TRYPTOPHAN METABOLISM IN ALCOHOLISM. TRYPTOPHAN BUT NOT EXCITATORY AMINO ACID AVAILABILITY TO THE BRAIN IS INCREASED BEFORE THE APPEARANCE OF THE ALCOHOL-WITHDRAWAL SYNDROME IN MEN

Tryptophan (Trp) metabolism and disposition and excitatory andother amino acid concentrations were determined in alcohol-dependentsubjects in relation to the alcohol-withdrawal syndrome (AWS).Parameters were examined in 12 alcohol-dependent male subjects,undergoing elective upper digestive tract tumour resection,and 12 age-, gender-, and medication-matched controls on threeoccasions: pre-operatively, post-operatively, and immediatelybefore (i.e. within 24 h of) the appearance of the AWS. No significantdifferences were observed between controls and alcoholic subjectson the first or second of these occasions. On the third occasion,within 24 h of the appearance of the AWS, alcoholics showeda dramatic elevation (117%) in free serum Tip concentrationand a consequent increase (111%) in the ratio of [free Trp]/[competingamino acids], which is an accurate predictor of Trp entry intothe brain. Increases were also observed on this third occasionin concentrations of total Trp (49%), cortisol (123%), and norharman(137%). Concentrations of glutamate, glycine, aspartate, serine,and taurine did not differ significantly within or between thecontrol and alcohol-dependent groups of subjects on any of thethree occasions. The possible significance of the Trp and relatedmetabolic changes in relation to the behavioural features ofthe AWS is discussed.     

一氧化氮在病毒性肝炎病理生理过程中的作用

一氧化氮参与肝细胞多项生理功能的调节,参与病毒性肝炎的病理生理过程.在病毒性肝炎发病机理方面,一氧化氮可能具有保护肝脏和导致肝损害的双重作用.此外,一氧化氮还具有潜在的抗病毒活性及增加肝细胞癌变的危险性.本文综述了一氧化氮在病毒性肝炎病理生理过程中的这些作用.     

THE INFLUENCE OF CHRONIC MODERATE ETHANOL ADMINISTRATION ON NADPH-DIAPHORASE (NITRIC OXIDE SYNTHASE) ACTIVITY IN RAT BRAIN

Nitric oxide synthase (NOS), the enzyme with reduced nicotinamide-adeninedinucleoude phosphate (NADPH)-diaphorase activity, generatesnitric oxide (NO) which is an important bioregulatory moleculein the nervous, immune, and cardiovascular systems. NOS is linkedto non-adrenergic non-cholinergic (NANC) neuronal pathways andmodulation of the N-methyl-D-aspartate receptors, yet its modificationby ethanol has been little explored. A possible modificationby chronic ethanol administration of activity and*or localizationof NADPH-diaphorase (NO-synthase) in rat brain may thus providethe pathogenic basis of alcohol-induced brain injury. When femaleWistar rats were treated chronically with ethanol for 50 days,the NADPH-diaphorase staining of granular neurons and neuronslocated in the molecular layer of the cerebral cortex was significantlyreduced. Chronic ethanol consumption led to a significant reductionin NADPH-diaphorase staining in the superficial layers of thesuperior colliculus. The number of NADPH-diaphorase-positiveneurons was significantly reduced (P < 0.001) in the stratumzonale and stratum griseum superficiale (by 42 3–65 6%of control values). This could alter synaptic processes in thehighly organized structures involved in oculomotor and somaticmotor coordination and thus contribute to the motor disturbanceswhich are associated with alcohol abuse.     

INACTIVATION OF CEREBELLAR NITRIC OXIDE SYNTHASE BY ETHANOL IN VITRO

The N-methyl-D-aspartate receptor/nitric oxide synthase (NOS)/guanylatecyclase pathway, which plays a crucial role in synaptic plasticityin the brain, is modulated by ethanol. We studied the effectof ethanol in vitro on NOS in rat cerebellum and showed thatethanol (25–200 mM) inactivated NOS in a dose-dependentmanner. This inactivation was prevented by the biopterin cofactortetrahydrobiopterin (BH⁴) as well as by L-arginine, a NOS substrate,but not by NADPH. These results suggest that ethanol reducesNOS activity by modulating the conformation of the enzyme andthereby its stability, probably by interacting with the bindingsites of BH⁴ and/or of L-arginine. Our data also suggest thatinactivation of NOS may contribute to the decrease in the cGMPlevel, and thus may play a role in the pharmacological actionsof ethanol in vivo.     

