内转

On the other hand, immunoGrB was found inactive in transient and stable transfection research until when translocating from endosome in HER-2 positive tumor cells, it turned into an active form.

另一方面，从瞬时转染和稳定转染角度研究证实，immunoGrB分子不具有杀伤活性，它只有当特异性内化入HER-2阳性肿瘤细胞后才转变为活性形式。

MSCs were cultured in a conditional medium in subculture including: dexamethasone, vitamin C, and β-glycerophosphate. At the same time, we constructed the eukaryotic expression vector containing VEGF165gene. VEGF165 gene was transfected into MSCs by means of LipofectAMINE?2000. The stably gene expressive cells were screened with G-418 for 14 days.

在脂质体介导下将构建成功的真核表达载体pcDNA3.1- VEGF165转染到诱导培养的MSCs，G-418筛选阳性细胞克隆，将阳性克隆细胞和未转染的MSCs共同接种于β-磷酸三钙上，体外培养7天后，植入原大鼠肌肉内。

The PSCA_3 fragment was selected for its superior expression level in eukaryotic cells.Then the sig-PSCA_3-Fc-GPI genetic fragment was cloned into pVAX1-neo-IRES-GM/B7 vector to construct the final immunological inhanced DNA vaccine pVAX1-PSCA_3-FcGB. Immunofluorescence and flow cytometry were used to confirm the expression of PSCA_3 fragment by transfected into Cos7 cell.Finally,the anti-tumor effect of pVAX1-PSCA_3-FcGB was tested in murine prostate cancer model generated by RM-1 cell line.The animal was immunized with pVAX1-PSCA_3-FcGB DNA vaccine by intramuscular injection plus electroporation,pVAX1 and pVAX1-PSCA_1-FcGB plasmid were used as control.The inhibitory effect of tumor was investigated by observion of forming time,volume and inhibition ratio of tumor.Results:DNA sequencing conformed that the heterological PSCA fusion antigen fragment which was synchronized by overlapping-extending-PCR,was consistent to design.Enzyme digestion analysis showed that the 1 to 4 copies heterological PSCA fusion antigen fragments were constructed successfully.

方法(1)检索GenBank，选择包含人主要T细胞抗原表位序列的人PSCA基因片段，应用异种化抗原设计技术，保留人T细胞抗原表位，设计异种化PSCA融合抗原片段；(2)根据核酸序列按中心模板法设计引物，应用重叠延伸PCR技术拼接合成异种化PSCA融合抗原片段基因，以PCR、限制性酶切和DNA序列测定法进行鉴定：(3)利用DNA限制性内切酶BssHⅡ和MluⅠ酶切后粘端互补的特点，采用同尾酶法构建1—4拷贝异种化PSCA融合抗原片段(PNCA_1-PSCA_4)，并将上述片段分别插入真核表达载体pCI-neo-Fc-GPI中，转染293T细胞，借助免疫荧光+流式细胞术考察插入片段表达效率，最终选定PSCA_3片段进行下一步研究；(4)将sig-PSCA_3-Fc-GPI基因片段自pCI-PSCA_3-Fc-GPI质粒上切下，插入pVAX1-neo-IRES—GM/B7载体中，构建免疫增效DNA疫苗pVAX1-PSCA_3-FcGB，并应用转染Cos7细胞+免疫荧光/流式细胞术方法鉴定其在真核细胞中的表达情况；(5)给8周龄雄性C57BL/6小鼠皮下种植RM-1细胞，制备小鼠前列腺癌模型，并采用股四头肌肌肉注射+电脉冲法(Electroporation，EP)接种DNA疫苗质粒pVAX1-PSCA_3-FcGB，同时接种pVAX1空载体质粒和pVAX1-PSCA_1-FcGB质粒作为对照，通过观察计算免疫动物的成瘤时间、肿瘤体积和抑瘤率，来评价该DNA疫苗在小鼠体内的抑瘤效果。

CaN Aβ gene silencing can reduce myocardial hypertrophy in cultured cells, si1280 (21bp) of CaN Aβ gene is the most effective target site for siRNA. The method of intrapericardial injection of plasmid, microbubbles and erzymes can improve transfection efficiency of non-viral plasmid with satisfying targeted transfection. But the scope of transfected myocytes is still limited. CaN Aβ shRNA expressing plasmid transfection in vivo by pericardial injection results in decreased CaN Aβ protein expression of small part of myocytes, and CaN Aβ mRNA only shows decreased trend. The dosage of non-viral vector and the parameters of ultrasound energy should be optimized in further study.

