动剂尼古丁仅在高浓度(≥10 mmol/L)时引起部分(7/32) VHCⅠ[Ca~(2+)]_i升高，在低浓度时对胞内钙离子浓度影响不明显，无钙外液中，10 mmol/L nicotine对胞内钙离子浓度影响不明显。

On the other hand, the decrease of Na-Ca exchange, because of the increase of intracellular Na〓 concentration resulted from depression in the sarcolemma Na, K-pump function, was responsible for exteranal Ca entry into the cell.

进一步的研究则提示，针刺是通过提高细胞膜Na〓通透性和调整体内正常细胞外液与损伤肌异常细胞外液间离子分布而完成这一作用。

Methods The peripheral blood lymphocytes were stimulated by Stimulator in order to enhance expression of cytokines .

先用刺激物刺激外周血T细胞，以增加细胞内细胞因子的表达，继用流式细胞仪分析特异性细胞因子表达水平。

Results: Tyroserleutide can significantly increase the life span of H22 tumor-bearing mice by 50-70% in dosages of 20ug/kg/d-80ug/kg/d,specially the high dosage of 80ug/ml can significantly increase the life span by 69.24%; Tyroserleutide can inhibit the growth of transplanted hepatocellular tumor BEL-7402 in nude mice,the rate of tumor inhibition was25-50% in dosages of 40-320ug/ml ,the inhibition rate of 160ng/ml was 44.03%; Tyroserleutide could inhibit the growth of H22 and BEL-7402 tumor in a dose-dependent manner. Simultaneously, tumoricidal activity of tyroserleutide against BEL-7402 cell line in vitro was observed hinger when compared with the control group(P.05).The inhibition effect of 72hrs was higher than 24hrs,48hrs,96hrs.And specially the high dosage of 160ug/ml can significantly inhibit growth of tumor cell by 19.36%. Tyroserleutide can activated PEM and marked enhance cytotoxicity andphagocytosis functions in vitro and in vivo. The OD values of cytotoxicity were observed hinger when compared with the control group(P.05).The cytotoxicity of macrophages activated by tyroserleutide against BEL-7402 and B16-F10 was 35.58%,61.2% in vitro and21.39%,47.63% in vivo. The cytotoxicity rate of nude mice PEM was 32.86%,73.07% in vivo. Furthermore, tyroserleutide alone could stimulated the production of IL-1B TNF- a and NO by M . Tyroserleutide and LPS could synergistically activated M producing more cytotoxicity effectors. Conclusion: Tyroserleutide had inhibition functions against hepatoma carcinoma .Its possible mechanisms were related to the affect that Tyroserleutide could inhibit tumor cell directively and induce tumor cells apoptosis or death effectively.

