COX．2介导的TH2类细胞因子与重症肝炎的关系研究

目的：探讨重症肝炎与COX-2的关系,为临床防治重症肝炎提供新的治疗理论和方法。方法：用FH643A-γ测量仪检测50例肝病患者和25例正常对照血清中PGs代谢产物TXB2、6-keto-PGF1α的水平。结果：各组肝病患者血清中TXB2和6-keto-PGFlet的表达水平均明显高于正常人（P〈0．001）,并且随着病情的好转,T／6-K比值明显下降,但肝病患者各组间细胞因子水平无显著性差异（p〉0．05）。结论：检测重症肝炎患者血清TXB2、6-keto—PGF1α的水平和T／6-K比值不仅对揭示肝病发生发展过程有一定的实用价值．而且对监测病情转归和预后有重要临床意义。     

缺血预适应对肢体缺血/再灌注骨骼肌、小肠和肺损伤的影响

目的:观察肢体缺血/再灌注(LI/R)后骨骼肌、小肠、肺功能损伤变化,并探讨缺血预适应(IPC)的保护效应及机制。方法:实验用雄性Wistar大鼠24只,随机分为3组(n=8):对照(Control)组,缺血/再灌注(I/R)组和缺血预适应(IPC+I/R)组。分别测定血浆乳酸脱氢酶(LDH)、肌酸激酶(CK)、活性氧(ROS)、丙二醛(MDA)、动脉血氧分压(PaO2)和二氧化碳分压(PaCO2),测定血浆血栓素B2(TXB2),6-酮-前列腺素F1α(6-keto-PGF1α)的含量以及TXB2/6-keto-PGF1α比值的变化;测定骨骼肌、小肠、肺组织髓过氧化物酶(MPO)含量,肺湿干比(W/D)及小肠组织DAO含量。观察骨骼肌组织的形态学变化。结果:IPC+I/R组血浆LDH、CK、ROS、MDA、TXB2/6-keto-PGF1α比值明显低于I/R组,PaO2较I/R组明显升高。IPC+I/R组肺湿干比(W/D),骨骼肌、肺、小肠组织MPO含量明显低于I/R组,而小肠DAO活性升高。骨骼肌组织病理学改变减轻。结论:缺血预适应减轻了缺血/再灌注后骨骼肌、小肠、肺功能的损伤,其机制可能与降低氧化损伤、改善TXB2/6-keto-PGF1α的平衡关系有关。     

塞来昔布对慢性低O_2高CO_2大鼠肺动脉高压的影响及其机制研究

目的：探讨塞来昔布对慢性低O2高CO2大鼠肺动脉高压的作用。方法：将SD大鼠分为正常对照组,慢性低O2高CO2组,慢性低O2高CO2＋塞来昔布组。用电镜、放免等方法,观察各组大鼠肺动脉平均压、颈动脉平均压、肺细小动脉显微结构、血浆和肺匀浆血栓素B2（TXB2）及6-酮-前列腺素F1α（6-keto-PGF1α）含量的变化。结果：①慢性低O2高CO2组平均肺动脉压（mPAP）比正常组显著升高,塞来昔布组的mPAP比慢性低O2高CO2组显著升高,3组间平均颈动脉压（mCAP）比较差异无显著性。②慢性低O2高CO2组与正常对照组相比血浆和肺匀浆TXB2浓度、TXB2/6-keto-PGF1α比值显著增高,6-keto-PGF1α浓度显著下降;塞来昔布组与慢性低O2高CO2组相比血浆和肺匀浆TXB2浓度无明显变化、TXB2/6-keto-PGF1α显著升高,6-keto-PGF1α显著下降。③光镜下慢性低O2高CO2组与正常组相比,肺细小动脉管壁面积/管总面积（WA/TA）和肺细小动脉中膜厚度（PAMT）均显著增高。塞来昔布组与慢性低O2高CO2组相比WA/TA和PAMT显著增高。④电镜下慢性低O2高CO2组大鼠肺细小动脉内皮细胞吞饮小泡增多,血管壁增厚,中膜平滑肌细胞增生,纤维细胞增多,肺泡II型上皮细胞微绒毛脱落;塞来昔布组中膜平滑肌细胞增大、增多,胞浆肌丝丰富,平滑肌细胞间隙增宽,肺泡隔胶原纤维增生明显。结论：塞来昔布可能有加重慢性低O2高CO2性肺动脉高压和肺血管结构重建倾向,过度抑制COX-2,使TXA2/PGI2比值升高可能是其作用机制之一。     

