细胞

Oogonium develops into early oocyte in the ovary, and then the oocyte leaves the ovary for the coelomic fluid in the form of single cell or cell mass followed by the rapid separation of the group of oocytes into individual ones. Oocyte enters into the nephridium after its maturation. The rupture of germinal vesicle marks the oocyte maturation. Oocyte in the coelom does not have follicle membrane and vitelline membrane is formed and developed by the oocyte itself. Smaller oocyte (0μm in diameter) is round, and larger ones (≥60μm in diameter) is ovate. The short and long diameters of a morphologically mature oocyte are about 115—120μm and 140—145μm respectively, and the vitelline membrane is 7—9μm thick.

卵原细胞在卵巢中发育至早期卵母细胞时期单个或成团脱离卵巢入体腔液中，卵母细胞团细胞很快分离为单个细胞；卵母细胞在体腔液中发育成熟后进入肾管；生发泡破裂是卵母细胞成熟的标志；体腔中卵母细胞无滤泡膜，卵黄膜的形成与发育靠卵母细胞本身；卵径小于60μm的卵母细胞呈圆形，卵径大于60μm 的卵母细胞为卵圆形，形态上成熟的卵母细胞短径约115—120μm、长径约140—145μm、卵黄膜厚7—9μm。

RESULT: 1.Ouabain act on lens sodium-pump,under the LM,lens anterior capsular membrane discontinuous,epithelium cells clustered,occluding zonule seperated,lens fiber layers fractured.Under the EM,cells totally hollowed,mitochondria swelling,myelin figure appeared.RT-PCR examine the expression condition of αsubunit of sodium pump on mRNA level,α1、α2 and α3-isoform are all decreased.2.Digoxin act on lens sodium-pump,under the LM,lens cell oedema,linkage distructed,extensive exfoliation.Under the EM,plasma appeared little half-transparant hollow region,mitochondria swelling and ridge disappeared. RT-PCR examine,α1、α2 and α3-isoform are all decreased.3.Amphotericin B act on lens sodium-pump,under the LM,lens epithelium cells linked tightly,arranged in-line,lens fiber layers arranged tightly and regularily.Under the EM,abbundant cellular organes,exuberant cells function indicated. RT-PCR examine the expression condition of αsubunit of sodium pump on mRNA level,α1 and α3-isoform are increased significantly,demonstrated isoform-specific action.4D-thyroxine act on lens sodium-pump,under the LM,lens plasmalemma integrated,cells arranged tightly and regularily.Under the EM,nucleus fission appeared,desmosome half-desmosome and tensile microfilaments linked the cells. RT-PCR examine,α2 and α3-isoform are increased, also demonstrated isoform-specific action.5.Vitamin E act on lens sodium-pump,under the LM,lens anterior capsular membrane continuous and smooth,epithelium cells tightly linked,lens fiber layers appearede hollow region occasionally.Under the EM,lateral membrane high density belt appeared,abundant nucleolus. RT-PCR examine,onlyα1-isoform are increased, demonstrated significantly isoform-specific action.6.DMSO act on lens sodium-pump,under the LM,lens anterior capsular membrane slightly thicker,cells linkage partly distructed.Under the EM,plasmalemma denaturation,mitochondria swelling.RT-PCR examine,α1、α2 and α3-isoform are all altered slightly and haven't significant meanning.

结果：1、哇巴因作用于晶状体钠泵后，光镜下晶状体前囊膜断裂、上皮细胞聚积、闭合连接分离、纤维板层裂隙，电镜下全层细胞空泡化、线粒体肿胀出现髓样结构，RT-PCR法检测晶状体钠泵α亚单位α1、α2及α3三种重整异构体在mRNA水平的表达均减弱。2、地高辛作用于晶状体钠泵后，光镜下晶状体细胞水肿、细胞连接破坏、广泛剥离，电镜下胞质见少许半透明空化区、线粒体肿胀嵴消失，RT-PCR法检测晶状体钠泵α亚单位α1、α2及α3三种重整异构体在mRNA水平的表达均减弱。3、两性霉素B作用于晶状体钠泵后，光镜下晶状体上皮细胞紧密连接、线状排列、纤维板层紧密规整，电镜下细胞器丰富、细胞功能旺盛，RT-PCR法检测α1及α3表达显著增强、具有一定的重整异构作用特异性。4、D甲状腺素作用于晶状体钠泵后，光镜下晶状体质膜完整、细胞排列紧密规整，电镜下胞核见分裂像、细胞间有桥粒、半桥粒及张力微丝，RT-PCR法检测α2及α3表达均增强、亦有一定的重整异构作用特异性。5、维生素E作用于晶状体钠泵后，光镜下晶状体前囊膜连续光滑、上皮细胞紧密连接、纤维板层偶见空化，电镜下囊侧膜内有高电子密度带、细胞核仁丰富，RT-PCR法检测仅有α1的表达显著增强、具有极强的重整异构作用特异性。6、二甲基亚砜作用于晶状体钠泵后，光镜下晶状体前囊膜轻度增厚、细胞连接部分破坏，电镜下质膜变性、线粒体肿胀，RT-PCR法检测晶状体钠泵α亚单位α1、α2及α3三种重整异构体在mRNA水平的表达无显著改变。

The height of bulliform cell groups reached 0.18 of leaves thickness, which was another important quantitative characteristic to achieve function of bulliform cells.

