后成质的

GFP gene eukaryotic express plasmids were transfected into ES cells. The cells were observed under the inverted fluorescent microscope. The clones which expressed the powerful green fluorescence were chosen to be named ES-GFP cells and cultured continuously. The examination of ES cell totipotent including Alkine phosphatase staining, embryonic body formation in vitro and teratomas formation in nude mice was implemented.

1。采用脂质体方法将GFP质粒转染ES-D3细胞株，经筛选后倒置荧光显微镜观察细胞形态及荧光状态，挑取荧光最强的克隆，命名为ES-GFP细胞，进一步扩增培养，观察细胞生长及荧光表达情况，行细胞分化全能性鉴定包括碱性磷酸酶（alkine Phosphtase，AKP）染色、体外胚胎体（embryonic body，EB）形成实验及裸鼠体内成瘤实验。2。

OPN Immuno-electron microscopy show the gold markers in thecraniopharyngioma cell of the ameloblast type be located on the endoplasmicreticulum that the swollen oncocyte afterbirth strach, from endoplasmic reticulumcreation behind secrete the outside of the afterbirth; But didnt see the gold markers inthe craniopharyngioma cell of the squamous type.

OPN免疫电镜显示成釉细胞型颅咽管瘤中金标OPN颗粒的产生位于肿瘤细胞胞浆的内质网上，由内质网产生后分泌到胞外；而在鳞状细胞型颅咽管瘤细胞中未见金标OPN颗粒，少量可见颗粒仅见于肿瘤间质中的血管内皮细胞上。

Use constituent piece to stick mural way depart to foster Niu Er to become fiber cell above all, after classics generation is rarefied, fat simple way will be contained double the skin of the 4th acting ox that outside conveying carrier to turn into body, the particularity of person bacteriolysis enzymatic mammary gland that chooses number fosters becomes fiber cell, g/mLG418 chooses 500 μ 2 weeks, g418 halve continues to develop 3 generation of 2 ～, catch the cell after choosing to undertake PCR detects to turning next reach nucleus analysis.

首先采用组织块贴壁法分离培养牛耳成纤维细胞，经传代纯化后，脂质法将含有双重筛选标记的人溶菌酶乳腺特异性表达载体转入体外培养的第4代牛皮肤成纤维细胞，500μg/mLG418筛选2周，G418减半继续培养2～3代，然后对转染筛选后的细胞进行PCR检测及核型分析。

Results Compared to those before treatment the volume of the implanted keloid of Group D began to decrease since 14 days after treatment time-dependently (all P 《 0.05), and the volumes of the other 3 groups continued to increase and peaked on days 21, 14, or 7 respectively (all P 《 0.05).Microscopy showed infiltration of a larger quantity of histiocyte in the keloid tissue, and more obvious collagen disorganization and apoptosis of fibroblasts in Group D than in the other 3 groups.The protein expression of Bcl-2 was more remarkable and the protein expression of BAX was less remarkable in Group D than iu the other 3 groups.

结果 用药前和用药后7、14、21、28、35 d，D组瘢痕疙瘩组织块体积(mm3)分别为173±5、172±5、147±5、125±6、112±7和84±9，从用药后14 d开始明显缩小(均P＜0.05)；而其他3组瘢痕疙瘩组织块体积均明显增大，从用约后7 d开始各时点测得的体积均明显大于D组(均P＜0.05)。D组瘢痕疙瘩组织中有大量小鼠组织细胞浸润，胶原结构破坏和成纤维细胞凋亡明显重于其他3组，Bcl-2蛋门质表达明显弱于而Bax蛋白质表达明显强于其他3组。

Results (1)ATP could inhibit the proliferation of U937 cells with inhibition rate over 43% after 48hour ATP treatment;(2)ATP treated U937 cells numbers increased obviously in G1 phase,and apoptosis peak appeared before G1 phase,apoptotic cell number was 3.4% for 24hour,and was 22.7% for 48hour of ATP treated U937 cells;(3) Nuclear chromatin condensed into sperical masses bound cytoplasma membrane formed apoptotic bodies which is shed from membrane surface into intercellular medium;(4) Apoptotic bodies were nigrosine staining negtive.

结果 (1)ATP对U937细胞的增殖有按摩明显的阻抑作用，加药48h后增殖的抑制率可达43%以上；(2)ATP处理的U937细胞周期发生改变，G1期细胞数按摩明显增多，ATP作用24h时G1期前出现亚二倍体峰——凋亡峰，凋亡细胞数为3.4%，作用48h时凋亡细胞数增多为22.7%；(3)ATP处理的U937细胞首先在核内染色质浓缩成半月形贴近核膜，逐渐向核膜外移动，进入胞浆内再移向质膜内外面，紧贴质膜外面再逐步脱离细胞体，进入细胞基质中，成为游离的凋亡小体。

Methods Cell culture,flow cytometry,HE staining,Nigrosine staining and electron microscopy were used. Results (1)ATP could inhibit the proliferation of U937 cells with inhibition rate over 43% after 48hour ATP treatment;(2)ATP treated U937 cells numbers increased obviously in G1 phase,and apoptosis peak appeared before G1 phase,apoptotic cell number was 3.4% for 24hour,and was 22.7% for 48hour of ATP treated U937 cells;(3) Nuclear chromatin condensed into sperical masses bound cytoplasma membrane formed apoptotic bodies which is shed from membrane surface into intercellular medium;(4) Apoptotic bodies were nigrosine staining negtive.

