敏感硫电极法在测定心血管组织细胞及血浆胱硫醚-γ-裂解酶/硫化氢的应用

目的:建立测定微量硫化氢(Hydrogen sulfide, H2S)的敏感硫电极法.方法:根据硫化氢的理化特性,应用化学反应将溶液中物理溶解和化学形式存在的硫化氢转变成硫离子(S2-),应用敏感硫电极检测微量S2-,换算出溶液中H2S,构建了敏感硫电极检测H2S的方法.并检测了大鼠及人血浆中H2S的浓度、大鼠心血管组织中内源性H2S的含量以及大鼠心血管组织和细胞胱硫醚-γ-裂解酶(cystathionine-γ-lyase, CSE)的活性.结果:敏感硫电极检测1～80 μmol/L的S2-有较好的指数相关关系,应用该方法检测到雄性和雌性大鼠血浆H2S的浓度分别为(40±4)和(41±5) μmol/L,差异无统计学意义,人类男性和女性静脉血血浆H2S浓度分别为(33±4) μmol/L 和(35±5) μmol/L,差异无统计学意义.雌、雄大鼠主动脉组织H2S的含量分别为每毫克蛋白(24±6)和(25±5) nmol,心肌组织含量分别为每毫克蛋白(19±4) 和(19±6) nmol,差异无统计学意义.采用敏感硫电极法测量主动脉组织CSE活性与传统方法测量结果差异无统计学意义,但可精确测量出血管平滑肌细胞CSE的活性.结论:敏感硫电极法可以应用于CSE/H2S信号通路的检测.     

新型气体信号分子硫化氢在大鼠肺纤维化发病中的作用

目的:在博来霉素(Bleomycin,BLM)诱导的肺纤维化大鼠模型上,观察内源性胱硫脒-γ-裂解酶/硫化氢(CSE/H2S)体系的变化以及应用外源性H2S供体硫氢化钠(NaHS)对肺纤维化的影响,以探讨内源性H2S在肺纤维化发病中的病理生理意义.方法:(1)雄性Wistar大鼠48只,随机分为对照组、肺纤维化组(经气管内滴入BLMA5 5 mg/kg)、肺纤维化 NaHS低剂量组(腹腔注射NaHS 1.4 μmol/kg,每日两次)和肺纤维化 NaHS高剂量组(腹腔注射NaHS 7 μmol/kg,每日两次).分别于第7天、第28天取肺组织行HE染色和Masson染色,用生化法测定丙二醛(MDA)、羟脯氨酸含量,采用敏感硫电极测定血浆H2S的生成量及亚甲基蓝法测定CSE活性,用半定量RT-PCR方法测定肺组织CSE的mRNA水平.(2)以羟自由基发生系统Fe2 (5 mmol/L)/H2O2(25 mmol/L)单独及与不同浓度NaHS(10-5、5×10-5、10-4、10-3mmol/L)共同孵育离体大鼠肺组织,测定MDA含量.结果:(1)第7天纤维化组大鼠血浆H2S含量较对照组下降46%,第28天时增加25%(均P＜0.01);肺组织CSE活性在第7天时比对照组降低26%(P＜0.01),第28天时CSE活性恢复,与对照组相比差异无统计学意义(P＞0.05);第7天和第28天纤维化组肺组织CSE mRNA含量较对照组分别升高34%和144%(P＜0.01);第7天及第28天纤维化组肺组织MDA、羟脯氨酸含量均较对照组明显升高(P＜0.01),NaHS高剂量组和低剂量组的MDA、羟脯氨酸含量均较纤维化组显著降低,高、低剂量组间差异无统计学意义(P＞0.05).(2)Fe2 /H2O2孵育组肺组织MDA含量较对照组明显升高,经给予不同浓度NaHS处理后,MDA含量均显著下降.结论:内源性CSE/H2S体系参与了大鼠肺纤维化的病理生理过程,外源性补充H2S可能通过减轻氧化应激抑制肺纤维化的发生.     

硫化氢的中枢心血管调节机制

硫化氢(H2S)是继一氧化氮(NO)和一氧化碳(CO)之后的第3种内源性气体信号分子.研究显示H2S对神经系统、呼吸系统、消化系统和心血管系统具有调节效应,在正常生理功能和病理生理过程的调控中发挥重要生物学作用.近年来,H2S的中枢心血管调节作用逐渐引起重视,并表现出复杂的调节效果.本文主要介绍H2S对血压调节中枢相关核团的作用特点,并分析H2S在中枢心血管功能调控中的意义及其进展.     

