TTNPB对人体外周血单核细胞和T淋巴细胞的影响

[目的]评价TTNPB对人体PBMC和T淋巴细胞的增殖的影响,确定剂量水平,建立一套通过受体途径研究维生素A功能的实验方法。[方法]用密度梯度分离法和磁性细胞分选法从健康人的外周血分离PBMC和T淋巴细胞,体外培养,采用MTT法,绘制人PBMC和T淋巴细胞的体外生长曲线,并测定在10-2mol/L,10-3mol/L,10-4mol/L,10-5mol/L,10-6mol/L,10-7mol/L6个不同浓度时的OD值,用方差检验分析各剂量对细胞的影响。[结果]TTNPB在10-2mol/L,PBMC的OD值为0.0600±0.0328(P﹤0.05),T淋巴细胞的OD值为0.0907±0.0110(P﹤0.05)。[结论]TTNPB在高浓度10-2mol/L对PBMC和T淋巴细胞增殖有显著的抑制作用;中等浓度有促进作用,但这还需要进一步的实验得出;低浓度时没有抑制或促进作用。     

TTNPB对体外培养的脐血T淋巴细胞分化的影响

[目的]研究视黄酸受体特异性激动剂TTNPB对体外培养的脐血T淋巴细胞表型分化的影响.[方法]采用体外人脐血淋巴细胞培养体系,以视黄酸受体(RARs)特异性激动剂TYNPB刺激细胞,TTNPB设立阴性对照组及10-4 mol/L、10-5 mol/L、10-6 mol/L、10-7 mol/L、10-8 mol/L 5个剂量组,培养72 h后收集淋巴细胞,采用流式细胞术分析T淋巴细胞表型分化情况.[结果]体外人脐血淋巴细胞培养体系能支持脐血T淋巴细胞的分化成熟.TTNPB剂量在10-4mol/L时,CDS+细胞比例为21.10%±0.10%(P=0.024),CD4+/CD8+比值为2.01±0.01(P=0.004).[结论]在体外TTNPB能抑制脐血CD4+CD8+胸腺细胞向CD8+细胞转化,TTNPB能升高CD4+/CD8+比值.视黄酸受体途径可能是维生素A调节免疫功能的机制之一.     

视黄酸受体特异性激动剂对脐血T淋巴细胞细胞因子的影响

[目的]研究体外培养中视黄酸受体(RAR)特异性激动剂四氢-四甲基萘丙烯苯甲酸(TTNPB)对脐血T淋巴细胞IFN-γ和IL-4产生的影响,初步探究其作用机制,为脐血运用于今后临床研究奠定基础.[方法]采用体外细胞培养体系,从足月正常分娩新生儿脐血中分离T淋巴细胞,加入不同浓度TTNPB(10-4 mol/L,100 mol/L,10-6 mol/L,10-7 mol/L,10-8 mol/L和空白对照组)体外培养,提取上清液,用EUSA法测定IFN-γ、IL-4的OD值,用单因素方差检验分析各剂量对IFN-γ、IL-4产生的影响.[结果]TTNPB在10-4 mol/L,IFN-γ的浓度为(12.11±5.57)pg/ml(P<0.05),IL-4的浓度为(2.937 1±0.076 6)pg/ml(P<0.05).[结论]TTNPB在10-4 mol/L下对IFN-γ有明显抑制作用,对IL-4有明显促进作用,这种调节可能是通过RAR选径起作用.     

锰对细胞免疫功能的影响

[目的 ]探讨锰对细胞免疫功能的影响。 [方法 ]利用体外培养的淋巴细胞 ,采用正交试验设计 ,观察不同染锰浓度和时间对淋巴细胞功能的影响。用溴化四氮唑盐检测法 (MTT法 )测定T淋巴细胞增殖功能和自然杀伤细胞 (NK细胞 )杀瘤细胞活性 ;用酶联免疫吸附试验 (ELISA)测定培养液中的白细胞介素 2 (IL 2 )含量。 [结果 ]随着培养液中的锰离子浓度由 1× 10 -6mol/L增大到 1× 10 -3 mol/L ,染锰时间由 2h延长到 4h ,T淋巴细胞增殖功能、NK细胞杀瘤细胞活性和IL 2含量均显著下降。 [结论 ]一定浓度的锰对淋巴细胞具有毒性作用     

