多核细胞

The stable clones are further identified by RT-PCR and Western blot; 6 MTT assay is used to investigate the effect of ZNRD1 on the cell growth of cells (AGS, SGC7901, MKN28, NIH3T3, GES-1); 7 Soft agar assay is used to investigate the effect of ZNRD1 on the clonality of cells (AGS, MKN28); 8 Nude mice assay is used to investigate the effect of ZNRD1 on the cell growth of gastric cancer cells (AGS, MKN28); 9 Flow cytometry is used to investigate the effect of ZNRD1 on the cell cycle distribution of cells (AGS, MKN28, NIH3T3, GES-1); 10 Flow cytometry is used to investigate the effect of ZNRD1 on the cell apoptosis of cells (AGS, MKN28, NIH3T3); 11 MTT assay is used to investigate the effect of ZNRD1 on the drug sensitivity of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR) in vitro; 12 SRCA is used to investigate the effect of ZNRD1 on the drug sensitivity of gastric cancer cells (SGC7901, SGC7901/VCR) in vivo; 13 Flow cytometry is used to investigate the effect of ZNRD1 on adriamycin accumulation of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR); 14 Transmission electron microscope is used to investigate the effect of ZNRD1 on the sensitivity of SGC7901 cells towards drug-induced apoptosis; 15 Flow cytometry and DNA ladder assay are used to investigate the effect of ZNRD1 on the sensitivity of cells (SGC7901, SGC7901/VCR, HL-60/VCR) towards drug-induced apoptosis; 16 Microarray is used to investigate the profiling of ZNRD1-responsive genes in gastric cancer cells (AGS, MKN28, SGC7901, SGC7901/VCR); 17 RT-PCR and Western blot are used to identify the results of microarray; 18 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of cyclin D1; 19 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of MDR1; 20 Kinase assay is used to investigate the effect of ZNRD1 on the activity of cyclin E-CDK2 kinase; 21 The antisensenucleic acids of p21 is used to inhibit the expression of p21, and flow cytometry is used to investigate the effect of p21 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 22 The antisensenucleic acids of p27 is used to inhibit the expression of p27, and flow cytometry is used to investigate the effect of p27 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 23 Liposome is used to up-regulate the expression of Skp2, and flow cytometry is used to investigate the effect of Skp2 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 24 Western blot is used to investigate the effect of ZNRD1 on the stability of Skp2 and p27 in gastric cancer cells; 25 MVD assay is used to investigate the effect of ZNRD1 on the angiopoietic activity of gastric cancer cells; 26 ELISA is used to investigate the effect of ZNRD1 on the expression of VEGF165 in gastric cancer cells; 27 The roles of DARPP-32 in MDR of gastric cancer cells are investigated using gene transfection, MTT assay, SRCA, flow cytometry and DNA ladder assay.