黄芪注射液对2型糖尿病动物模型KKAy小鼠脑微血管病变的影响

目的:观察黄芪注射液对2型糖尿病动物模型KKAy小鼠脑微血管病变的影响,探讨黄芪注射液对糖尿病脑血管病变的保护作用.方法:饲养至14周龄的雄性KKAy小鼠随机分成模型组和黄芪注射液治疗组(每日腹腔给药,剂量为3 mL/kg),同龄雄性C57BL/6J小鼠作为对照组.血糖仪测量20、24、28周龄时各组小鼠的空腹血糖水平.28周龄时处死各组小鼠,放射免疫法检测血清6-酮-前列腺素-F1α( 6-Keto-PGF1α)和血栓素B2(TXB2)的含量.透射电子显微镜观察脑组织超微结构变化.结果:模型组KKAy小鼠从20周龄开始血糖水平明显高于正常组小鼠(P＜0.01);黄芪治疗组小鼠从20周龄开始血糖水平明显高于正常组小鼠(P＜0.01),但低于模型组小鼠(P＜0.05或P＜0.01).模型组小鼠血清6-Keto-PGF1α水平较正常组降低(P＜0.01),TXB2含量增高(P＜0.01);与模型组相比,黄芪注射液治疗组小鼠6-Keto-PGF1α水平升高(P＜0.01),TXB2含量下降(P＜0.01).透射电镜显示模型组小鼠神经细胞胞核染色质疏松,线粒体肿胀,粗面内质网缩小,核糖体减少;治疗组小鼠以上病变明显改善.结论:黄芪注射液可以有效改善2型糖尿病动物模型KKAy小鼠脑微血管病变,保护神经细胞结构.     

大鼠脑缺血-再灌注前后血浆TXB2及6-Keto-PGF1α含量的变化

目的:本实验旨在揭示脑缺血-再灌注损伤前后大鼠血浆血栓素B2(TXB2)及6-酮前列腺环素F1(6-Keto-PGF1α)的动态变化.方法:制作脑缺血-再灌注大鼠模型,45只健康SD大鼠随机分为正常对照组、模型组和假手术组大鼠.在术后1天、3天、5天和7天分别观察大鼠血浆TXB2和6-Keto-PGF1α含量.结果:线栓大脑中动脉后造成脑缺血,血浆TXB2含量和TXB2/6-Keto-PGF1α比值明显高于假手术组和正常对照组(p＜0.05或0.01),在缺血第1天最显著.结论:脑缺血-再灌注损伤后,血浆TXB2和TXB2/6-Keto-PGF1α的变化规律可为临床缺血性脑损伤治疗提供重要参考.     