P,P’-DDD对人乳腺癌细胞的增殖作用

[目的]观察对,对-二氯二苯基二氯乙烷（P,P’-DDD）对人乳腺癌（MCF-7）细胞增殖的影响。[方法]采用体外培养MCF-7细胞增殖试验检测DDD的类雌激素活性,并用生长曲线对其作用机制进行初步探讨。[结果]DDD染毒剂量在3×10^-7mol／L、3×10^-6mol／L时,存在刺激MCF-7细胞增殖的类雌激素活性（P〈0．05）,在3×10^-6mol／L剂量时其增殖效应最大。细胞生长曲线方面,DDD组在第3、5、7天各时间段细胞数均明显高于溶剂对照组（P〈0．05）。[结论]DDD在3×10^-7mol／L、3×10^-6mol／L具有刺激MCF．7细胞增殖的类雌激素活性。     

他莫昔芬对小鼠淋巴细胞功能的影响

[目的]研究他莫昔芬对小鼠淋巴细胞功能的影响.[方法]通过体外培养淋巴细胞,用MTT法分析他莫昔芬对小鼠淋巴细胞增殖及NK细胞杀伤活性的影响;用溶血空斑试验检测他莫昔芬对小鼠B淋巴细胞功能的影响. [结果]1、10、100、1 000 nmol/L他莫昔芬对小鼠T、B淋巴细胞增殖活性无明显增强或抑制作用(P＞0.05);1、10、100、1 000 nmol/L他莫昔芬显著增加小鼠溶血空斑形成细胞的数量(P<0.05);10、100、1 000 nmol/L他莫昔芬对小鼠NK细胞的杀伤活性有明显增强作用(P<0.01). [结论]他莫昔芬能增加小鼠溶血空斑形成细胞的数量和提高NK细胞的杀伤活性,对免疫功能有不同程度的调节作用.     

精氨酸通过肝脏调节免疫效应的作用及其机制

目的 :　观察精氨酸 (arginine,Arg)的免疫促进作用是否部分通过肝脏并探讨其机制。方法 :　3 H- Td R掺入法测定 Arg对 T淋巴细胞增殖的直接效应 ;制备大鼠肝细胞进行原代无血清培养 (DMEM/ F1 2 ) ,用不同浓度的 Arg(μmol/ L:0 ,7.5 ,75 ,75 0 ,75 0 0 )处理肝细胞 ,收集 0、2 4、48、72 h培养上清液。将上述上清液加入脾细胞悬液中 ,测定 T淋巴细胞增殖反应 ,NK细胞活性、IL- 2活性 ,淋巴细胞活化时 [Ca2 + ]i变化。结果 :　完全 RPMI1 6 4 0培养基中直接补充精氨酸不能促进脾淋巴细胞增殖 ;Arg处理大鼠肝细胞后 ,其培养上清液对淋巴细胞 [Ca2 + ]i、IL- 2活性和T细胞增殖有明显的促进作用 ,其中 75 0 0μmol/ L Arg最强。结论 :　在体外 ,Arg可通过刺激肝细胞分泌有关因子促进 T淋巴细胞增殖。     