应用杂交瘤技术制备ZNRD1的首个单克隆抗体；2）利用RT-PCR、Western blot和免疫组化检测ZNRD1在胃癌组织、胃炎组织、正常胃上皮组织、胃癌细胞和正常胃组织上皮细胞中的表达；3）构建ZNRD1的小干扰RNA载体，并测序鉴定；4）利用脂质体将ZNRD1的真核表达载体及其空载体转染胃癌细胞（AGS、SGC7901、MKN28）和小鼠成纤维细胞（NIH3T3），G418筛选后进行鉴定；5）利用脂质体将ZNRD1的小干扰RNA载体及其空载体转染药敏胃癌细胞（SGC7901）、正常胃组织上皮细胞（GES-1）、对长春新碱耐药的胃癌细胞（SGC7901/VCR）、药敏白血病细胞（HL-60）、对长春新碱耐药的白血病细胞（HL-60/VCR），G418筛选后进行鉴定；6）利用MTT实验检测ZNRD1高/低表达对细胞（AGS、SGC7901、MKN28、NIH3T3、GES-1）生长的影响；7）通过软琼脂克隆形成实验检测上调ZNRD1对AGS、MKN28细胞克隆形成能力的影响；8）通过裸鼠成瘤实验检测上调ZNRD1对AGS、MKN28细胞体内成瘤性的影响；9）通过流式细胞仪分析ZNRD1高/低表达对细胞（AGS、MKN28、NIH3T3、GES-1）的细胞周期的影响；10）通过流式细胞仪分析上调ZNRD1对细胞（AGS、MKN28、NIH3T3）的凋亡的影响；11）通过MTT实验检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60、HL-60/VCR）体外药物敏感性的影响；12）通过肾包膜下移植法检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR）体内药物敏感性的影响；13）通过流式细胞仪分析ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60、HL-60/VCR）内阿霉素蓄积和泵出的影响；14）通过透射电镜检测上调ZNRD1对SGC7901细胞凋亡敏感性的影响；15）通过流式细胞仪和DNA梯度试验检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60）凋亡敏感性的影响；16）通过基因芯片检测ZNRD1高/低表达对胃癌细胞内基因表达谱的影响；17）利用RT-PCR、Western blot对基因芯片的结果进行鉴定；18）利用报告基因实验检测ZNRD1对cyclin D1的启动子活性的调节作用；19）利用报告基因实验检测ZNRD1高/低表达对MDR1的启动子活性的调节作用；20）利用激酶试验检测ZNRD1对cyclin E-CDK2 激酶活力的影响；21）利用反义核酸技术抑制p21的表达；通过流式细胞仪检测抑制p21对ZNRD1介导的细胞周期阻滞的影响；22）利用反义核酸技术抑制p27的表达；通过流式细胞仪检测抑制p27对ZNRD1介导的细胞周期阻滞的影响；23）利用脂质体转染法上调Skp2的表达；通过流式细胞仪检测上调Skp2对ZNRD1介导的细胞周期阻滞的影响；24）利用Western blot检测ZNRD1对p27和Skp2的蛋白稳定性的影响；25）利用微血管密度实验检测ZNRD1对AGS、MKN28细胞裸鼠移植瘤微血管形成的影响；26）利用ELISA检测ZNRD1对AGS、MKN28细胞培养上清和移植瘤匀浆中VEGF165含量的影响；27）利用脂质体转染法、MTT实验、肾包膜下移植法、流式细胞仪和DNA梯度试验检测新耐药相关分子DARPP-32对细胞（SGC7901、SGC7901/VCR、对阿霉素耐药的胃癌细胞SGC7901/ADR）多药耐药表型的影响；利用脂质体转染法和MTT实验检测下调ZNRD1对DARPP-32介导的胃癌多药耐药的调控作用。

Grew on the normal temperature, it was shown that the deposits of calcium antimonate being the indicator for Ca(superscript 2+) localization mainly concentrated within the vacuoles and intercellular spaces and there was also some Ca(superscript 2+) deposits in cell walls. But when Garyota urens L. was treated by the temperature of 2℃ for 48 h, the level of Ca(superscript 2+) increased in cytoplasm and plasma membrane, but decreased in vacuoles and intercellular spaces considerably. At the same time, the ultrastructure of chloroplasts suffered from chilling: the membrane of chloroplasts had been damaged, the layer of thylakoids was exiguous and unclear, the photosynthetic rate decreased evidently. And when Garyota urens L. was treated by the temperature of 2℃ for 120 h, the deposits of Ca(superscript 2+) mainly concentrated within the cytoplasm, nucleus and plasma membrane and there was also some Ca(superscript 2+) deposits in vacuoles, and the ultrastructure of some cells was simultaneously damaged severely: Chloroplasts structure, vacuole membrane and nuclear membrane had been damaged fully, the structure within the cell had become unclear, and the cell only have respiration.

研究结果表明，未经低温处理的董棕幼苗叶肉细胞，焦锑酸钙沉淀颗粒大量出现在液泡和细胞间隙中，细胞壁中也可见少量沉淀，而细胞基质中则看不到焦锑酸钙沉淀；经2℃ 48 h低温处理后，细胞基质和细胞膜上焦锑酸钙沉淀增加，而液泡和细胞间隙中的焦锑酸钙沉淀则显著减少，并且超微结构已初步显示出寒害的特徵，叶绿体外膜部分破损，类囊体片层稀疏且排列不规则，光合速率明显下降等；经2℃ 120 h低温处理后，细胞间隙内的焦锑酸钙沉淀极少，有的也紧贴在细胞外壁上，而细胞基质和细胞膜上则分布有非常多的焦锑酸钙沉淀，在核基质和液泡中也可见到少量的焦锑酸钙沉淀，并且超微结构遭到了显著破坏，叶绿体结构完全被破坏，核膜与液泡膜严重破损，内部结构模糊，细胞只表现为呼吸作用，不进行光合作用。

The ultrastructure characters of pollen grain in Brassica napus are:1There are vacuoles in cytoplasm but no starch grain in uniceelluar pollengrain.2The veget- ative nucleus of bicellular pollen grain is generally spheroidal;while the generative cell is spinde-shaped,with a big nucleus,thin layer of cytoplasmand few cell orgens,no cell wall.3In three-celled pollen grain the sperm cell is separated from the cytoplasm of vegetative cell by two layers of plasmic membrances.

