Effects of hydrogen-rich saline treatment on polymicrobial.pdf 脓毒症（ sepsis ）是指由感染引起的全身炎症反应综合征（ systemic inflammatory response syndrome, SIRS ），临床上证实有细菌存在或有高度可疑感染灶。虽然脓毒症是由感染引起，但是一旦发生后，其发生发展遵循其自身的病理过程和规律，故从本质上讲脓毒症是机体对感染性因素的反应。脓毒症曾经被称为败血症。从严重程度上分为 3 种类型：脓毒症、严重脓毒症（ severe sepsis ）和脓毒性休克（ septic shock ）。脓毒症发生率高，全球每年有超过 1800 万严重脓毒症病例，美国每年有 75 万例脓毒症患者，并且这一数字还以每年 1.5% ～ 8.0% 的速度上升。脓毒症病死率高，全球每天约 1.4 万人死于其并发症。流行病学调查显示，脓毒症病死率已经超过心肌梗死，成为重症监护病房内非心脏病人死亡的主要原因。脓毒症治疗花费高，医疗资源消耗大，严重影响人类的生活质量，已经对人类健康造成巨大威胁。 治疗和预防脓毒症最有效的方法是以脓毒症的发病机制为基础进行治疗和预防，但是遗憾的是目前脓毒症的发病机制仍未完全阐明，在这种情况下，针对发病原因应做好临床各方面的预防工作，努力降低诱发感染的危险因素对脓毒症的治疗和预防有着重要作用。 最近来自南京总医院麻醉科的一项研究在线发表在《外科研究杂志》上，证明氢气生理盐水对脓毒症的治疗作用。 2009 年天津医科大学谢克亮等的研究证明呼吸氢气可以对脓毒症具有治疗作用，随后他们用不同的模型证明了同样的治疗效果，最重要的发现是首次在世界上报道了氢气在治疗疾病的同时具有促进内源性抗氧化酶活性的作用，并发现对 HMGB1 具有影响。 氢气可以选择性中和毒性自由基羟基自由基和亚硝酸阴离子，对许多疾病和病理状态具有治疗效果，呼吸氢气对脓毒症的治疗作用早就有报道，本研究是观察氢气生理盐水对脓毒症的治疗作用。 SD 大鼠盲肠结扎和穿孔制备脓毒症疾病模型， 24 只动物平均分成假手术、脓毒症和氢气生理盐水等 3 组。在模型制备后 0 、 6 和 18 小时 3 次给动物腹腔注射 5ml/Kg 氢气生理盐水或无氢气正常生理盐水。手术后 24 小时分别检测 MDA （脂质过氧化指标）、 SOD （抗氧化酶）、炎症介质、肺组织 NO 、 MPO （中性粒细胞）、干湿重（肺水肿程度）、组织病理学指数（病理变化）、细胞凋亡分析、丙氨酸转氨酶（肝细胞损伤）、肌酐和血尿素氮（肾脏功能）。手术后 7 天的动物生存率也进行了详细记录。结果发现氢气生理盐水可以使血清内 HMGB1 下降（这是最近几年研究比较热门的炎症因子，正常情况下分布在细胞内，在细胞核内和 DNA 结合发挥调节基因表达的作用，当细胞死亡或损伤时，可以被动或主动释放到细胞外，和一些重要炎症受体 RAGE TLR 结合，并启动炎症反应）。另外转氨酶、肌酐、尿素氮水平均显著下降。肺组织白介素 6 、 HMGB1 、 NO 、 MDA ；肺干湿比、组织学变化指数和细胞凋亡阳性细胞等君明显改善。 SOD 明显升高。但动物生存率无明显改变。研究结果证明氢气生理盐水可以对脓毒症具有治疗作用。 Effects of hydrogen-rich saline treatment on polymicrobial sepsis Guo-min Li, MDa, 1, a Department of Anesthesiology and Intensive Care, Jintan Hospital, Jiangsu University, Changzhou, China Received 18 April 2012. Revised 18 June 2012. Accepted 22 June 2012. Available online 7 July 2012. http://dx.doi.org/10.1016/j.jss.2012.06.058, How to Cite or Link Using DOI Background Hydrogen has been reported to selectively reduce hydroxyl radicals and peroxynitrite anion in many pathologic processes. This study aimed to test the hypothesis that hydrogen-rich saline (HRS) may ameliorate organ dysfunction in a rat model of polymicrobial sepsis. Methods Sepsis was induced in male Sprague-Dawley rats by cecal ligation and puncture (CLP). Twenty-four rats were equally assigned to Sham group, CLP group, and CLP + HRS group (n = 8). At 0, 6, and 18 h after CLP or sham operation, rats received an intraperitoneal injection of HRS (5 mL/kg) or the same volume of normal saline. Malondialdehyde, superoxide dismutase activities, inflammatory mediators, pulmonary nitric oxide, myeloperoxidase activities, wet-to-dry weight ratio, histologic scores, apoptotic analysis, alanine aminotransferase, creatinine, and blood urea nitrogen were assessed at 24 h after operation. The 7-d survival rate was also recorded. Results HRS administration significantly reduced the serum high-mobility group box, alanine aminotransferase, creatinine, and blood urea nitrogen levels; the pulmonary interleukin 6, high-mobility group box, nitric oxide, and malondialdehyde levels; and the wet-to-dry weight ratio, total histologic scores, and TUNEL-positive cells, whereas it increased the superoxide dismutase activities 24 h after CLP when compared with the CLP group. However, there was no significant difference in survival rate between the CLP + HRS and CLP groups. Conclusions HRS has potential protective effects against sepsis by decreasing proinflammatory responses, oxidative stress, and apoptosis in a rat model of polymicrobial sepsis.
氢气生物学研究目前进展迅速，虽然有大量的动物实验证明对许多疾病具有治疗作用，但如果没有严格的随机对照临床试验的证据，则无法获得临床治疗特别是国家医药管理局等的最终许可，也就是说无法获得官方的正式批准用于临床的使用。开展严格对照的临床研究是氢气医学发展最重要的任务和手段。也是将来深入研究氢气生物学效应最重要的环境保障和研究目的。 但国际上在临床试验方面进展并不快，到目前为止，临床试验的报道基本都来自日本，这里是从世界卫生组织临床试验注册的信息中检索到的临床试验注册信息也说明这个问题，这些信息显示出日本在神经系统损伤治疗方面的关注度比较大，例如 6 项注册试验中有 3 项属于神经系统损伤治疗效果的研究，分别是中风、巴金森病和中度认知障碍的研究。比较有意思的是，最早报道氢气生物学效应的日本医科大学没有注册临床试验的信息，是他们没有信心，还是没有获得研究经费的资助。因为他们曾经获得来自商业公司的赞助，并成立氢气医学中心。从这一点上看，似乎没有这些问题。但内情不清楚。 第一项：氢气水治疗巴金森病 第二项：氢气水对正常人的抗氧化效果评价 第三项 氢气生理盐水注射对脑缺血的治疗效果评价 第四项 氢气水对中度认知障碍治疗效果的研究 第五项 氢气水治疗间质性膀胱炎 第六项 氢气水对糖尿病的治疗效果评价 所有信息可从世界卫生组织的临床试验注册网上免费检索，更详细的信息可从 http://apps.who.int/trialsearch/AdvSearch.aspx 检索。 建议检索词为 ： hydrogen 。 Recruitment status 选择 ALL 。否则无法获得全面的信息。 详细信息 第一项：氢气水治疗巴金森病 2012 年 3 月 14 日注册的用“氢气水治疗巴金森病”开始实验 2010 年 1 月 1 日顺天堂大学附属医院神经外科，联系人 Asako Yoritaka 。日本学者曾经报道使用氢气水治疗巴金森病的动物实验效果，发表论文 3 篇。全部使用自由饮用氢气水。治疗设计 The subjects should make 1000 ml of molecular hydrogen water which contains 1.6 ppm dissolved hydrogen by Aquerable, and consume for 48 weeks . Placebo water which is not contained molecular hydrogen water made from pseudo-machine. The subjects consume for48 weeks. 第二项：氢气水对正常人的抗氧化效果评价 Studies on in vivo effects of drinking a water product dissolving hydrogen gas as an in vitro antioxidant additive 杏林大学 Atsushi Hiraoka 自 2009 年 5 月 1 日开始的针对健康人的一项研究，排除肝脏肾脏功能异常和月经期女性。 Ingestion of 500ml per day of hydrogen gas-dissolving water for 1 week.Ingestion of 500ml per day of control water without hydrogen gas for 1 week.. 观察指标 the levels of serum LPO and urine 8-OHdG in subjects immediately before and after 1-week drinking period for 500ml per day of tap water with or without dissolved hydrogen gas at 0.34mg/l and 1-week before and after the drinking per day. 第三项 氢气生理盐水注射对脑缺血的治疗效果评价 日本国防医科大学神经外科 Hiroshi Nawashiro 于 2011/06/01 开始的 Molecular hydrogen for ischemic stroke 。选择诊断后症状发生 24 小时内脑缺血患者 Patients were eligible for enrollment if they were 18 years or older and had a clinical diagnosis of acute ischemic stroke within 24 hours of symptom onset. They had to score at least 6 on the National Institutes of Health Stroke Scale (NIHSS) with at least 2 points for limb weakness. All patients received appropriate routine stroke care as per local treatment practices, including alteplase for eligible patients presenting 3 hours from onset; patients receiving alteplase had to commence the study drug before the alteplase infusion.Exclusion criteria: Patients with acute ischemic stroke beyond 24 hours of symptom onset. 治疗方法为静脉点滴注射氢气生理盐水。