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Results AQP1 is expressed at the apical and basolateral membrane of the microvascular endothelium; AQP3 was detected at basal cells of both the bronchiole epithelium and submucosal gland acinus; AQP4 is present in the basolateral membrane of columnar cells in bronchiole; while AQP5 is expressed in the apical membrane of type Ⅰ pneumocytes, and also at the apical of columnar cells of superficial epithelium and submucosal gland acinar cells.

结果本研究发现AQPs基因在羊肺中的表达分布与人相似,AQP1在肺内的毛细血管内皮细胞表达;AQP3在小支气管黏膜上皮的基底细胞的基侧膜表达,AQP4存在于小支气管黏膜上皮的柱状纤毛细胞的基侧膜;AQP5存在于Ⅰ型肺泡上皮细胞的顶质膜,存在于小支气管黏膜上皮柱状纤毛细胞,以及在气道黏膜下腺的腺细胞的顶质膜表达。

One hand mechanical obstruct led to the increase of veinous resistance and the obstacle of microcirculation, the other hand the adhesive PMN was activated in excess, the white blood cells released a lot of enzymes, in which PMN-elastase can decompose the components of cell and many albumens, inclusive of immunoglobulin、alexin and fibrication. These components induced the injury of the pancreatic capillary vessels and cell and lysosome enzy made the tissue protein hydrolyze and produced unsaturated fatty acids, which destroyed the structure and function of cellar membrane. The inflammatory cellar factors activate other immunocytes to produce the injury and necrosis of tissue, which aggravated the pathological injury and led to shock、pyaemia and MODS. So ICAM-1 and LFA-1 played an important role in SAP. Frossard found that the expression of ICAM-1 in the rat model, especially in serum、pancreas and lung. All these showed ICAM-1 is an important factor in AP and concomitant lung injury.

胰腺小叶组织局部血管EC首先被激活,ICAM-1表达升高,与被激活的PMN表面表达的LFA-1相结合,"PMN-EC"相互作用加剧,一方面机械性阻塞毛细血管导致静脉阻力增加、微循环障碍;另一方面粘附的PMN过度吞噬或激活,当白细胞吞噬的颗粒不能被封闭隔离,连同细胞内的酶被释放出来,其中的PMN-elastase能够降解细胞基质中各种成分,水解多种蛋白,加重胰腺的毛细血管内皮细胞和腺泡的损伤;释放的溶酶体酶使组织蛋白水解,产生的不饱和脂肪酸引发脂质过氧化方应,破坏细胞膜的结构和功能;释放的炎性细胞因子,激发其他的免疫细胞的功能,导致进一步的组织损伤和坏死,加重SAP的病理损伤,最终导致休克、脓毒血症及多器官功能障碍等严重后果。

Objectives:(1) to introduce immunetolerance in mice sciatic nerve allograft by intrathymic injection of allogene.(2) to compare the effect of fresh sciatic nerve allotransplantion and cold preserved sciatic nerve allotransplantation with different diameter.

研究目的:(1)探讨小鼠胸腺内注射异基因抗原在同种异体异基因坐骨神经移植免疫耐受中的作用;(2)比较不同直径的新鲜自体神经和冷冻异体神经移植的效果,探讨神经直径对神经移植效果的影响。

In the normal uterus, Cytokeratins immunolabelling were detected in glandular cell, luminal epithelial cell, Vimentin immunolabelling were detected in stromal cell and endoblastic cell; CK7 immunolabelling were not detected in any tissue of the yak utenus.

研究结果显示:未妊娠时,泛角蛋白在子宫内膜腺上皮细胞、腔上皮细胞内表达,波形蛋白在子宫内膜基质细胞内表达,平滑肌肌动蛋白在子宫平滑肌和血管平滑肌内表达,牦牛子宫任何部位均不表达角蛋白7;妊娠30天左右时,泛角蛋白在子宫内膜腺上皮细胞、子宫内膜腔上皮细胞、滋养层细胞、内胚层细胞和尿囊细胞内表达,波形蛋白在子宫内膜基质细胞和内胚层细胞内表达,平滑肌肌动蛋白在子宫平滑肌和血管平滑肌内表达,角蛋白7在尿囊细胞内表达,偶尔在腔上皮细胞的细胞核边缘表达;消化法进行原代培养时,组织经胶原酶消化并通过100目和400目筛网组合可以有效地分离原代子宫内膜基质细胞和子宫内膜腺上皮细胞;分离得到的子宫内膜基质细胞活率达90%以上,并可在体外传代7次以上;分离得到的子宫内膜腺上皮细胞活率可达85%以上,并可在体外传代5次以上;RPMI1640培养基最适合子宫内膜基质细胞和子宫内膜腺上皮细胞的生长,维持子宫内膜基质细胞正常生长的FBS添加量为20%,维持子宫内膜腺上皮细胞正常生长的胎牛血清添加量为30%。

