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①Rat MSC and VSMC were cultured and identified, respectively. MSC were labeled with DAPI firstly, and then co-cultivated with VSMC. The changes of morphology and ultrastructure of co-cultured cells were observed. Immunfluorescence analysis was performed by using monoclonal antibodies against specific antigen.②We established the regulatable system in two steps: a stable MSC line expressing rtTA has been constructed and characterized firstly by transfected with pUHD 17-1hyg and then selected by hygromycin B; in a second step, this line was used for trandfer the AT2R gene to MSC to get the well establishing double stable MSC lines;③The expression of AT2R regulated by doxycycline was evaluated by western blot;④The MSCs were transduced into rat carotid arteries with regulatable AT2R gene after the establishment of rat carotid balloon injury restenosis model. The intimal/medial area ratio were measured by digital analysis system.

研究方法:(1)密度梯度离心法及胶原酶消化法分别培养原代大鼠MSC及VSMC,细胞共培养并行免疫荧光化学染色和透射电镜观察超微结构;(2)组成受Dox调控的哺乳动物表达系统的四种成分的转化、扩增及提纯并酶切鉴定;(3)采用常规分子生物学方法连续两个回合转染体外培养的MSC,并分别采用发光计检测不同细胞克隆萤光素酶活性改变以及RT-PCR方法检测AT2R目的基因mRNA表达情况,根据各个细胞克隆受Dox调控表达的程度,选择低背景、高诱导表达AT2R的细胞系,作为双重稳定MSC细胞系;蛋白免疫印迹法观察该细胞系在Dox调控下AT2R表达的时相性、持续性及在不同浓度Dox调控下的表达情况;(4)建立大鼠颈动脉球囊损伤动物模型,将双重稳定MSC在术中导入血管,分别于14 d、28 d进行病理切片,检测可调控AT2R对新生内膜增生的影响;采用RT-PCR免疫组织化学免疫荧光等技术观察AT2R基因在新生内膜中的表达以及细胞外基质成分表达的改变,TUNEL法检测血管组织中细胞凋亡的变化情况。

METHODS: Female BALB/c mice were treated with CCK-8 and keyhole limpet haemocyanin, splenocytes were acquired and incubated in vitro with KLH restimulation, the productions of Th1 cytokines, interferon-γ, interleukin-2 and Th2 cytokines, IL-4, IL-5 in cell culture supernatants were detected by enzyme-linked immunosorbnent assay. The mRNA expressions of IFN-γ, IL-2, IL-4 and IL-5 in splenocytes were detected by reverse transcription-polymerase chain reaction. The levels of Th1 antibody IgG2a and Th2 antibodies IgG1 in serum were also detected by ELISA.

给予BALB/c小鼠钥孔戚血蓝蛋白免疫同时体内给予不同剂量的CCK-8,酶联免疫吸附试验检测其脾细胞培养上清中Th1型细胞因子γ-干扰素、白细胞介素-2(IL-2)和Th2型细胞因子白细胞介素-4(IL-4)、白细胞介素-5(IL-5)水平,逆转录聚合酶链式反应法检测脾细胞中IFN-γ、IL-2、IL-4、IL-5 mRNA表达;ELISA法检测血清中Th1型抗KLH抗体IgG2a和Th2型抗KLH抗体IgG1水平。

Results: The cultured chondrocytes were polygonal cells. There were many rough endoplasmic reticula and mitochondria in cytoplasm, and a lot of secretory vesicles under cell membrane and in the cytoplasm. When cultured for 10 days, some small and white nodules were formed on the bottom of the culture dished, and volcanic-mouth-like structures were formed when cultured for 20 days. Both these nodules and structures contained GAG-positive substances were demonstrated by safranine-O staining.