黄芪丹参不同配伍对气虚血瘀模型大鼠血液流变学和血管内皮因子的影响

目的:研究黄芪丹参不同配伍对气虚血瘀证大鼠血液流变学和血管内皮因子的影响。方法:将SD大鼠随机分为对照组、模型组、丹参组、黄芪组、黄芪丹参1:1组、2:1组、4:1组,采用限食、游泳、皮下注射肾上腺素方法建立气虚血瘀证大鼠模型,对照组与模型组给予蒸馏水,用药组分别给予不同剂量的丹参、黄芪及不同比例黄芪丹参配伍灌胃给药治疗,连续27日。测定血液流变学指标,内皮素-1(ET-1)、一氧化氮(NO)、血栓素B2(TXB2)和6-酮-前列腺素-F1α(6-keto-PGF1α)含量。结果:模型组大鼠血液流变学参数和血管内皮因子与对照组比较,差异具有统计学意义(P0.01)。与模型组比较,黄芪丹参1:1组、2:1组、4:1组血液流变学参数显著降低(P0.05或P0.01),NO、6-keto-PGF1α、6-keto-PGF1α/TXB2明显升高(P0.01),ET-1、TXB2明显降低(P0.01)。与1:1组比较,2:1组血液流变学参数显著下降(P0.05或P0.01),NO、6-keto-PGF1α、6-keto-PGF1α/TXB2显著升高(P0.05或P0.01),TXB2明显降低(P0.05)。2:1组大鼠血液流变学参数和血管内皮因子与4:1组比较,差异具有统计学意义(P0.05或P0.01)。结论:黄芪丹参配伍对气虚血瘀证大鼠模型具有改善血液流变及保护血管内皮作用,其中以黄芪丹参2:1组最佳。     

0期诊断技术对军事训练所致椎间盘源性下腰痛患者血清IL-1β, 6-keto-PGF1α及TNF-α水平的影响及其意义

目的:探讨0期诊断技术对军事训练所致椎间盘源性下腰痛患者血清白介素1β(IL-1β)、6酮前列腺素F1α(6-keto-PGF1α)、肿瘤坏死因子α(TNF-α)水平的影响及其意义。方法:随机抽取某部2014年度新兵一营306名新兵为实验组,新兵二营290名新兵为对照组,在入伍复检时为每名士兵抽血检测相关血清学指标,并建立健康档案。由经过技术培训的医师对实验组采用0期诊断技术,分别在新兵训练开始后的第2、4、6、8、10、12周进行0期椎间盘源性下腰痛的筛查,并实施相应的防治措施,同时,按照"军标"对所有实验对象进行下腰痛的常规诊治及登统计,对于诊断为0期椎间盘源性下腰痛阳性和椎间盘源性下腰痛阳性的士兵及时抽血检测IL-1β、6-keto-PGF1α、TNF-α血清值。结果:实验组椎间盘源性下腰痛总发生率(2.94%)明显低于对照组(9.66%)(x~2=11.527,P0.001)。在新训开始后的第8、10、12周,实验组椎间盘源性下腰痛的发生率均明显低于对照组(P0.05);实验组和对照组阳性士兵血清IL-1β、6-keto-PGF1α、TNF-α水平比较无差异,0期阳性和阳性士兵血清IL-1β、6-keto-PGF1α、TNF-α水平与训练前相比均明显升高(P0.05),0期阳性士兵血清IL-1β、6-keto-PGF1α、TNF-α值均明显低于本组和对照组阳性士兵(P0.05)。结论:0期诊断技术预防军事训练所致新兵椎间盘源性下腰痛的效果明显,通过监测IL-1β、6-keto-PGF1α、TNF-α血清的水平变化也进一步证实0期诊断技术用于预防军事训练所致椎间盘源性下腰痛的可行性。     

瘦素上调动脉内皮细胞环氧合酶-2 及其产物的实验研究

目的:研究瘦素对大鼠主动脉内皮细胞(RAECs)环氧化酶-2(COX-2)及其下游产物前列环素和血栓素A2表达的影响。方法:酶消化法分离RAECs;免疫荧光检测细胞Ⅷ因子的表达;实时荧光定量PCR检测不同浓度瘦素(10~(-10)M、10~(-9)M、10~(-8)M)和时间(36h、48 h)处理后对RAECs COX-2 mRNA表达的影响;Western blot检测各组COX-2蛋白的改变;酶联免疫吸附测定(ELISA)检测各组细胞上清液中6-酮前列腺素F1α(6-Keto PGF1α)和血栓素B2(TXB2)的水平。结果:酶消化法6天后见细胞呈长梭形或多边形,培养10天后细胞呈典型的"铺路石"样改变,免疫荧光检测发现Ⅷ因子在胞浆中特异性表达,内皮细胞纯度达90%。与对照组(PBS处理)相比较,瘦素处理后,各组大鼠主动脉内皮细胞COX-2 m RNA及蛋白表达量均有升高趋势,两者表达量均随浓度增加和处理时间延长而升高(P0.05)。而6-Keto PGF1α水平升高及TXB2/6-Keto PGF1α比例降低仅见于相对高浓度瘦素(10-9、10-8M)处理后(P0.05),但瘦素对RAECs TXB2表达无影响(P0.05)。结论:瘦素增加RAECs炎性标志物COX-2及其下游舒张血管产物6-Keto PGF1α的表达,降低TXB2(收缩血管物质)/6-Keto PGF1α比例。COX-2可能在瘦素介导的高血压发病中起重要作用,但因其下游产物则有利于血管舒张,故选择性COX-2抑制剂可能会增加高血压风险。     

