华蟾素抑制NIH／3T3细胞增殖对防治器官组织纤维化的作用

目的：观察华蟾素对NIH／3T3细胞的抑制作用，为治疗纤维化有关疾病提供实验依据。方法：体外培养NIH／3T3细胞株，加入不同浓度的华蟾素，24h后观察细胞形态学，以碱性磷酸酶为指标观察细胞毒性，用四甲基偶氮唑盐法检测其增殖活性。结果：不同浓度的华蟾素均能改变成纤维细胞形态，抑制细胞增殖，浓度在10mg／L以下无明显的细胞毒作用。结论：华蟾素对NIH／3T3细胞具有抑制作用，并呈剂量依赖关系，具有防治纤维化有关疾病的应用前景。     

斑蝥素抑制NIH/3T3细胞增殖对防治器官组织纤维化的作用

目的 观察斑蝥素对 NIH/3T3细胞的抑制作用,为治疗纤维化有关疾病提供实验依据. 方法 体外培养 NIH/3T3细胞株,加入 1.000 0, 0.500 0, 0.250 0, 0.125 0, 0.062 5, 0.031 3 mg/L浓度的斑蝥素, 24 h后观察细胞形态学,以乳酸脱氢酶为指标观察细胞毒性,用 MTT法检测其增殖活性. 结果 不同浓度的斑蝥素均能改变成纤维细胞形态,抑制细胞增殖,浓度在 1 mg/L以下无明显的细胞毒作用. 结论 斑蝥素对 NIH/3T3细胞具有抑制作用,并呈剂量依赖关系,具有防治纤维化有关疾病的应用前景.     

斑蝥素抑制NIH／3T3细胞增殖对防治器官组织纤维化的作用

目的：观察斑蝥素对NIH／3T3细胞的抑制作用，为治疗纤维化有关疾病提供实验依据。方法：体外培养NIH／3T3细胞株，加入1．0000，0．5000，0．2500，0．1250，0．0625，0．0313mg／L浓度的斑蝥素，24h后观察细胞形态学，以乳酸脱氢酶为指标观察细胞毒性，用MTT法检测其增殖活性。结果：不同浓度的斑蝥素均能改变成纤维细胞形态，抑制细胞增殖，浓度在1mg／L以下无明显的细胞毒作用。结论：斑蝥素对NIH／3T3细胞具有抑制作用，并呈剂量依赖关系，具有防治纤维化有关疾病的应用前景。     