气体信号分子在心血管疾病发病中的意义

气体分子一氧化氮(NO)的发现开创了气体信号分子这一新型研究领域,目前已发现3种气体信号分子:NO、一氧化碳(CO)和硫化氢(H2S).他们在体内内源性生成,发挥广泛的生物学效应,本文仅就3种气体信号分子在心血管系统中的意义进行简要阐述.在心血管系统中内源性气体信号分子NO、CO和H2S分别与其相应的合成酶一氧化氮合酶(NOS)、血红素加氧酶(HO)和胱硫醚-γ-裂解酶(CSE)形成独立而又相互关联的体系(NO/NOS体系、CO/HO体系、H2S/CSE体系),不仅参与心血管系统生理状态下功能和结构的维持,而且在高血压、肺动脉高压、感染性休克、动脉粥样硬化等心血管疾病发病中发挥重要的病理生理学作用.     

脐血浆中硫化氢与羊水污染的相关性研究

目的 研究脐血浆中硫化氢(H2S)和羊水污染的相关性.方法 收集孕产妇脐血浆60份,根据分娩时羊水是否污染分为2组.用敏感硫电极检测脐血浆中硫离子的浓度,并换算出H2S的含量;免疫组织化学方法检测胎盘中胱硫醚-γ-裂解酶(CSE)和胱硫醚-β-合成酶(CBS)的表达程度.结果 羊水污染组中H2S含量高于羊水清组,差异有显著性.免疫组化显示:CSE表达于胎盘绒毛的细胞滋养层胞浆中,羊水污染组CSE呈现高表达.CBS在任何组中均未见有表达.结论 胎盘绒毛的细胞滋养层胞浆中含有CSE,不舍CBS;脐血浆中H2S由细胞滋养层细胞参与调控生成;孕产妇脐血浆中H2S与羊水污染有密切关系.     

胱硫醚-γ-裂解酶在小鼠肺脏中的表达与血中H2S含量研究

目的 探讨外源性硫化氢(H2S)对小鼠肺脏胱硫醚-γ-裂解酶(CSE)和血H2S的影响.方法 取4周龄健康昆明小鼠20只,随机分为生理盐水对照组,NaHS作用24h,72h,5d实验组,每组5只,均采用腹腔注射给药.在3个不同时间点处死小鼠,生理盐水对照组与24h组一并处死,取肺脏进行免疫组织化学染色与图像分析,敏感硫电极检测循环血中H2S含量.结果 ①免疫组织化学见CSE主要表达在支气管上皮细胞和肺泡隔中毛细血管内皮细胞浆及核膜上,与对照组比较,24h实验组表达最强,图像分析相对含量显著升高(P<0.01),72h实验组也有升高(P<0.05);实验组之间比较,5d实验组与72h,24h实验组比较差异非常显著(P<0.01),而5d实验组与对照组间差异无统计学意义(P>0.05).②各实验组小鼠外周血中H2S含量均高于对照组(P<0.01),其含量随时间延长而降低,5d实验组最低.结论 小鼠肺脏中确有CSE表达.外源性H2S可提高小鼠肺脏CSE表达及循环血中H2S的含量,随时间延长,CSE表达和H2S含量逐渐减少.     

硫化氢在心血管系统中的研究进展

继一氧化氮 (NO)和一氧化碳 (CO)这两种气体信号分子被发现之后 ,在 2 0世纪 90年代中期 ,又发现半胱氨酸代谢生成气体分子硫化氢 (H2 S) ,对神经系统特别是海马的功能具有调节作用 ,并可以调节消化道和血管平滑肌的张力 ,而其作用特点有别于NO及CO这两种气体信号分子 ,H2 S的信号转导途径一直未能阐明 ,直到最近研究证实 ,内源性H2 S直接作用于K(ATP)通道实现对血管的调节作用 ;并抑制平滑肌细胞的增殖。越来越多的证据表明 ,内源性H2 S是一种新的气体信号分子 ,对其研究是当前生物学领域的崭新课题 ,具有重要的理论和临床意义。本…     

H2S/CBS体系在大鼠癫痫发病中的变化及意义

目的 探讨大鼠癫痫发病过程中H2S/CBS的变化及意义.方法 雄性SD大鼠随机分为正常对照组(NC组)和癫痫模型组(EP组),每组大鼠又分为2h、24 h、7d和30d 4个时间点,氯化锂-匹罗卡品腹腔注射复制癫痫模型,出现Ⅳ级及以上无明显间隔的癫痫持续状态(status epilepticus,SE)为模型复制成功 ;敏感硫电极法测定大鼠血浆中H2S含量 ;免疫印迹(WB)检测大鼠海马中CBS蛋白的表达情况 ;荧光定量PCR检测海马中的CBS mRNA表达变化.结果 癫痫模型组大鼠癫痫发作急性期(2h和24 h)血浆H2S水平、海马CBS mRNA和蛋白表达水平明显高于正常对照组(P＜0.05) ;随后在7d和30 d时H2S/CBS水平降低,显著低于正常对照组(P＜0.05).结论 大鼠癫痫发作急性期H2S和CBS表达水平升高,慢性自发性痫性发作期间H2S和CBS表达水平下降,提示H2S/CBS参与癫痫的发病过程.     