Brassica napus的花粉粒，在不同发育时期超微结构的特征如下：1单胞花粉粒有一个球形的细胞核和明显的核仁，细胞质内出现液泡，缺少典型的淀粉粒。2双胞花粉粒的营养核多为球形。生殖细胞纺锤形，无细胞壁，细胞核比例大，细胞质呈薄层，细胞器少。3三胞花粉粒有一个裂片状的营养核和两个纺锤形的精细胞，精细胞无壁，以两层质膜与营养细胞的细胞质相隔。

1,After transfected the mutated mtDNA of colorectal carcinoma,the mtDNA D-loop region of the transfected cells displays new mutation points.2,The external source pieces of the mutated mtDNA can integrate to nuclear genome after transfection.3,There's no differences in apoptosis between combinations after transfected the mutation of mtDNA in NIH3T3 and LST cells.4,The mutated mtDNA may affect the action mechanism of occurrence and development in colorectal carcinoma through affecting its mtDNA mutation or integrating exogenetic mtDNA to its nuclear which may cause the abnormal expression of oncogene or anti-oncogene.

（1）转染突变的大肠癌细胞mtDNA后转染细胞的mtDNA均可发生多处的突变位点。（2）通过转染后突变的外源性的mtDNA可以整合到核基因组内。（3）突变的mtDNA转染LST细胞及NIH3T3细胞后，不影响转染细胞的凋亡改变。（4）mtDNA的突变可能通过影响体细胞mtDNA的突变和通过外源性mtDNA在核内的整合从而影响癌基因或抑癌基因的表达异常，从而参与肿瘤的发生发展。

1,After transfected the mutated mtDNA of colorectal carcinoma,the mtDNA D-loop region of the transfected cells displays new mutation points.2,The external source pieces of the mutated mtDNA can integrate to nuclear genome after transfection.3,There's no differences in apoptosis between combinations after transfected the mutation of mtDNA in NIH3T3 and LST cells.4,The mutated mtDNA may affect the action mechanism of occurrence and development in colorectal carcinoma through affecting its mtDNA mutation or integrating exogenetic mtDNA to its nuclear which may cause the abnormal expression of oncogene or anti-oncogene.

（1）转染突变的大肠癌细胞mtDNA后转染细胞的mtDNA均可发生多处的突变位点。（2）通过转染后突变的外源性的mtDNA可以整合到核基因组内。（3）突变的mtDNA转染LST细胞及NIH3T3细胞后，不影响转染细胞的凋亡改变。（4）mtDNA的突变可能通过影响体细胞mtDNA的突变和通过外源性mtDNA在核内的整合从而影响癌基因或抑癌基因的表达异常，从而参与肿瘤的发生发展。线粒体DNA；D―环区；突变；质粒；pcDNA3.1；转染

ResultsThe glandular cell in the endometrial implant after therapy with middle or high-dose of (10,15 gkg-1d-1) XiaoChaiHu Decoction showed characteristic features of apoptosis which displayed by the cell decreased in size, karyopyknosis , cytoplasmic and nuclear chromatin condensation , density increased and a lot of apoptotic bodies among cell. Whereas some stromal cell displayed degeneration and necrosis. The protein expression of Fas and Caspase-3 in endometriotic tissue of XiaoChaiHu Decoction group was higher than that in the endometrium.