The results of study show:1. White pulp and red pulp in parenchyma of spleen of embryo can be obviously discerned after 18 days. Periarterial lymphoid sheath and ellipsoid periarterial lymphoid sheath also can be obviously discerned in spleen of 4 days chicken. T, B lymphocytes in appendix basement of embryo emerge after 20 days. It is the initial shape of cecal tonsil. Crypt structure of conjunction of esophago and stomachus glandularis form obviously at 4 days. It is the initial shape of esophago tonsil. The germinal center firstly emerges in these three organs at 14 days. With the increase of day age, the characteristic structure peripheral immune organs gradually develop mature. Spleen achieve mature at 21 days and cecal tonsil at 35 days.2. IgM~+ and IgA~+ cells in spleen of embryo emerge at 15 days. IgG~+ cell, CD3~+ and CD8~+T lymphocytes of embryo emerge at 20 days. CD3~+, CD8~+ and IgM~+ cells in cecal tonsil of embryo emerge at 20 days. However CD4~+, IgG~+ and IgA~+ cells all emerge in 1 day age chicken out of crust. CD3~+, CD4~+, CD8~+, IgM~+ and IgG~+ cells in esophago tonsil of embryo all emerge at 20 days. However IgA~+ cells emerge in 1 day age chicken out of crust.3. The amount of T, B lymphocytes in peripheral immune organs increase follow with the increase of day age, and hold an upgrade tendency. The amount of T, B lymphocytes in spleen achieved stabilization at 21 days, and in tonsil of esophago and appendix at 35 days.

研究结果表明:1、在组织结构方面,脾脏实质内的白髓与红髓在胚胎18日龄后明显可辨,4日龄雏鸡脾脏中形成明显可辨的动脉周围淋巴鞘和椭球周围淋巴鞘;盲肠基部T、B淋巴细胞在胚胎20日龄时开始出现,即初步形成盲肠扁桃体;食管与腺胃结合处在4日龄时形成明显的隐窝结构,即食管扁桃体初步形成;14日龄时,三种器官中首次出现生发中心;随着日龄的增长,外周免疫器官特征结构不断发育成熟,脾脏在21日龄时达到成熟水平,盲肠扁桃体和食管扁桃体在35日龄时达到成熟水平。2、在T、B淋巴细胞出现时间方面,脾脏中IgM~+和IgA~+细胞在胚胎15日龄时开始出现,IgG~+细胞、CD3~+和CD8~+T淋巴细胞在胚胎18日龄时出现,CD4~+细胞在胚胎20日龄时出现;盲肠扁桃体中CD3~+、CD8~+和IgM~+细胞在胚胎20日龄时开始出现,而CD4~+、IgG~+和IgA~+细胞均在雏鸡出壳后1日龄时出现;食管扁桃体中CD3~+、CD4~+、CD8~+、IgM~+和IgG~+细胞均在胚胎20日龄时开始出现,而IgA~+细胞则在雏鸡出壳后1日龄时出现。3、在T、B淋巴细胞数量变化方面,外周免疫器官中T、B淋巴细胞的数量随日龄增长,整体均呈上升趋势。

Results:There is a close positive correlation between myoepithelial cell differentiation and glandular cell maturation and the ratio of myoepithelial cells to glandular endpieces area was decreased.