锌对染铅大鼠海马一氧化氮合酶的保护作用

目的 :　探讨铅对海马长时程增强 (LTP)的影响与海马不同亚区一氧化氮合酶(NOS)变化的关系以及锌的拮抗作用。方法 :　采用反映学习记忆功能的 Y迷宫法测试大鼠神经行为的改变 ;用 NADPH-黄递酶 (NADPH- d)组化法和神经元型 NOS(n NOS)的免疫组化法研究大鼠海马不同亚区 NOS的活性及表达情况。结果 :　染铅组大鼠的学习记忆能力比铅锌组和对照组明显下降 (P<0 .0 5 ) ,铅锌组与对照组之间无差别 ;组化及免疫组化显示 :染铅组大鼠海马CA1区和齿状回的 NOS和 n NOS阳性神经元明显少于铅锌组和对照组 (P<0 .0 5 ) ,在 CA3区无差别 ,铅锌组与对照组各亚区均无差别。结论 :　铅可损伤大鼠学习记忆能力 ,鉴于 NOS参与LTP这一代表学习记忆的电生理指标的形成和维持 ,推测铅对学习记忆和海马 LTP的影响可能与染铅后海马各区 NOS的不同变化有关。锌对铅引起的学习记忆损伤和 NOS的影响有拮抗作用。     

一氧化氮对小鼠胚胎种植的影响

目的：评价一氧化氮对小鼠胚胎种植的影响。方法：受试昆明小鼠于妊娠第d3～d5分别腹腔注射NO供体硝普钠（SNP）1．0mg／kg、5．0mg／kg和10．0mg／kg，对照组小鼠注射等量的生理盐水；妊娠第6d颈椎脱臼法处死小鼠，肉眼观察每一侧子宫角孕囊数，形态计量学方法分析子宫内膜组织学变化，明胶酶谱法检测子宫内膜基质金属蛋白酶的表达。结果：①1．0mg／kgSNP组小鼠子宫角孕囊数与对照组比较无显著性差异（P〉0．05），5．0mg／kgSNP组小鼠宫角未观察到孕囊（P〈0．001），10．0mg/kg SNP组小鼠在注射药物20min后全部死亡；②1．0mg／kgSNP组与对照组小鼠子宫内膜均观察到明显的蜕膜样改变，丽组间腺体面积和间质面积比较无显著性差异（P〉0．05），5．0mg／kgSNP组小鼠子宫内膜未观察到蜕膜样改变，子宫内膜腺体面积明显低于对照组和1．0mg／kgSNP组（P〈0．001），间质面积明显高于对照组和1．0mg／kgSNP组（P〈0．001）；③1．0mg／kgSNP组与对照组小鼠子宫内膜均检测到酶原型MMP-2、激活型MMP-2和酶原型MMP-9的表达，5．0mg／kgSNP组小鼠子宫内膜仅检测到酶原型MMP-2的表达。结论：子宫内膜较高浓度的NO对胚胎种植具有明显的抑制效应，这种作用可能是通过影响MMP-2和MMP-9的表达及其活化以及干扰子宫内膜的蜕膜化实现的。     

宝宝不“听话”?方法出了错!

情景回放 1 贝贝今年3岁了,总是不吃青菜,却特别爱吃巧克力.刚开始的时候,奶奶总是会夹着青菜哄着贝贝吃,有的时候甚至会追在贝贝身后,但她依旧不爱吃青菜. 贝贝妈在一次吃饭的时候爆发了:”贝贝,如果你今天不吃青菜,就什么都不许吃.” 奶奶实在是心疼贝贝,就说:“贝贝妈,也不用这么吓唬宝宝呀,不吃就不吃吧.“贝贝妈好不容易下定决心要惩罚贝贝,结果奶奶的这番话让贝贝顿时有了后援,贝贝的脸上写着“奶奶都说了我可以不吃”. 贝贝妈一下子就发起火来,说:”妈,您能不能不要添乱?” 奶奶说:”宝宝这么小,干吗总要求这么多,她不想吃你就不要逼她吃嘛,这怎么是我添乱呢?你这么说我就不爱听了.“ 饭桌上的氛围顿时尴尬起来,贝贝妈继续说:”贝贝,我说话你听见了没有,今天你不吃青菜,就不要吃饭,什么都不许吃.”贝贝听到这儿就大哭起来,奶奶见状赶忙抱起贝贝回房间去了.     

