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Primary culture of goat mammary gland cells could be derived by outgrowth of migrating cells from fragment of tissue. Fibroblast and epithelial cells could be pured according to their different sensibility to trypsin. Complex cultures treated by 0.25% trypsin in Hanks' for 5~7min at 37℃, the dispersed cells were mainly fibroblasts,then the rest treated by 0.15% trypsin-0.02% EDTA in Hanks'for 5~8min at 37℃, the majority cells harvested were mammary epithelial cells.

用组织块培养法可获得良好的山羊乳腺细胞原代培养物；培养的山羊乳腺成纤维细胞比上皮细胞对胰蛋白酶更敏感，据此可在传代过程中将二者分离纯化，获得纯细胞系；混合培养物先用0.25%胰蛋白酶在37℃消化5~7min 所收集的细胞主要为成纤维细胞；再加0.15%胰蛋白酶-0.02鞹A 消化液在37℃继续消化5~6min 所回收的细胞绝大多数为上皮细胞，经过2~3 代，即可得到纯化的乳腺上皮细胞系。

Primary culture of goat mammary gland cells could be derived by outgrowth of migrating cells from fragment of tissue. Fibroblast and epithelial cells could be pured according to their different sensibility to trypsin. Complex cultures treated by 0.25% trypsin in Hanks' for 5~7min at 37℃, the dispersed cells were mainly fibroblasts,then the rest treated by 0.15% trypsin-0.02% EDTA in Hanks'for 5~8min at 37℃, the majority cells harvested were mammary epithelial cells.

用组织块培养法可获得良好的山羊乳腺细胞原代培养物；培养的山羊乳腺成纤维细胞比上皮细胞对胰蛋白酶更敏感，据此可在传代过程中将二者分离纯化，获得纯细胞系；混合培养物先用0.25%胰蛋白酶在37℃消化5~7min 所收集的细胞主要为成纤维细胞；再加0.15%胰蛋白酶-0.02%EDTA 消化液在37℃继续消化5~6min 所回收的细胞绝大多数为上皮细胞，经过2~3 代，即可得到纯化的乳腺上皮细胞系。

After being induced to differentiate in vitro, cells with various morphologies showed β-gal activity, including nerve cells, neuroglial cells, epithelial cells. We also detected β-gal activity in a wide variety of tissue elements in solid tumors made by injecting the MC15 and MA34 cells into syngenic mice. Then we produced chimeric embryos by injecting the MC15 cells into Kunming blastocysts and recovering the embryos at various developmental stages.

SiCMVIE(Simian Cytomegalovirus Immediately Early)启动子是一个强启动子，本文首次将SCMVIE启动子应用到ES细胞途径上制作嵌合鼠，并通过lacZ基因在ES细胞和早期嵌合胚胎中的表达，表明该启动子是一个能在小鼠多数组织表达的启动子，这与1993年Peppel等人用原核显微注射法获得的转基因小鼠上所得的有关结果是基本上是一致的。

D. The function of stimulating xenogenous lymphocyte proliferation was the same between peripheral DCs and ascites PCs. E. The percentage of CD3〓CD56〓 cells was the same in CIK cells co-culture with DCs transfected with SKOV3 RNA, CIK cells co-culture with DCs, and CIK cells. F. The expansion rate of CIK cells can be accelerated by co-culturing with loaded or unloaded DCs. However, the expansion rate between loaded or unloaded group is the same. F. The strongest cytotoxicity against SKOV3 cell line was achieved by CIK cells co-cultured with DCs loaded with SKOV3 lysate.

结果：1、腹水可获得0.83±0.24×10〓个AMC/ml，单核细胞有0.74±0.25×10〓个/ml；2、卵巢癌患者外周血可获得0.87±0.20×10〓个AMC/ml，单核细胞有0.92±0.17×10〓个/ml；3、除CD86外周血单核细胞来源DC表达较高以外，其他表面分子在不同来源DC间没有统计学差异；4、不同来源DC的异基因刺激能力没有差异；5、与负载或未负载卵巢癌抗原的DC共培养并不能提高CIK细胞群中CD3〓CD56〓细胞的数量；6、CIK细胞增殖显著，培养14天时可扩增19.18±4.70倍，培养21天时可扩增35.82±4.36倍；7、与未负载或负载DC共培养的CIK细胞在培养第14天后增殖速率大于单纯CIK细胞。

RESULTS: NK cells represent a unique subset of lymphocytes that have no restriction by MHC antigens and have the ability to lyses certain tumor cells without the requirement for prior immune sensitization of the host. In addition, NK cells can also produce cytokines and regulate the function of other immune cells. So, it is emerging to apply NK cells as therapeutic agents against a broad range of malignancies. In this respect, the anti-tumor activity of NK cells is the key factor of NK-cell-based immunotherapy.