结果:实验结果提示,小鼠泪腺肌上皮细胞从出现到成熟的形态变化,与腺细胞内的分泌颗粒从出现到增多是同期发育的,泪腺腺房与肌上皮细胞的面积比率呈减小趋势。

One hand mechanical obstruct led to the increase of veinous resistance and the obstacle of microcirculation, the other hand the adhesive PMN was activated in excess, the white blood cells released a lot of enzymes, in which PMN-elastase can decompose the components of cell and many albumens, inclusive of immunoglobulin、alexin and fibrication. These components induced the injury of the pancreatic capillary vessels and cell and lysosome enzy made the tissue protein hydrolyze and produced unsaturated fatty acids, which destroyed the structure and function of cellar membrane. The inflammatory cellar factors activate other immunocytes to produce the injury and necrosis of tissue, which aggravated the pathological injury and led to shock、pyaemia and MODS. So ICAM-1 and LFA-1 played an important role in SAP. Frossard found that the expression of ICAM-1 in the rat model, especially in serum、pancreas and lung. All these showed ICAM-1 is an important factor in AP and concomitant lung injury.

胰腺小叶组织局部血管EC首先被激活,ICAM-1表达升高,与被激活的PMN表面表达的LFA-1相结合,&PMN-EC&相互作用加剧,一方面机械性阻塞毛细血管导致静脉阻力增加、微循环障碍;另一方面粘附的PMN过度吞噬或激活,当白细胞吞噬的颗粒不能被封闭隔离,连同细胞内的酶被释放出来,其中的PMN-elastase能够降解细胞基质中各种成分,水解多种蛋白,加重胰腺的毛细血管内皮细胞和腺泡的损伤;释放的溶酶体酶使组织蛋白水解,产生的不饱和脂肪酸引发脂质过氧化方应,破坏细胞膜的结构和功能;释放的炎性细胞因子,激发其他的免疫细胞的功能,导致进一步的组织损伤和坏死,加重SAP的病理损伤,最终导致休克、脓毒血症及多器官功能障碍等严重后果。

Objective: To investigate the effect of IFN-γ transgene expression on allergen-induced pulmonary eosinophil infiltration in murine asthmatic model .

目的:探讨腺病毒介导的小鼠γ-干扰素在小鼠哮喘模型肺上皮细胞内的转基因表达对过敏原所致的嗜酸性粒细胞浸润的作用。

The effects and mechanism of GABAergic neurons, NOergic neurons, opioid peptide and cyclic adenosine monophosphate in the nucleus reticularis thalami on sleep-wakefulness cycle of rats and the effects and mechanism of the 5-HTergic nerve fibers project from the nucleus raphes dorsalis to RT on sleep-wakefulness cycle of rats were investigated with the methods of brain stereotaxic, nucleus spile, microinjection and polysomngraphy.1. The effects of GABAergic neurons in RT on sleep-wakefulness cycle of rats1.1 Microinjection of 3-mercaptopropionic acid (3-MP, a kind of glutamate decarboxylase inhibitor) into RT. On the day of microinjection, sleep only decreased a litter. On the second day, sleep marked decreased and wakefulness marked increased. On the third and fourth day, sleep and wakefulness stages resumed to normal.1.2 Microinjection of gamma-amino butyric acid (GABA 1.0μg) into RT enhanced sleep and reduced wakefulness compared with control; while microinjection of L-glutamate (L-Glu, 0.2μg) decreased sleep and increased wakefulness; microinjection of bicuculline (BIC, 1.0μg), a GABAA receptor antagonist, enhanced wakefulness and reduced sleep; microinjection of baclofen (BAC, 1.0μg), GABAB receptor agonist, had the same effects as GABA.2. The effects of NOergic neurons in RT on sleep-wakefulness cycle of rats2.1 Microinjection of L-arginine (L-Arg, 0.5μg) into RT decreased sleep compared with control, but there were on statistaical difference between L-Arg group and control; while microinjection of sodium nitroprusside (SNP, 0.2μg), a NO donor into RT, sleep marked decreased and wakefulness marked increased. Microinjection of nitric oxide synthase inhibitor, N-nitro-L-arginine (L-NNA, 2.0μg) into RT enhanced sleep and reduced wakefulness.2.2 After simultaneous microinjection of L-NNA (2.0μg) and SNP (0.2μg) into RT, SNP abolished the sleep-promoting effect of L-NNA compared with L-NNA group; after simultaneous microinjection of L-NNA (2.0μg) and L-Arg(0.5μg) into RT, we found that L-NNA could not blocked the wakefulness-promoting effect of L-Arg.3. The effects of opioid peptide in RT on sleep-wakefulness cycle of rats3.1 Microinjection of morphine sulfate (MOR, 1.0μg) into RT increased wakefulness and decreased sleep compared with control; while microinjection of naloxone hydrochloride (NAL, 1.0μg), the antagonist of opiate receptors, into RT, enhanced sleep and reduced wakefulness.3.2 After simultaneous microinjection of MOR (1.0μg) and NAL (1.0μg) into RT, the wakefulness-promoting effect of MOR and the sleep-promoting effect of NAL were not observed compared with control.4. The effects of cAMP in RT on sleep-wakefulness cycle of rats Microinjection of cAMP (1.0μg) into RT increased sleep and decreased wakefulness compared with control; microinjection of methylene blue (MB,1.0μg) into RT enhanced sleep and reduced wakefulness compared with control.5. The effects of the 5-HTergic nerve fibers project from DRN to RT on sleep-wakefulness cycle of rats5.1 When L-Glu (0.2μg) was microinjected into DRN and normal sodium (NS,1.0μg) was microinjected into bilateral RT. We found that sleep was decreased and wakefulness was increased compared with control; when L-Glu (0.2μg) was microinjected into DRN and methysergide (MS,1.0μg), a non-selective 5-HT antagonist, was microinjected into bilateral RT, We found that sleep was enhanced and wakefulness was reduced compared with L-Glu group.5.2 When p-chlorophenylalanine (PCPA, 10μg) was microinjected into DRN and NS (1.0μg) was microinjected into bilateral RT, We found that sleep was increased and wakefulness was decreased compared with control; microinjection of 5-hydroxytryptaphan (5-HTP, 1.0μg), which can convert to 5-HT by the enzyme tryptophane hydroxylase and enhance 5-HT into bilateral RT, could block the effect of microinjection of PCPA into DRN on sleep-wakefulness cycle.