结果:体外培养的软骨细胞为多角形,透射电镜下见胞浆内有丰富的粗面内质网系统及线粒体,胞膜下及胞浆中有较多分泌泡;连续培养10天时,培养瓶底部出现肉眼可见的白色结节,蕃红-O染色显示白色结节含大量氨基葡聚糖阳性物质,20天时形成火山口样结构,也被蕃红-O染成深红色。

Objetive:To investigate the histopathological features and detect the mutation of JAK2V617F in PV、ET and IMF;to analyze the effects of the stages of myelofibrosis and the mutation on the number of leucocyte in PV and ET patients;Methods: the morphological features of 69 patients who dignosed in the first time were studied by HGF and Gomori stains ; the mutation of JAK2V617F was detected in 38 blood or marrow samples of PV、ET and IMF patients by real-time PCR;the effects of the stages of myelofibrosis and the mutation on leucocytes were analyzed;Results: Different stages of reticulin were observed in 15 PV and 15 ET patiens'marrow sections,and the cytosis、the pleomorphism and the bone trabecula side location of megakaryocytes were observed in all PV、ET and IMF patients;the means of leucocyte in patients with and without JAK2V617F were 20.4×109/Land 11.4×109/L respectively;the P value was 0.367 as comparing the leucocytes among different stages of reticulin increasing.

目的:探讨PV、ET、IMF患者的骨髓病理特点并且检测JAK2V617F突变的发生率,分析骨髓中纤维增生程度以及JAK2V617F对PV、ET患者白细胞数的影响;方法:69例初诊患者的骨髓切片经HGF染色以及Gomori染色后进行形态学研究;并利用real-time PCR法测38例患者的骨髓或者外周血的JAK2V617F突变;分析骨髓切片中纤维组织增生程度以及JAK2突变对患者白细胞数的影响;结果:15例PV和15例ET患者的骨髓切片中可以观察到不同程度的纤维组织增生,所有PV、ET、IMF患者骨髓病理中可见到巨核细胞增多、巨核细胞多形性改变以及巨核细胞定位于骨小梁旁;12例PV、4例ET、4例IMF中检测到JAK2V617F;有突变和无突变患者发病时白细胞数均值分别为20.4×109/L和11.4×109/L,比较不同纤维增生程度的PV、ET患者的白细胞数均值,P值为0.367。

In this study, samples were taken during the period November 2005 to March 2007. Of total of 1109 dogs sampled, 168 Salmonella spp. were isolated, 16.0% were positive for Salmonella spp. of stray dogs; 7.4% of household dogs were positive for salmonellae. Salmonella Newport (16.7%) and Enteritids (10.7%) was the most prevalent serotype. Additionally, the result of antimicrobial susceptibility test showed that 99.4% of isolated strains (167/168) were resist to one or more than one antibiotic.

因此为了探讨台湾中部地区犬只沙门氏菌感染的情形,本次调查於2005年11月至2007年3月针对中部地区5个县市的收容所犬只及家犬进行调查,共采集样本1109个样本,分离出168株沙门氏菌,其中收容所犬只之总分离率为16.0%;家犬部分分离率为7.4%,血清型部份则以Newport(16.7%)及Enteritids(10.7%)为主;此外,沙门氏菌的药物敏感性试验结果显示高达99.4%(167/168)的菌株对1种或1种以上的抗菌剂产生抗药性。

The synthetic technologies of three hindered amine light stabilizers were established, the reason for the insoluble side products formed in the production of polymerized hindered amine light stabilizers were studied, and a series of novel hindered amine light stabilizers with antibacterial properties were designed and synthesized in this present paper.1. Based on the investigation of the transesterification between dimethyl sebacate and 1,2,2,6,6-pentamethyl-4-piperinol, the synthetic technologies of Tinuvin292 and Tinuvin 765 were established. The two kinds of hindered amine light stabilizers were obtained in the yield of 93.3% and 92.4% respectively and qualified as the advanced level of the kindred overseas products.2. The synthetic technology of Tinuvin123 was studied and established.