镉通过类雌激素样作用促进人乳腺癌MCF-7细胞增殖

[目的]研究镉对人乳腺癌MCF-7细胞增殖及雌激素受体(estrogen receptor,ER)蛋白表达水平的影响及其可能机制。[方法]用1×10~(-6)、1×10~(-7)、1×10~(-8)、1×10~(-9)、1×10~(-10)、1×10~(-11) mol/L的17β-雌二醇(后称雌激素)培养不同来源的A、B两株MCF-7细胞4 d,应用CCK8法筛选敏感细胞株并确定雌激素促细胞增殖作用最强的浓度。用1×10~(-6)、1×10~(-7)、1×10~(-8)、1×10~(-9)、10~(-10)、1×10~(-11) mol/L的镉溶液染毒敏感细胞株,确定镉促进细胞增殖的最大浓度。设置阴性对照组(去雌激素培养液)、实验组(1×10~(-8) mol/L镉)、阳性对照组(1×10~(-9) mol/L雌激素),通过克隆形成实验检测镉对MCF-7细胞集落形成能力的影响。利用ER抑制剂(ICI182780)初步探讨镉致细胞增殖的可能机制,增设实验抑制剂组(1×10~(-8) mol/L镉+1×10~(-7) mol/L ER抑制剂)、阳性抑制剂组(1×10~(-9) mol/L雌激素+1×10~(-7) mol/L ER抑制剂),通过CCK8法、流式细胞术和Western blot分别检测各组的细胞增殖率、细胞周期S期比例和ER表达水平。[结果]实验所用B株MCF-7细胞为雌激素敏感细胞株,且当雌激素浓度为1×10~(-9) mol/L时,对MCF-7细胞的促进增殖作用[(203.55±36.65)%]最大(P0.001)。与阴性对照组(100%)相比,1×10~(-8)~1×10~(-10) mol/L镉溶液可促进MCF-7细胞增殖[(163.78±31.90)%~(176.88±10.06)%](P0.001)。1×10~(-8) mol/L镉可促进细胞集落形成(P0.05),增加细胞S期比例(P0.001),提高细胞ER蛋白表达水平(P0.001)。与实验组相比,加入ER抑制剂能够抑制镉对MCF-7细胞的促增殖作用[由(135.17±23.96)%降至(107.66±7.64)%](P0.01),抑制镉诱导的细胞S期比例增加[由(24.17±0.53)%降至(12.36±0.43)%](P0.001),拮抗镉诱导的ER表达增加[由(56.19±3.67)%降至(38.84±1.04)%](P0.05)。[结论]镉可以促进人乳腺癌细胞MCF-7增殖和ER表达,这种作用可被ER抑制剂所抑制,提示镉可能具有雌激素样作用。     

过氧化氢和环磷酰胺对辐射损伤小鼠脾淋巴细胞增殖的作用

目的：探讨化学因子对辐射损伤小鼠脾淋巴细胞增殖的影响。方法：采用^3H—TdR掺入法观察小鼠脾淋巴细胞增殖的变化。结果：与刀豆球蛋白A(Concanavalin A，Con A)同时作用于小鼠脾淋巴细胞的10^-6mol/L，和10^-5mol/L的H2O2对Con A刺激的小鼠脾淋巴细胞增殖有明显的促进作用，H2O2在Con A作用于小鼠脾淋巴细胞56h加入，H2O2对小鼠脾淋巴细胞增殖表现为抑制效应.当环磷酰胺(cyclophosphamide，CP)浓度≥10^-3mol/L和≥10^-3mol／L时，CP对脂多糖(lipopolysaccharide，LPS)和Con A刺激的小鼠脾淋巴细胞增殖分别表现为明显的抑制作用。当H2O2浓度≥10^-6mol／L和CP浓度≥10^-3mol／L时，H2O2和CP对辐射损伤的小鼠脾淋巴细胞增殖具有明显的抑制作用。结论：低浓度H2O2可以诱导小鼠脾淋巴细胞增殖的适应性反应，而H2O2联合CP不能诱导对辐射损伤小鼠脾淋巴细胞增殖的抗性．     

促肾上腺皮质激素释放因子对镉的淋巴细胞免疫毒性的体内外双向调节作用

目的 探讨体内外镉的免疫功能抑制过程中促肾上腺皮质激素释放因子（CRF）的修饰调节使用。方法 用垂体前叶细胞培养法测定中枢下丘脑和外周血浆CRF水平,用[^3H-TdR]掺入法分别检测体内、体外染毒的鼠脾T、B淋巴细胞增殖转化功能。结果 （1）整体实验动物各镉剂量组中枢下丘脑组织CRF水平均升高,有丝分裂原诱导脾T、B淋巴细胞增殖转化的刺激指数均低于对照组,差异有显著性（P＜0．05）;下丘脑CRF水平与T、B淋巴细胞增殖指数之间呈负相关关系（r＝－0．6750、－0．7560,P＜0．05）;外周血浆CRF水平各组间差异无显著性（P＜＞0．05。（2）体外染毒时,镉明显抑制大鼠脾淋巴细胞的增殖转化;1．0-100．0nmol/LCRF明显增强T、B淋巴细胞增殖转化能力;当以1．0nmol/LCRF与5－20μmol/L氯化镉联合作用对各实验组两种免疫组织[^3H-TdR]掺入率依然高于对照组,差异均有显著性（P＜0．05）。体内染镉时存在中枢组织CRF介导的免疫功能抑制性调节作用,但体外CRF却能增强免疫细胞活性并阻断镉对鼠脾淋巴细胞的直接免疫功能抑制。CRF对镉的淋巴细胞免疫毒性具有双向调节作用。     