本研究采用脑立体定位、核团插管、微量注射、多导睡眠描记等方法,研究丘脑网状核(nucleus reticularis thalami,RT)中γ-氨基丁酸(gamma-amino butyric acid ,GABA)能神经元、一氧化氮(nitrogen monoxidum,NO)能神经元、阿片肽类神经递质、环一磷酸腺苷(cyclic adenosine monophosphate,cAMP)及中缝背核(nucleus raphes dorsalis,DRN)至RT的5-羟色胺(5-hydroxytryptamine,5-HT)能神经纤维投射对大鼠睡眠-觉醒周期的影响及其作用机制。1 RT内GABA能神经元对大鼠睡眠-觉醒周期的影响1.1大鼠RT内微量注射GABA合成关键酶抑制剂3-巯基丙酸(3-MP,5μg),注射当天睡眠时间略有减少,第二日睡眠时间显著减少,觉醒时间明显增多,第三、四日睡眠和觉醒时间逐渐恢复至正常。1.2大鼠RT内微量注射GABA受体激动剂GABA( 1.0μg)后,与生理盐水组比较,睡眠时间增加,觉醒时间减少;而RT内微量注射L-谷氨酸(glutamic acid, L-Glu, 0.2μg)后,睡眠时间减少,觉醒时间增加;RT内微量注射GABAA受体阻断剂荷包牡丹碱(bicuculline,BIC,1.0μg)后,睡眠时间减少,觉醒时间增加;RT内微量注射GABAB受体激动剂氯苯氨丁酸(baclofen,BAC,1.0μg)后,产生了与GABA相似的促睡眠效果。2 RT内NO能神经元对大鼠睡眠-觉醒周期的影响2.1大鼠RT内微量注射NO的前体L-精氨酸(L-Arg,0.5μg)后,与生理盐水组对比,睡眠时间略有减少,但无显著性意义;而RT内微量注射NO的供体硝普钠(Sodium Nitroprusside,SNP,0.2μg)后可明显增加觉醒时间,缩短睡眠时间;微量注射一氧化氮合酶抑制剂L-硝基精氨酸(L-arginine,L-NNA,2.0μg)后,引起睡眠时间增多,觉醒时间减少。2.2大鼠RT内同时微量注射L-NNA(2.0μg)和SNP(0.2μg)后与L-NNA组比较发现SNP逆转了L-NNA的促睡眠作用;RT内同时微量注射L-NNA(2.0μg)和L-Arg(0.5μg)后,与L-NNA(2.0μg)组比较发现L-Arg可以增加觉醒而缩短睡眠,其促觉醒作用未能被NOS的抑制剂L-NNA所逆转。3 RT内阿片肽对大鼠睡眠-觉醒周期的影响3.1大鼠RT内微量注射硫酸吗啡(morphine sulfate,MOR,1.0μg)后与生理盐水组对比,睡眠时间减少而觉醒时间增加; RT内微量注射阿片肽受体拮抗剂盐酸纳洛酮(naloxone hydrochloride,NAL,1.0μg)后与生理盐水组比较,睡眠时间增加而觉醒时间减少。3.2大鼠RT内同时微量注射MOR(1.0μg)和NAL(1.0μg)后,与生理盐水组对比,原有的MOR促觉醒效果和NAL的促睡眠效果都没有表现。4 RT内环一磷酸腺苷信使对大鼠睡眠-觉醒周期的影响大鼠RT内微量注射cAMP(1.0μg)后与NS(1.0μg)组比较,睡眠时间增多而觉醒时间减少;RT内微量注射亚甲蓝(methylene blue,MB,1.0μg)后,与NS组比较,睡眠时间增多而觉醒时间减少。5中缝背核投射到丘脑网状核的5-羟色胺能神经纤维对大鼠睡眠-觉醒周期的影响5.1大鼠DRN内微量注射L-Glu(0.2μg),同时在双侧RT内微量注射NS (1.0μg)后,与对照组(DRN和双侧RT注射NS, 0.2μg)比较,睡眠时间减少,觉醒时间增多;大鼠DRN内微量注射L-Glu(0.2μg),同时在双侧RT内微量注射二甲基麦角新碱(methysergide, MS, 1.0μg )后,与对照组(DRN注射L-Glu 0.2μg,双侧RT注射NS 1.0μg)比较,睡眠时间增多,觉醒时间减少。5.2大鼠DRN内微量注射对氯苯丙氨酸(p-chlorophenylalanine,PCPA,10μg),同时在双侧RT内微量注射NS (1.0μg)后,与对照组(DRN和双侧RT注射NS, 1.0μg)比较,睡眠时间增多,觉醒时间减少;大鼠DRN内微量注射PCPA(10μg),产生睡眠增多效应后,在双侧RT内微量注射5-羟色胺酸(5-hydroxytryptaphan , 5-HTP, 1.0μg )后,与对照组(DRN注射PCPA 10μg,双侧RT注射NS 1.0μg)比较,睡眠时间减少,觉醒时间增多。

