细胞

Under normal temperature, anthers of not fertility restoration stopped developing in the stage of archesporical cell; 1-2 comers of a few anther could differentiate and form archesporical cell, sporogenous cells and microspore mother cell, but microspore mother cell disintegrated on the first meiotic division.

常温下，未发生育性恢复的花药在孢原细胞期就停止发育；少数花药1~2个角隅处可分化形成孢原细胞，造孢细胞，花粉母细胞，但花粉母细胞在减数分裂的第一次分裂时解体。

Objective: To explore the mechanisms resulting in the recurrence of urethral scar which make urethral strictures difficult to be cured, a series experiments were conducted to find potential effective factors involved in urethral scar formation and degradation, including the studies of extracellular matrix component of urethral stricture scar, the characteristics of urethral scar fibroblast, and the effects of urine on urethral fibroblast in vitro, as well as the studies to compare the difference of collagenase activity, type Ⅰ collagen and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the tissues and cultured fibroblasts from normal urethra and strictured urethra respectively, and the studies to investigate the effect of antisense TIMP-1 oligodeoxyonucleotide on cell proliferation and collagenase activity of urethral scar fibroblast.

中文题名尿道瘢痕基础研究副题名胶原酶活性，TIMP-1的表达及其反义基因治疗外文题名 Experimental study on urethral scar-activity of collagenase，expression of TIMP-1，and antisense TIMP-1 gene transfection of urethral scar fibroblast 论文作者黄翔导师杨宇如教授学科专业外科学研究领域\研究方向学位级别博士学位授予单位四川大学学位授予日期2002 论文页码总数104页关键词尿道手术瘢痕胶原酶成纤维细胞尿道瘢痕馆藏号BSLW /2003 /R699 /12 目的：研究尿道瘢痕的细胞外基质的组成，尿道瘢痕成纤维细胞的生物学特性以及尿液对其生长的影响；比较胶原酶活性，金属蛋白酶组织抑制因子-1（TIMP-1）以及Ⅰ型胶原含量在尿道瘢痕和正常尿道组织及体外培养的成纤维细胞中的差异；研究反义TIMP-1寡核苷酸对尿道瘢痕成纤维细胞增殖以及胶原酶活性的影响。

It is suggested that the algae symbiot can affect host organellar function,quantity and location, stimulate the food uptake and digestive activity, restrain the functional activity of the organelle, change the gene expression and cause the change of genome structiure in the P. bursaria-chlorellae system.

据结果推测，纤毛虫中的藻类共生体可影响宿主细胞中胞器的功能、数量和分布，可影响细胞核核仁的数量和分布；可影响宿主细胞的形态、生长、抗氧化能力及胞质糖含量；可影响宿主细胞的食物摄取及相关胞器的消化活性，抑制宿主胞器的功能活动；并可改变宿主细胞基因的表达，导致细胞基因组结构的变化。

Methods:Human CD80 cDNA was transfected into B16 melanoma cells by lipofectin-mediated gene transfer before the expansion of tumor cells were tested by MTT method in vitro;C57BL/6 mice were inculated subcutaneously either mock-transfected B16 cells (B16-neo)or CD80-transfected B16 cells (B16-CD80),then the latency,survival times and tumor mass growth were investigated.The lymphocytes were examined for both proliferation indices and specific cytotoxic activity by MTT method and improved LDH-releasing method,after syngenic Mixed Lymphocyte tumor cultures.

通过脂质体介导将CD80基因导入小鼠黑色素瘤B16细胞后，利用SABC法检测CD80的表达；MTT法测定肿瘤细胞体外增殖能力；将肿瘤细胞接种至同源小鼠皮下后观察成瘤期、荷瘤小鼠存活期及肿瘤结节生长速度；在同源淋巴细胞肿瘤细胞混合培养后，通过测定淋巴细胞增殖指数和CTL杀伤活性检测肿瘤细胞的免疫原性。

Part II The experiment study on anti-tumour effect of dendritic cells derived from peripheral blood monocyte of patients with oral cancer after pulsed by tumour antigen in different ways. Collected 50ml peripheral blood from each patients and volunteers to induced DC in vitro. We parted the test group and control group into three subgroups respectively, untreated group A, freezed and thawn group B, RNA transfected group C. To group A, after induced DCs for five days , we added 200U/mlTNF-α in each hole and cultured on. To group B, we added protein antigen (DC/ tumor cells = 1 : 1 ,each hole) for 4h at 5d, and then added 200U/mlTNF-α in each hole and cultured on. To group C, transfected immature DC with tumor cell RNA(DC:RNA=1×105/ml: 5ug)at 5d, operated completely with the specification of invitrogen DMRIE-C reagents. After transfection we replaced transfection agent with complete medium, and then added 200U/mlTNF-α in each hole and cultured on.

第二部分口腔癌患者外周血来源的树突状细胞体外负载抗原后抑瘤作用的实验研究无菌采集20例口腔癌患者和20例健康志愿者外周血50ml，分离单个核细胞体外诱导培养DC，并将实验组和对照组细胞各分为三组，A组为未处理组，B组为冻融组，C组为RNA转染组。A组DC在培养5d后，每孔加入TNF-α200U/ml；B组DC培养第5d后，每孔中加入癌细胞蛋白抗原，使DC/肿瘤细胞=1：1，继续培养4h后，每孔加入TNF-α200U/ml；C组将培养至第5d的未成熟DC用肿瘤细胞RNA进行转染(DC：RNA=1×105/ml： 5ug，按照invitrogen DMRIE-C试剂说明书进行操作)，转染结束后用完全培基取代转染剂，每孔加入TNF-α200U/ml。

Results 500fold enrichments were observed by tittering phages in the cell lysate after five rounds of selection. A signal significantly greater than background binding was observed from the third to the fifth round of selection on CHOEGFRGFP1 and CHOK1 cell, but a part of polyclonal scFv bound EGFR specially. About 45% of the selected clones contained a fullsized insert of 1 kb. One unique human antiEGFR scFv (F4scFv) was isolated by analyzing with cell ELISA and DNA sequencing.