广西壮族自治区艾滋病抗病毒治疗患者CD4+T淋巴细胞动态趋势及影响因素分析

目的 了解广西壮族自治区(广西)抗病毒治疗患者CD₄⁺T淋巴细胞动态趋势及影响因素。方法 选择2013年1月1日后首次开始抗病毒治疗的成年艾滋病患者,对基线、治疗后6个月和12个月时间段的CD₄⁺T淋巴细胞计数结果进行回顾分析,采用一般线性模型重复测量方差分析描述和分析患者CD₄⁺T淋巴细胞动态趋势及影响因素。结果 4 082例患者基线、治疗后6个月和12个月的CD₄⁺T淋巴细胞计数均值分别为(195.3±155.7)cells/mm³、(331.9±202.6) cells/mm³ 和 (380.9±221.3) cells/mm³, 仅有时间效应时,总体差异有统计学意义(F=3 161.124,P=0.000)。治疗后CD₄⁺T淋巴细胞计数随时间推移而提升,主要影响因素为性别、年龄、基线CD₄⁺T淋巴细胞计数、治疗方案、漏服药物和停药。受性别、年龄、治疗方案、漏服药物、停药等因素影响,治疗后CD₄⁺T淋巴细胞计数随时间推移而呈线性提升趋势。受基线CD₄⁺T淋巴细胞计数、停药等因素影响,治疗后CD₄⁺T淋巴细胞计数提升趋势符合二次方曲线方程。结论 广西艾滋病患者抗病毒治疗后CD₄⁺T淋巴细胞计数提升速度受多种因素影响。针对不同患者适时进行抗病毒治疗可以获得良好预后结果。     

Ethanol-induced alterations in the function of cerebral GABAA receptor complex: effect on GABA-dependent 36Cl- influx into cerebral membrane vesicles.

Effect of addition in vitro of ethanol on the functions of the GABAA receptor complex was investigated using synaptic membrane preparation obtained from mouse brain. Ethanol (50-200 mM) had no significant effect on the specific bindings of [3H]muscimol to GABAA receptor and [3H]flunitrazepam to benzodiazepine receptor in cerebral particulate preparations. However, ethanol induced an inhibition of [3H]TBOB binding to Cl- channel and a suppression of flunitrazepam-induced enhancement of [3H]muscimol binding and of salsolinol-induced accentuation of [3H]flunitrazepam binding to cerebral particulate fraction. In contrast, ethanol facilitated GABA-dependent 36Cl- influx but eliminated the stimulatory effects of flunitrazepam and salsolinol on GABA-dependent 36Cl- influx into cerebral membrane vesicles. These results suggest that ethanol may facilitate the function of GABA-gated chloride channel in spite of inducing deteriorations of antagonist binding capacity of chloride channel as well as of the functional coupling between GABAA receptor and benzodiazepine receptor in the brain.     

Ionotropic excitatory amino acid receptor ligands. Synthesis and pharmacology of a new amino acid AMPA antagonist

We have previously described the potent and selective (RS)-2-amino-3-(3-hydroxy-5-methyl-4-isoxazolyl)propionic acid (AMPA) receptor agonist, (RS)-2-amino-3-(3-carboxy-5-methyl-4-isoxazolyl)propionic acid (ACPA), and the AMPA receptor antagonist (RS)-2-amino-3-[3-(carboxymethoxy)-5-methyl-4-isoxazolyl]propionic acid (AMOA). Using these AMPA receptor ligands as leads, a series of compounds have been developed as tools for further elucidation of the structural requirements for activation and blockade of AMPA receptors. The synthesized compounds have been tested for activity at ionotropic excitatory amino acid (EAA) receptors using receptor binding and electrophysiological techniques, and for activity at metabotropic EAA receptors using second messenger assays. Compounds 1 and 4 were essentially inactive. (RS)-2-Amino-3-[3-(2-carboxyethyl)-5-methyl-4-isoxazolyl]propionic acid (ACMP, 2), on the other hand, was shown to be a selective AMPA receptor antagonist (IC(50) = 73 microM), more potent in electrophysiological experiments than AMOA (IC(50) = 320 microM). The isomeric analogue of 2, compound 5, did not show AMPA antagonist effects, but was a weak NMDA receptor antagonist (IC(50) = 540 microM). Finally, compound 3, which is an isomer of ACPA, turned out to be a very weak NMDA antagonist, and an AMPA receptor agonist approximately 1000 times weaker than ACPA. None of the compounds showed agonist or antagonist effects at metabotropic EAA receptors.     