效果评价 modified Rankin scale (mRS) (days 7, 30, and 90), the NIHSS (days 7 and 90), and the Barthel Index (days 7, 30, and 90) Safety Assessments Vital signs were recorded at enrollment and at specified times throughout the infusion and follow-up periods. Routine laboratory data and ECGs were performed at the time of enrollment, at 24 and 72 hours, and on day 7 and were analyzed centrally (ECGs at day 7 were performed only if abnormal at 72 hours). To assess any effect of hydrogen on hemorrhagic transformation after alteplase administration, brain imaging was repeated after 72 hours in patients who were receiving concomitant treatment with alteplase. Symptomatic hemorrhagic transformation was defined as an increase in the NIHSS score of at least 4 points within 36 hours, plus evidence of any blood on neuroimaging after treatment with alteplase. Patients meeting criteria for progressive stroke (NIHSS increase of 4 points within 72 hours) or new stroke in the first week were also reimaged. 第四项 氢气水对中度认知障碍治疗效果的研究 筑波医科大学临床医学研究所神经精神学系 Takashi Asada2009/07/01 开始的 A randomized trial to assess the effects of hydrogen-ride dissolution water for the patients with mild cognitive impairment (MCI). 中度认知障碍的研究。 招募对象： Inclusion criteria: 1) Participants of the Tone project. 2) Being able to give a written informed consent to the participation in the present study. 3) Having diagnosis of the mild cognitive impairment. 4) Being able to observe the following requirement: good compliance with the water; participation in the scheduled examinations for assessment; keeping a log-diary recording the consumption of the water. 5) Having a modified Hachinski Ischemic score of 4 or less. 6) Having the 15-item Geriatric Depression Scale score of 6 or less. Exclusion criteria: 1) Meeting DSM-IV TR criteria for dementing illnesses. 2) Having serious or unstable illnesses. 3) Having a history within past 5 years of serious infectious disease affecting the brain and/or malignant diseases. 4) Having a history of alcohol or drug abuse or dependence (on DSM-IV TR) within the past 5 years. 5) Receiving any types of anti-Alzheimer drugs. 6) Recent (within 4 weeks) initiation of medications that affect the central nervous system.Age minimum: 67years-old.Age maximum: Not applicable Gender: Male and Female 研究内容： mild cognitive impairment 治疗手段 The patients of hydrogen group will be intervened with 500ml hydrogen dissolution water every-day for 2 years. The patients of placebo group will be intervened with 500ml ordinary water every-day for 2 years. 效果评价 Score in Japanese version of ADAS-Cog and Mini Mental State Examination. Scores in Japanese version of ADCS-ADL, MRI and SPCET imaging, and Geriatric Depression Scale. 第五项 氢气水治疗间质性膀胱炎 Koushinkai Hospital 的 Comprehensive Support Project for Clinical Research Office 于 2008/07/01 开始的 A randomized trial to asses the effects of hydrogen-rich dissolution water in patients with interstitial cystitis 。至少现在没有氢气在间质性膀胱炎方面的研究报道，无论是动物实验还是临床报道。 研究对象标准： Inclusion criteria: 1) Patients who are able to give written informed consent 2) Patients who has the characteristic finding under hydraulic distension of the bladder in interstitial cystitis by cystoscope 3) It has taken more than 12 weeks after patients took the hydraulic distension of the bladder, and symptom of patients are in stable. 4) More than 7 marks in total of Interstitial Cystitis Score in registration 5) More than 4 marks in Q4 (degree of bladder pain) in Interstitial Symptom Score 6) Age is over 20 years and less than 80 years 7) Patients are able to do the following things in this trial; getting good compliance with intaking investigating food and coming to hospital, and writing the diary and the questionnaire accurately by themselvesExclusion criteria: 1) More than 200ml of an average voided volume at a time before the registration 2) Patients with active infection of urinary tract 3) Patients with bacterial cystitis within 12 weeks before registration 4) Patients with vaginosis 5) Patients with calculus of lower urinary tract or urethral diverticulum 6) Patients with nephrosis syndrome 7) Patients with active genital herpes 8) Patients who have operated the surgery in pelvis or its circumference and it has not taken more than 24 hours after the surgery 9) Patients with cerebrospinal disease 10) Patients with the follow disease or suspected disease; neurogenic bladder, cystitis radiation, tuberculous cystitis, cystitis with BCG, drug associated cystitis 11) Start, stop, or change of the dose of the following drugs within 4 weeks after the registration; (a) Antiphlogistic analgetic (b) Antidepressant (c) Anticholinergic drug (d) Antihistamine drug (e) Ataractic drug (f) Drug treatment for frequent urination and acraturesis (g) Steroid drug 12) Start or stop new bladder training or diet therapy within 4 weeks befor registration 13) Patients who has received bladder instillation therapy, electrical stimulation therapy, or acupuncture and moxibustion within 12 weeks before registration 14) Patients with serious hepatic or kidney damage 15) Patients with serious heart disease 16) Patients with malignant tumors which effect their general status or survival time 17) Patients with the history of serious drug-induced adverse effect 18) Patients who are in pregnancy, while breast-feeding, or have possibilities of them, or desire pregnancy in test period 19) Patients who have taken part in the her clinical research within 12 weeks 20) Patients who have taken part in the her clinical research within 12 weeks 21) Patients who are inadequate, which their physicians assessed itAge minimum: 20years-old Age maximum: 80years-oldGender: Male and Female 治疗方法 The patients will be intervened with hydrogen dissolution water group (hydrogen group) 200ml every three times in a day in 2 months (56days) . After that, the patients in hydrogen dissolution water group will be transferred to the additional intervention term after the end of intervention.And after that, the patients will be randomized to withdrawal terms for more 1 month with hydrogen dissolution water or with placebo water. The proportion of the patients who has been assessed "success" at the end of the intervention Secondary Outcome(s) 1) Changes of the Symptom Score in Interstitial Cystitis Symptom Index(ICSI) 2) Changes of the Problem Score in Interstitial Cystitis Problem Index(ICPI) 3) An Average frequency of urination per day 4) An Average voided volume at a time 5) Degree of urge to urinate; PUF symptom score 6) Degree of bladder pain 7) Impression by patients with GRA (Global Response Assessment) 8) Urine Test; 8-OHdG in urine 9) Adverse Events (we cannot deny the association between the food and the event) 第六项 氢气水对糖尿病的治疗效果评价 研究京都大学医学院 Comprehensive Support Project for Clinical Research Office 于 21/08/2008 开始的 A Randomized trial to assess the effects of hydrogen-rich dissolution water for patients with impaired glucose tolerance or impaired fasting glucose 。