结果：酪丝亮肽能显著延长腹水型肝癌H_(22)小鼠的生存时间，给药剂量为80μg／kg／d时疗效最显著，达到69.24％，在20μg／kg／d-80μg／kg／d剂量范围内生命延长率为50-70％，给药剂量与荷瘤鼠生存时间呈现一定量效关系；酪丝亮肽能显著抑制人肝癌BEL-7402移植瘤裸鼠的肿瘤生长，给药剂量为160μg／kg／d时疗效最显著，抑制率为44.03％，并且在40-320μg／kg／d剂量范围内抑制率为25-50％，给药剂量与肿瘤抑制率呈现一定量效关系；酪丝亮肽体外对人肝癌BEL-7402细胞生长有一定的抑制作用，在作用72hrs时各浓度酪丝亮肽对肿瘤细胞的抑制作用较24hrs、48hrs、96hrs明显，其中浓度为100μg／ml时抑制率达19.36％；酪丝亮肽体内外均能增强小鼠腹腔巨噬细胞对肿瘤细胞的杀伤：体外作用中巨噬细胞对BEL-7402、B16-F10的杀伤功能明显增强，与效应细胞对照组相比有显著性差异(P＜0.05)杀伤率分别达到35.58％、61.2％；体内作用中巨噬细胞对BEL-7402、B16-F10的杀伤功能明显增强，与生理盐水对照组相比有显著性差异(P 。05)，杀伤率分别达到21.39%、47.63%；裸鼠腹腔巨噬细胞经酪丝亮肤作用后对BEL一7402、B 16一F10杀伤功能明显增强，与生理盐水对照组相比有显著性差异(P.05)，最高杀伤率分别达到32.86%、73.07%；酪丝亮肤能增强单核巨噬细胞系统的吞噬功能，吞噬指数与生理盐水组比较有显著性差异(P.05)；酪丝亮肤体外作用能促进小鼠腹腔巨噬细胞分泌合成细胞毒效应分子IL一lp、TNF一Q和NO，与效应细胞对照组相比有显著性差异(P.05)；酪丝亮肤体内作用能促进小鼠腹腔巨噬细胞分泌合成细胞毒效应分子IL一lp、TNF一Q和NO，与生理盐水对照组相比有显著性差异(P.05)；酪丝亮肤能促进鼠巨噬细胞株R戌W264.7分泌合成IL一1p和NO，IL一1日、NO水平分别在酪丝亮肤作用24hrs、12hrs时达到高峰，酪丝亮肤单独应用能提高巨噬细胞的分泌合成功能，而且酪丝亮肤能与LPS协同作用刺激巨噬细胞的细胞毒效应分子分泌合成。

Results: NKA-I cells was first found in the duodenum mucose on the 14th day of fetus, earlier than in the jejunum ( 16th day ) and ileum ( 17th day ), and its Positive reaction and the number of cells increased with further development, after birth the 30th day, it became quite identical to that of adult rat; NKA-I cells were seen in the colon mucosa on the 17th day of fetus, later on the number kept decreasing till grown up stage; No change was observed in the stomach mucosa except that few cells showed up on the 15th day of fetus; No positive cell was found on the oesphagus wall.

结果： NKA-I细胞首先见于胚胎14 d的十二指肠粘膜，早于空肠(胚胎16 d)和回肠(胚胎17 d)，其阳性反应强度及细胞量随发育而增加，30 d时与成年鼠相似；结肠粘膜内于胚胎17 d见细胞，但此后数量较少直至成年；胃粘膜于胚胎15 d见个别细胞外，以后无明显变化；食道壁始终未见阳性细胞。

Results: NKA-I cells was first found in the duodenum mucose on the 14th day of fetus, earlier than in the jejunum ( 16th day ) and ileum ( 17th day ), and its Positive reaction and the number of cells increased with further development, after birth the 30th day, it became quite identical to that of adult rat; NKA-I cells were seen in the colon mucosa on the 17th day of fetus, later on the number kept decreasing till grown up stage; No change was observed in the stomach mucosa except that few cells showed up on the 15th day of fetus; No positive cell was found on the oesphagus wall.

结果：NKA-I细胞首先见于胚胎14 d的十二指肠粘膜，早于空肠(胚胎16 d)和回肠(胚胎17 d)，其阳性反应强度及细胞数量随发育而增加，30 d时与成年鼠相似；结肠粘膜内于胚胎17 d见多个细胞，但此后数量较少直至成年；胃粘膜于胚胎15 d见个别细胞外，以后无明显变化；食道壁始终未见阳性细胞。

And 4 h of the HI group were 107±9, 91±10, 76±6, 37±11, respectively, compared to that of the control group of 116±8, showing a tendency of time-dependent decreasing manner (all P 《 0. 05). Conclusion During HI, the dowuregnlation of outward potassium conductance may prevent the emission of interacellularly accumulated K+ ions, thus resulting in osmotically drived water influx into astrocytes via aquaporin-4 and then cell swelling.