DETECTION AND BIOIMAGING OF NITRIC OXIDE IN BOVINE OOCYTES AND SPERM CELLS

Mammalian gametes contain constitutive nitric oxide synthases (NOS) to synthesize nitric oxide (NO). The detection and bioimaging of NO in bovine gametes is important to determine the regulatory roles of NO during the different events of fertilization. Diaminofluoresceins, new fluorescence indicators for NO, were applied to detect the release of NO from bovine gametes. These compounds yield green fluorescent triazolofluoresceins, which provide the advantages of high specificity, sensitivity, and simplicity for the detection of NO. In this study, we mapped the expression of NOS in the bovine sperm and ova. NOS activity in sperm first appeared in the acrosome, then 60?min later in the head, middle piece, cytoplasmic droplet, and tail. Cow ova had high NO activity in the cytoplasm and in the surrounding corona cells, but not in the zona pellucida. These results show that for bovine gametes, the synthesis NO by the NOS system presents clear patterns of time and spatial distribution that may be important for the different events of fertilization.     

胃癌患者一氧化氮、超氧化物歧化酶及脂质过氧化物的变化

目的:研究胃癌患者一氧化氮、超氧化物歧化酶及脂质过氧化物的变化。方法:检测51 例胃癌患者( 患者组)和50 例健康成人( 对照组) 血浆一氧化氮(P- NO) 含量、红细胞超氧化物歧化酶(E- SOD) 活性、血浆过氧化脂质(P- LPO) 及红细胞过氧化脂质(E- LPO) 。结果:与对照组相比,患者组的P- NO、P- LPO 及E- LPO 平均值均显著升高( P< 0 .001) ,E- SOD 值显著降低( P< 0 .001) 。结论:提示胃癌患者体内的NO 代谢异常,脂质过氧化反应病理性加剧。     

SPERM DNA FRAGMENTATION IS INCREASED IN COUPLES WITH UNEXPLAINED RECURRENT PREGNANCY LOSS

Previous studies have indicated that sperm quality may be related to unexplained recurrent pregnancy loss. This study evaluated the degree of sperm DNA fragmentation using the TUNEL assay on sperm from 24 couples with unexplained recurrent pregnancy loss (RPL) compared to sperm from 2 control groups: donors of known fertility and unscreened men from the general population. The percentage of sperm staining positive for DNA fragmentation was increased ( p <.001) in the RPL group (38±4.2) compared to the donor (11.9±1.0) or general population (22±2.0) control groups. In the RPL group, no correlation was observed between semen quality parameters and the TUNEL data. These data indicate that some RPL patients have a significant increase of sperm DNA fragmentation, which may be causative of pregnancy loss in some patients.     

ACTIVATION OF ENDOTHELIAL NITRIC OXIDE SYNTHASE IN CONTRALATERAL TESTIS DURING UNILATERAL TESTICULAR TORSION IN RATS

There are controversies about the injury of the contralateral testis during unilateral testicular torsion (UTT). An autonomic reflex arc between bilateral testes has been proposed. The authors focused on the involvement of nitric oxide (NO) in the contralateral testis during UTT. Eight-week-old male Wistar rats underwent unilateral torsion (1 h)-detorsion (up to 24 h). NO synthase (NOS) activity was detected as NADPH-diaphorase activity after fixation by paraformaldehyde. N -nitro- L -Arginine methyl ester (L-NAME, 20 mg/kg) was injected intravenously to the other group of rats. To evaluate the testicular injury, proteolysis of &#102 -fodrin production was detected by Western blotting. Apoptosis of the germ cells was evaluated by TUNEL. Long-term effect on spermatogenesis was evaluated by flow cytometry at 60 days after UTT. Transient activation of NOS was detected following the proteolysis of &#102 -fodrin in the contralateral testis. L-NAME inhibited these alterations. NADPH-diaphorase activity and eNOS immunoreactivity were co-localized in the endothelial cells. These reactions were not observed in other organs. There was neither enhanced apoptosis nor deteriorated spermatogenesis in the contralateral testis during and 60 days after UTT. In the contralateral testis, eNOS-derived NO regulates the vasomotor function against unilateral testicular torsion, whereas it acts slightly cytotoxic. These results suggest the possible involvement of a testis-specific neurovasomotor reflex between the bilateral testes.     