泡状细胞的周长面积比仅占普通表皮细胞的0.723，是泡状细胞功能实现的重要数量特征；(2)以细胞面积、周长、等效圆直径、每平方公厘细胞数、周长面积比，可以建立小麦叶片泡状细胞Fisher's线形判别函数，判别正确率达99%；(3)泡状细胞组高占叶厚的比例达到了0.18，是泡状细胞功能实现的又一重要数量特征；(4)相关分析表明，泡状细胞组长、泡状细胞组高、泡状细胞组面积均与泡状细胞距离叶缘的位置呈极显著负相关。

Higher Ca distributed in bulliformcell than in mesophyll cell and bundle sheath cell of dune reed, higher Mg distributedin mesophyll cell and higher K, Na and Cl distributed in sheath cell HigherNa and Mg distributed in mesophyll than in bulliform cell and bundle sheathcell of light salt meadow reed, and higher K, Ca and Cl distributed in itsbundle sheath cell. Higher Na and Mg distributed in bulliform cell than in mesophyllcell and bundle sheath cell of heavy salt meadow reed, higher K, Ca and Cl distributedin its mesophyll cell. This paper discussed the distribution conditions of theabove five ions in leaf cell of the four reed ecotypes and the meaning ofphysiological adaptation to habitat in detail.

沙丘芦苇的泡状细胞内Ca分布较叶肉细胞和鞘细胞高，叶细胞内Mg分布较高，在鞘细胞内K，Na和Cl布较高；轻度盐化草甸芦苇叶肉细胞内分布了较多的Na和Mg，在鞘细胞内K，Ca和C1分布较叶肉细胞和泡状细胞高；而重度盐化草甸芦苇泡状细胞内分布了较多的Na和Mg，叶肉细胞分布了较多的K，Ca和Cl；详细讨论了以上五种离子在不同生态型芦苇叶片内不同细胞类型的分布状况与环境适应的意义。

CD8~+ cell is the main T lymphocyte subset in spleen, and B lymphocyte mainly is IgG~+ cell, moreover the amount of these B lymphocytes could exceed CD3~+ T lymphocyte subset after 7 days. CD8~+ cell is the main T lymphocyte subset in tonsil of appendix, and B lymphocyte is IgM~+ cell, and the amount could exceed CD3~+ T lymphocytes after 35 days. After 21 days, B lymphocytes in esophago tonsil are the main IgA~+ cells and the amount exceeds CD3~+ lymphocytes. The amount of CD4~+ lymphocytes is more than CD8+ lymphocytes.4. CD3~+、CD4~+ and CD8~+ T lymphocytes in spleen mainly distribute in periarterial lymphoid sheath. However IgM~+、IgG~+ and IgA~+ cells mainly distribute in ellipsoid periarterial lymphoid sheath and germinal center. T lymphocytes in appendix tonsil mainly distribute in middle and inferior part of mucous and the B lymphocytes mainly in middle and mucous between 4~7 days. Whereafter T, B lymphocytes equably distribute in mucous. CD4~+ cells arrange tightly and mainly occupy the central part in aggregates of T lymphocytes in esophago tonsil and CD8~+ lymphocytes mainly distribute in periphery. Meanwhile B lymphocytes encircle the periphery of aggregates of T lymphocytes. The aggregates of B lymphocytes is mainly the germinal center with lots of IgM~+、IgG~+ and IgA~+ cells. Meanwihle T lymphocytes encircle the periphery of aggregates of B lymphocytes.5. There is an intimate relationship between the development of tissue structure of peripheral immune organs and lymphcytopoiesis. The maturation of tissue structure is stimulated by the immigration of lymphocytes and the mature tissue structure provides place where lymphocytes grow mature and functionate.

