组织细胞

The cell biological features were observed by inverted phase contrast microscope, l ight microscope, electron microscope, cell vital ity assay, cell growth curve and cells staining after harvest and during the periods of culturing the primary, the 1st passage and 2nd passage.

分别在取材后、原代、第1 代、第2 代细胞培养期间，进行髓核细胞活力测定；爬片培养后进行甲苯胺蓝、HE、聚集蛋白聚糖番红O、Ⅰ型及Ⅱ型胶原免疫组织化学染色观察；MTT 法绘制髓核细胞生长曲线，并行原代及第2 代细胞透射电镜观察，对体外细胞的生物学特性进行研究。

There was significant difference in signal intensity between the cell concentration group and unlabelled cell group during 2 weeks. And significant difference occurred in the 5×105 SPIO-labeled cells per milliliter medium group and unlabelled cell group during 3 weeks.3. In kidneys of ischemia-reperfusion injury,the labeled BMSCs were demonstrated as signal intensity loss in renal medulla on T2*WI sequence in days 1,3,and5.The signal intensity loss was visualized up to 8 days after transplantation,and the signal intensity in the third day was the lowest. Histological analyses showed that most Prussian blue staining-positive cells were well correlated with the area where a signal intensity loss was observed in MRI.

在此细胞浓度的基础上，标记后细胞信号在T2*WI序列的降低在2w后失去统计学意义；而在细胞浓度为5×105/ml时，标记3w后信号在T2*WI序列的降低才失去统计学意义。3缺血再灌注肾损伤家兔细胞移植后第1、3、5天肾脏髓质区信号强度明显下降，至移植后第8天信号改变不明显，其中第3天信号改变最为明显；组织学分析见绝大多数普鲁士蓝染阳性标记细胞分布于肾小管周围，与MRI信号强度下降区域基本一致。

By using electroporation transfects,the TGF〓expression vector were introduced into HL-60 cells.Thecells were allowed to grow for 1 day before beingsubjected to selection for the ability to grow in mediumcontaining 600ug/ml geneticin.Stable cell clones resistantto G418 sulfate were obtained after 2-3 WK growth withdrug-containing medium.Southern blot hybridization wascarried out.RNA slot have higher expression of TGF〓mRNA in HL-60 transfected.All results followed suggestedthat TGF〓 have suppressed the maliganant phenotype of HL-60 cells.

利用电击法将〓表达载体导入HL-60细胞中，经G418筛选培养20天后再行Southern杂交检测外源〓整合情况，利用RNA狭线杂交法证明了在转录水平上表达有所提高，进一步进行了流式细胞仪测定DNA含量，使细胞在〓期产生生长阻滞作用，细胞生长速度，在软琼脂中成集落数、在组织中的浸润性及在裸鼠中的成瘤性均发生变化，我们的实验结果表明TGF〓对HL-60细胞具有抑制恶性表型的作用。

Objective To establish a competitive RT-PCR method for quantitative determination of IFN-γ mRNA and IL-4 mRNA and explore the optimal inductive condition in vitro. Methods IFN-γ mRNA and IL-4 mRNA in PBMC induced by PMA and calcium ionophore of a healthy volunteer and in unstimulated PBMC of 11 HVs and 13 asthma patients were determined quantitatively with the established competitive RT-PCR method using self-prepared internal standard as competitive templets.

Subject words 】 Competitive templet； IFN-γ； IL-4； mRNA； RT-PCR 检测细胞因子最早使用生物学或免疫学测定法，这两类检测可反映某种细胞因子分泌、吸附、消耗和降解后的净含量〔1〕，但不能为细胞因子产生过程中基因转录、翻译及翻译后调控机制提供必要的信息，也不能反映细胞或组织内细胞因子基因表达水平。

In this essay, the culture methods of corneal epithelial cell and keratocytes have been studied. The method of rabbit corneal epithelial cells has been improved in order to solve the problem that the cells moved slowly and could not form monolayer when the traditional tissue nubbles cultivation was adopted.

本文对兔角膜上皮细胞和基质细胞的培养方法进行了研究，解决了传统角膜上皮细胞组织块贴壁培养时细胞迁出慢，不易形成单层的难题，建立了兔角膜上皮细胞原代培养的方法。

The methods of sticking tissues piece to culture primary hFCECs and mouse 3T3 fibroblast conditioned medium to culture passage hFCECs were established;2.The Proliferative capacity of corneal limbus hFCECs is better than central hFCECs.3.3T3 conditioned medium could markedly promote the proliferation of epithelial cells

结论一、组织块贴壁法原代培养人胎儿角膜上皮细胞以及小鼠3T3成纤维细胞条件培养液传代培养方法基本建立；二、人胎儿角膜缘部上皮细胞增殖能力优于中央部；三、3T3细胞条件培养液能促进角膜上皮细胞的增殖。

In order to confirm this suspect, we choosed a cultured rat granulation tissue fibroblasts model in vitro and applied RT-PCR and cell-ELISA technology to detect the changes of firbroblasts intrinsic EGF, bFGF, TGFβ-1 with their receptors gene expressions and protein synthesises after stimulated by SP. Our purpose was to explore the possible effects and patterns imposed by SP on fibroblasts intrinsic growth factors and their receptors expressions, which maybe offer theoretical basis for promoting wound healing via improving nervous functions and regulating neuropeptides secretions.