在细胞缺氧缺血时，细胞外向性钾通道下调，可能阻止细胞内堆积的钾离子流出细胞外，引起渗透性改变而导致水通过AQP4流入细胞内，从而出现细胞水肿。

The PCR products were examined by agarose gel electrophoresis. The target gene fragments were purified by gel extraction kit and ligated to cloning vector pMD18-T. The recombinant vectors were transformed into host strain E. coli K802 by lithium chloride method, screened and identified with PCR and restrictive enzymatic digestion. Their sequences were confirmed by DNA sequencing.(2) sTWEAK1 gene was subcloned into expression vector pProEx HTb and transformed into E. coli BL21. sTWEAK2 gene was subcloned into expression vector pMAL-C2x and transformed into E. coli TB1. The recombinant vectors were screened and identified with PCR and restrictive enzymatic digestion. The recombinant fusion proteins were induced to express with IPTG, detected by coomassie brilliant blue-stained SDS-polyacrylamide gel electrophoresis , and confirmed by Western blot analysis.(3) The sTWEAK1 fusion protein was purified with Ni-NTA Spin Kit.(4) The biological activity was assayed on transformed and tumor cells by microplate photometer after crystal violet or sulfur rodamine B staining.(5) The contents of IL-8 in the supernatant of 1990 cell cultures were determined by ELISA.(6) The morphological changes of the sensitive cells were observed by light and transmission electron microscopies.(7) The cell cycle and apoptotic rate were assayed by flow cytometry in 1990 and M85 cells.(8) The effect of fusion proteins on induction of NF-κB in 1990 and LOVO cells was detected with Dual-Luciferase Reporter Assay system.(9) The TWEAK gene was subcloned into Adeno-X Viral DNA with pShuttle vector and transfected into HEK293 cells by lipofectamine method.

(1)本研究用RT-PCR方法，从人组织细胞总RNA中扩增可溶性TWEAK胞外区（sTWEAK1和sTWEAK2）的cDNA序列及全长编码序列，用琼脂糖凝胶电泳分析PCR产物，胶回收目的基因片段，连接到pMD18-T克隆载体中，转化大肠杆菌K802，PCR和酶切筛选阳性克隆，全自动DNA测序验证序列；(2)sTWEAK1和sTWEAK2分别亚克隆到pProEx HTb和pMAL-C2x表达载体中，分别转化大肠杆菌BL21和TB1，PCR筛选和酶切鉴定，阳性克隆用IPTG诱导表达，表达产物用SDS-PAGE分析和Western blot验证融合蛋白；(3)用NTA-Ni Spin试剂盒初步分离纯化sTWEAK1融合蛋白；(4)用体外培养的肿瘤细胞和正常对表达产物进行活性检测，贴壁细胞用结晶紫染色法，悬浮细胞用磺酰罗丹明B染色法，酶标仪检测OD值；(5)敏感细胞用ELISA法检测细胞培养上清中IL-8的含量；(6)用光镜和电镜观察敏感细胞死亡和细胞凋亡情况；(7)用流式细胞仪分析表达产物对敏感细胞凋亡率和细胞周期的影响；(8)用双荧光素酶报告基因检测法，测定表达产物对敏感细胞NF-κB的影响；(9)用pShuttle穿梭质粒将TWEAK重组到腺病毒载体上，用脂质体转染法转染HEK293细胞，PCR鉴定重组质粒。

However, as the name（read－only memory）implies, CD disks cannot be written onorchanged in any way.

然而，正如其名字所指出的那样，CD盘不能写，也不能用任何方式改变其内容。

Galvanizes steel pallet is mainly export which suits standard packing of European Union, the North America. galvanizes steel pallet is suitable to heavy rack. Pallet surface can design plate type, corrugated and the gap form, satisfies the different requirements.

镀锌钢托盘多用于出口，替代木托盘，免薰蒸，符合欧盟、北美各国对出口货物包装材料的法令要求；喷涂钢托盘适用于重载上货架之用，托盘表面根据需要制作成平板状、波纹状及间隔形式，满足不同的使用要求。