脾脏在21日龄时达到稳定，食管扁桃体和盲肠扁桃体均在35日龄时达到稳定；脾脏中T淋巴细胞亚群以CD8~+细胞为主，B淋巴细胞则以IgG~+细胞为主，并在7日龄后数量超过CD3~+T淋巴细胞；盲肠扁桃体中T淋巴细胞亚群以CD8~+细胞为主，B淋巴细胞以IgM~+细胞为主，并在35日龄后数量超过CD3~+T淋巴细胞；21日龄后，食管扁桃体中B淋巴细胞以IgA~+细胞为主，数量超过CD3~+细胞，CD4~+细胞的数量多于CD8~+细胞。4、在T、B淋巴细胞组织定位方面，脾脏中CD3~+、CD4~+和CD8~+T淋巴细胞主要分布在动脉周围淋巴鞘中，而IgM~+、IgG~+和IgA~+细胞主要分布在椭球周围淋巴鞘和生发中心中；4～7日龄时，盲肠扁桃体中T淋巴细胞主要分布在粘膜固有层的中下部区域，而B淋巴细胞则主要分布在中上部区域，随后各日龄T、B淋巴细胞均匀地分布在粘膜固有层中；在食管扁桃体的T淋巴细胞聚集物中，CD4~+细胞紧密排列，主要占据中央部位，CD8~+细胞主要散布在外周，同时B淋巴细胞又环绕在整个T淋巴细胞聚集物的外周；B淋巴细胞聚集物主要为生发中心，其中存在大量IgM~+、IgG~+和IgA~+细胞，同时T淋巴细胞又环绕在整个B淋巴细胞聚集物的外周。5、外周免疫器官的组织结构发育和淋巴细胞发生之间存在密切的关系，淋巴细胞迁入淋巴器官刺激组织结构的发育成熟，同时成熟的组织结构又为淋巴细胞发育成熟并行使功能活动提供场所。

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。

Many factors may be involve in the course. To investigate the regulation activity of mesenchymal cells to differentiation of epithelial cells from hair follicle and to study its differentiation property, mesenchymal cells gel was made by nubby dermal papilla cells, free dermal papilla cells, skin fibroblasts. Skin keratinocytes and epithelial cells from hair follicle were inoculated on the gel surface and cultured in air-liquid interface. Three-dimensional model of DPC using to induce epithelial cells differentiation is built in vitro.

为了进一步研究毛囊细胞间的相互作用，探讨毛囊间质细胞对毛囊上皮细胞分化的调节作用，研究毛囊上皮细胞的分化特性，我们利用团块状的毛乳头细胞，游离分散的毛乳头细胞或皮肤成纤维细胞制成间质细胞胶原凝胶，表面接种皮肤角质形成细胞或毛囊上皮细胞，进行气-液界面培养，在体外建立了毛乳头细胞诱导毛囊上皮细胞分化的立体模型。

Restoration of ODC expression in the HL60-ODC cells resulted in rescue of rottlerin-induced apoptosis of HL60 cells. Simultaneously, phosphothreonine-ODC of rottlerin-treated HL60-ODC cells was highly elevated comparing with HL60-pcDNA3 cells. Taken together, we suggested that rottlerin, a PKCd inhibitor, might decrease phosphothreonine-ODC to reduce both protein stability and enzyme activity of ornithine decarbooxylase and finally resulted in cell apoptosis.

为了侦测鸟胺酸去羧化是否能够负调节rottlerin所引发的细胞凋亡，我们将具有鸟胺酸去羧化cDNA载体稳定转染至HL60细胞内，此细胞称为HL60-ODC细胞，另转染载体的细胞称为HL60-pcDNA3细胞；实验发现HL60-ODC细胞中的鸟胺酸去羧化蛋白为HL60-pcDNA3细胞与HL60细胞表现之2.5倍，同时HL60-ODC细胞提升鸟胺酸去羧化蛋白表现，能够减少rottlerin造成之细胞凋亡现象，并且此蛋白在HL60-ODC细胞中酥胺酸磷酸化较HL60-pcDNA3细胞表现高。

The EC_(50) of QKL against the formation of syncytia of MT-2 induced by HIV-1_ was 1/198.02 with a SI of 5.36. By detecting the survival rate of co-culturing H9/HIV-1_ cells and MT-2 cells or MT-2 cell direct infected by HIV-1_ QKL was found to be protective to cells. The EC_(50) was 1/166.67 and 1/144.93 with SI of 4.51 and 3.92 respectively. The EC_(50) of QKL against P24 antigen production was 1/175.44 with SI of 4.75. The drug serum of QKL was also found to be effective to inhibit the cell fusion and protect cells infected by HIV-1_.

结果：QKL对H9／HIV-1_细胞和MT-2细胞的CC_(50)分别为1／50.76和1／36.97；抑制H9／HIV-1_细胞和MT-2细胞早期融合的EC_(50)=1／235.29，SI=6.36；抑制HIV-1_细胞诱导MT-2细胞形成合胞体的EC_(50)=1／198.02，SI=5.36；H9／HIV-1_细胞和MT-2细胞混合培养和HIV-1_感染MT-2细胞时，QKL保护病毒感染细胞免于死亡的EC_(50)分别为1／166.67和1／144.93，SI分别为4.51和3.92；抑制p24抗原产生的EC_(50)=1／175.44，SI=4.75；QKL药物血清也可抑制H9／HIV-1_细胞和MT-2细胞的早期融合，保护细胞免于死亡。

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。

I didn't watch TV last night, because it .

昨晚我没有看电视，因为电视机坏了。

Since this year, in a lot of villages of Beijing, TV of elevator liquid crystal was removed.

今年以来，在北京的很多小区里，电梯液晶电视被撤了下来。