该研究已经发表论文。 研究对象标准： Inclusion criteria: 1) Patients who are abele to give written informed consent 2) FBS is over 100mg/dl and under 126mg/dl in registration 3) Age is over 20 years and less than 80 years 4) Type of practice: outpatient department 5) Patients are able to do the following things in this trial - getting good compliance with consuming investigational food and coming to hospital, and writing the diary and the questionnaire accurately by themselvesExclusion criteria: 1) Patients who have receive drug treatment for diabetes 2) Patients with the diseases which have possibility with impaired glucose tolerance 3) Patients with serious liver or kidney damage 4) Patients with serious heart disease or cerebrovascular disorders 5) Patients with serious disease in pancreas or blood disease 6) Patients with malignant tumors which effect their general status or survival time 7) Patients who are in pregnancy, while breast-feeding, have possibilities of them, or desire pregnancy in test period 8) Patients with alcohol abuse 9) Patients who have taken part in the her clinical research within 12 weeks 10) Patients who have taken part in the her clinical research within 12 weeks 11) Patients who are inadequate, which their physicians assessed it Age minimum: 20years-oldAge maximum: 70years-oldGender: Male and Female 研究内容 Impaired glucose tolerance or impaired fasting glucose 治疗手段 The patients will be intervened with hydrogen dissolution water group (hydrogen group) 200ml every three times in a day in 3 months (84days) . The patients will be intervened with normal water group (placebo group) 200ml every three times in a day in 3 months (84days) 效果评价方法 1)75gOGTT(glucose);0 minute (in the fasting state),30,60,90 minutes later after loading2)Delta AUC(0-120min);The difference of the area under the plasma glucose concentration before and after administration
Hydrogen saline is protective for acute lung.pdf 四川大学华西医院 麻醉系刘进教授课题组 2011 年在《外科研究杂志》上发表了氢气治疗兔肺缺血再灌注损伤的文章。 该实验室是国际上最早开展氢气研究的单位之一 。 最近 上海 胸科医院发表类似论文，证明注射氢气生理盐水可以治疗大鼠肺缺血再灌注损伤，该文章发表在 Heart, Lung and Circulation 杂志上，该研究证明注射氢气生理盐水（先连续提前 3 天，然后在缺血再灌注时注射 0.5 ml/kg ）可以治疗肺缺血再灌注组织损伤，作者分别采用肺组织干湿比（可以分析肺水肿程度）、肺动脉体外收缩和舒张功能（分别用乙酰胆碱和肾上腺素诱导血管的扩张和收缩功能）、 HE 染色（肺组织病理损伤程度）、 TUNEL 染色和 caspase-3 活性（显示肺组织细胞凋亡的程度）、 MDA 和 8-OHdG （组织氧化损伤程度，分别代表脂质和核酸氧化损伤的程度）以及血液白细胞 CRP 等生物标志。等研究手段，证明氢气盐水注射对大鼠肺缺血再灌注损伤有治疗作用。 疑问： 1 、为什么要提前使用药物，什么原因？提前抗氧化有什么价值？如果有，为什么没有和单独损伤后进行对比，至少过去没有见过类似给药物的方法。 2 、氢气生理盐水剂量为什么采用 0.5 ml/kg ，有什么依据？ 3 、我前面提到 2011 年就有四川华西医院发表氢气生理盐水对兔肺缺血再灌注损伤有治疗作用，本文为什么没有引用？本文投稿日期是 2012 年 2 月，应该可以看到该文献，从文献类似度上考虑，这是一篇绝对必须引用的文献。 Hydrogen saline is protective for acute lung ischaemia/reperfusion injuries in rats Jianxin Shi1, Department of Thoracic Surgery, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai 200030, People's Republic of China Protective effects of saturated hydrogen (H2) saline on cardiac ischaemia–reperfusion (I/R) injury have been demonstrated previously. This study was designed to show that hydrogen-rich saline is protective in preventing lung I/R injury in rats. Adult male Sprague-Dawley rats underwent 45 min occlusion of the right lung roots and 120 min reperfusion. Rats were divided randomly into three groups: sham-operated control group, I/R plus saline treatment, and I/R plus hydrogen-rich saline treatment (0.6 mmol/L, 0.5 ml/kg/d). Three days of intraperitoneal injection of hydrogen-rich saline before the reperfusion combined with immediate administration of hydrogen-rich saline after the reperfusion were performed. Following reperfusion, the lung tissue and the pulmonary artery was immediately obtained and the W/D ratio, pulmonary artery contraction and relaxation ability, H–E staining, TUNEL staining, caspase-3, MDA, 8-OHdG content and measurement of such biomarkers as WBC, CRP were measured or carried out. Results Hydrogen saline significantly protected vasoactivity of the pulmonary artery, reduced pulmonary oedema, decreased lung malondialdehyde (MDA), 8-OHdG concentration, alleviated lung epithelial cell apoptosis and lowered the level of such biomarkers as WBC, CRP, ALT and TBiL. Conclusions It is concluded that hydrogen-rich saline is a novel, simple, safe and effective method to attenuate pulmonary I/R injury. Hydrogen-Rich Saline Attenuates Lung Ischemia-Reperfusion Injury in Rabbits.pdf
Hydrogen-rich saline reduces the oxidative stress and relieves the severity of t.pdf 胰腺炎是胰腺因胰蛋白酶的自身消化作用而引起的疾病。常见类型有急性胆源性胰腺炎、蛔虫性急性胰腺炎、胆源性急性胰腺炎、急性出血坏死型胰腺炎等急性胰腺炎，自身免疫性胰腺炎、胆源性慢性胰腺炎等慢性胰腺炎。治疗原则是手术和对症支持治疗。 胰腺是人体第二大消化腺体，是消化作用最强的器官。胰腺分泌的胰液是人体最重要的消化液。正常情况下，胰液在其腺体组织中含有不活动即无活性的胰酶原。胰液沿胰腺管道不断地经胆总管奥狄氏括约肌流入十二指肠，由于十二指肠内有胆汁存在，加上十二指肠壁粘膜分泌一种肠激酶，在二者的作用下，胰酶原开始转变成活性很强的消化酶。一般情况下，胰管和胆管虽然都经过一条通道流入十二指肠，但由于胰管内的压力高于胆管内的压力，胆汁不会反流入胰管内。只有当奥狄氏括约肌痉挛或胆管内压力升高，如结石，肿瘤阻塞，胆汁才会反流入胰管并进入胰腺组织，此时，胆汁内所含的卵磷脂被胰液内所含的卵磷脂酶 A 分解为溶血卵磷脂，可对胰腺产生毒害作用。或者胆道感染时，细菌可释放出激酶将胰酶激活，同样可变成能损害和溶解胰腺组织的活性物质。这些物质将胰液中所含的胰酶原转化成胰蛋白酶，此酶消化活性强，渗透入胰腺组织引起自身消化，亦可引起胰腺炎。 胰腺炎发病原因有胆道系统疾病、酒精或药物引起、传染病并发胰腺炎，家族性高脂血症患者、动脉粥样硬化及结节性动脉周围炎、十二指肠克罗恩病波及胰腺容易发生胰腺炎，胰管阻塞，胰管结石、狭窄、肿瘤等可引起胰液分泌旺盛，胰管内压增高，胰管小分支和胰腺腺泡破裂，胰液与消化酶渗入间质，引起急性胰腺炎。低蛋白饮食可导致慢性胰腺炎，多见于东南亚、非洲及拉丁美洲各国。遗传性胰腺炎 (hereditary pancreatitis) 较少见，属染色体显性遗传。 外伤与手术是急性胰腺炎的常见原因，只有在创伤严重或损伤主胰管后方可能引起慢性胰腺炎。 胰腺炎该病主要由胰腺组织受胰蛋白酶的自身消化作用。胰腺炎时因某些因素（下述）激活了胰蛋白酶，后者又激活了其它酶反应，如弹性硬蛋白酶（ elastase ）及磷脂酶 A （ phospholipaseA ），对胰腺发生自身消化作用，促进了其坏死溶解。另外，胰蛋白酶对由脂蛋白构成的细胞膜及线粒体膜并无作用，而胰液中的磷脂酶 A 被脱氧胆酸激活后，作用于细胞膜和线粒体膜的甘油磷脂，使之分解变为脱脂酸卵磷脂，亦称溶血卵磷脂（ lysolecithin ），后者对细胞膜有强烈的溶解作用，可溶解、破坏胰腺细胞膜和线粒体膜的脂蛋白结构，致细胞坏死。