结果 经过5轮筛选，细胞裂解液中洗脱出噬菌体效价有500倍以上增长；细胞ELISA结果显示多克隆pscFv与CHOEGFRGFP1细胞和CHOK1细胞均有明显结合，但有部分特异性结合EGFR；菌落PCR显示约45％克隆中含有完整的1kb scFv片断；经细胞ELISA、 DNA测序检测共获得1株EGFR特异性单链抗体，命名为F4scFv。

Reactive oxygen species causing DNA oxidative damage comes from two kinds of ways:one is from cellular normal physiological metabolism;the other is from outer environment.Redox-sensitive green fluorescent protein was expressed in Saccharomyces cerevisiae.Recombinant cells were evaluated in monitoring the changes in the redox state of living cells when challenged with toxicologically relevant metal ions NaAsO_2 or Pb(NO_3)_2 by measuring emission intensity at 510 nm with a Hitachi F6500 fluorescence spectrophotometer,roGFP expressed in yeast responded not only to typical membrane-permeant oxidants H_2O_2 and reductants DTT,but also to toxicological metal ion-induced intracellular redox changes in a dose-dependent manner.Moreover,exposure of yeast cells to NaAsO_2 or Pb(NO_3)_2 at concentrations that induced redox changes reported by roGFP caused up to 2～3 fold increases in DNA mutation frequency.This mutagenic effect was largely caused by oxidative stress since blocking the production of hydryl radicals with thiourea significantly reduced the mutation rate as well as delayed the cell death.

本文将对氧化还原状态变化敏感的绿色荧光蛋白roGFP1-R12，在酵母细胞中实现了多拷贝强表达；荧光扫描经强氧化剂H_2O_2和还原剂DTT以及环境中重金属NaAsO_2或Pb(NO_3)_2处理后的酵母细胞悬液，测定510 nm处的荧光发射强度结果显示，表达的绿色荧光蛋白对氧化还原水平敏感，且在510 nm处的荧光强度与一定的重金属浓度呈正相关，即roGFP1-R12在510nm处的荧光发射值随重金属浓度的增高而增强，从而说明重金属对细胞的毒性在一定程度上很可能是通过破坏细胞内的氧化还原平衡发生作用；同时通过该绿色荧光蛋白对胞内氧化还原状态变化的响应情况可以来实时检测环境中的重金属；遗传学的点突变频率及致死率实验数据表明，重金属能导致菌体的点突变频率和致死率升高，且活性氧的清除剂巯基脲能明显降低这种点突变和致死率，说明由重金属引发的这种点突变和致死效应在很大程度上是依赖于重金属对细胞诱导产生的氧化胁迫。

Conclusions1、E_2 may induce the apoptosis and Fas,FasL expressions of Jurkat cell,its effect positively related to the dose and treating time of E_2.2、There is intrinsic co-expression of ERαand ERβon Jurkat cell,which can be upregulated by E_2,positively related to E_2 doses.3、E_2 may act as an apoptosis inducer of Jurkat cell via ERβ.4、E_2 can promote the transcriptive activity of NF-κB in Jurkat cell.The increased NF-κB activity may lead to increased apoptosis of Jurkat cell.

结论1、E_2可增加Jurkat细胞的凋亡，并增加Fas、FasL的蛋白表达，且其作用与E_2的剂量和处理时间呈正相关。2、Jurkat细胞上存在ERα、ERβ的表达，且E_2可上调这两种受体的表达，E_2剂量越大，ER表达量越高。3、E_2可能通过ERβ发挥其促进Jurkat细胞凋亡的作用。4、E_2能增加Jurkat细胞中NF-κB的转录活性，Jurkat细胞中NF-κB活性增加有促进凋亡的作用。

At 4 wk, the cells in the liver bud migrated into the domain of transversum mesenchyme, where endothelial cells began to form vascular structures, and branched to form a number of hepatic cords.

结果：3wk时靠近肝芽的间充质中的血管内皮细胞多，而肝芽内未见血管内皮细胞及造血细胞：4wk时，肝索形成处可见较多正在形成的新生血管增多，肝索间开始出现少量的造血细胞：5wk时肝索间出现原始的肝血窦结构，血窦内造血细胞的数量较4wk明显增多。

Methods: Viable cell numbers and viability were counted by means of trypan－blue exclusion.

在台盼蓝排除法计数活细胞和细胞活力的基础上，通过细胞形态学和流式细胞仪检测细胞凋亡。

With Death guitarist Schuldiner adopting vocal duties, the band made a major impact on the scene.

随着死亡的吉他手Schuldiner接受主唱的职务，乐队在现实中树立了重要的影响。

But he could still end up breakfasting on Swiss-government issue muesli because all six are accused of nicking around 45 million pounds they should have paid to FIFA.

不过他最后仍有可能沦为瑞士政府&议事餐桌&上的一道早餐，因为这所有六个人都被指控把本应支付给国际足联的大约4500万英镑骗了个精光。