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大鼠

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Results:(1) Progesterone-treated models showed intact nasal mucosa, regular ciliary lining and inactive glands;(2) Untreated or normal saline treated models showed disrupted mucosa, inverted cilia and massive mucosal infiltration of neutrophils;(3) Smearing of nasal discharge revealed limited vs abundant number of neutrophils in SD models treated with progesterone vs untreated or treated with normal saline.(4) Hoechst stain: significantly fewer apoptotic cells per field were found in progesterone-treated models (1.583 ± 0.28) compared with untreated or normal saline treated models (2.85 ± 0.285 and 4.8 ± 0.715, respectively).

结果:(1)用黄体酮滴鼻液治疗的SD大鼠鼻腔黏膜上皮完整,纤毛整齐,腺体开放不多;(2)用或不用生理盐水滴鼻液治疗的SD大鼠鼻腔黏膜上皮纤毛倒伏,上皮结构散乱,上皮层大量中性粒细胞浸润;(3)鼻腔分泌物涂片示黄体酮滴鼻液治疗的SD大鼠有少许中性粒细胞,而用或不用生理盐水滴鼻液治疗的SD大鼠鼻腔分泌物含有大量中性粒细胞;(4)Hoechst染色:用黄体酮滴鼻液治疗的SD大鼠鼻腔黏膜上皮凋亡细胞数[(1.583±0.28)/视野]明显少于用或不用生理盐水滴鼻液治疗的SD大鼠[分别是(2.85±0.285)/视野和(4.8±0.715)/视野]。

The results show that: This tablet can decrease the rat"s footpad swell induced by Freud"s, inhibit the permeability of the blood capillary in inflammatory joint and decrease the 11-2 level in serum while increasing the rat"s body weight,(2) It can lower the rat"s temperature and the permeability of the blood capillary in inflammatory joint induced by collagen and inhibit the delayed type hypersensitivity,It can lower the stickiness of full blood and the accumulation of erythrocyte obviously inblood stasis model rats,lt can inhibit the formation of hemolysin and decrease the index of carbon granule clearance in mice, It can lower the permeability of celiac blood capillary and inhibit the granulomatous hyperplasia by sc cotton ,It can inhibit the pain induced by acetic acid and raise the pain threshold value of heat stimulation.

结果表明:关络通片能明显降低佐剂性关节炎大鼠左右足趾肿胀度,降低大鼠炎症关节的通透性,抑制其IL-2含量,对大鼠体重增长缓慢有一定的改善作用;能改善Ⅱ型胶原诱导性关节炎大鼠的表征指标及炎症关节的通透性,对其迟发型超敏反应有一定的抑制作用;对血瘀模型大鼠,能显著性降低其全血粘度、血浆粘度、红细胞压积;能抑制小鼠的溶血素抗体生成,抑制小鼠网状内皮系统的吞噬能力,具有一定的抑制机体免疫功能的作用,能抑制小鼠腹腔毛细血管通透性增高,抑制大鼠棉球肉芽组织增生,具有一定的抗炎作用;能抑制小鼠醋酸扭体反应、提高小鼠热板痛阈值,具有一定的镇痛作用。

To choose SD rat as experiment obiect, which was divided in to 5 groups: normal control group, model group, methylaminopterin-treat group, tripterygiumglycosides-treat group, and GuiZhiShaoYaoZhiMuTang-treat group, establishing immunityinflammation rat model., through measuring rat voix pedis intumesce degree, the content ofIL-6, TNF-A in peripheral blood, and rats weight as well as organ index, we can observe theeffect of GuiZhiShaoYaoZhiMuTang for treating adjuvant-induced arthritis rat and theby-effect.

以SD大鼠为实验对象,将大鼠随机分为5组:正常对照组、模型对照组、甲氨喋吟治疗组、雷公藤多甙治疗组、桂枝芍药知母汤治疗组。建立免疫性炎症关节炎大鼠模型。通过测量大鼠足跖肿胀程度,外周血IL-6、TNF-α及大鼠的体重和脏器指数,来观察桂枝芍药知母汤对佐剂性关节炎大鼠的治疗作用和副作用。

Rats with CHF were divided randomly into two groups, one group was injected dexamethasone(1mg /kg) intramuscularly at the first and forth day respectively,the other group was injected equi-volume saline likewise, and observing the changes of symptoms of rats with CHF; measuring rats hemodynamics index including blood pressure, heart rate and left ventricular end-diastolic pressure 4 days later. Part 2: To measure every group rats myocardialα1、β1、β2、β3-AR density by immunohistochemical method ,and to understand the AR density changes .Taking the first part rats'myocardium tissues for testing myocardialα1、β1、β2、β3-AR density and 5 normal rats for normo-contrast group and taking gray scale of slice as AR density. Part 3: The study enrolled 35 patients with DCHF. Patients were included if they had orthopnea and refractory edema due to acute DCHF that was severe and had taken more 1 week hospitalization and intravenous therapy in addition to diuretics.