本文研究建立了三个受阻胺类光稳定剂的合成工艺,探索了聚合型受阻胺类光稳定剂生产过程中不溶性副产物产生的原因,并设计、合成了一系列具有抑菌活性的新型受阻胺类光稳定剂。1、在研究癸二酸二甲酯与1,2,2,6,6-五甲基哌啶醇的酯交换反应的基础上,建立了受阻胺类光稳定剂Tinuvin292、Tinuvin765的合成工艺,分别以93.3%和92.4%的收率得到了光稳定剂Tinuvin292、Tinuvin765,所得产品质量达到了国外同类产品的水平。2、研究建立了受阻胺类光稳定剂Tinuvin123的合成工艺。

In this paper, arrangement scheme of a novel multi-star phase shifted pole changing coil was given and the methods to determine coil split ratio were analyzed. According to electromagnetic theory, 2D electromagnetic mathematical model in which stator windings were spitted coil was developed, and magnetic field distribution of the novel motor at starting was solved by finite element method, starting performance of motor was calculated. Based on energy method, rotor slot leakage reactance under different poles and different loads were calculated, and the variation of rotor slot leakage reactance with loads was gotten. Based on time-step finite element method, distribution of branch current in stator windings and operating performance were calculated and analyzed when branch was unbalance, the method to calculate circulating loss in stator windings of novel multi-star phase shifted pole changing motor was given, and the effects of the loads variation and degree of unequilibrium on circulating loss in stator windings were calculated and analyzed.

论文系统的给出了采用分裂线圈法新型多星形移相变极绕组的排列方案,进一步分析线圈分裂比的确定方法;根据电磁场理论,建立了定子为分裂线圈绕组的电机二维电磁场数学模型,并采用场路耦合法求解了新型多星形移相变极电机起动时的磁场分布,计算了电机起动性能和基于能量法计算了不同极数下起动时转子槽漏抗和不同负载时转子槽漏抗,得出转子槽漏抗随负载变化的规律;采用时步有限元法计算分析了定子绕组支路不平衡时支路电流的分布情况及运行性能,给出了新型多星形移相变极电机定子绕组环流损耗的计算方法,分析比较了负载的变化和支路不平衡度大小对定子绕组环流损耗的影响。

METHODS: Bone marrow was sterilely separated from human. After heparinization, human BMSCs were harvested using density gradient centrifugation and adherence method. At the fifth passage, BMSCs at 1×108/L were incubated in the 6-well plate and divided into 2 groups. BMSCs in the edaravone group were 50% confluent and incubated in L-DMEM containing basic fibroblast growth factor and fetal bovine serum for 24 hours. After washing in PBS, these BMSCs were incubated in serum-free L-DMEM containing 20 mg/L edaravone for 24 hours. BMSCs in the blank control group were incubated in L-DMEM, supplemented with 10% fetal bovine serum.

无菌抽取的骨髓经肝素化后,采用密度梯度离心法及贴壁筛选法分离获得人骨髓间充质干细胞,传至第5代按1× 108 L-1接种于6孔板内,设立2组,依达拉奉组细胞达50%融合时用含碱性成纤维生长因子、胎牛血清的L-DMEM预诱导24 h,PBS洗涤后再用20 mg/L依达拉奉无血清L-DMEM诱导24 h;空白对照组始终用含体积分数为10%胎牛血清的L-DMEM培养,不加任何预诱导剂和诱导剂。

24 New Zealand white rabbits were divided into control group,Saline group and Mailuoning group,on the latter two groups the models of Hemifacial spasm were made by the temporal superficial artery closely contacting the main trunk of facial nerve at stylomastoid foramen.From the 5th week,the Saline and Mailuoning were injected intravenously into ear margin for 2 weeks on Saline and Mailuoning group respectively.At the 7th week,the MDA and SOD in serum were measured,meawhile the microstructure and ultrastructure of facial nerve were observed on 3 animal groups.