Caveolin-1与内皮型一氧化氮合成酶在门静脉高压症性血管病变中的表达

目的检测肝硬化门静脉高压症病人的脾血管中小凹蛋白caveolin-1、内皮型一氧化氮合成酶(endothelial nitric oxide synthase,eNOS)的mRNA及蛋白表达水平的变化,探讨caveolin-1的表达对eNOS的影响。方法逆转录-聚合酶链反应(RT-PCR)方法检测27例肝硬化门静脉高压症病人脾静脉组织和20例脾外伤病人正常脾静脉血管中caveolin-1和eNOS的mRNA;Western Blot检测caveolin-1和eNOS的蛋白表达水平变化。结果肝硬化门静脉高压症组脾静脉Caveolin-1 mRNA与对照组脾静脉组织表达分别为(0.73±0.18)、(0.38±0.12),两组比较差异有显著性(p<0.05);肝硬化门静脉高压症组脾静脉eNOS mRNA为(0.23±0.11),显著低于对照组内脾静脉组织eNOS mRNA的表达(0.47±0.15)(p<0.05)。Western Blot检测caveolin-1在肝硬化门静脉高压症组脾静脉中较正常脾静脉中表达明显增强;eNOS在肝硬化门静脉高压症组脾静脉中呈低水平表达,较正常脾静脉中表达明显减少。结论肝硬化门静脉高压症组脾静脉中caveolin-1的过量表达及eNOS表达降低,导致NO合成减少,脾静脉血管阻力持续增加,及caveolin-1致静脉血管病理改变作用,共同作用致脾静脉出现对门静脉高压失代偿改变。     

艾滋病不是不治之症

艾滋病是患者感染了人类免疫缺陷病毒[HIV]失去了正常免疫功能而感染各种疾病,最终导致死亡,现己发展成全球性的瘟疫。按现代医学的”鸡尾酒疗法”只能缓解病情,用对症疗法可以治好一些机会感染、而无法治愈艾滋病。医学界以为疫苗是可以防治艾滋病,各国竟相投入大量人力和物力进行研究,结果全部失败告终。医学界断言艾滋病无法治愈为时过早,其实,在医学界以上路子不通,不等于所有路子都不通,天无绝人之路嘛。笔者观察到在非洲有几十名妓女”对艾滋病病毒感而不染”,而且,在国内外已有3-5例没有用药或骨髓移植艾滋病患者痊愈。所以,笔者坚信艾滋病用元素医学食疗法有可能被治愈,在艾滋病疫区河南上蔡文楼村经16个月努力终于2005年11月底有2例完全痊愈,说明艾滋病不是不治之症。已超越卫生部国家中医药管理局推荐《中医药治疗艾滋痞临床技术方案[试行]》”目前中医对艾滋病治疗的主要目标是提高免疫功能、控制机会性感染,改善生存质量,使患者带毒生存”的要求,正在向完全攻克艾滋病方向的迈进。     

艾滋病不是不治之症

艾滋病是患者感染了人类免疫缺陷病毒[HIV]失去了正常免疫功能而感染各种疾病，最终导致死亡，现己发展成全球性的瘟疫。按现代医学的“鸡尾酒疗法”只能缓解病情，用对症疗法可以治好一些机会感染、而无法治愈艾滋病。医学界以为疫苗是可以防治艾滋病，各国竟相投入大量人力和物力进行研究，结果全部失败告终。医学界断言艾滋病无法治愈为时过早，其实，在医学界以上路子不通，不等于所有路子都不通，天无绝人之路嘛。笔者观察到在非洲有几十名妓女“对艾滋病病毒感而不染”，而且，在国内外已有3—5例没有用药或骨髓移植艾滋病患者痊愈。所以，笔者坚信艾滋病用元素医学食疗法有可能被治愈，在艾滋病疫区河南上蔡文楼村经16个月努力终于2005年11月底有2例完全痊愈，说明艾滋病不是不治之症。已超越卫生部国家中医药管理局推荐《中医药治疗艾滋病临床技术方案（试行]》“目前中医对艾滋病治疗的主要目标是提高免疫功能、控制机会性感染，改善生存质量，使患者带毒生存”的要求，正在向完全攻克艾滋病方向的迈进。     

艾滋病不是不治之症

艾滋病是患者感染了人类免疫缺陷病毒（HIV）失去了正常免疫功能而感染各种疾病,最终导致死亡,现己发展成全球性的瘟疫。按现代医学的“鸡尾酒疗法”只能缓解病情,用对症疗法可以治好一些机会感染、而无法治愈艾滋病。医学界以为疫苗是可以防治艾滋病,各国竞相投入大量人力和物力进行研究,结果全部失败告终。医学界断言艾滋病无法治愈为时过早,其实,在医学界以上路子不通,不等于所有路子都不通,天无绝人之路嘛。笔者观察到在非洲有几十名妓女“对艾滋病病毒感而不染”,而且,在国内外已有3．5例没有用药或骨髓移植艾滋病患者痊愈。所以,笔者坚信艾滋病用元素医学食疗法有可能被治愈,在艾滋病疫区河南上蔡文楼村经16个月努力终于2005年11月底有2例完全痊愈,说明艾滋病不是不治之症。已超越卫生部国家中医药管理局推荐《中医药治疗艾滋痞临床技术方案（试行）》“目前中医对艾滋病治疗的主要目标是提高免疫功能、控制机会性感染,改善生存质量,使患者带毒生存”的要求,正在向完全攻克艾滋病方向的迈进。     