具G418抗性NIH3T3细胞饲养层的制备

背景:建立具有抗药(新霉素或潮霉素)性的饲养层细胞是转染外源目的基因胚胎干细胞阳性克隆的筛选必备条件。建成的具有抗药性的NIH3T3细胞系可用于胚胎干细胞其他目的基因的筛选。目的:建立具有G418抗性的NIH3T3细胞系,用于转染pTet-on基因的胚胎干细胞阳性细胞克隆筛选的饲养层。设计:细胞培养观察及DNA检测。单位:中国医科大学实验动物部。材料:NIH3T3细胞由中国医科大学细胞生物教研室惠赠,pWL/neo质粒为美国哈佛大学医学院金壮教授惠赠。G418、白血病抑制因子(胚胎干细胞GRO106U/mL)、DMEM为GIBCOBRL产品,丝裂霉素-C、DIG标记及检测试剂盒为Roche产品,脂质体为Invitrogen产品,均购自沈阳联星生物技术有限公司。方法:①通过脂质体转染的方法,将含有neo基因的质粒pWL/neo导入NIH3T3细胞中。②将细胞传至25mL培养瓶中,加入梯度浓度的G418继续培养,通过观察细胞生长及死亡情况测定G418对NIH3T3细胞最小致死量。③将转染的细胞进行传代,挑取单个细胞的克隆至24孔培养板继续进行筛选扩增,选用正常的NIH3T3细胞加入相同剂量的G418的选择性培养基作为筛选的阴性对照。④采用较低的细胞密度铺板,加入含最小致死量G418的DMEM培养基进行再次筛选;同时选用正常的NIH3T3细胞为对照,使用含有最小致死量G418及未加G418的DMEM培养基进行培养,采用光学显微镜对G418抗性NIH3T3细胞进行观察。⑤选用具有G418抗性的NIH3T3细胞和小鼠胚胎成纤维细胞分别作为饲养层细胞进行传代,采用光学显微镜对培养后胚胎干细胞-D3细胞进行观察,并制备细胞DNA,对其进行PCR和southernblot鉴定。主要观察指标:①G418对NIH3T3细胞最小致死量的测定结果。②G418抗性NIH3T3细胞的筛选结果。③G418抗性NIH3T3细胞的生长观察结果。④胚胎干细胞-D3细胞生长观察结果及G418抗性NIH3T3细胞的鉴定。结果:①G418对NIH3T3细胞最小致死量为500mg/L。②经过500mg/LG418压力筛选后,获得了抗G418细胞克隆。③抗性NIH3T3细胞的形态和生长速度与正常NIH3T3细胞没有差异。④饲养层培养的胚胎干细胞呈集落形状生长,边缘光滑,保持未分化的状态。用特异性核苷酸引物检测抗性细胞基因组DNA,可以扩增出对应的核苷酸片段,Southernblot鉴定结果表明neo基因片段已整合入G418抗性NIH3T3细胞中。结论:成功地培育了G418抗性NIH3T3细胞,生长在G418抗性NIH3T3细胞饲养层上胚胎干细胞细胞基本保持正常胚胎干细胞细胞特征。     

互隔交链孢酚对NIH/3T3细胞中DNA聚合酶β的致突变作用

背景:近年来研究发现食管癌细胞中存在DNA聚合酶β的突变。目的:观察互隔交链孢酚对NIH/3T3细胞中DNA聚合酶β基因序列的影响,研究互隔交链孢酚致细胞变异的机制。方法:用不同浓度(2,4,6,8μmol/L)的互隔交链孢酚作用于NIH/3T3细胞,并设置对照组。用RT-PCR法从细胞总RNA中扩增出DNA聚合酶β基因cDNA序列,应用T-A克隆技术,克隆至pGEM-T载体后进行测序,分析其序列变化。结果与结论:2μmol/L互隔交链孢酚处理后的细胞内DNA聚合酶β基因序列无任何变化。而4μmol/L组中DNA聚合酶β基因有1个位点发生突变,8μmol/L组和16μmol/L组中各有2个位点发生突变,此3组的突变存在相同的位点。结果证实,互隔交链孢酚可导致NIH/3T3细胞中的DNA聚合酶β发生点突变,互隔交链孢酚浓度越高,点突变的数量越多,且存在突变活跃位点。     