红花注射液对慢性低O2高CO2肺动脉高压的影响

目的：探讨红花注射液对大鼠在慢性低O2高CO2下肺动脉高压的抑制作用。方法：将SD大鼠分为对照组,慢性低O2高CO2组,慢性低O2高CO2＋红花注射液组。用电镜、放免等方法,观察各组大鼠肺动脉平均压、颈动脉平均压、肺细小动脉显微结构、血浆和肺匀浆TXB2及6-keto-PGF1a含量的变化。结果：①慢性低O2高CO2组mPAP比对照组显著增高,红花注射液组的mPAP比慢性低O2高CO2组显著降低,3组间mCAP比较差异无显著性。②慢性低O2高CO2组与对照组相比血浆和肺匀浆TXB2浓度、TXB2／6-keto-PGF1a比值显著增高,6-keto-PGF1a浓度显著下降;红花注射液组与慢性低P2高CO2相比血浆和肺匀浆TXB2浓度、TXB2／6-keto-PGF1a显著下降,6-keto-PGF1a显著升高。③光镜下慢性低O2高CO2组与对照组相比,肺细小动脉管壁面积／管总面积（WA／TA）和肺细小动脉中膜厚度（PAMT）均显著增高。红花注射液组WA／TA和PAMT显著降低。④电镜下慢性低O2高CO2组大鼠肺细小动脉内皮细胞吞饮小泡增多,血管壁增厚,中膜平滑肌细胞增生,纤维细胞增多,肺泡Ⅱ型上皮细胞微绒毛脱落;红花注射液组肺细小动脉中膜平滑肌细胞增生减轻,纤维细胞少,胶原纤维减少,肺泡Ⅱ型上皮细胞微绒毛丰富、结构清。结论：红花注射液有减轻慢性低O2高CO2性肺动脉高压和肺血管结构重建的作用,可能与抑制TXA2的合成,保护血管内皮细胞,使TXA2/PGI2比值降低有关.     

阿斯匹林对大鼠在低O2高CO2下肺动脉高压的作用

目的：研究阿斯匹林(ASA)对慢性低O2高CO2性肺动脉高压的抑制作用。方法：将SD大鼠分为正常对照组,慢性低O2高CO2组,慢性低O2高CO2 阿斯匹林组。用光镜、放射免疫等方法,观察各组大鼠肺动脉平均压(mPAP)、颈动脉平均压(mCAP)、肺细小动脉显微结构、血浆和肺匀浆TXB2及6-keto-PGF1α含量的变化。结果：①低O2高CO2组mPAP比正常组显著增高,ASA组的mPAP比低O2高CO2组显著降低,3组间mCAP比较差异无显著性。②光镜下低O2高CO2组与正常组相比,肺细小动脉管壁面积／管总面积(WA／TA)和肺细小动脉中膜厚度(PAMT)均显著增高。ASA组WA／TA和PAMT显著降低。③低O2高CO2组血浆和肺匀浆TXB2、6-keto-PGF1α浓度以及TXB2／6-keto-PGF1α比正常组显著增高,而ASA组与低O2高CO2组相比显著降低。结论：ASA有抑制慢性低O2高CO2性肺动脉高压和肺血管重构的作用。     