脂肪坏死也同样先由胰液中的脱脂酸卵磷脂溶解、破坏了脂肪细胞膜后，胰脂酶才能发挥作用。 急性胰腺炎是胰酶消化胰腺及其周围组织所引起的急性炎症，主要表现为胰腺呈炎性水肿、出血及坏死，故又称急性出血性胰腺坏死（ acutehemorrhagicnecrosisofpancreas ），好发于中年男性，发作前多有暴饮暴食或胆道疾病史。临床表现为突然发作的上腹部剧烈疼痛并可出现休克。按病变表现不同，可将本病分为急性水肿性（或间质性）胰腺炎及急性出血性胰腺炎二型。坏死出血型较少见，但病情严重，死亡率高。 急性胰腺炎非手术治疗除常规支持疗法，如禁食鼻胃管减压、补充体液防治休克、解痉止痛、抗生素、中药和腹腔渗出液的处理外，比较特异的方法是抑制胰腺外分泌及胰酶抑制剂，胃管减压、 H2 受体阻滞剂 { 如西咪替丁 ) 、抗胆碱能药 ( 如山莨菪碱、阿托品 ) 、生长抑素等，但后者价格昂贵，一般用于病情比较严重的病人。胰蛋白酶抑制剂如抑肽酶、加贝酯等具有 — 定的抑制胰蛋白酶的作用。 曾经有报道氢气生理盐水可以治疗精氨酸诱导的急性胰腺炎，最近 J Trauma Acute Care Surg. 发表来自成都军区总医院的文章，证明氢气生理盐水注射可以治疗创伤性急性胰腺炎，研究发现氢气生理盐水注射后，尽管创伤性急性胰腺炎血浆中淀粉酶、脂酶活性未见明显改变，但治疗组动物死亡率明显下降，血浆炎症因子、胰腺组织 丙二醛显著下降， 胰腺组织谷胱甘肽和超氧化物歧化酶（ SOD ）显著增高。研究结果表明，氢气对创伤性急性胰腺炎后全身炎症反应、局部组织抗氧化能力和氧化应激等均有改善作用。提示氢气对创伤性急性胰腺炎具有治疗价值。 Ren J, Luo Z, Tian F, Wang Q, Li K, Wang C. Hydrogen-rich saline reduces the oxidative stress and relieves the severity of trauma-induced acute pancreatitis in rats. J Trauma Acute Care Surg. 2012 Jun;72(6):1555-61. Currently, little evidence exists to support whether the therapeutic approaches for treating ordinary acute pancreatitis (AP) are effective in trauma-induced pancreatitis. Hydrogen-rich (H2) saline is an antioxidant treatment capable of ameliorating the severity of L-arginine-induced AP. In this study, we attempted to validate its protective role against traumatic pancreatitis (TP). A previously established experimental rat model of TP was generated by controlled delivery of high pressure air impact. The protective effects of H2 saline against TP were evaluated in this model system by measuring survival rate and determining changes in histopathology, plasma enzymes, cytokines, and oxidative stress-associated molecules. Intraperitoneal administration of H2-rich saline produced a pronounced protection against TP in rats. Significant improvements were observed in survival rate and histopathological findings. In addition, plasma cytokines concentrations were reduced in H2 saline-treated TP rats. Although no marked inhibitory effect on plasma amylase and lipase activities was observed, H2 saline caused considerable suppression of pancreatic malondialdehyde level and recruitment of endogenous pancreatic antioxidants, such as glutathione and superoxide dismutase. H2-rich saline has beneficial effects on TP, presumably because of its detoxification activities against excessive reactive oxygen species. Our findings highlight the potential of H2-rich saline as a therapeutic agent of trauma-induced AP.
基因治疗从概念上来讲很简单，就是用含有正确序列的新基因拷贝来取代或补充致病的突变基因。从理论上来说，基因治疗策略不光仅仅能够用以治疗，而且可以从根本上治愈人类遗传病。然而，在实际应用中要实现安全有效的基因治疗，从来就不是理论上设想的那么简单。 从 1989 年首次应用于人类，基因治疗临床试验至今已走过了二十余载跌宕起伏的路程。如果不是最近几年以来相继报道的基因治疗在肿瘤（实体肿瘤和白血病）、血友病和神经遗传病等领域的突变性进展，人们对基因治疗的信心恐怕远不会像今天这样乐观。 6 月 1 日 《科学家》杂志发表《 Targeting DNA 》一文，以访谈形式系统总结了基因治疗的历史与现状。 Targeting DNA By Jef Akst | June 1, 2012 T he concept is simple: if a mutated gene is causing a problem, replace or supplement it with a new, accurate copy. In theory, such a strategy could not just treat, but cure countless human genetic diseases. In practice, however, developing safe and effective gene therapies has not been easy. Even when identifying a disorder’s genetic basis is fairly straightforward, finding the appropriate delivery vector to target the diseased tissues in the body, while avoiding unintended consequences, has challenged would-be gene therapists for more than 20 years. But more and more researchers are convinced that the technique is on the brink of becoming a common medical practice. “It’s an incredibly exciting time for the field,” says researcher and medical oncologist David Kirn, founder, president, and chief medical officer at Jennerex, Inc., a San Francisco-based biotherapeutics company that develops and commercializes oncolytic drugs. In the last year alone, he says, major breakthroughs have been published for the use of gene therapy in patients with hemophilia, solid tumors, and leukemia, not to mention the dozens of trials yielding positive results for gene therapies to treat various types of blindness. “It’s just remarkable,” he says. “These decades of work are suddenly really paying off.” Fits and starts It hasn’t always been such high times for gene therapy, however. The field was booming in its early days, with approvals for gene therapy clinical trials rising exponentially from the first one in 1989 to 116 in 1999. But that year, gene therapy trial participant Jessie Gelsinger, a relatively healthy 18-year-old who had an unusually mild form of liver disease caused by mutations in a gene on the X chromosome, died 4 days after receiving an injection of an adenovirus carrying an unmutated copy of the gene meant to correct his condition. The viral vector apparently triggered a massive immune response that caused multiple organ failure and brain death. Then, starting in 2002, reports from Paris and London told of patients developing a leukemia-like disease following treatment in clinical trials for a rare autoimmune disorder called severe combined immunodeficiency (SCID), or “bubble-boy” disease. SCID patients lack a functioning immune system, and thus must live in highly sterile conditions to prevent life-threatening infections. The studies started out extremely well: most of the infant boys were able to live relatively normal lives, no longer confined to their “bubbles.” The trials were hailed as the first unequivocal gene therapy success. But in the years that followed, 5 of the 20 trial patients developed a leukemia-like disease—an effect that was traced to the retroviral vector used to deliver the corrective gene to bone marrow cells ex vivo. The vector had inappropriately inserted the gene into the babies’ genomes close to a proto-oncogene involved in white blood cell proliferation, activating the gene and triggering a flood of T cells. After the second child fell ill, the US Food and Drug Administration suspended 30 US trials using the same retrovirus, or about 15 percent of the 200 gene therapy trials under way at that time—a move the agency called a precautionary measure. Of the five patients that developed leukemia, one died; the rest are in remission. Events like these had “a