结果中、高剂量小柴胡汤（10，15 gkg-1d-1）治疗后，异位内膜有较多腺上皮细胞出现凋亡的特征，表现为细胞体积变小，核固缩，胞浆和核染色质凝集，密度增高，细胞间凋亡小体，同时间质细胞中可见一些坏死细胞；异位内膜Fas蛋白、Caspase-3蛋白的表达水平明显高于其在位内膜。

TyPe II collagen induced arthritisln the rat ank1e joint andoVathumin as antigen induced arthritis WA in the rabbit knee joint wereestab1ish2 Qualitative evaluation of me in skin, muscle, synovium, cedilagearound joint and blood was performed by OMA3 The CIA rats were treated on day 7 after hind paw swelling and erythemaAnimals were injected intravenously with ase at a dose of 10mg/kg,tWenty minuots 1ater, one ankle of the rats random1y assigned was exPosedlaser irradiation at l00J/cm fOr l000 seconds, and another ankle wasM grouP wihout laser The other two groups is unmanipulatedcontrol group and untreated CIA group Bimaleolar ankle widthmeasuremellts were taken in all animals every tWo days using amicrometer The histopathology of the ank1e Joint was assessed at day 21after disease onset4 The pro1iferating cell nuclear antigen WCNA of CIA treated by PDT andthe HMME group without laser was doterdrined by immunohistochemiStry5 The AfA rabbits were treated on day 7 after knee swelling and erythemaThe theraPy invo1ved lntravenous injection of l0mg/kg HMME, fOl1owedby 20 minues period in dim light, and transdermal light treatment with\l00 J/cm2 fOr l000 seconds The inner sides of the treated Anees wereirradiated at first, and then the outer side did 24 hours later, the synovialtissue of the Anees joint were removed and in situ cel1 aPoptosis wasdetCCted With tednal deoxync1eotidyl transferase-mediated dUTP nickend labelingR6suIt8:l The pathologic changes of CIA and AIA include subsynovial inflammation,opovial hyPerplasia, pannus formation, cartilage and bone destructionresemble RA.2 The studies demonstrated that there are different uptake of HMME withinskin, muscle, synovium, cartilage and b1ood, and the synovium cou1draPidly uPtake more ase than skin and cartilage at the firSt 30 minuesaller intravenous injection of HMME3 The bimaleolar anke width had no different among PDT treated group,H group withollt 1aser and untreated CIA group But hlstologicalevaluation showed statiStical1y significallt reductions in synovialhyperplasia, pannus formation and cart1lage reosion, bone destruction andtotal score in PDT treated group4 Image analysis showed that the ratlo bforeen the areas of the coufltedobect to that of the entire area in PDTtreated grOup is lower than that in conirol group, but the integrated oPticaldensity had no different between the two groups5 Imape analysis showed that the ratio between the area of the countedobject to that of the e

治疗组在大鼠出现踝关节红肿后1周，炎症达到高峰时进行PDT治疗。随机治疗大鼠一侧的踝关节，另。2。一一侧作单纯HMME 对照。治疗方法是大鼠麻醉后尾静脉注入 HMME10ngkg，20分钟后踝关节照光，激光波长627.sum，功率密度 100mwcm'，照射时间1000秒，能量密度100）／。治疗后避光喂养72 小时。隔日一次测量大鼠的踝关节左右横径，治疗后两周取关节进行病理d 观察。 4。大鼠CIA模型用上述方法进行PDT治疗后，治疗组和单纯HMME 组用兔疫组化SP法检测石蜡切片的核增殖抗原。 5。兔AIA模型在关节炎出现第七天进行PDT治疗，随机治疗一侧膝关节，另一侧作自身对照。兔耳静脉注入I'arrainrelomg／Kg，20分钟后，膝关节用金蒸气激光照射，激光能量密度100）儿旷。24 ／J'时后取膝关节滑膜作病理检查，并用脱氧核昔酸末端转移酶介导的缺口末端标记法原位检测凋亡细胞。结果： 1。模型观察：CIA大鼠炎症高峰期滑膜下炎细胞浸润明显，滑膜细胞明显增殖，炎症达到高峰后二周，血管缀形成，并侵蚀和破坏软骨和骨， CIA模型病理改变与人类RA相似。兔AIA模型膝关节滑膜病理可见滑膜细胞增生，滑膜下炎细胞浸润，也与人类RA滑膜改变相似。 2。关节周围组织中光敏剂含量的测定结果表明，各组织对HMME 的吸收速度和吸收量不同，荧光值一时间曲线不同，滑膜组织比皮肤和软骨对 HMME的吸收多，在 2 0分钟时即有明显差异。 3.PDT对CIA模型的治疗结果表明：PDT治疗后关节炎组、单纯 HMME组和治疗组踝关节左右横径统计学检验差异没有显著性，但病理评分PDT治疗组滑膜增生、血管资形成及软骨破坏、骨破坏和总分比关节炎对照组和HMME对照组好，统计学检验差异有显著性。。3＿军医进修学院硕士学位论文中文摘要 4.PDT治疗组PCNA阳性细胞较对照组少，图像分析结果表明面密度（阳性染色的面积总和与统计视野面积的比值）治疗组小于对照组，统计学检验差异有显著性。。 5.PDT治疗组凋亡阳性细胞较对照组明显增多，图像分析结果单位视野内阳性细胞数和面密度PDT治疗组高于对照组，统计学检验差异有显著性。凋亡细胞核直径PDT治疗组较小，与对照组相比，统计学检验差异有显著性。结论：二。CIA、AIA的病理改变类似人类RA，可作为研究RA病因、发病机制、检查及治疗方法的模型。 2。各组织对HMME的吸收速度和吸收量不同，滑膜组织比皮。