BCL2-CISD2

CISD2, an ER BCL2-associated autophagy regulator also known as NAF-1, is responsible for the human degenerative disorder Wolfram Syndrome 2. In order to interrogate the physiological role of CISD2 we generated and characterized the Cisd2 gene deletion in mice. Cisd2 null mice manifest significant degeneration in skeletal muscle tissues, which is accompanied with augmented autophagy, dysregulated Ca²⁺ homeostasis and elongated mitochondria. Our findings describe a novel role for BCL2-CISD2 in the homeostatic maintenance of skeletal muscle. It remains to be elucidated how and if the antagonism of the BECN1 autophagy-initiating complex and modulation of ER Ca²⁺ homeostasis by BCL2-CISD2 are interconnected.     

Synthesis of [11-2H2], [8-2H2], [7-2H2], [6-2H2], [5-2H2], [4-2H2] and [3-2H2] cis-9-octadecenoates

Deuterated oleates have been synthesized by semihydrogenation of acetylenic intermediates. [11-²H₂]Oleate was prepared by two-carbon chain extension of the C₁₆ alcohol obtained from [1-²H₂]octyl bromide and 7-octyn-1-ol. [8-²H₂] and [7-²H₂]oleates were both prepared from dimethyl suberate, tetradeutero intermediate C₁₆ alcohols were synthesized from [1,8-²H₄] and [2,7-²H₄]octane diols by monobromination, conversion to deuterated 9-decyn-1-ols and reaction with octyl bromide. Oxidation gave [8-²H₂]-9-octadecynoate and [2,7-²H₂]-9-octadecynoate, after semihydrogenation of the latter, deuterons at C-2 were removed by exchange with aqueous alkali. [6-²H₂] and [5-²H₂]oleates were obtained from methyl 5-tetradecynoate, semihydrogenation, deuterium exchange at C-2 and two malonate extensions gave [6-²H₂]oleate; reduction with lithium aluminum deuteride, two malonate extensions and semihydrogenation gave the [5-²H₂] ester. [4-²H₂] and [3-²H₂]oleates were both obtained from methyl 7-cis-hexadecenoate, exchange of the α protons and chain extension gave the [4-²H₂] ester and reduction with lithium aluminum deuteride and chain extension gave the [3-²H₂] ester.     

Synthesis and physicochemical properties of 2'-deoxy-2',2'-difluoro-beta-D-ribofuranosyl and 2'-deoxy-2',2'-difluoro-alpha-D-ribofuranosyl oligonucleotides

We present procedures for nucleoside and oligonucleotide synthesis, binding affinity (Tm) and structural analysis (CD spectra) of 2'-deoxy-2',2'-difluoro-alpha-D-ribofuranosyl and 2'-deoxy-2',2'-difluoro-beta-D-ribofuranosyl oligothymidylates. Possible reasons for the thermal instability of duplexes formed between these compounds and RNA or DNA targets are discussed.     

Stereospecific 2'-amination and 2'-chlorination of adenosine by Actinomadura in the biosynthesis of 2'-amino-2'-deoxyadenosine and 2'-chloro-2'-deoxycoformycin