big negative impact in the field,” recalls molecular cell biologist Mien-Chie Hung of the University of Texas MD Anderson Cancer Center. Interest in gene therapy started to wane, and treatments that might have been expected to hit the market years ago are still plugging through the clinical trial process. But things are looking up. Just last year, for example, researchers published long-term survival data for two UK gene therapy trials for SCID: the original London trial for X-linked SCID (SCID-X1) and a second trial for adenosine deaminase (ADA) SCID. 1 , 2 In both trials, the researchers had extracted the patients’ bone marrow, inserted a functioning copy of the disease-causing gene, and infused the altered cells back into the patients. The impressive bottom line: up to 9 years after treatment, 14 of the 16 children treated have had their immune systems restored and have been able to live relatively normal lives free of any bubbles. “These kids were living in bubbles with a life expectancy of less than 20 years; they had no quality of life,” says Kirn. “And now many of them are essentially cured. I mean, it’s a medical miracle.” Many other gene therapy trials are currently underway—and yielding positive results—for numerous other diseases, including various forms of hereditary blindness, HIV, hemophilia, neurodegenerative diseases, and a variety of cancers. Though no gene therapies have yet received FDA approval, nearly 2,000 clinical trials have been initiated in the last 5 years alone, according to clinicaltrials.gov, many with seemingly miraculous results and—thanks to improved vectors and techniques—none of the devastating side effects that plagued the field in its earlier days. “There is a lot of exciting information coming out right now,” says Howard Hughes Medical Institute investigator Katherine High of the University of Pennsylvania School of Medicine and the Children’s Hospital of Philadelphia, who coauthored a report published last December of a successful gene therapy for hemophilia B. 3 Loading up an adenovirus-associated virus with a gene encoding a functional version of the clotting agent known as factor IX, researchers in the United Kingdom infused the vector into six men with severe hemophilia B. A single treatment was enough to increase clotting factor IX to levels that, while still well below normal, enabled sufficient clotting to allow four of the patients to discontinue factor IX replacement therapy altogether, and the other two to receive factor IX injections less frequently. “To me, clearly is going to be a therapeutic pathway forward for a whole range of diseases,” predicts High. An eye on gene therapy Nowhere has gene therapy made more of a splash than in blindness research. Because the eye is an immune-privileged site, injecting viruses is unlikely to result in the sorts of immune complications that killed Gelsinger in 1999. Indeed, there are some 23 completed and ongoing clinical trials for various types of blindness disorders, and no serious side effects have been reported. “It’ll be a few years” before these therapies reach the market, says University of Florida molecular virologist William Hauswirth. But the results so far are “bordering on spectacular as far as improving vision in the patients,” he adds. In a recent, 3-year follow up on 15 patients with Leber congenital amaurosis (LCA), a degenerative retinal disease that causes childhood blindness, Hauswirth and his colleagues found that within the area of the retina they are targeting for treatment, “patients have gained light sensitivity from as little as to 200-fold to as much as 60,000-fold,” he says. Twelve of the patients have also demonstrated significant improvement in visual acuity, reading an extra three lines lower on an eye chart, and in 13 of the patients, their pupils constricted when exposed to light as much as 100-fold dimmer, a more objective measure of light sensitivity. 4 This particular group of patients was treated with a modified adeno-associated virus (AAV) carrying the gene RPE65 , which helps metabolize a form of vitamin A that allows rods and cones to function. The modified virus was injected behind the eye, directly under the retina, where the corrective gene entered some 15 to 20 percent of cells in the retinal pigment epithelium (RPE), the nourishing cell layer just beneath the retinal visual cells. The AAV is a popular choice for gene therapies now in development. It is a nonpathogenic virus that usually elicits no noticeable immune response, and does not integrate into the host genome, meaning there is little risk of triggering disease-causing mutations, via the activation of an oncogene, for example. The virus simply delivers the gene to the cell’s nucleus, where it forms small circles of DNA called episomes that can be expressed under the control of promoters, also delivered by the virus. “AAV is by far the most successful vector for many applications in disease so far,” says Hauswirth. The disadvantage to this strategy is that if the replacement gene is not integrated into the cell’s DNA, it will be lost when the cell divides, as circular DNA is not replicated with the rest of the nuclear genome. But because retinal cells are extremely long-lived, a nonintegrating virus is perfectly suitable for treating eye diseases. The main factor limiting the utility of AAVs as gene vectors is their small size—no larger than a nanoparticle. This means they can only carry about 4.7 kilobases of DNA, and that must include any promoters needed to regulate the expression of the therapeutic DNA. This works just fine for Hauswirth’s RPE65 replacement therapy, but many other ocular diseases, as well as diseases of other body systems, result from mutations in genes that are quite a bit larger. For therapies to correct these disorders, researchers must turn to other options. Most early experiments in gene therapy for eye diseases used adenoviral vectors, which, like AAVs, are nonintegrating vectors, but, with a 36-kilobase genome, provide much more space for therapeutic DNA. However, because many humans