具G418抗性NIH3T3细胞饲养层的制备

背景：建立具有抗药（新霉素或潮霉素）性的饲养层细胞是转染外源目的基因胚胎干细胞阳性克隆的筛选必备条件。建成的具有抗药性的NIH3T3细胞系可用于胚胎干细胞其他目的基因的筛选。 目的：建立具有G418抗性的NIH3T3细胞系，用于转染pTet—on基因的胚胎干细胞阳性细胞克隆筛选的饲养层。 设计：细胞培养观察及DNA检测。 单位：中国医科大学实验动物部。 材料：NIH3T3细胞由中国医科大学细胞生物教研室惠赠，pWL／neo质粒为美国哈佛大学医学院金壮教授惠赠。G418、白血病抑制因子（胚胎干细胞GRO 106U／mL）、DMEM为GIBCOBRL产品，丝裂霉素-C、DIG标记及检测试剂盒为Roche产品，脂质体为Invitrogen产品，均购自沈阳联星生物技术有限公司。 方法：①通过脂质体转染的方法，将含有neo基因的质粒pWL／neo导入NIH3T3细胞中。②将细胞传至25mL培养瓶中，加入梯度浓度的G418继续培养，通过观察细胞生长及死亡情况测定G418对NIH3T3细胞最小致死量。③将转染的细胞进行传代，挑取单个细胞的克隆至24孔培养板继续进行筛选扩增，选用正常的NIH3T3细胞加入相同剂量的G418的选择性培养基作为筛选的阴性对照。④采用较低的细胞密度铺板，加入含最小致死量G418的DMEM培养基进行再次筛选；同时选用正常的NIH3T3细胞为对照，使用含有最小致死量G418及未加G418的DMEM培养基进行培养，采用光学显微镜对G418抗性NIH3T3细胞进行观察。⑤选用具有G418抗性的NIH3T3细胞和小鼠胚胎成纤维细胞分别作为饲养层细胞进行传代，采用光学显微镜对培养后胚胎干细胞-D3细胞进行观察，并制备细胞DNA，对其进行PCR和southern blot鉴定。 主要观察指标：①G418对NIH3T3细胞最小致死量的测定结果。②G418抗性NIH3T3细胞的筛选结果。③G418抗性NIH3T3细胞的生长观察结果。④胚胎干细胞-D3细胞生长观察结果及G418抗性NIH3T3细胞的鉴定。 结果：①G418对NIH3T3细胞最小致死量为500mg／L。②经过500mg/L G418压力筛选后，获得了抗G418细胞克隆。③抗性NIH3T3细胞的形态和生长速度与正常NIH3T3细胞没有差异。④饲养层培养的胚胎干细胞呈集落形状生长，边缘光滑，保持未分化的状态。用特异性核苷酸引物检测抗性细胞基因组DNA，可以扩增出对应的核苷酸片段，Southern blot鉴定结果表明neo基因片段已整合入G418抗性NIH3T3细胞中。 结论：成功地培育了G418抗性NIH3T3细胞，生长在G418抗性NIH3T3细胞饲养层上胚胎干细胞细胞基本保持正常胚胎干细胞细胞特征：     

OGP对NIH3T3细胞作用机理的研究

目的：研究成骨生长肽(OGP)对NIH3T3细胞的作用机理。方法：细胞计数法测定不同浓度的OGP14肽及5肽对NIH3T3细胞的促增殖作用，以流式细胞仪测定细胞膜上OGP的表达。观察0GP抗体对OGP促增殖作用的影响。结果：OGP14肽及5肽在10^-11mol／L时对NIH3T3细胞有促增殖作用，同时细胞膜上OGP表达增加；OGP抗体可抑制OGP的促增殖作用，同时使细胞膜上OGP表达下降。结论：OGP14肽及5肽可以通过对OGP的正反馈来调节NIH3T3细胞的增殖。     

芒果甙对鼻咽癌细胞系CNE3生长抑制与细胞周期阻滞的研究

目的 研究芒果甙在体外对人鼻咽癌细胞系CNE3细胞的生长抑制与细胞周期阻滞作用.方法 采用MTT法观察不同浓度芒果甙对CNE3细胞的生长抑制作用,采用流式细胞术检测不同浓度芒果甙诱导CNE3细胞周期阻滞作用.结果 ①不同浓度的芒果甙在不同时间对CNE3细胞有明显的生长抑制作用,且随着芒果甙浓度的升高和作用时间的延长而增高,芒果甙作用48,72 h后各浓度组较前一组抑制率差异均具有统计学显著性意义(P<0.01);②芒果甙作用72 h后,随芒果甙浓度的增加,各药物浓度组与空白对照组相比,S期与G2/M期细胞比例明显增多,出现S期与G2/M期阻滞,其中S期阻滞呈现剂量依赖性(P<0.05).结论 芒果甙能明显抑制CNE3细胞的增殖,并具有S期与G2期阻滞效应,具有潜在的抗肿瘤作用.     