Enhancement of Ketone Supplements-Evoked Effect on Absence Epileptic Activity by Co-Administration of Uridine in Wistar Albino Glaxo Rijswijk Rats

Both uridine and exogenous ketone supplements decreased the number of spike-wave discharges (SWDs) in a rat model of human absence epilepsy Wistar Albino Glaxo/Rijswijk (WAG/Rij) rats. It has been suggested that alleviating influence of both uridine and ketone supplements on absence epileptic activity may be modulated by A₁ type adenosine receptors (A₁Rs). The first aim was to determine whether intraperitoneal (i.p.) administration of a specific A₁R antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX; 0.2 mg/kg) and a selective adenosine A_2A receptor antagonist (7-(2-phenylethyl)-5-amino-2-(2-furyl)-pyrazolo-[4,3-e]-1,2,4-triazolo [1,5-c]pyrimidine) (SCH 58261; 0.5 mg/kg) have a modulatory influence on i.p. 1000 mg/kg uridine-evoked effects on SWD number in WAG/Rij rats. The second aim was to assess efficacy of a sub-effective dose of uridine (i.p. 250 mg/kg) combined with beta-hydroxybutyrate salt + medium chain triglyceride (KSMCT; 2.5 g/kg, gavage) on absence epilepsy. DPCPX completely abolished the i.p. 1000 mg/kg uridine-evoked alleviating effect on SWD number whereas SCH 58261 was ineffective, confirming the A₁R mechanism. Moreover, the sub-effective dose of uridine markedly enhanced the effect of KSMCT (2.5 g/kg, gavage) on absence epileptic activity. These results demonstrate the anti-epilepsy benefits of co-administrating uridine and exogenous ketone supplements as a means to treat absence epilepsy.     

食品中单核细胞增生李斯特菌PCR快速检测方法

[目的]建立食品中单核细胞增生李斯特菌的快速、敏感、特异的PCR检测方法。[方法]选择Hly基因作为靶序列设计一对引物，用该引物对单增李斯特菌和10株非单增李斯特菌进行PCR扩增，并且用此方法对30份食品进行检测。[结果]扩增片断表现出极好的单增李斯特菌特异性，最低检出限为190cfu/ml。[结论]本实验建立的PCR检测方法为一种简便、快速、敏感、特异的单增李斯特菌检测方法。     

Importance of extracellular Ca2+ and intracellular Ca2+ release in ethanol-induced contraction of cerebral arterial smooth muscle.

The present study was designed to investigate the roles of extracellular Ca2+ ([Ca2+]0) influx and intracellular free Ca2+ ([Ca2+]i) release in ethanol-induced contractions of isolated canine cerebral arteries and primary cultured, cerebral vascular smooth muscle cells. Ethanol (20-200 mM) produced significant contractions in isolated canine basilar arterial rings in a concentration-dependent manner. Removal of [Ca2+]0 and pretreatment of canine basilar arterial rings with verapamil (an antagonist of voltage-gated Ca2+ channels), thapsigargin (a selective antagonist of the sarcoplasmic reticulum Ca2+ pump), caffeine plus ryanodine (a specific antagonist of ryanodine-sensitive Ca2+ release), or heparin (an inositol 1,4,5,-trisphosphate [InsP3]-mediated Ca2+ release antagonist) markedly attenuated (approximately 50%-80%) ethanol-induced contractions. The absence of [Ca2+]0 and preincubation of primary single smooth muscle cells obtained from canine basilar arteries with verapamil, thapsigargin, heparin, or caffeine plus ryanodine markedly attenuated (approximately 50%-80%) the transient and sustained elevations in [Ca2+]i induced by ethanol. Results of the present study suggest to us that both Ca2+ influx through voltage-gated Ca2+ channels and Ca2+ release from intracellular stores (both InsP3 sensitive and ryanodine sensitive) are required for ethanol-induced contractions of isolated canine basilar arteries.     