为进一步验证这一推测，同时排除在体多因素干扰，我们采用了一种大鼠肉芽组织成纤维细胞体外培养模型，采用RT-PCR与细胞ELISA技术，检测SP刺激成纤维细胞后，成纤维细胞内源性EGF、bFGF、TGFβ-1及其受体基因表达和蛋白合成的改变情况，探讨SP对成纤维细胞内源性生长因子及其受体表达的影响以及方式，以期为经由改善神经功能、调节神经肽分泌途径促进伤口愈合提供理论依据。

The animal model of this method system have, the characteristics of the high fat, high leptin, high Ins, and match with the characteristic of clinical and the pathologic of simple obesity. This studies showed: The acupuncture can obviously reduce the vaule of intaking food and water of the fat rats, and lower the body weight and fat; The acupuncture can urge the diameter, area, physical volume of white fat cell contract, Brown fat tissue of fat rat group surroundings vascular containing large quantity of white fat cell, the acupuncture group was less. this showed: The acupuncture enhanced the white fat cell metabolism, and promotesed the white fat cell to convert to brown fat cell; The frequency of spontaneous electric of nerve cell of pvn of the fat rat is high, the acupuncture counld depress the spontaneous electric of nerve cell of pvn, This showed: the acupuncture can depress the appetite passing to adjust function of pvn nerve; The monoaminergic neurotransmitter of inside of the fat rat brain is disfunction, the acupuncture can rectify this disfunction, and adjust level of 5-HT, and affect appetite, and improve metabolism; The level of leptin and Ins of fat rat increased high, The level of it of acupuncture group decreased, compared with the fat group p.001, This showed: the fat rats have the defeat of leptin and Ins in the body, The acupuncture can lowering this defeat, The level of leptin and Ins of the fat rat in the brain is less than the normal group, The level of leptin of the acupuncture group is high than the fat group, This showed: the acupuncture can increase the transportation of leptin from blood to brain.

本方法所制肥胖大鼠模型具有高体重、高脂、高leptin和高Ins血症的特点，符合人类单纯性肥胖病的临床和病理特征；实验研究显示：针灸能明显减少肥胖大鼠的摄水、摄食量及降低其体重、体脂；针灸能促使白色脂肪细胞直径、面积、体积缩小，减少肥胖大鼠棕色脂肪组织血管周围白色脂肪细胞数量，显示：针灸增强了白色脂肪细胞的代谢，促进了白色脂肪细胞向棕色脂肪细胞的转化：肥胖大鼠PVN神经细胞自发放电频率增加，脑内单胺递质紊乱，针灸通过抑制PVN亢奋的自发放电，调整5-HT水平，达到影响食欲，改善代谢的作用；肥胖大鼠血中leptin及Ins水平增高，与正常组大鼠比有明显差异，提示大鼠体内存在leptin及Ins抵抗，针灸后肥胖大鼠随着体重体脂的下降，血中leptin、Ins水平降低，与肥胖组比均有显著性差异，而针灸后大鼠脑内leptin增加，与肥胖组比P.05，提示：针灸具有改善leptin、Ins抵抗及增加leptin脑内转运的作用。

There were many acidophil corpuscles in hepatic sinus.

Subacute severe hepatitis 1 肝细胞水肿，肝细胞亚大块坏死，纤维组织紊乱，肝窦内见许多嗜酸性小体，肝细胞及毛细胆管淤胆明显，肝细胞部分脂肪变性，汇管区充血，有炎症细胞浸润

There were many acidophil corpuscles in hepatic sinus.

Subacute severe hepatitis 1 肝细胞水肿，肝细胞亚大块坏死，纤维组织紊乱，肝窦内见许多嗜酸性小体，肝细胞及毛细胆管淤胆明显，肝细胞部分脂肪变性，汇管区充血，有炎症颈椎病细胞浸润

The objective is to subjugate and discourage the people, because that allows the elite to continue to rule unopposed.

其目的是压制和打击人民的积极性，因为这可以让实权派继续统治不会沦为反对派。

GOD,this is the second time you vanquished me!

天啊，这是第二次你打败了我！