观察心衰大鼠的症状,4天后对两组大鼠进行血流动力学检测,检测指标包括血压、心率和左室舒张末压;第二部分:取第一部分实验动物大鼠的心肌组织,并取5只正常大鼠的心肌组织作为正常对照,用免疫组化法检测大鼠心衰前后及糖皮质激素治疗前后大鼠心肌组织肾上腺素能受体α1、β1、β2、β3受体的变化,取镜下切片的灰度值反应受体的密度;第三部分:研究对象为临床难治性心衰病人35例,病人的选择标准包括由于急性失代偿的心力衰竭出现的端坐呼吸,难治性水肿,患者病情严重,住院超过1周,包括利尿剂之外药物的静脉治疗。

Results:(1)Among 40 rats, 36 rats were successfully established and the rate of success is 90 percent;(2)All the successfully established models demonstrated polydipsia, polyuria and the body weight was not increased. 6 rats suffered cataract after 3 months, and 4 rats died in 6 months;(3)There was an approximately 61% loss of retinal ganglion cells in the central retina and the thickness of retina thinner under microscope ( P 0.01 ).(4) Electroscope changes include the thinner of retina, disorganization of the membranous disc of the rod cells and the thickness of basal membrane of vessal.(5)In normal group, 1 month dibetes mellitus and 1 month treatmen group, there was no expression of ERK1/ERK2 on the retina tissues. In 3 month diabetes mellitus group, the expression of ERK1/ERK2 was positive.

结果:①40只建模大鼠中36只建模成功,建模成功率为90%;②建模成功的大鼠都表现出多饮、多尿、消瘦、体重不增的表现,有6只大鼠在3个月后出现白内障,有4只大鼠在喂养接近至6个月时死亡;③HE染色光镜下6个月大鼠后极部视网膜节细胞层细胞数明显减少,减少约61%(P.01),后极部视网膜明显变薄(P.01);④电镜观察,视网膜变薄,视杆细胞膜盘紊乱,血管基底膜增厚等表现;⑤正常组、糖尿病组和治疗组1个月大鼠视网膜中未见ERK1/ERK2的表达,糖尿病组视网膜组织中3个月时可见少量表达,ERK1/ERK2表达部位为神经节细胞层和内核层;6个月时表达强阳性,部位表达不仅见于内核层、神经节细胞层,色素上皮层也见表达。

The SMMC-7721 hepatoma cell line of experimental groups and control groups were cultured with the addition of sodium selenite, potassium iodide or triiodothyronine (T3). The cell quantity was checked by the MTT assay , and the cell cycle was observed by flow cytometry and levels of AFP and ALB in the culture solution were determined by RIA.The SD rats were divided into positive control group, negative control group, T3 group and mixture group (the mixture of sodium selenite, KI and T3). After two weeks of the administration of sodium selenite、 KI and T3,Walker256 cells were transplanted into the abdominal cavity. The administration was kept on for another two weeks, then observed the morbility and mortality of the rats with ascites carcinoma. At the end, the ascites were for the cells counting and the cell cycles analysising.

方 法:在体外,应用细胞培养的方法,培养SMMC-7721肝癌细胞系,分为试验组和对照组,试验组加入不同浓度Se、I或T3干预,培养一段时间后,通过噻唑蓝试验、流式细胞仪,观察细胞数量及细胞周期的变化,测定细胞培养上清液中甲胎蛋白、白蛋白的水平,观察细胞分化情况;在体内,将试验用SD大鼠分成阴性对照组、阳性对照组、T3组和混合组(为Se、I和T3混合物),首先预防性用药2周,然后给试验组大鼠和阳性对照组大鼠腹腔注射Walker256细胞,制造腹水癌大鼠模型;继续用药2周,观察各组大鼠发病率及死亡率,测量不同时期大鼠血清FT3含量,通过流式细胞仪测量大鼠腹水细胞数及细胞周期变化。

This research was composed of three parts:in the first part, we observed the effect of CTA on small intestine peristalsis;in the sencond part, we observed the effect of sweet stimulability and bitter stimulability on discharge of vagus efferent never after CTA;in the three part,effect of CTA on c-Fos expressions in the brain nerve nucleus was observed.