Rh-SAA对3T3-L1脂肪细胞胰岛素敏感性的影响

目的:通过检测不同浓度和不同时间点Rh-SAA干预的3T3-LI脂肪细胞对葡萄糖的转运情况,了解Rh-SAA对脂肪细胞胰岛素敏感性的影响.方法:采用不同浓度的Rh-SAA(1、10、20 μg/mL)干预3T3-LI脂肪细胞48 h及20 μg/mL Rh-SAA分别干预细胞6、12、24、48 h,采用3H-2-DG摄入法检测细胞对葡萄糖的转运率.结果:Rh-SAA显著减少3T3-L1脂肪细胞在胰岛素刺激下的葡萄糖摄取,呈剂量和时间依赖效应.结论:Rh-SAA能降低3T3-L1脂肪细胞对胰岛素的敏感性,提示炎症与胰岛素抵抗密切相关.     

强力霉素调控小鼠E1A激活基因阻遏子表达NIH3T3细胞系的建立

背景:前期研究发现,E1A激活基因阻遏子蛋白调控人血管平滑肌细胞的增殖及表型转化,但其表达调控细胞增殖、分化、凋亡的量效关系有待深入探讨。目的:建立强力霉素调控小鼠E1A激活基因阻遏子表达的NIH3T3细胞系,拟为定量观察E1A激活基因阻遏子的生物学功能提供研究基础。设计、时间及地点:对比观察实验,于2006-05/2008-03在沈阳军区总医院全军心血管病研究所实验室完成。材料:强力霉素调控基因表达系统,3T3细胞系。方法:通过反转录-聚合酶链反应方法获得小鼠E1A激活基因阻遏子(mCREG)cDNA序列;将mCREGcDNA序列亚克隆入pRev-TRE载体中,构建强力霉素调控E1A激活基因阻遏子表达pRev-TRE-mCREG载体;分别经G418(新霉素)及潮霉素筛选表达pRev-Tet-On、pRev-TRE-mCREG的NIH3T3细胞克隆;反转录-聚合酶链反应鉴定转染细胞克隆中抗性基因和插入基因的表达。主要观察指标:Westernblotting检测不同浓度的强力霉素诱导后NIH3T3细胞中mCREG的表达。结果:成功构建了强力霉素可调控表达的pRev-TRE-mCREG重组载体;筛选出稳定表达双抗性基因的NIH3T3-mCREG细胞克隆;反转录-聚合酶链反应检测证实细胞克隆中G418及插入基因表达均为阳性;Western blotting检测发现随强力霉素诱导剂量的增加(0,0.01,0.1,1mg/L),NIH3T3-mCREG细胞中mCREG表达呈剂量依赖性增加。结论:成功建立强力霉素调控E1A激活基因阻遏子表达的小鼠NIH3T3细胞系。     

3-hydroxy-3-methylglutaric aciduria: Deficiency of 3-hydroxy-3-methylglutaryl coenzyme a lyase

A marked deficiency of 3-hydroxy-3-methylglutaryl coenzyme A lyase activity is present in cultured skin fibroblasts from a baby with 3-hydroxy-3-methylglutaric aciduria.     

Pharmacokinetics of 3''-fluoro-3''-deoxythymidine and 3''-deoxy-2'',3''-didehydrothymidine in rats.

Concentrations of 3'-fluoro-3'-deoxythymidine (FDT) and 3'-deoxy-2',3'-didehydrothymidine (D4T) in plasma declined in a biexponential fashion. Total clearance of D4T (1.75 +/- 0.22 liters/h/kg; mean +/- standard deviation) was significantly greater than that of FDT (1.19 +/- 0.19 liters/h/kg) owing to greater renal and nonrenal clearances of the former. Steady-state volumes of distribution of FDT (1.20 +/- 0.12 liters/kg) and D4T (1.07 +/- 0.15 liters/kg) were similar.     