To study the effects on the body of animals after incorporation of 3H-TdR and its mechanism, the laws of distribution A retention and absorbed dose in blood , thymus, spleen, femur, heart, lung,kidney, liver, brain , spermary , small intestine and muscle were detected by liquid scintillation counter after 3H-TdR was injected into the tail vein of mouse; the process of dynamic incorporation of in the lymphocytic cell nucleus of blood , thymus , spleen and the nucleus of femur was monitored; the absorbed dose in the cell nucleus was estimated; the influence to RNA"s synthesizing and cell"s survival was observed using the technology of CLSM; A segment of Egr-1 gene was cloned with reverse transcriptase- polymerase chained reaction, the influence to the expression of Egr-1 gene in the spleen cells was observed.

为了探讨有机结合氚内污染对生物机体的影响及其作用机理,本课题应用均相液体闪烁测量技术研究了氚标记胸腺嘧啶核苷(~3H-TdR)经尾静脉注入小鼠机体后,在血液、脑、胸腺、心脏、肺脏、肝脏、脾脏、肾脏、小肠、睾丸、肌肉和股骨共12种组织器官的生物动力学分布规律及其吸收剂量;应用纸片法液体闪烁测量技术研究了~3H-TdR内污染在外周血、胸腺和脾脏淋巴细胞及骨髓细胞的动态滞留规律,估算了细胞核的吸收剂量;应用共聚焦显微镜观察了~3H-TdR内污染对脾细胞核酸合成及其存活率的作用,应用反转录PCR技术研究了~3H-TdR内污染对脾细胞早期生长反应基因-1(Egr-1)转录表达的影响。

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