结果 (1)ATP对U937细胞的增殖有明显的阻抑作用，加药48h后增殖的抑制率可达43%以上；(2)ATP处理的U937细胞周期发生改变，G1期细胞数明显增多，ATP作用24h时G1期前出现亚二倍体峰——凋亡峰，凋亡细胞数为3.4%，作用48h时凋亡细胞数增多为22.7%；(3)ATP处理的U937细胞首先在核内染色质浓缩成半月形贴近核膜，逐渐向核膜外移动，进入胞浆内再移向质膜内外面，紧贴质膜外面再逐步脱离细胞体，进入细胞基质中，成为游离的凋亡小体。

Histological structures of bursas of Fabricius were observed at the different developmental stages from chicken embryoes to chickens. The results were as follows. The bursa of Fabricius began to develop on the 13th day of post-hatching and its cell aggregated into the nodosity clique underneath the epithelium mucosae. On the 15th day of post-hatching, those cell cliques were surrounded by the flat epithelium and formed the basic structure of folliculus lymphaticus. The number of lymphocytes in the folliculus lymphaticus increased gradually, and the medulla and cortex of a majority of folliculus were not discriminated obviously. Until on the 7th day for chickens, the medulla and cortex of folliculus formed completely and the number of folliculus increased significantly. From 21 to 35 days old, the folliculus lymphaticus transformed from polygon into rectangle. Mucosa epithelial cells increased gradually and their cytoblasts arranged compactly into aline in the basilar part on the 35th day.

本研究观察了从鸡胚到出壳后的不同生长发育阶段法氏囊的组织结构，结果表明：胚胎13日龄时，法氏囊开始发育，粘膜上皮下形成结节状细胞集团；胚胎15日龄，聚集的细胞集团形成初始的淋巴滤泡；胚胎18日龄到出壳后1日龄，滤泡内淋巴细胞数量增多，多数滤泡髓质和皮质界限仍不明显；7日龄雏鸡法氏囊髓质皮质分界清晰，滤泡数量显著增多；21日龄到35日龄，滤泡由多边形逐渐转变成长方形；在35日龄时，粘膜上皮细胞逐渐增多，排列紧密，细胞核紧密排列成直线位于基底部。

RESULTS: After differentiation of human adherent myoblasts by neural induction medium, no cells with a neural cell morphology (ie., small, refractile cell body with dendritic cell extensions) were seen. All remaining myoblasts cultured with neural induction medium, myoblasts with proliferation medium and myotubes with differentiation medium containing 20 mL/L HS were positive for β Tubulin Ⅲ. C2C12 myoblasts were negative for β Tubulin Ⅲ. In contrast, all the above cells were negative for the markers Neurofilament Mr 68×103 and GFAP.

结果：用诱导分化液作用后，未见小的、伴有突起的放射状形态的神经细胞；抗β Tubulin Ⅲ对经神经元胶质细胞诱导分化液作用后的人成肌细胞、增殖培养液培养后的各代人成肌细胞及仅含20 mL/L HS分化液分化形成的肌管细胞染色均为阳性； C2C12细胞β Tubulin Ⅲ抗体染色阴性；上述所有细胞抗Neurofilament Mr 68×103和抗GFAP染色均为阴性。

Objective: To verificate culture、amplification、induction anddifferentiation of myoblast and Mesenchymal stem cells of themouse in vitro; to transplant the myoblast and MSCs to mdx mice, themodel animal of DMD, by vena caudalis; To observe the growth andfusion of myoblast and MSCs in the host after transplantation.

目的：研究成肌细胞及骨髓间充质干细胞体外培养、扩增、诱导分化；利用成肌细胞及骨髓间充质干细胞经尾静脉注射移植治疗DMD模型鼠mdx小鼠；观察移植后的细胞在宿主体内的生长及融合情况；探讨基因治疗DMD的方法及可行性。

Following successful modeling, rats of bFGF group were intratracheally injected with 400 U bFGF and rats of VEGF group with 2 μg VEGF, once a week for three times. MSCs group was injected 1 mL suspension of 4×109/L MSCs into tail vein. MSCs+VEGF group was injected MSCs into tail vein and intratracheally injected VEGF (2 ug, three times) at the same time. Model control and normal control groups were intratracheally injected with equal volume of sodium chloride.

成功造模后，碱性成纤维细胞生长因子组气管内注入400 U碱性成纤维细胞生长因子，血管内皮生长因子组气管内注入2 μg血管内皮生长因子，1次/周，共3次；单纯细胞移植组于尾静脉注入4×109 L-1骨髓间充质干细胞悬液1 mL；血管内皮生长因子+细胞移植组气管内注入血管内皮生长因子的同时，尾静脉注入骨髓间充质干细胞；模型对照组、正常对照组给予相同体积的生理盐水。

The split between the two groups can hardly be papered over.

这两个团体间的分歧难以掩饰。

This approach not only encourages a greater number of responses, but minimizes the likelihood of stale groupthink.

这种做法不仅鼓励了更多的反应，而且减少跟风的可能性。