Pharmacokinetics of 3''-fluoro-3''-deoxythymidine and 3''-deoxy-2'',3''-didehydrothymidine in rhesus monkeys.

3'-Fluoro-3'-deoxythymidine and 3'-deoxy-2',3'-didehydrothymidine are nucleoside analogs which inhibit human and simian immunodeficiency virus in vitro. The pharmacokinetic properties of these compounds in rhesus monkeys after intravenous, oral, and subcutaneous administration of the drug were compared. Half-lives, total clearances, and steady-state volumes of distribution of the two drugs were determined. The half-lives for the drugs by the different routes were between 0.58 and 1.4 h. Oral bioavailability of 3'-deoxy-2',3'-didehydrothymidine was incomplete, with an average of 42% +/- 15% of the dose reaching the systemic circulation. Absorption of 3'-fluoro-3'-deoxythymidine after oral administration was variable, with bioavailability ranging from 21 to 95%. Bioavailability after subcutaneous administration ranged from 59 to 77% for 3'-deoxy-2',3'-didehydrothymidine and from 52 to 59% for 3'-fluoro-3'-deoxythymidine. The ratio of concentrations in cerebrospinal fluid and serum for the drugs was about 0.15 at 1 h after drug administration and was independent of the route of administration, suggesting that a nucleoside carrier-mediated process is involved in the transport of these compounds to the central nervous system. Because of the similar metabolism of nucleoside analogs in monkeys and humans, the potential glucuronide formation was assessed. Whereas the glucuronide of 3'-fluoro-3'-deoxythymidine was readily detected in urine, the amount of 3'-deoxy-2',3'-didehydrothymidine glucuronidated was small or not detectable in one-half of the urine samples. Pharmacokinetic parameters for the two drugs were similar to each other and analogous to those for 3'-azido-3'-deoxythymidine in monkeys, suggesting that the same dose and scheduling of the drug can be used for all three compounds in prophylactic and therapeutic efficacy drug studies in rhesus monkeys.     

Antiviral Activity of 3-Deazaguanine, 3-Deazaguanosine, and 3-Deazaguanylic Acid

3-Deazaguanine (ICN 4221), 3-deazaguanosine (ICN 4793), and 3-deazaguanylic acid (ICN 5412) represent a new class of synthetic guanine analogs having antiviral activity. In vitro, nine ribonucleic acid and seven deoxyribonucleic acid viruses were inhibited, including influenza, parainfluenza, rhino-, vesicular stomatitis, adeno-, herpes-, cytomegalo-, vaccinia, pseudorabies, and myxoma viruses. They were effective orally against influenza types A and B and parainfluenza type 1 (Sendai) virus infections in mice, with a therapeutic index of 16 against the latter two viruses. The course of herpes encephalitis was altered only when the drugs were applied directly into the brain. In addition, these drugs were effective inhibitors of Friend leukemia virus-induced splenomegaly in mice; treatment also produced extensions of life in these animals.     

3-Methylcrotonylglycine excretion in 3-hydroxy-3-methylglutaric aciduria

1. 3-Methylcrotonylglycine was identified in urine from an infant with 3-hydroxy-3-methylglutaric aciduria. 2. The concentration of 3-methylcrotonylglycine in urine was approximately one sixth of that of the other metabolite of 3-methylcrotonyl-CoA, 3-hydroxyisovaleric acid. 3. The presence of both metabolites in the infant's urine indicates an inhibition of 3-methylcrotonyl-CoA carboxylase activity in tissues of the infant.     