already carry antibodies to adenovirus, the great drawback of this approach is the risk of immunotoxicity, which can disable the therapeutic vector or cause side effects in the host. It was an extreme reaction to an adenoviral vector that killed Jesse Gelsinger. Because adenoviruses are efficient at entering many cell types and delivering the goods, they are targets of intensive research to make them safer for the treatment of cancer, diabetes, HIV, and genetic diseases. Lentiviruses, a type of RNA retrovirus, also have a sizeable carrying capacity: some 9 kilobases of genetic material. “You only have to delete a few genes to get lentivirus to carry twice as much as AAVs,” Hauswirth says. And in contrast to adenoviruses and AAVs, they do integrate into the genome, making them efficient at delivering and establishing stable high levels of transgene expression in both dividing and nondividing cells. While they cannot yet be targeted to integrate at specific sequences, they don’t gravitate to oncogenes or growth-related genes as some other retroviral vectors do, and are at the forefront of the race to market. Oxford BioMedica, for example, is using its integrating lentiviral vector, called LentiVector, to deliver treatments for a variety of eye diseases, including wet age-related macular degeneration, Stargardt disease, Usher syndrome, and corneal graft rejection. All of these therapies are in phase I/II development in partnership with Sanofi, and “the results we have seen in our clinical trials to date have been encouraging,” says the company’s chief scientific officer, Stuart Naylor. “We believe it is only a matter of time before a gene therapy is approved for the market.” “The field is running at the speed of light,” agrees veterinary ophthalmologist and basic vision scientist Gustavo Aguirre of the University of Pennsylvania. “We’re beyond anything that we thought in the ’90s.” Targeting cancer: pluses and minuses As with gene therapies for other diseases, the technique’s use in treating cancer is gathering steam. Oncolytic viruses that target and destroy tumor cells are being combined with gene therapy techniques to provide tools to jack up those viral vectors with more potent genetic loads. “The concepts have been around for centuries—the fact that viruses can certainly destroy cancer cells,” says Jennerex’s Kirn. Now, “in addition to replicating and expressing viral genes, we also express therapeutic transgenes and imaging genes.” Jennerex is developing a vaccinia virus vector called JX-594, for example, to deliver genes that activate the epidermal growth factor receptor (EGFR)/Ras pathway in cancer cells, resulting in cell lysis and increased anticancer immunity. In multiple phase I and II trials involving numerous cancer types, including liver, colon, kidney, lung, and melanoma, JX-594 has shrunk tumors and is well-tolerated by patients. Last November, Jennerex researchers announced that advanced liver cancer patients receiving JX-594 in a phase II trial had a 60 percent decreased risk of death after 1 year as compared to controls. Another engineered oncolytic virus nearing clinical approval is OncoVEX, developed by Massachusetts-based biotech BioVex, which last year partnered with Amgen in a deal that could be worth $1 billion. The drug, named last year as one of FierceBiotech’s 10 promising late-stage cancer drugs, is a special strain of the Herpes simplex type 1 (HSV-1) virus that carries an immune-boosting component. Results of a phase II trial for metastatic melanoma announced in 2009 showed that 26 percent of 50 patients responded to treatment, with 8 completely recovering, 5 and the treatment is now in phase III trials. OncoVex has also shown activity against breast and pancreatic cancers in phase I trials. And these successful examples are not the outliers. “We’re entering a golden age here of genetic therapies and viral therapies,” says Kirn. Getting to this point took some careful planning, however, to avoid the destruction of healthy tissue. “The most important thing is you need a specific target,” says MD Anderson’s Hung. Most oncolytic viruses are designed to target a receptor or surface protein that is overexpressed on cancer cells, to increase the chance that the viruses enter and kill only diseased tissue. But this, of course, requires that tumors have unique surface antigens. “That’s the ideal situation—to find something on tumor cells that’s not on normal cells,” says hematologist-oncologist David Porter of the University of Pennsylvania Medical Center. “That’s not possible in most tumors,” he says, which may be part of “the reason this field has been so slow to develop.” So Hung is taking a slightly different approach. Instead of focusing on viruses that selectively target cancer cells, or rigging them to do so, Hung and his colleagues have designed a vector, dubbed VISA, that goes everywhere, but whose package is only activated in cancer cells. “DNA by itself does not cause side effects—every cell has DNA,” says Hung. “It the gene product.” The VISA vector is designed to amplify expression of the genes it carries under the control of a promoter that is expressed at higher levels in tumors. “Then whatever DNA we inject into the bloodstream may go anywhere, but will express only in cancer cells,” Hung explains. In 2007, the researchers used VISA, equipped with a modified apoptosis promoter, called BikDD , to knock down pancreatic cancer in mice. 6 The protocol is currently being tested for safety in a phase I trial, and Hung and his colleagues are now applying the VISA vector to breast cancer, with promising preclinical results coming out just last year, 7 as well as to lung, ovarian, and liver cancers. Yet another gene therapy strategy for fighting cancer involves the collection of immune cells from patients, insertion of genetic material that essentially trains the cells to target and kill cancer, and the infusion of those cells back into the patients. “The idea is to somehow modify a person’s own T cells so they can now recognize and attack tumor cells that they otherwise aren’t able to kill,” says Porter. “By definition, if someone has a tumor, their T cells aren’t able to kill .” In 2011, Porter, along with Carl June and other colleagues at Penn Medicine, reported that they had engineered a patient’s T cells ex vivo to target chronic lymphocytic leukemia (CLL) cells, marking the first gene therapy success for advanced cancer. The researchers used a lentivirus to insert a chimeric antigen receptor targeting CD-19—a molecule found on “all CLL cells but only a small subset of normal cells,” Porter says—as well as two potent signaling domains, which help the T cells survive longer and activate strongly at the appropriate time. Last August, the researchers published the results of a phase I trial, in which two participants had been in complete remission for up to a year, and a third was showing signs of a strong antitumor response. 