体素培养

Finally the high expression and low degradation of hirudin were achieved.

在分离纯化并鉴定出水蛭素的四种活性异构体后，通过恒化培养的研究探索影响水蛭素降解的因素，最终在分批发酵中实现水蛭素的低降解高表达。

With the optimization of the cultivated conditions, the recombinant fusion protein staphylokinase-hirudin was over-expressed in E.coli BL21(DE3), and the final cell density reach 115g wet cell weight per liter, the target protein is about 1.1~1.2g/L accounting for more than 30% of the total protein. The culture process in 5L fermentor was performed on 40L fermentor successfully, and indicated that the process was easy scalable.

通过对培养条件的不断优化，重组葡激酶-水蛭素融合蛋白在大肠杆菌BL21(DE3)里得到了高效表达，菌体密度最终达到115g/L以上，可溶性重组融合蛋白占菌体总蛋白的30%以上，含量约为1.1~1.2g/L.5L发酵罐的发酵工艺参数在40L发酵罐中进行了放大培养，结果表明该工艺能有效的放大，可适用于工业生产。

As antioxidants, the vitamin E or quercetin (10μg/mL) significantly attenuated the toxic effects of PCBs on the cultured hepatocytes. The above results indicated that vitamin E or quercetin might antagonize the toxic effect of PCBs via antioxidizing effects on in vitro cultured chicken embryo hepatocytes.

结果表明离体培养的鸡胚肝细胞可作为肝损伤的细胞模型，PCB对培养的肝细胞的损伤及抗氧化剂维生素E和槲皮素对它的缓解作用提示， PCB对肝细胞的损伤可能是通过诱导细胞膜发生脂质过氧化而导致细胞的死亡。

Some patients have ''monomicrobial bacterascites'' in which cultures are positive but there is normal ascitic neutrophil count.138 Such infections are thought to occur relatively commonly, and the majority are eradicated by the body's natural defence mechanisms (for example, opsonic and complement mediated bactericidal activity).138 139 When a positive culture is obtained, a further ascitic tap with a neutrophil count should be obtained.

有些患者有着&单一微生物细菌性腹水&，其腹水培养为阳性但腹水白细胞计数正常。138这些感染被认为较为常见，大部分可被机体天然的防御机制（如调理素和补体介导的杀菌活性）所根除。138，139当腹水培养为阳性时，应进一步进行腹穿并行嗜中性粒细胞计数。

The results indicated that the highest tetraploid induction rate could reach 23.4% when the leaves precultured on the optimal organogenetic MS medium for 12d and treated with 10 mg/L colchione on the double layer media for 24h in vitro.

结果表明，预培养12d的兰考泡桐叶片在秋水仙素浓度为10mg/L的双层培养基上预培养处理24h时，四倍体诱导率最高达到23.4%。

We have, therefore, investigated the direct effects of adiponectin on primary cultures of osteoblastic and osteoclastic cells in vitro and determined its integrated effects in vivo by characterizing the bone phenotype of adiponectin-deficient mice.

为此，我们研究了脂联素对离体培养的原代成骨细胞和破骨细胞的直接影响，并通过对脂联素缺陷小鼠的骨基因表型的研究明确其在体内的综合作用。

Methods: Seven male patients with kidney stones were admitted. Abdominal subcutaneous adipose tissue and perirenal adipose tissue were collected from the operating-patients, and were cultivated with different concentrations of curcumin (10 and 100 μg/ml) in vitro.

用体外组织培养法培养手术后离体人脂肪组织，分别予10和100μg/ml姜黄素干预6h和24h，用夹心酶联免疫吸附测定法检测各脂肪组织培养液中APN和IL-6的含量。

Results: The hairy root was successfully induced and the hairy root culture system was set up; The opines assay was positive for mannopine and agropine; The result of determination showed that there were a lot of anthroquinone constituents in the rhubarb hairy root and the contents of rhein, aloe-emodin, emodin or 8-O-methylchrysophanol were respectively higher than those in wild plant root.