结果:1给予蔗糖刺激时,CTA组大鼠小肠推进率与对照组相比差异显著,小肠推进率降低;在CTA组大鼠中,给予糖刺激的大鼠小肠推进率明显低于给予水刺激的大鼠。2给予正常大鼠蔗糖刺激时,切断迷走神经的大鼠的小肠推进率明显低于假手术组的。3建立CTA后的大鼠,口腔内给予蔗糖刺激或奎宁刺激时,迷走神经传出放电在刺激后即刻、刺激后30min、刺激后60min都比正常组有所增加。

Each male ratwas paired with 2 female rats in proestrus when administration terminated. Femalerats were examined the next morning for the presence of sperm in vaginal smears andunderwent a cesarean section on day 13 of gestation. Then the reproductive indiceswere calculated as follows: copulation index, pregnancy index, and fertility index,which can evaluate male fertility directly. The sperm motility and morphology wereevaluated by computer-assisted sperm analysis, moreover, the caudaepididymal sperm ultramicrostructure were analyzed by transmission electronmicroscope, which can evaluate the influence on sperm maturation bydutasteride ; sperm survival rate was assessed by SYBR-14 and propidium iodidefluorescent staining; serumal dihydrotestosterone and testosterone of ratswere detected by enzyme-labeled immunoassay, which can evaluate the effecton androgenic levels; the weights of testes and epididymides, were determined byprecise electronic balance; histological examination of above tissues were evaluatedby HE staining and TEM, which can evaluate treatment on histology of reproductiveorgans by dutasteride.

给药结束后雄鼠与处于动情前期的雌鼠按2:1合笼,计算雌鼠的交配指数、受孕指数和生育指数,直接评价雄鼠生育力;采用计算机辅助精子分析系统分析大鼠附睾精子活力和形态,在此基础上合并使用透射电镜技术对各组附睾尾部精子进行透射电镜分析,评价度他雄胺对大鼠精子成熟的影响;采用SYBR-14和propidium iodide双重荧光染色计算精子存活率;采用Elisa法测定大鼠睾酮(testosterone, T)和双氢睾酮(dihydrotestosterone, DHT)血清浓度,评价度他雄胺对雄鼠雄激素水平的影响;采用精密电子天平对各组睾丸、附睾、前列腺和精囊进行称重,评价度他雄胺对大鼠生殖器官重量的影响;采用HE染色法对各组睾丸和附睾进行组织学分析,同时采用透射电镜技术对附睾上皮细胞超微结构进行分析,评价度他雄胺对大鼠主要生殖器官组织学的影响。

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。

The animal model of this method system have, the characteristics of the high fat, high leptin, high Ins, and match with the characteristic of clinical and the pathologic of simple obesity. This studies showed: The acupuncture can obviously reduce the vaule of intaking food and water of the fat rats, and lower the body weight and fat; The acupuncture can urge the diameter, area, physical volume of white fat cell contract, Brown fat tissue of fat rat group surroundings vascular containing large quantity of white fat cell, the acupuncture group was less. this showed: The acupuncture enhanced the white fat cell metabolism, and promotesed the white fat cell to convert to brown fat cell; The frequency of spontaneous electric of nerve cell of pvn of the fat rat is high, the acupuncture counld depress the spontaneous electric of nerve cell of pvn, This showed: the acupuncture can depress the appetite passing to adjust function of pvn nerve; The monoaminergic neurotransmitter of inside of the fat rat brain is disfunction, the acupuncture can rectify this disfunction, and adjust level of 5-HT, and affect appetite, and improve metabolism; The level of leptin and Ins of fat rat increased high, The level of it of acupuncture group decreased, compared with the fat group p.001, This showed: the fat rats have the defeat of leptin and Ins in the body, The acupuncture can lowering this defeat, The level of leptin and Ins of the fat rat in the brain is less than the normal group, The level of leptin of the acupuncture group is high than the fat group, This showed: the acupuncture can increase the transportation of leptin from blood to brain.

本方法所制肥胖大鼠模型具有高体重、高脂、高leptin和高Ins血症的特点,符合人类单纯性肥胖病的临床和病理特征;实验研究显示:针灸能明显减少肥胖大鼠的摄水、摄食量及降低其体重、体脂;针灸能促使白色脂肪细胞直径、面积、体积缩小,减少肥胖大鼠棕色脂肪组织血管周围白色脂肪细胞数量,显示:针灸增强了白色脂肪细胞的代谢,促进了白色脂肪细胞向棕色脂肪细胞的转化:肥胖大鼠PVN神经细胞自发放电频率增加,脑内单胺递质紊乱,针灸通过抑制PVN亢奋的自发放电,调整5-HT水平,达到影响食欲,改善代谢的作用;肥胖大鼠血中leptin及Ins水平增高,与正常组大鼠比有明显差异,提示大鼠体内存在leptin及Ins抵抗,针灸后肥胖大鼠随着体重体脂的下降,血中leptin、Ins水平降低,与肥胖组比均有显著性差异,而针灸后大鼠脑内leptin增加,与肥胖组比P.05,提示:针灸具有改善leptin、Ins抵抗及增加leptin脑内转运的作用。

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