8 , 9 In addition, some of the cells persisted as memory T-cells, primed to attack in the event of a recurrence. However, researchers had to weigh the fact that the engineered T cells could also attack healthy tissue. CLL is characterized by the abnormal proliferation of B cells, but it’s not just the malignant B cells that express CD-19; normal B cells also carry the antigen on their surface. As a result of the lentiviral therapy, patients experienced a loss of B cells down to undetectable levels. “ potentially makes them susceptible to infections,” Porter notes. But he is not worried. No patients have contracted any unusual infections in a year and a half of follow up, and antibody replacement is always an option. “We think it is possible to live without B cells,” he says. But it’s an important concern when developing such therapies, he adds. “If you were to target a cell that was also on the lining of your heart or your lungs or your intestines, that wouldn’t be safe.” This strategy is now being applied to diverse cancer types, including mesothelioma (a type of lung cancer), breast cancer, ovarian cancer, and melanoma, all in early-stage trials. With the improved ability to grow T cells in the lab and make better-targeted viral vectors, “the field is really growing,” Porter says. “You can now use this technology to target really almost anything you can identify as a unique target.” The gene’s the limit In 2007, Timothy Brown, a 40-year-old American living in Berlin, had a relapse of acute myeloid leukemia and received a bone-marrow transplant to boost his immune function. Because he was also living with HIV, his doctor chose a donor with a mutation in both copies of the CCR5 gene, which encodes an HIV co-receptor carried on the surface of T cells to which HIV usually must bind in order to enter the cells. People with mutations in both copies of the CCR5 gene are resistant to HIV infection. A year later, Brown relapsed again, and once again received a stem-cell transplant from the CCR5 -mutant donor. Finally, he beat his cancer, and as of 2010, his HIV was still at undetectable levels, despite his having discontinued immune-suppressive treatment. 10 By most accounts, Brown is the first person to have been cured of an HIV infection. The impressive result validated Sangamo Biosciences’ efforts to design a gene therapy to modify the CCR5 gene of HIV patients’ T cells. This is done ex vivo, using an adenovirus vector that delivers its package to T cells carrying the glycoprotein receptor CD4, after which the cells are infused back into the patients. This strategy is different from corrective gene therapies in that it’s taking a normal gene and replacing it with a defective copy. In this case, it’s important to not just supplement the existing copies of the gene, which would continue to produce normal CCR5 receptors, but to render the gene defective so the cells present no CCR5 receptors on their surfaces. To do this, Sangamo researchers are using zinc-finger nucleases, which act as molecular scissors to edit the existing gene. “What we’re doing is . . . actually editing the gene in a way that changes its nucleic acid sequence,” says Sangamo CEO Edward Lanphier. “It’s quite different from classical gene therapy, where you’re just essentially putting in the coding region of a gene.” Preliminary results of these trials, presented at the Conference on Retroviruses and Opportunistic Infections (CROI) last March, have shown that all 21 HIV patients treated with one infusion of their own modified T cells have tolerated the treatment well, and show increased CD4 + T cell counts more than a year later. And when six of the patients took a planned 12-week hiatus from antiretroviral treatment, they had lowered HIV viral loads that correlated with the levels of circulating modified CD4 + T cells. One patient, who naturally carried one copy of the defective CCR5 gene, had undetectable viral levels. The results are promising, says June, who is heading up one of two phase I trials of the therapy, and who hopes that such gene-editing techniques will be applicable to a wide range of diseases. “We’re trying to make a good gene bad in the case of the HIV, so that the cells aren’t infected,” he says, “but you can do the inverse, which is to take a bad gene and fix it.” Such genome editing would be permanent, and leaves the gene under the control of natural promoters, making it more therapeutically appealing, says Lanphier. “The downstream biology of changing the endogenous gene is fundamentally different—and quite frankly, from a therapeutic outcome, superior—than conventional gene therapy approaches,” he says. Other novel gene therapy strategies are also in development, such as employing transiently expressed RNA to confer a therapeutic benefit, and using cell- and protein-based delivery systems instead of viral vectors. With these and other techniques, researchers have recently demonstrated successes in treatments for hemophilia, with promising trial results released last December; numerous neurodegenerative diseases, such as Parkinson’s and Huntington’s; and autoimmune disorders, such as SCID, among others. And importantly, there have been no more safety