2'-Amino-2'-deoxyadenosine and 2'-chloro-2'-deoxycoformycin (2'-CldCF) are two nucleoside antibiotics produced by Actinomadura. The biosynthesis of these two nucleoside antibiotics has been studied by the addition of [U-14C]adenosine with or without unlabeled adenine to cultures of Actinomadura. By this experimental approach, it is possible to demonstrate that adenosine is the direct precursor for the biosynthesis of 2'-amino-2'-deoxyadenosine and 2'-CldCF. These conclusions are based on the observation that the percentage distribution of 14C in the aglyconic and pentofuranosyl moieties of 2'-amino-2'-deoxyadenosine and 2'-CldCF were similar to the distribution of 14C in the adenine and ribosyl moieties of the [U-14C]adenosine (i.e., 48:52) added to cultures of Actinomadura. Experimentally, the percentage distribution of 14C in the (i) adenine:2-amino-2-deoxy-beta-D-ribofuranose of 2'-amino-2'-deoxyadenosine is 51:49; (ii) 8-(R)-3,6,7,8-tetrahydroimidazo[4,5-d]-[1,3-diazepin-8-o1]:2 -chloro-2- beta-D-ribofuranose of 2'-CldCF is 45:55; and (iii) adenine:ribose of the adenosine isolated from the RNA of Actinomadura is 42:58. Further proof that adenosine is the direct precursor for the biosynthesis 2'-amino-2'-deoxyadenosine and 2'-CldCF was demonstrated by the addition of 75 mumol of unlabeled adenine together with [U-14C]adenosine to nucleoside-producing cultures of Actinomadura. The percentage distribution of 14C in the aglycon and the sugar moieties of 2'-amino-2'-deoxyadenosine and 2'-CldCF were 46:54 and 47:53, respectively; the percentage distribution of 14C in the adenine and ribose moieties of the adenosine isolated from the RNA of Actinomadura was 51:49. These data show that the hydroxyl on C-2' of the ribosyl moiety of adenosine undergoes a replacement by a 2'-amino or a 2'-chloro group to form 2'-amino-2'-deoxyadenosine or 2'-CldCF with retention of stereconfiguration at C-2'. Finally, Actinomadura can utilize inorganic chloride from the medium as demonstrated by the isolation of [36Cl]2'-CldCF following the addition of [36Cl]chloride to the culture medium. Mechanisms for the regioselective modification of the C-2' hydroxyl group and stereospecific insertion of the amino and chloro groups are discussed.     

Stereospecific formation of alpha-hydroxycarboxylato oxo peroxo complexes of vanadium(V). Crystal structure of (NBu4)2[V2O2(O2)2(L-lact)2] x 2H2O and (NBu4)2[V2O2(O2)2(D-lact)(L-lact)] x 2H2O

An overview of structurally characterized alpha-hydroxycarboxylatodioxo- and alpha-hydroxycarboxylatooxoperoxovanadates(V) is presented and the geometric parameters of the V2O2 bridging core are discussed. The first case of a stereospecific formation of oxoperoxovanadates(V) is reported: The crystal structures of the isomeric compounds (NBu4)2[V2O2(O2)2(L-lact)2] x 2H2O and (NBu4)2[V2O2(O2)2(D-lact)(L-lact)] x 2H2O (lact = C3H4O3(2-), the anion of the lactic acid) differ mainly in the arrangement of the V2O2 core and in mutual orientation of the V=O bonds. The complexes with achiral ligands adopt the same structural type as the complexes formed from a racemic mixture of a chiral ligand, while the structure obtained using an enantiopure L,L-hydroxycarboxylate is different.     

Convenient synthesis of 2'-deoxy-2-fluoroadenosine from 2-fluoroadenine

A convenient synthesis of 2'-deoxy-2-fluoroadenosine from commercially available 2-fluoroadenine is described. The coupling reaction of silylated 2-fluoroadenine with phenyl 3,5-bis[O-(t-butyldimethylsilyl)]-2-deoxy-1-thio-D-erythro-pentofuranoside gave the corresponding 2-fluoro-2'-deoxyadenosine derivative (alpha/beta = 1:1) in good yield. The alpha- and beta-anomers were separated by chromatography, and then desilylated to give compounds 1a and 1b.     

Stereoselective Synthesis of 2-Amino-2-deoxy-d-arabinose and 2-Deoxy-d-ribose

An efficient method for the stereoselective synthesis of 2-amino-2-deoxy-d-arabinose and 2-deoxy-d-ribose is described.

The key step in this method was accomplished by the nucleophilic addition of methyl isocyanoacetate to 2,3-O-isopropylidene-d-glyceraldehyde with high erythro-selectivity (nearly 100%).

Subsequent intermolecular cyclization predominantly gave the desired oxazoline derivative (trans-form), in which two new chiral centers were formed. The oxazoline derivative was efficiently converted to both 2-amino-2-deoxy-d-arabinose and 2-deoxy-d-ribose.