结果：诱导出了天山大黄发根，并建立了发根离体培养系；从发根中检测到甘露碱、农杆碱的存在；定量分析发现，天山大黄发根中含有多种蒽醌类成分，其中大黄酸、芦荟大黄素、大黄素、大黄酚-8-甲醚的含量分别高于原植物根中的含量。

The PCR products were examined by agarose gel electrophoresis. The target gene fragments were purified by gel extraction kit and ligated to cloning vector pMD18-T. The recombinant vectors were transformed into host strain E. coli K802 by lithium chloride method, screened and identified with PCR and restrictive enzymatic digestion. Their sequences were confirmed by DNA sequencing.(2) sTWEAK1 gene was subcloned into expression vector pProEx HTb and transformed into E. coli BL21. sTWEAK2 gene was subcloned into expression vector pMAL-C2x and transformed into E. coli TB1. The recombinant vectors were screened and identified with PCR and restrictive enzymatic digestion. The recombinant fusion proteins were induced to express with IPTG, detected by coomassie brilliant blue-stained SDS-polyacrylamide gel electrophoresis , and confirmed by Western blot analysis.(3) The sTWEAK1 fusion protein was purified with Ni-NTA Spin Kit.(4) The biological activity was assayed on transformed and tumor cells by microplate photometer after crystal violet or sulfur rodamine B staining.(5) The contents of IL-8 in the supernatant of 1990 cell cultures were determined by ELISA.(6) The morphological changes of the sensitive cells were observed by light and transmission electron microscopies.(7) The cell cycle and apoptotic rate were assayed by flow cytometry in 1990 and M85 cells.(8) The effect of fusion proteins on induction of NF-κB in 1990 and LOVO cells was detected with Dual-Luciferase Reporter Assay system.(9) The TWEAK gene was subcloned into Adeno-X Viral DNA with pShuttle vector and transfected into HEK293 cells by lipofectamine method.

(1)本研究用RT-PCR方法，从人组织细胞总RNA中扩增可溶性TWEAK胞外区（sTWEAK1和sTWEAK2）的cDNA序列及全长编码序列，用琼脂糖凝胶电泳分析PCR产物，胶回收目的基因片段，连接到pMD18-T克隆载体中，转化大肠杆菌K802，PCR和酶切筛选阳性克隆，全自动DNA测序验证序列；(2)sTWEAK1和sTWEAK2分别亚克隆到pProEx HTb和pMAL-C2x表达载体中，分别转化大肠杆菌BL21和TB1，PCR筛选和酶切鉴定，阳性克隆用IPTG诱导表达，表达产物用SDS-PAGE分析和Western blot验证融合蛋白；(3)用NTA-Ni Spin试剂盒初步分离纯化sTWEAK1融合蛋白；(4)用体外培养的肿瘤细胞和正常对表达产物进行活性检测，贴壁细胞用结晶紫染色法，悬浮细胞用磺酰罗丹明B染色法，酶标仪检测OD值；(5)敏感细胞用ELISA法检测细胞培养上清中IL-8的含量；(6)用光镜和电镜观察敏感细胞死亡和细胞凋亡情况；(7)用流式细胞仪分析表达产物对敏感细胞凋亡率和细胞周期的影响；(8)用双荧光素酶报告基因检测法，测定表达产物对敏感细胞NF-κB的影响；(9)用pShuttle穿梭质粒将TWEAK重组到腺病毒载体上，用脂质体转染法转染HEK293细胞，PCR鉴定重组质粒。

Lamblia were cultivated axenically with modified TYI-S-33 medium contained dihydroartemisinin (0.002 mg/L) for 12 h and then stained by rhodanmine phalloidin and paclitaxel (P22310). The cytoskeletons of the organisms were detected and analyzed by flow cytometry, and then observed by confocal microscopy.RESULTS: The cytoskeleton of trophozoites was markedly damaged after treatment with 0.002 mg/L dihydroartemisinin for 12 h.

将用含双氢青蒿素的改良TYI-S-33培养基培养后的蓝氏贾第鞭毛虫滋养体，用专一性结合f-肌动蛋白的荧光染料罗丹明－鬼笔环肽(rhodanmine phalloidin， RDP)标记微丝，结合微管的紫杉醇(Paclitaxel， Oregen Green 488， P22310)标记微管，用流式细胞术检测、分析，用激光共聚焦显微镜(confocal microscopy， CFM)观察滋养体微丝和微管的变化。

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。