结果：NK细胞是一种独特的淋巴细胞，对靶细胞的识别无MHC限制性，无需预先致敏即可直接杀伤肿瘤细胞，也可分泌细胞因子调节其他免疫细胞的功能，是机体天然免疫的主要承担者，也是获得性免疫的核心调节细胞，故NK细胞在抗肿瘤免疫中的地位越来越受到重视，随之增强NK细胞肿瘤杀伤活性的研究也逐步深入。

Some of transplanted cells expressed titin and the assembled titin could be detected in the cells adjacent to host myocardium. Cx43 could be observed between these cells. But the size and structure of transplanted cells were different from normal contractile myocardium.3. Most of the grafted cells were isolated by fibrous tissue. Part of these cells differentiated into fibrocytes. The structure of host myocardium was slightly disturbed.

MSCs植入正常心肌组织后能存活至12周，部分移植细胞表达titin，且与宿主心肌细胞相邻的细胞表达titin组装程度更高，随时间延长表达逐渐明显，其后进入平台期，但细胞体积及胞浆发育均不及正常心肌细胞；在移植细胞与宿主细胞之间可检测到Cx43的表达。

Total RNAs from KAx-3 cells and AK127 cells(developed for 14h) were isolated. After the reverse transcription and PCR reaction, two distinct differential fragments were acquired., fragment A was from KAx-3 cells and fragment C was from AK127 cells. After retriving and reamplifying the differentially expressed fragments, white-blue plaqueselection, the fragments were purified. Northern blot proved that fragment A was from KAx-3 cells and fragment C was from AK127 cells. The results of sequencing and researching for NCBI database have been showed: part sequence of fragment A shows 91% similarity to the gene encoding DhkA, 92% similarity to the gene encoding DhkF, 91% similarity to the gene encoding STATc, 97% similarity to the homoeobox gene encoding protein. These genes play important part in controlling cell differentiation and cell proportion in Dictyostelium discoideum.

本研究通过提取盘基网柄菌发育14小时的野生型KAx-3细胞和突变型AK127细胞的总RNA，运用mRNA差异显示法分离出了两条明显的差异表达片段，其中片段A来自野生型KAX-3细胞，片段C来自突变型Ak127细胞；并通过凝胶回收差异片段、对差异片段进行再次PCR、蓝白斑筛选克隆、提取质粒、酶切电泳纯化差异片段；接着进行Northern杂交的结果表明，片段A只与野生型KAx-3细胞的总RNA有杂交信号，片段C只与突变型AK127细胞的总RNA有杂交信号，这就排除了差异片段假阳性的可能；最后通过测序，搜索NCBI BLAST数据库发现：片段A的小部分序列与编码组氨酸激酶DhkA基因中一段序列的相似性高达91%，与编码组氨酸激酶DhkF基因中的一段序列相似性高达92%，与编码STATc蛋白基因的一段序列相似性达91%，以及与编码同源框蛋白的基因中的一段序列相似性达97%，这些基因在盘基网柄菌细胞分化和细胞比例调控过程中起着相当重要的作用，这些数据进一步说明了突变细胞不能完成发育的原因。