scares like the ones that rattled the field a decade ago. “The history of medicine says every new technology starts with a great idea and then requires hard work and optimization,” says Kirn. “And I think that’s exactly what’s happened with gene therapy. Hurdles were identified—and there’s always hurdles once you get into a complex human disease situation—and they’ve been addressed.” “The concepts aren’t that much different than they were early on, but the tools are much better,” agrees June. “Now is actually fulfilling the promise that people said it would have.” References H.B. Gaspar et al., “Long-term persistence of a polyclonal T cell repertoire after gene therapy for X-linked severe combined immunodeficiency,” Sci Transl Med , 3:97ra79, 2011. ↩ H.B. Gaspar et al., “Hematopoietic stem cell gene therapy for adenosine deaminase–deficient severe combined immunodeficiency leads to long-term immunological recovery and metabolic correction,” Sci Transl Med , 3:97ra80, 2011. ↩ A.C. Nathwani et al., “Adenovirus-associated virus vector-mediated gene transfer in hemophilia B,” N Engl J Med , 365:2357-65, 2011. ↩ S.G. Jacobson et al., “Gene therapy for Leber congenital amaurosis caused by RPE65 mutations,” Arch Ophthalmol , 130:9-24, 2012. ↩ H.L. Kaufman et al., “Local and distant immunity induced by intralesional vaccination with an oncolytic herpes virus encoding GM-CSF in patients with stage IIIc and IV melanoma,” Annals of Surgical Oncology , 17:718-30, 2010. ↩ X.M. Xie et al., “Targeted expression of BikDD eradicates pancreatic tumors in noninvasive imaging models,” Cancer Cell , 12:52-65, 2007. ↩ J.-Y. Lang, “BikDD eliminates breast cancer initiating cells and synergizes with lapatinib for breast cancer treatment,” Cancer Cell , 20:341-56, 2011. ↩ D.L. Porter et al., “Chimeric antigen receptor-modified T cells in chronic lymphoid leukemia,” N Engl J Med , 365:725-33, 2011. ↩ M. Kalos et al., “T cells with chimeric antigen receptors have potent antitumor effects and can establish memory in patients with advanced leukemia,” Sci Transl Med , 3:95ra73, 2011. ↩ G. Hütter et al., “Long-term control of HIV by CCR5 Delta32/Delta32 stem-cell transplantation,” N Engl J Med , 360:692-98, 2009. ↩ http://the-scientist.com/2012/06/01/targeting-dna/
Treatment of hydrogen molecule abates oxidative stress and alleviates bone loss .pdf 太空飞行因为失重可导致航天员一系列的器官功能改变，特别是骨骼钙丢失和肌肉萎缩等，这些改变不仅对航天员的健康产生危害，而且会影响他们的运动能力，容易使他们在执行太空任务中遇到麻烦，因此开展对这些改变的机制和对抗治疗手段的研究非常重要。对一些希望正在发展太空飞行的国家尤其重要。 氢气是最近几年发现的一种非常特殊的抗氧化物质，虽然目前人们仍没有完全弄清楚氢气治疗疾病的具体分子过程和机制，但大量的临床和动物实验结果表明，氢气对许多类型的炎症、氧化损伤和细胞凋亡等具有非常显著的治疗效果，给人们试图使用氢气治疗许多重要疾病带来了许多联想。 当然民航和太空飞行还会遇到太空辐射的危害，美国航空航天局已经开展了氢气对太空辐射损伤的研究，初步研究证明氢气对太空辐射具有理想的预防效果，并提出将来用氢气作为预防太空辐射的手段。辐射损伤的最重要机制是辐射引起的自由基增加，这是自由基生物学最经典的研究结果，由于氢气具有抗氧化损伤的作用，因此使用氢气作为治疗太空飞行辐射损伤是非常容易从理论上猜测到的。 本研究者根据文献分析认为，太空飞行过程因为示众导致的骨骼肌萎缩和钙丢失和氧化损伤关系密切，采用细胞学和动物模型两类手段,证明使用氢气可以对抗失重导致的动物骨质丢失,并证明这种效应和关键信号分子例如erk、NfkB和iNOS等关系密切。由于骨质疏松不仅在太空飞行，也经常见于临床长期卧床患者的情况，长期卧床由于长期不使用肌肉运动和承受重力，非常容易发生肌肉萎缩和骨质丢失，这种情况和太空飞行存在类似性，因此本研究也提示，氢气可能对那些长期卧床患者的骨质丢失也值得尝试。曾经有研究发现，氢气对废用性肌肉萎缩有一定治疗作用，因此从这个角度考虑，氢气对那些卧床患者可能有对肌肉和骨骼的双重价值。 本研究来自北京 301 医院急救科，文章发表在《国际骨质疏松》杂志上。文章的题目作者和摘要如下： Sun Y , Shuang F , Chen DM , Zhou RB . Treatment of hydrogen molecule abates oxidative stress and alleviates bone loss induced by modeled microgravity in rats. Osteoporos Int. 2012 May 31. Abstract Treatment with molecular hydrogen alleviates microgravity-induced bone loss through abating oxidative stress, restoring osteoblastic differentiation, and suppressing osteoclast differentiation and osteoclastogenesis. INTRODUCTION: Recently, it has been suggested that hydrogen gas exerts a therapeutic antioxidant activity by selectively reducing cytotoxic reactive oxygen species (ROS). The aim of the present study was to elucidate whether treatment with molecular hydrogen alleviated bone loss induced by modeled microgravity in rats. METHODS: Hindlimb suspension (HLS) and rotary wall vessel bioreactor were used to model microgravity in vivo and in vitro, respectively. Sprague-Dawley rats were exposed to HLS for 6 weeks to induced bone loss and simultaneously administrated with hydrogen water (HW). Then, we investigated the effects of incubation with hydrogen-rich medium (HRM) on MC3T3-E1 and RAW264.7 cells exposed to modeled microgravity. RESULTS: Treatment with HW alleviated HLS-induced reduction of bone mineral density, ultimate load, stiffness, and energy in femur and lumbar vertebra. Treatment with HW alleviated HLS-induced augmentation of malondialdehyde content and peroxynitrite content and reduction of total sulfhydryl content in femur and lumbar vertebra. In cultured MC3T3-E1 cells, incubation with HRM inhibited modeled microgravity-induced ROS formation, reduction of osteoblastic differentiation, increase of ratio of receptor activator of nuclear factor kappa B ligand to osteoprotegerin, inducible nitric oxide synthetase upregulation, and Erk1/2 phosphorylation. In cultured RAW264.7, incubation with HRM aggravated modeled microgravity-induced ROS formation, osteoclastic differentiation, and osteoclastogenesis. CONCLUSION: Treatment with molecular hydrogen alleviates microgravity-induced bone loss in rats. Molecular hydrogen could thus be envisaged as a nutritional countermeasure for spaceflight but remains to be tested in humans. PMID:22648000 细胞学研究方法,日本学者是将培养瓶或板放在一个密封的湿盒内,用三个气源根据比例通气,并通过一个排气空气体排到房间外,如果先混合在灌入培养瓶,如何操作? Over a 2-h period, we dissolved H2 into DMEM under 0.4 MPa pressure based on the method described by Ohsawa et al. . We dissolved O2 into a second medium by bubbling O2 gas at the saturated level, and CO2 into a third medium by bubbling CO2 gas. All three media were maintained at atmospheric pressure. Then, we combined the three media (H2 medium/O2 medium/CO2 medium) in the proportion 75:20:5 % (vol/vol/vol) and added fetal bovine serum to achieve a final concentration of 1 %. For culture, we put the combined medium into a culture flask. Then, we filled the culture flask with mixed gas consisting of 75 %H2, 20 % O2, and 5 % CO2 (vol/vol/vol) and cultured cells in the closed culture flask.
标签: 治疗