新型气体信号分子--硫化氢心血管作用的研究进展

硫化氢(hydrogen sulfide,H2S)一直被认为是污染环境的毒性气体,是造成大气和水以及造成某些职业病的主要危害物质之一.超过生理剂量的H2S对活体器官的毒性体现在对中枢神经系统以及对呼吸系统的抑制作用[1～3].继内源性气体分子一氧化氮(nitric oxide,NO)、一氧化碳(carbonmonoxide,CO)被证实为气体信号分子后,20世纪90年代,发现半胱氨酸代谢产生的气体H2S,对神经系统特别是海马的功能具有调节作用.H2S通过刺激神经细胞,增加细胞内环腺苷酸(cyclicadenosine monophosphate,cAMP)水平,提高受体介导的兴奋性突触后电位,诱导海马的长时程增强效应[4,5].H2S可以调节消化道和血管平滑肌的张力,还可以舒张血管和抑制血管平滑肌细胞增殖[4～6].近年来研究发现,H2S还可以调节生理及病理状态下心脏的功能.本文就H2S在心血管系统中的生理学作用及其机制进行综述.     

红细胞内源性硫化氢的测定

目的:建立测定红细胞内源性硫化氢(hydrogen sulfide, H2S)的方法.方法:根据红细胞硫化氢生成酶的理化特性,给予酶的底物β-巯基丙酮酸,并根据亚甲基蓝法测定释放硫化氢的量.结果:应用RT-PCR方法检测到红细胞内存在巯基丙酮酸转硫酶(3-mercaptopyruvate sulfurtransferase, MPST)的表达;红细胞内源性硫化氢的产率高达每108个细胞(22.76±1.53) μmol/min,约是肝和肾组织内硫化氢生成率的5倍.而检测L-半胱氢酸(L-cysteine, L-cys)途径内源性硫化氢的产率,并以肝和肾作为阳性对照组织,发现红细胞内几乎检测不到硫化氢的产生.结论:红细胞内源性硫化氢的产生主要依赖MPST途径,本研究构建的方法可应用于红细胞生成硫化氢的检测.     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

A preliminary screening study on the associated proteins in human psoriasis vulgaris by serum proteomics technologies

Objective：To investigate the optimum screening conditions of associated proteins in human psoriasis vulgaris by serum proteomics technique, and to screen the different expression proteins related with psoriasis vulgaris. Methods：Serum samples of peripheral blood were collected from newly diagnosed psoriasis vulgaris patients in the clinic, and 20 matched healthy persons. Serum albumin IgG was removed by filtering with ProteoExtract Albumin/IgG. After comparative proteomics analysis the different protein spots were identified using 2-DE and MS. Results ：Electrophoresis figures with high resolution and reproducibility were obtained. Three different expression proteins were found only in the serum from psoriasis vulgaris patients, while nine other different proteins expressing from healthy volunteers. Conclusion：The protein expression was different in the serum between the psoriasis vulgaris patients and healthy volunteers. It was hoped that we could find the biomarkers related to psoriasis vulgaris by using proteomics.     

A CASE OF HUGE GASTRIC DISOPYROBEZOAR

PYTOBEZOARS are common bezoars in gastrontestinal tract, commonly seen in stomach and mall intestine,1but huge disopyrobezoars are     

Acupuncture Treatment of Chest Bi Syndrome

CASE HISTORY A male,a retired officer of 61 years old,paid his first visit on May 12,2006.The patient stated that 2 weeks before he suddenly got paroxysmal precordial choking pain,with irregular attacks.Each attack would last several seconds to several minutes.The attack was accompanied with heavy oppressed sensation in the precordial region,palpitation,lassitude,and in severe cases,sweating,which was once diagnosed by a hospital as'coronary heart disease'.Recalling the case history,the patient said that several days before he got angry with others because of some affairs in work,and he was not so used to the life pattern since he had just retired from his busy working post.At usual times,he was quick tempered and peevish.In recent days,he had poor appetite,and loose stool.     

Instructions to Contributors

The English edition of Chinese Journal of Integrative Medicine is edited and published by the Chinese Journal of Integrated Traditional and Western Medicine Press.Original scientific papers of advanced clinical and experimental medicine in the field of complementary/alternative medicine or integrative medicine are expected.