Histologically, the seminiferous tubules contained numerous Sertoli cells and more Sertoli-spermatozoa complexes, accompanied by the depletion of Leydig cells with deeply stained nuclei. Mature spermatozoa were stored up in the epididymis, but only a few in the efferent ducts. In the second place was testicular atrophy（32/120; 26.7%）. The seminiferous tubules showed moderate to severe inactivity of spermiogenesis with evidence of only spermatogonia, spermatocytes and Sertoli cells. The Leydig cells were obviously decreased in numbers associated with decrease of lipid droplets in their cytoplasms. Testicular hypoplasia was the third disorders（22/120; 18.3%）. Only a few spermatogonia and Sertoli cells appeared without any spermiogenesis. The associated changes was decreased in Leydig cells and fibrous hyperplasia in the interstitium.Epididymal stones were sometimes found（12/120; 10%）. Grossly, yellowish-white nodules with various sizes and firm in consistency were observed in the epididymis and the front efferent ducts. Microscopically, the epididymal ducts were dilated with voluminous spermatozoa storage, even showed calcification in severe cases. The deposited calcium salts were stained positively by Von Kossa and Alizarin red methods.Amyloidosis was also detected in 10 roosters（8.3%）. Eosinophilic, homogeneous, amyloid-like substances were deposited mainly in the testicular interstitium and the periphery of blood vessels. These substances showed positive reaction by Congo red staining. Five roosters（4.2%）had Marek's lesions in the testis, epididymis and peripheral nerves with infiltration of pleomorphic lymphocytes. Only one case showed epithelial necrosis of seminiferous tubules accompanied by fibrous proliferation in the interstitium.

结果发现，在总共搜集的120个病例中，其中因年老所导致的产精力不佳为最多，占38例（31.7%），於镜下可见大量精虫黏附於Sertoli cell的表面，并可见Sertoli cell数量明显增多而Leydig cell明显减少，且其细胞核呈现浓染的现象，而在其副睪中仍可见到成熟精虫蓄留於管腔中，但在其输精管内却只有少量精虫存在；其次为睪丸萎缩，占32例（26.7%），镜下可见中度至重度无造精作用，其生精小管中只见到精母细胞、精原细胞及Sertoli cell存在，但Leydig cell数量明显减少且其细胞质内的脂质也明显减少；睪丸发育不全，占22例（18.3%），於生精小管内只见到精母细胞及少量Sertoli cell存在，不见造精细胞分化，於生精小管间质可见Leydig cell减少并伴随结缔组织增生；副睪结石，占12例（10%），肉眼下可在副睪及输精管前段见到黄白色大小不一的结节，触感坚硬，於镜下可见副睪管扩张并有大量成熟精虫蓄积，严重时可见钙化现象，以Von Kossa及茜素红染色均呈阳性反应；类淀粉沉著症，占10例（8.3%），镜下在睪丸间质及血管周围可见粉红均质样的物质沉积，以刚果红染色成阳性反应；马立克病，占5例（4.2%），镜下可在睪丸、副睪实质及周边神经内均可见到嗜碱性大小不一的淋巴样细胞浸润；睪丸坏死，占1例（0.8%），镜下可见生精小管上皮细胞坏死脱落及间质结缔组织增生。

CTRL+Displays the Delete dialog box to delete the selected cells.

显示删除删除对话框来删除选定的单元格。

Our results demonstrate: The structures of the organs are normal, and the shapes of cells are clearly visible. There are lots of positive brown granules in Chief cells and Parietal cells in abomasum as well as the mucosa epithelial cells and gland cells of duodenum. Three bands with a molecular mass close to 120KDa、110KD and 98KDa were identified by Western Blot. The Ob-R levels of 120KDa in abomasum were significantly higher than that of in small intestine. The levels of 110KDa were similar in the two organs. The expression of 98KDa Ob-R was weak.

HE染色结果显示，各组织结构正常，细胞形态清晰可见；免疫组织化学SABC染色显示，在皱胃胃体部固有层胃底腺的主细胞和壁细胞及十二指肠黏膜上皮细胞和固有层肠腺的柱状细胞中均可见大小数量不等的棕黄色颗粒；western Blot 实验发现，在胃和小肠均检测到120KDa、110KDa和98KDa三条带。120KDa长型瘦素受体蛋白在胃中表达量显著高于小肠中的表达；110KDa的短型瘦素受体蛋白，在小肠和皱胃中表达量接近。98KDa短型受体蛋白在胃和小肠表达均较弱。

Some of them were ligated to the plasmid vector pCR 2.1 andsequenced,respectively.

将所获DNA片段克隆到pCR2.1载体进行测序，并与基因组数据库进行同源性比较。

Thinking since Google owns Blogger, I would be able to escalate this issue to the support team.

自谷歌的思考拥有Blogger的，我将能够缓解这一问题的支持团队。