结论RNAi技术抑制CaN Aβ基因表达可有效减轻Ald诱导的心肌细胞肥大程度；CaN Aβ基因中si1280(21bp)片段为实现RNAi的更有效片段；微泡+酶类心包腔内注射超声导入的方法可有效改善非病毒载体在体心肌的转染效率，同时具有一定的靶向性，但总的转染范围仍然有限；采用这一方法进一步进行&一肾一夹&心肌肥大模型大鼠在体心肌细胞的CaNAβ的RNAi干预，发现心肌肥大大鼠心外膜下局部心肌细胞CaN Aβ蛋白水平降低，CaN AβmRNA水平虽有下降趋势，但无统计学差异，提示质粒的用量及超声导入的参数有待进一步研究使其优化。

Immunofluorescence staining shows that, the CaN Aβ protein expression of intrapericardial injection group decreased in some myocytes under epicardium, while other groups show no decreased fluorescence.

三在体心肌CaN AβsiRNA表达质粒转染后，转染各组与心肌肥大模型组CaN AβmRNA水平比较无差异(P＞0.05)，但免疫荧光结果显示心包腔内途径CaN Aβ干扰组较心肌肥大模型组心外膜下部分心肌荧光减弱。

Although nuclear cascades in ethylene signal transduction pathway from EIN3 to ERF1 to PR proteins has been established in Arabidopsis thaliana, a similar route from EIN3 to EREBPs to fruit ripening and senescence has not been elucidated in Lycopersicon esculentum .

乙烯对植物的生长发育起重要的调控作用，随着乙烯信号转导研究的不断深入，许多乙烯信号转导组分已被分离和鉴定，并在模式植物拟南芥中已建立乙烯信号传递过程中的核内级联反应：EIN3→ERF1→病原相关蛋白，但在番茄中还没有建立EIN3→EREBPs→果实成熟相关基因的相似路径。

All internal fixators were removed and re-fixed with half ring sculcus external fixator. For 32 cases with soft tissue defect, 10 were repaired with gastrocnemius musculocutaneous flap, 6 with medial head of gastrocnemius and 16 with fascial flap.

所有病例均取出失效内固定后采用半环槽式外固定治疗。32 例软组织缺损和骨外露，10例经腓肠肌皮瓣转位覆盖，6例用腓肠肌内侧头转位加植皮消除创面，16例采用筋膜皮瓣修复。

Restoration of ODC expression in the HL60-ODC cells resulted in rescue of rottlerin-induced apoptosis of HL60 cells. Simultaneously, phosphothreonine-ODC of rottlerin-treated HL60-ODC cells was highly elevated comparing with HL60-pcDNA3 cells. Taken together, we suggested that rottlerin, a PKCd inhibitor, might decrease phosphothreonine-ODC to reduce both protein stability and enzyme activity of ornithine decarbooxylase and finally resulted in cell apoptosis.

为了侦测鸟胺酸去羧化是否能够负调节rottlerin所引发的细胞凋亡，我们将具有鸟胺酸去羧化cDNA载体稳定转染至HL60细胞内，此细胞称为HL60-ODC细胞，另转染载体的细胞称为HL60-pcDNA3细胞；实验发现HL60-ODC细胞中的鸟胺酸去羧化蛋白为HL60-pcDNA3细胞与HL60细胞表现之2.5倍，同时HL60-ODC细胞提升鸟胺酸去羧化蛋白表现，能够减少rottlerin造成之细胞凋亡现象，并且此蛋白在HL60-ODC细胞中酥胺酸磷酸化较HL60-pcDNA3细胞表现高。

Hepatitis B virus is the smallest DNA virus which is a member of the viral family Hepadnavirida. It uses its DNA as template and the RNA polymerase II of the host cells to transcribe its viral transcripts. There are four major RNAs transcribed from HBV genome. All of the four major transcripts are unspliced, terminated at a common site, polyadenylated at the 3' terminal and capped at the 5' terminal.

B 型肝炎病毒属於肝病毒科，为目前已知最小的去氧核醣核酸病毒，利用其病毒的去氧核醣核酸以及宿主细胞内的核醣核酸聚合酶可转录出四种主要的核醣核酸产物，此四种主要的转录产物均未经剪接并终止於核心开放编阅序列的共同polyadenylation 讯号，且在其5'端均经过capping修饰。

In the early stage of its development, once induced by the morphogen,transcription factors such as SHH, WNT and BMP, are specifically expressed in neuronalcells to trigger the downstream effects, and determine the cell fate finally.

在神经系统发育的早期，在SHH，WNT，BMP等形态发生因子的调控下，神经细胞内的转录因子特异性表达，并启动一系列的转录事件，最终决定神经细胞分化方向。

In the negative and interrogative forms, of course, this is identical to the non-emphatic forms.

。但是，在否定句或疑问句里，这种带有"do"的方法表达的效果却没有什么强调的意思。

Go down on one's knees;kneel down

屈膝跪下。。。下跪祈祷