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The efficiency of the nuclear transfer was 63.6%(63/99).Forty-seven clone embryos were transferred into 5 recipient goats. After embryo transfer, two recipient goats have not been seen oestrous within 2 oestrum, and one of the two recipient goats was examined positive by gravid testing, which testified the clone embryos were viable in the uterus of recipient goats.

阳性细胞克隆中的2株(M9,M10)已经进行了首次细胞核移植,核移植成功率为63.6%(63/99),将47个发育较好的克隆胚胎进行了胚胎移植,共用受体羊5只,2只受体羊2个情期未见返情,在没有返情的2只受体山羊中有1只妊娠检测阳性,说明转基因克隆胚胚胎移植后在受体山羊体内发育良好。

Cobra venom factor was given to prevent hyperacute rejection and FK506 was given to prevent cellular rejection. The xenograft recipients were killed and cardiac xenografts harvested 4, 8, 12, 16, 24 hours after transplantation. Histologic examination was used to demostrate the degree of rejection and coagulation while revers transcriptase-polymerase chain reaction was used to detect the expression of tissue factor mRNA, and the guinea pigs hearts, harvested but not transplanted, were studied as controls in all samples.

建立豚鼠-大鼠异种异位心脏移植动物模型,使用CVF抑制超急性排异反应,FK506抑制细胞免疫排斥反应,分别在移植术后4、8、12、16、24h切取移植心脏,通过病理切片观察排异反应和凝血强度,同时通过RT-PCR方法检测组织因子mRNA表达强度,未移植豚鼠心脏作对照。

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。

Results The long term results of the vascularized fibular graft varied in different types of bone defects. Nine patients with congenital pseudoarthrosis of tibia had the limb function restored in 65% and one had nonunion of fibular graft, 3 fractures(7 times), and 5 angular deformities. Three patients with congenital radial deficiency had 62% restoration of the limb function. All of them showed early closure of epiphyses of the grafted fibulae. Five cases with osteomyelitis and bone defects combined soft tissue deficiency had 94% restoration of the limb function, and only one occurred fracture of fibular graft. One case with soft tissue defect and concomitant ulnar defect, ulnar nerve and median nerve injury, 67% function of limb was recovered, and union of fibula graft was good. 100% function of limb was recovered in 1 case with cavernoma of radius and without occurring valgus deformity of donor site.

结果应用带血管腓骨移植修复骨缺损的远期效果因缺损类型的不同而存在较大差异。9例先天性胫骨假关节患者,术后肢体功能恢复65%,其中移植腓骨骨不连1例、骨折3例(7次)、成角畸形5例;3例先天性桡骨缺如患者,术后肢体功能恢复62%,3例术后均出现移植腓骨骨骺线早闭,畸形复发;5例骨髓炎、骨外露、骨缺损并软组织缺损患者,术后肢体功能恢复94%,仅1例发生移植腓骨骨折;1例前臂软组织缺损并尺骨缺损,尺神经、正中神经损伤者,术后肢体功能恢复67%,骨愈合顺利;1例桡骨海绵状血管瘤者,术后肢体功能恢复100%,骨愈合顺利。

Allogenetic hemapoietic stem cell transplantation is considered to be the curative therapeutic method for CML.

异基因造血干细胞移植是CML的唯一根治方法,在CML慢性期移植可以获得较理想的疗效,但加速期及急变期移植的相关死亡率和移植后复发率明显升高。

The allograft was harvested at 6h posttransplantation. Scrambled ODN group: The donor kidney was stored for 6h in ECs containing 1 u M Liposome/scrambled ODN complexes under hypothermal and hypoxia condition, then the donor kidney was transplanted into a SD recipient, and at 6h posttransplantation the allograft was harvested.

五、试验分四组:(1) Control组假手术组,Wister大鼠麻醉后,开腹后,即将伤口缝合,6h后切取左肾;(2) ECs组供肾经ECs(不含脂质体/ODN复合物)低温缺氧保存6h,移植到受体大鼠6h后切取移植肾;(3)deeoy onN组供肾经含1 pM的deeoy onN脂质体复合物的ECs 低温缺氧保存6h,移植到受体大鼠6h切取移植肾;(4)sc~。

But after transplantation, the occurrence rate of incomplete kidneyfunction is still higher, the happen of the renal transplant complications afteroperation is the mainest cause lead to the function of transplanted kidneydamage even lose, and it is the key to improve survival rate of the TK if diagnosingaccurately in early phase and giving the rational iatrotechnics.

然而移植术后肾功能不全的发生率仍较高,肾移植术后并发症的发生是导致移植肾功能损害乃至功能丧失的最主要原因,如能早期、准确的诊断并予以合理的治疗术后并发症是提高移植肾存活率的关键。

Methods RI and PI of main renal artery and interlobar artery were obtained by color Doppler ultrasonography within the first week of transplantation.Then they were correlated with allograft glumerular filtration rate at 1 year.

在100例无并发症肾移植患者中,应用彩色多普勒超声,于肾移植术后1周内,测量移植肾肾动脉和叶间动脉的阻力指数和搏动指数,并与术后1年的移植肾肾小球滤过率进行相关分析。

PartⅢExperimental study of autologous BM-MNCs by intramyocardial transplantation into ischemic myocardium of AMI in swines Objective: To evaluate the effectiveness and safety of BM-MNCs by intramyocardial transplantation. Methods: 11 swines were divided into two groups: BM-MNCs group (n=6) and control group (n=5), total 2×10~8 cells.

实验动物分为心肌内移植MNCs组(n=6)及心肌内盐水对照组(n=5),心肌内移植细胞数2×10~8;移植后4周观察移植细胞归巢情况、小血管密度、心功能及冠脉侧枝血管形成情况及可能发生的副作用。

Finally,Tumorigenecity of KB-Neo,KB-PF4 and KBp17-70 cells wasassayed with xenograft tumor growth in isogenous nude mice by examiningthe growth speed of xenograft,survival span,and histochemistry ofxenograft nude mice.

最后,对重组的PF4和p17-70基因进行体内实验,建立裸鼠移植瘤模型,观察移植瘤的生长速度、移植瘤的体积、荷瘤鼠的生存时间、移植瘤瘤体组织中的血管密度。

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