Ac-D-NAL(2)1,4FD-Phe2,D-Trp3,D-Arg6]-LHRH, a potent antagonist of LHRH, produces transient edema and behavioral changes in rats

Acute toxicity studies of [Ac-D-NAL(2)1,4FD-Phe2,D-Trp3,D-Arg6]-LHRH (LHRH-A), a potent antagonist of LHRH were performed. Subcutaneous administration of this peptide to rats induced transient edema of the face and extremities. This effect was maximal 3-5 h after peptide administration and subsided by 24 h. These effects were not seen with an LHRH agonist or two other antagonists. This side effects of LHRH-A was peculiar to rats and not observed in mice, rabbits and rhesus monkeys. Intravenous administration led within minutes to depression of spontaneous activity in rats and monkeys. We conclude that some LHRH antagonists produce species specific effects on vascular permeability and spontaneous activity.     

Site-directed mutagenesis of the rat beta1-adrenoceptor. Involvement of Tyr356 (7.43) in (+/-)cyanopindolol but not (+/-)[125Iodo]cyanopindolol binding

To determine the role played by Tyr(356 (7.43)) in the rat beta(1)-adrenoceptor in binding the antagonists (+/-)cyanopindolol (4-[3-(t-butylamino]-3-(2'-cyano-indoloxy)-2-propanolol) and its iodinated analogue (+/-)[(125)Iodo]cyanopindolol (1-(t-butylamino]-3-(2'-cyano-3'-iodo-indoloxy)-2-propanolol), Tyr(356 (7.43)) was mutated to either Phe or Ala and binding affinities determined for wild type and mutant rat beta(1)-adrenoceptors. Our results indicate that Tyr(356 (7.43)) is important for (+/-)cyanopindolol, but not (+/-)[(125)Iodo]cyanopindolol, binding and that (+/-)cyanopindolol adopts a "reverse" binding orientation whereas (+/-)[(125)Iodo]cyanopindolol cannot be accommodated in this binding mode. We define a "reverse" antagonist binding mode as one where the aryloxy moiety interacts with residues on transmembrane helices 1, 2, 3 and 7. The beta(1)-adrenoceptor site-directed mutagenesis results are the first to support a "reverse" antagonist binding orientation and the involvement of Tyr(356 (7.43)) in this binding mode.     

Cell mediated immune responses in children with iron deficiency and combined iron and zinc deficiency

This study has been carried out to evaluated cell-mediated immune response in children with iron deficiency alone and iron deficiency associated with zinc deficiency, comparatively. Thirty patients were studied, with thirteen having iron and zinc deficiency. Absolute lymphocyte counts were found to be above 1200/cu mm whereas, E-rosette forming T-Lymphocytes were significantly lower than the control subjects in both groups, with much lower values for combined iron and zinc deficient children. Although delayed hypersensitivity responses were impaired in both groups of patients, it was found to be much lower in combined ironzinc deficient group. It is concluded that both iron deficiency and iron-and zinc deficiency may impair cell mediated immune responses, however zinc may have a greater influence.     

Synthesis and evaluation of 7-amino-2-(2(3)-furyl)-5-phenylethylamino-oxazolo[5,4-d]pyrimidines as potential A2A adenosine receptor antagonists for positron emission tomography (PET)

The brain A2A adenosine receptor (A2AAR) participates with the dopamine D2 receptor in the control of movement and also might influence behavior. Because PET is an important tool for studying the roles of receptors in disease, a ligand for imaging the brain A2AAR is desirable. This report describes the synthesis and A2AAR antagonist activities of a panel of phenyl-substituted 7-amino-2-(2-furyl)-5-phenylethylamino-oxazolo[5,4-d]pyrimidines, 11aa-af, and their 3-furyl congeners, 11ba-bd. In competitive binding studies all compounds displaced [3H]CGS21680 from the A2AAR with Ki values of 14-33 nM with selectivity for the A2AAR over the A1AR of 5- to 94-fold. Autoradiography of brain sections showed a high level of unspecific binding that obscured specific binding. Thus, these compounds are not promising PET ligands.