细胞

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。

Furthermore, flow cytometry showed that the number of cells in G1 phase increased and the number of cells in S phase decreased, the number of cells in G2/M phases relatively increased. The changes of subcell structure could be seen, such as cavernous cells, cytoplasm agglutination, increasing apoptosis.

结果：0、10、20和40mgL^(-1)，姜黄素对Hela细胞增殖的抑制率分别为3.0%、21.4%、32.8%和49.2%，且呈剂量依赖性；流式细胞术显示，G1期细胞增多，S期细胞减少，G2/M期细胞相对增多；细胞结构显示，细胞空化、染色质凝集、凋亡小体增多。

Results Curcuma （0,10,20 and 40 mg·L-1）had obvious inhibitory effects on the Hela cell proliferation in a dose-dependant manner,the inhibitory rates were 3.0%,21.4%,32.8% and 49.2%,respectively. Furthermore, flow cytometry showed that the number of cells in G1 phase increased and the number of cells in S phase decreased, the number of cells in G2／M phases relatively increased. The changes of subcell structure could be seen, such as cavernous cells, cytoplasm agglutination, increasing apoptosis.

结果：0、10、20和40 mg·L-1姜黄素对Hela细胞增殖的抑制率分别为3.0%、21.4%、32.8%和49.2%，且呈剂量依赖性；流式细胞术显示，G1期细胞增多，S期细胞减少，G 2／M期细胞相对增多；细胞结构显示，细胞空化、染色质凝集、凋亡小体增多。

No tapetal cells were formed or tapetam collapsed at very early stage in some anther chamber during the microsporogenesis; Pollen mother cells in another chamber were not developed and microspore vacuolized; Tapetum cells developed abnormally, giantized and crammed the anther chamber; Some tapetum cells developed normally, but PMCs developed abnomally, which deformed at sporogeneous stage, and then were abhensive each other and finally degenerated. Nearly no PMCs could complete the process of meiosis, and form the tetrad, leading to the failure of normal microspore development.

药室的绒毡层细胞不形成或提前解体；药室内的花粉母细胞不发育或小孢子液泡化；绒毡层细胞发育异常，出现巨型化而挤满整个药室；绒毡层细胞虽发育正常，但花粉母细胞发育异常，在造孢细胞时期即开始出现变形现象，随后发生粘连退化；绝大部分花粉母细胞不能完成正常的减数分裂过程形成四分体，导致小孢子退化。

To test the hypothesis, neontal rat cadiomyocytes Treat with heat shock(HS,42℃,2h) to induce the expression of HSP70,then teated with 0.5mmol/L H2O2. We first found H2O2 can reduced cadiomyocytes apoptosis and HSP70 only over-expression in the HS group, other two group only express HSP70 a little in the test of expression of HSP70.Then we tested the concentration of Ca2+, the [Ca2+ ] of CON was 192.224+6.654, the [Ca2+ ] of H2O2 group was 290.6918+8.922and the [Ca2+]of HS group was 214.2633+4.484.To further determine how HSP70 repaired the Ca2+ homestasis, we measured calcium transient of each group.

实验中应用MTT和流式细胞技术检测细胞的凋亡，发现损伤组的存活率明显低于其他2组，热休克组(Heat shock group，HS group)细胞存活率略低于正常组细胞；免疫组化结果表明只有在HS组才检测到阳性反应，即是HSP70的表达，其他2组均为弱阳性结果；应用离子成像技术对细胞内钙离子浓度进行测定，发现损伤组290.6918+8.922明显高于正常组192.224+6.654和HS组214.2633+4.484，HS组细胞内的钙离子浓度略高于正常组；为了进一步探讨HSP70对于ROS引起的心肌凋亡过程中钙离子的调控机制，又对各组的心肌细胞进行了钙瞬变的测定。

The tibiae of the rats were kept for the samples of decalcification, IGF-1 in bone tissue was measured by immunohistochemistry and Enzyme linked-immuno-sorbent assay was used to detect the TGF- P 1, IL-6, IL-1 P and TNF- a in serum.

结果 成骨细胞呈线状排列于骨小梁的边缘，细胞因子IGF-1表达于成骨细胞胞浆，骨细胞、软骨细胞及骨基质中也存在IGF-1。不同部位的成骨细胞IGF-1的表达量并不一致，卵巢切除及PTH注射对不同部位成骨细胞IGF-1表达的影响也不一样。

The efficiency of selection was monitored by comparing the number of phage recovered from the acid elution and cell lysate in each round,and by testing EGFR binding specificity of polyclonal phagescFv on CHOEGFRGFP1 and CHOK1 cell with cell ELISA. Bacterial PCR was used to select clones containing a 1 kb insert. Cell ELISA was used to determine EGFR binding specificity of monoclonal pscFv on EGFR positive and negative cell. The number of individual EGFRbinding clones was determined with nucleotide sequencing. Results 500fold enrichments were observed by tittering phages in the cell lysate after five rounds of selection.