乙型肝炎病毒前C区变异患者血清Ⅳ型胶原/层粘连蛋白和一氧化氮的分析

目的：分析乙型肝炎病毒（ＨＢＶ）前Ｃ区基因变异在肝纤维化形成中的作用,探讨肝纤维化与一氧化氮的关系。方法：利用错配ＰＣＲ限制片段多态性分析,检测ＨＢＶ前Ｃ区１８９６位点突变,并检测慢性乙肝患者血清Ⅳ型胶原／层粘连蛋白（ＣⅣ／ＬＮ）和ＮＯ。结果：Ａ组３１例前Ｃ区变异或变异优势患者ＣⅣ（１２７．３５±２４．３３）μｇ／Ｌ,ＬＮ（１６９．１０±２８．２３）μｇ／Ｌ明显高于Ｂ组３０例无前Ｃ区基因变异患者ＣⅣ（８６．８４±１２．６６）μｇ／Ｌ,ＬＮ（９４．２０±１５．２９）μｇ／Ｌ（Ｐ＜０．００１）,且Ａ组ＮＯ－２／ＮＯ－３（４８．５５±４．０３）μｍｏｌ／Ｌ明显高于Ｂ组ＮＯ－２／ＮＯ－３（３６．６０±４．０２）μｍｏｌ／Ｌ（Ｐ＜０．００１）。结论：ＨＢＶ前Ｃ区变异与肝纤维化、肝硬化发生发展有关系,且血清ＮＯ随ＣⅣ和ＬＮ增高而增高。     

冷暴露大鼠血浆肾上腺素及血管内皮细胞 NOS活性的变化

目的观察冷暴露大鼠血浆肾上腺素(Adr)水平及血管内皮细胞NOS活性的变化,为探寻防治冷损伤的措施提供理论依据。方法使用L-精氨酸(L-Arg)调节主动脉内皮细胞NOS活性。大鼠随机分为5组,即对照组(室温)、冷暴露Ⅰ组(-15℃,1 h)、冷暴露Ⅱ组(-15℃,1.5 h)、L-Arg组(室温,L-Arg 2 g.kg-1灌服)、冷暴露 L-Arg组(-15℃,1 h;L-Arg 2 g.kg-1灌服)。冷暴露后6 h再取血样。以酶联免疫法测定血浆肾上腺素水平;以硝酸酶还原法测定血管内皮细胞NOS活性;以紫外分光光度计检测血清和血管内皮细胞培养液中的LDH活性;反转录PCR方法检测血管内皮细胞NOS mRNA表达水平。结果对照组、冷暴露Ⅰ组与Ⅱ组血浆中Adr分别为(10.81±1.85)、(31.37±4.48)、(39.35±5.59)nmol.L-1,冷暴露大鼠血浆中Adr水平明显地高于对照组(P<0.05);上述3组血管内皮细胞NOS活性分别为(16.13±3.68)、(9.19±1.87)、(5.94±1.05)μmol.L-1;冷暴露大鼠血管内皮细胞NOS活性明显低于对照组(P<0.05),使用L-Arg可使冷暴露大鼠血管内皮细胞NOS活性上调到(12.87±1.60)μmol.L-1(P<0.01)。同时,血浆中LDH水平也明显降低(P<0.01)。结论冷暴露血浆高浓度肾上腺素可抑制主动脉内皮细胞NOS活性;L-Arg可上调血管内皮细胞NOS活性,对减轻机体冷损伤程度有一定作用。     

一氧化氮对体外保存血小板活化的影响研究

目的研究一氧化氮(NO)溶液对体外保存的手工分离血小板活化的影响。方法用高纯铜与稀硝酸反应,制备NO气体,然后溶解于蒸馏水中制备成NO饱和溶液。手工制备富含血小板血浆(PRP),在滤过白细胞前注入1/10PRP体积的不同浓度NO溶液,将血小板最大回收率时的NO溶液浓度定为实验的最适浓度,观察含此NO浓度的血小板保存5d内各项相关指标的变化,并进行统计学分析。结果NO饱和溶液10-2稀释时,血小板的回收率最大。加入NO组和对照组比较,血小板pH、MPV、PDW三项指标无统计学差异,但保存1天后CD62p再表达率有显著性差异(p<0.01)。结论NO可减少体外保存血小板的活化。