Results: Of all the semen samples, 8 were HCMV positive, 4 HSV-Ⅱ positive, but none were both HCMV and HSV-Ⅱ positive. HCMV late antigens were positively and HCMV early antigens negatively expressed in the spermatogenic cells of the 8 HCMV positive cases. In the 4 HSV-Ⅱ positive cases, 3 were positively and 1 weakly positively expressed. In the semen of the 12 positive cases were found large numbers of immature spermatogenic cells, with different manifestations of apoptosis, such as chromatin pycnosis, vacuoles, damaged nuclear membrane, and apoptotic bodies, but without virus infection-induced specific morphological alteration.

结果：83例精液样本PCR检测HCMV阳性8例，HSV-Ⅱ阳性4例，未发现2种病毒同时阳性病例。8例HCMV阳性病例ICC标记生精细胞HCMV晚期抗原均见阳性表达，HCMV早期抗原均为阴性表达；4例HSV-Ⅱ阳性病例ICC标记生精细胞3例阳性表达，1例弱阳性。12例病毒阳性病例均发现较多未成熟生精细胞，并有不同程度的核染色质固缩、出现空泡、核膜破损、核崩解、凋亡小体等凋亡现象，但未能找到病毒感染的特异性形态学改变。

There were no statistically differences between the 8th~ 14thday, the 15th - 21th day post-BC and the control group. During the rt~24tkday post-BC , pycnosis degeneration or necrosis neurons of the locus cerebral cortex, dorsal hippocampus, dentate fornix were significantly increased, then decreased gradually, but recovered to normal by the 24thday after BC, and especially in parietal cortex and piriform cortex the necrosis neurons were significantly increased than temporal cortex , and there were no statistically difference berween the left and the right side. Pycnosis degeneration or necrosis neurons in the brainstem reticular formation were markedly increased in the 4thday after BC, and there were no statistically difference among the other groups and the control group.

与对照组相比，BC后在大脑皮质、背侧海马和齿状回部位，固缩变性和不完全坏死细胞数先显著增加(P＜0.05)，然后逐渐减少，至24d基本恢复正常；顶叶、梨状皮质比颞叶皮质变性坏死细胞多，差异有高度显著性(P＜0.01)；大脑左侧比右侧变性坏死细胞稍少但差异无显著性；在BC后4d，脑桥核、斜方体核平面的脑干网状结构中固缩变性和不完全坏死细胞数明显增多(P＜0.05)，其它组间无显著性差异。

A lot of organelles such as mitochondria, rough endoplasmic reticula and Golgi bodies were observed in fiber cell. Then, double karyotheca disappeared, and the organelles disintegrated. Multivesicular bodies appeared in fiber cytoplasm. With the further development of fiber, fiber wall underwent continual thickening with aging, and polylaminate structure gradually appeared. While the agglutinated nucleus, transfer vesicles, plasma membrane and plasmodesmata still presented.

研究发现，次生壁形成早期，细胞核具有双层核膜，染色质凝聚，可见大量的线粒体、粗面内质网和高尔基体等细胞器存在于纤维细胞中；随后，双层核膜消失，细胞器将逐渐解体，多泡体开始出现在纤维细胞的细胞质；随着年龄的增加，纤维细胞壁逐渐增厚，并出现多层结构现象，而运输小泡、细胞膜、胞间连丝和凝聚的染色质将持续存在。

In the negative and interrogative forms, of course, this is identical to the non-emphatic forms.

。但是，在否定句或疑问句里，这种带有"do"的方法表达的效果却没有什么强调的意思。

Go down on one's knees;kneel down

屈膝跪下。。。下跪祈祷