以稳定转染的CHOEGFRGFP1细胞和未转染的CHOK1细胞分别作为EGFR阳性和阴性细胞，采用负筛选的方法进行筛选；通过比较每轮投入及洗脱出噬菌体的效价比以及细胞ELISA检测多克隆pscFv与阳性、阴性细胞结合情况对筛选过程进行监测；采用菌落PCR挑选含有全长scFv片断的菌落，进一步用细胞ELISA检测单克隆pscFv与EGFR阳性及阴性细胞结合特异性；挑选EGFR特异性单克隆pscFv采用DNA测序法确定克隆多样性。

Cell ELISA was used to determine EGFR binding specificity of monoclonal pscFv on EGFR positive and negative cell. The number of individual EGFRbinding clones was determined with nucleotide sequencing. Results 500fold enrichments were observed by tittering phages in the cell lysate after five rounds of selection.

以稳定转染的CHOEGFRGFP1细胞和未转染的CHOK1细胞分别作为EGFR阳性和阴性细胞，采用负筛选的方法进行筛选；通过比较每轮投入及洗脱出噬菌体的效价比以及细胞ELISA检测多克隆pscFv与阳性、阴性细胞结合情况对筛选过程进行监测；采用菌落PCR挑选含有全长scFv片断的菌落，进一步用细胞ELISA检测单克隆pscFv与EGFR阳性及阴性细胞结合特异性；挑选EGFR特异性单克隆pscFv采用DNA测序法确定克隆多样性。

Kangdai Ⅰ has protective function to the damaged neurons and astrocytes: Main results:(1) It has direct protective function to the damaged neurons. It can increase the activity and survival rate, decrease the mortality and the transudation rate of LDH in cultured medium and the strong positive cell count of NOS expression of injured neurons.(2) It has also directly protective function to the damaged astrocytes. It can increase the activity and survival rate and protein content in conditioned medium.(3) It can strengthen the ability of BDNF, GDNF, bFGF, HSP and IL-6expression in damaged astrocytes.(4) It can also strengthen obviously the expressions of NSE, bFGF-receptor and bc1-2, lower the expression of bax and caspase-3.(5) It can indirectly protect and restore the damaged neurons by astrocytes. Because the effect of ACMK (ACM interfered by Kangdai Ⅰ) is stronger than ACM+K (ACM associated with Kangdai Ⅰ).

抗呆Ⅰ号对受损的神经元和星形胶质细胞均具有保护作用：主要表现为：(1)对受损神经元具有直接的保护作用，可提高受损神经元的活性和存活率，降低细胞培养液LDH的漏出率、细胞死亡率和NOS染色强阳性细胞的表达量；(2)对受损的星形胶质细胞也有直接的保护作用，可提高其活性、存活率以及培养液蛋白质的含量；(3)能增强受损星形胶质细胞分泌BDNF、GDNF、bFGF、HSP和IL-6的能力；(4)可明显增强受损神经元对NSE、bFGF的受体和bc1-2的表达，降低受损神经元对bax和caspase-3的表达；(5)抗呆Ⅰ号可通过星形胶质细胞间接地保护和修复受损的神经元，因为在多数实验组中经抗呆Ⅰ号作用的ACM的作用远大于ACM与抗呆Ⅰ号联合应用的作用，统计学上具有显著性差异。

Furthermore, preliminary work also performed to examine whether PI3K/AKT signal transduction pathway was activated in the process of refractory leukemia development. Materials and methods An immortalized human bone marrow stromal cell line, HS-5, was introduced to establish a bi-phase culture system for the cultivation of B-lineage precursor leukemia cells. ELISA and RT-PCR were used to investigate the expression of VEGF and its receptors in the leukemia cell lines and primary childhood leukemia cells in different treated groups. Flow cytometory method and immunofluorescent staining were employed to examine the apoptosis signals both in the VP16 treated and untreated leukemia cells. Western blot was utilized to explore the PI3K/AKT activated status in the drug induced or uninduced leukemia cells and lymphocytes from healthy donors.

材料和方法使用来源于人类骨髓基质细胞的细胞株HS-5作为滋养层细胞进行急性淋巴细胞性白血病细胞的体外培养，通过细胞生物学和免疫学方法评估培养体系并鉴定出难治性白血病细胞克隆；以ELISA和RT-PCR方法检测急性白血病细胞株和患儿白血病细胞VEGF及其受体的表达，了解不同治疗阶段VEGF及其受体的表达状况，并结合临床指标进行分析，明确VEGF及其受体在白血病发生过程中的作用；流式细胞仪和免疫荧光染色法对正常健康儿童、初发白血病患儿、复发白血病患儿及缓解后患儿进行凋亡因子检测和分析，初步阐明难治性白血病抗凋亡形成的原因；蛋白印记分析检测PI3K/AKT信号传导通路在健康儿童、初发白血病和复发白血病患儿的表达，初步了解难治性白血病形成的分子生物学机制。

With Death guitarist Schuldiner adopting vocal duties, the band made a major impact on the scene.

随着死亡的吉他手Schuldiner接受主唱的职务，乐队在现实中树立了重要的影响。

But he could still end up breakfasting on Swiss-government issue muesli because all six are accused of nicking around 45 million pounds they should have paid to FIFA.

不过他最后仍有可能沦为瑞士政府&议事餐桌&上的一道早餐，因为这所有六个人都被指控把本应支付给国际足联的大约4500万英镑骗了个精光。