亚急性甲状腺炎33例临床分析

目的 探讨亚急性甲状腺炎的病因、临床表现、诊断及治疗。方法 对33例患者的病史，临床表现，实验室资料及治疗进行总结分析。结果 患病的女性多见，占78．7％，病前无上怂病史记载：所有病例均有甲状腺肿大并触痛，彩超均有甲状腺肿大伴低回声，所有患者经口服泼尼松治疗效果好。结论 为减少漏诊、误诊，对咽痛伴发热病例，按上感治疗无效且持续时间较长应警惕本病。注意触诊甲状腺有无肿痛并进行必要的辅助检查。肾上腺糖皮质激素是治疗本病唯一确切有效药物。     

In-plane stimulated emission of polycrystalline CH3NH3PbBr3 perovskite thin films

Hybrid organic–inorganic lead halide perovskites have been investigated extensively within the last decades, for its great potential in efficient solar cells and as an ideal light source. Among the studies on stimulated emission (SE), the emission is either out-of-plane for polycrystalline films or in-plane with randomly aligned single microcrystals and nanowires. In this work, we revealed in-plane propagation of SE from bromine-based perovskite polycrystalline thin films (CH₃NH₃PbBr₃, or MAPbBr₃). The output from in-plane SE is an order higher than the out-of-plane emission. It is proposed that large crystalline flakes in the films lead to the in-plane lasing phenomena. The output coupling can be found at grain boundaries, intergrain gaps, and artificial structures. Simulative results support the experimental phenomenon that large crystalline grains are profitable for in-plane propagation and over 90% photons can be sufficiently outcoupled when the gap is larger than a micron. Considering the fabrication and handling convenience, we propose that the MAPbBr₃ thin films can be easily integrated for in-plane applications as the light source for photonic chips etc.

MAPbBr₃ perovskite thin film contains large crystal flakes, which support the in-plane stimulated emission and its propagation within these polycrystalline films. The emission scatters at the natural or artificial edge of the film.     

3-Hydroxy-3-methylglutaric aciduria: a new assay of 3-hydroxy-3-methylglutaryl-CoA lyase using high performance liquid chromatography

A new assay has been developed for 3-hydroxy-3-methylglutaryl-CoA lyase, the final enzyme in the leucine degradative pathway. The assay was performed by incubating lysates of fibroblasts with [glutaryl-3-¹⁴C](d,l)-3-hydroxy-3-methylglutaryl coenzyme A. The products were analysed by high performance liquid chromatography with continuous liquid scintillation counting. This provided simultaneous identification and quantification of one of the enzymatic products, [3-¹⁴C] acetoacetic acid. The mean 3-hydroxy-3-methylglutaryl-CoA lyase activity in fibroblasts from five controls was 732 ± 81 (SD) pmol/min · mg protein. Using this assay, we have studied skin fibroblasts cultured from a patient with 3-hydroxy-3-methylglutaric aciduria and found 3% of normal 3-hydroxy-3-methylglutaryl-CoA lyase activity. The activities in skin fibroblasts cultured from the parents were 46 and 53% of control activity which is consistent with heterozygocity. Kinetic studies of 3-hydroxy-3-methylglutaryl-CoA lyase in skin fibroblasts cultured from two normal subjects yielded K_m values of 14.4 and 18.8 μmol/l for 3-hydroxy-3-methylglutaryl-CoA.     

Low temperature scintillation performance of a Br-doped CH3NH3PbCl3 single-crystalline perovskite

Time response and light yield are two of the most important features of a scintillation detector, and are mostly determined by the luminescence properties of the scintillator. Here we have investigated the radioluminescence (RL) characteristics of a single-crystalline hybrid lead halide perovskite at both room temperature and low temperature. A dual-channel single photon correlation (DCSPC) system with a vacuum chamber is employed for the measurement. A rise time faster than 100 ps and several times enhancement of the crystal scintillation performances at low temperature have been observed. These behaviors demonstrated that bulk solution-grown single crystals of hybrid lead halide perovskites (MAPbCl₃ and Br-doped MAPbBr_0.08Cl_2.92, where MA = CH₃NH₃) can serve as stable scintillating materials for pulsed gamma detectors. In addition, this work provides a pathway for perovskite application and also attracts attention to investigating low-temperature scintillators.

Time response and light yield are two of the most important features of a scintillation detector, and are mostly determined by the luminescence properties of the scintillator.