将24只新西兰大白兔随机分为对照组、脉络宁组和生理盐水组,在手术显微镜下分离出其茎乳孔处的面神经主干和颞浅动脉,使二者密切接触,以制备半面痉挛动物模型;术后第5周起对脉络宁组和生理盐水组分别自耳缘静脉推注脉络宁注射液和生理盐水持续2周;第7周取3组动物血清测定丙二醛和超氧化物歧化酶,并处死取材行面神经光、电镜观察。

There were no significant differences for the firing rates in the site of contralateral TNC neurons among during pre-CSD,CSD,and post-CSD (P>0.05).For flunarizine group,the firing rates in the site of ipsilateral TNC neurons during pre-CSD were higher as compared with during CSD(P<0.05).2.1 There were statistical differences on palasma levels of CGRP and SP among the three groups(P<0.05).The levels of CGRP and SP in CSD group were higher than control group(P<0.05).No significant differences on the levels of CGRP and SP in ipsilateral trigeminal ganglia were found among the three groups(P>0.05).2 The number of neurons with positive CGRP and SP immunoreactivity was statistically different in right-sided trigeminal ganglia among the three groups (P<0.05).The number in fight-sided trigeminal ganglia in CSD group was higher as compared with control group(P<0.05).The number in right-sided trigeminal ganglia was statistically higher than that in left-sided trigeminal ganglion in CSD group(P<0.05).3.1 Altered ReHo in ipsilateral pons and other brain regions response to pain such as basal nuclei,thalamus,cingulated gyms and prefrontal cortex was detected during the acute spontaneous attack as compared with during headache remission(P<0.05,corrected by Monte Carlo simulation). 2 Positive functional connectivity was detected between ipsilateral pons and other brain regions related to pain within pain state and within non-pain state (P<0.05,corrected by false discovery rate,FDR).Increased functional correlation between ipsilateral pons and other pain-related brain regions such as ipsilateral prefrontal cortex and contralateral subcallosal gyrus was detected during the acute spontaneous attack as compared with during headache remission(P<0.05,corrected by Monte Carlo simulation).

结果1。对照组未发现CSD;同侧TNC放电频率,CSD中>CSD后>CSD前P<0.05对侧TNC放电频率,CSD前、中、后无统计学差异(P>0.05氟桂利嗪组同侧TNC放电频率,CSD前>CSD中(P<0.05),CSD前与CSD后及CSD中与CSD后之间无统计学差异(P>0.05)。2.1关于放免测定,各组血浆CGRP、SP水平有统计学差异(P<0.05),CSD组高于对照组(P<0.05),CSD组与氟桂利嗪组、对照组与氟桂利嗪组之间均无统计学差异P>0.05各组之间同侧三叉神经节中CGRP、SP水平未见变化(P>0.05.2关于免疫组化研究,右侧三叉神经节CGRP、SP免疫阳性细胞数三组之间有统计学差异(P<0.05),多重两两比较结果CSD组大于对照组(P<0.05),CSD组与氟桂利嗪组之间、对照组与氟桂利嗪组之间无统计学差异P>0.05左侧三叉神经节CGRP、SP免疫阳性细胞数三组之间无统计学差异(P>0.05CSD组中右侧三叉神经节CGRP、SP免疫反应阳性细胞数大于左侧(P<0.05)。3.1局部一致性分析发现两组患者头痛疼痛状态较非疼痛状态脑活动发生变化的脑区有同侧脑桥以及其他疼痛相关脑区如基底节区、丘脑、扣带回、前额叶皮层等(P<0.05,蒙特卡罗模拟校正)。2功能连接分析发现疼痛状态与非疼痛状态下主要疼痛相关脑区均与同侧脑桥有功能联系P<0.05,false discovery rate,FDR校正疼痛状态与非疼痛状态比较,同侧前额叶皮层、对侧胼胝下回等疼痛相关脑区与同侧脑桥之间功能联系增强(P<0.05,蒙特卡罗模拟校正。

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推荐网络例句

Through comparing with the results by simulation to study the effects of theprojectile"s final velocity, the angle of rotation and the ballistic trajectory"s migration withdifferent projectile"s rotating speeds, different target"s moving speeds and differentpenetration angles.

通过比较数值模拟的结果来研究不同弹头转速、目标速度、侵彻角对侵彻过程中弹头最终速度、翻转角度和弹道偏移的影响。

I love stationery and all the accoutrement of writing.

我爱文具以及所有的书写的工具装备。

Just loll there: quiet dusk: let everything rip.

只消懒洋洋地享受这宁静的黄昏,一切全听其自然。