Hydrogen, a non-cytotoxic molecule, is one of nature’s most simple elements [1,2]. Recent studies revealed that intraperitoneal injection of hydrogen-rich saline has surprising anti-inflammation, anti-oxidant, anti-apoptosis effects and protected organism against polymicrobial sepsis injury, acute peritonitis injury both by reducing oxidative stress and via decreasing mass proinflammatory responses. It is also well known that the majority of viral -induced tissue damage and discomfort are mainly caused by an inflammatory cytokine storm and oxidative stress rather than by virus itself [3-5]. Studies have shown that suppressing the cytokine storm and reducing oxidative stress can significantly alleviate the symptoms of influenza and other severe viral infections diseases [3-7]. However, none of the studies have been focused on the solution as an anti-virus infection therapy yet. Therefore, we hypothesize that hydrogen-rich solution therapy may be a safe, reliable, and effective treatment for Multiple Organ Dysfunction Syndrome (MODS) induced by influenza and other viral infectious diseases.

Keywords: Hydrogen Medicine; Anti-Oxidant; Anti-Inflammation; Inflammatory Cytokine Storm; Molecular Hydrogen; Multiple Organ Dysfunction Syndrome (MODS)

Hydrogen is one of nature’s most simple elements. As a gas (H₂), it is a colorless, tasteless, odorless, highly flammable diatomic molecule which has been used for fossil fuel processing and ammonia production. In the past decade, molecular hydrogen was considered a surprising agent, which can significantly reduce oxidative stress by selectively reducing hydroxyl radical (•OH) and Peroxynitrite (ONOO-) [1,8-10]. It has recently been revealed that hydrogen can both down-regulate expression of oxidative-related genes and pro-inflammatory cytokine genes directly and indirectly [2-5,11,12]. Oxidative stress and systemic inflammatory response syndrome have been confirmed to play critical roles in tissue and organ damages after polymicrobial sepsis injury, acute peritonitis injury, and peritonitis, which can develop into lethal sepsis with inappropriate treatment [13,14]. In spite of some available antibiotic therapies for the some stages of sepsis, Multiple Organ Dysfunction Syndrome (MODS) induced by sepsis is still the leading cause of death in the Intensive Care Unit (ICU) [15,16]. Recent studies reveal potential protective effects of hydrogen against sepsis and acute peritonitis by decreasing proinflammatory responses, oxidative stress, and apoptosis, which indicate hydrogen medicine as a new no-toxic therapy for bacterial infections [13,14]. Other researchers have also demonstrated that chronic hepatitis B, acute pancreatitis, and sepsis can also be alleviated by treatment of hydrogen medicine [10,14,17]. However, none of the research ever investigated the therapeutic effect of hydrogen gas for MODS induced by influenza and other viral infectious diseases in which inflammation and oxidative stress also play pivotal roles.

Viruses that cause influenza, Ebola, Severe Acute Respiratory Syndrome (SARS), and Middle East Respiratory Syndrome (MERS) are emerging as infectious pathogens in this century that are extremely difficult to control effectively, triggering MODS [18-21]. The trends, spread, scope, and development speed of these emerging infectious diseases cannot be estimated, as their routes of transmission and patterns of spread are extensive and different. Diseases induced by these viruses also differ from each other. To be specific, influenza virus can cause flu, whereby the patient has a stuffy nose, cough, sore throat, runny nose, headache, muscle pain and discomfort symptoms [22]. SARS induces diffuse alveolar damage, acute lung injury, leading to Acute Respiratory Distress Syndrome (ARDS), hypoxemia, and high mortality rate [20,23-25]. Like SARS, MERS, a new type of corona virus, can cause symptoms even with many complications including renal failure [18,19].

The body will be in a state of stress that is caused by the excitement of the sympathetic system after invasion of these pathogens [26,27]. Therefore, oxidative stress increases the release of catecholamine [28]. As auto-oxidation of catecholamine occurs, a large number of free radicals can be produced, accelerating formation of oxidative stress [1]. Meanwhile, oxidative stress activates the complement system, producing a variety of chemotactic substances, such as C3 fragments, leukotrienes etc, which attract and activate neutrophils ultimately [29]. Thus, inflammatory infiltrates develops in many corresponding organs. Furthermore, all these pathogens also stimulate the immune system continuously to launch an inflammatory response flaring out of control [30]. Proinflammatory cytokines are secreted throughout the body; these cytokines also initiate activation of inflammation cells like neutrophils, eosinophils, basophils, lymphocytes, and monocytes to produce more proinflammatory cytokines [5,30]. Those cytokines and inflammation cells are reciprocal causation, developing into a cytokine storm(systemic inflammatory response syndrome)[30,31]. The term cytokine storm is used to accommodate the observation that multiple excessive inflammatory causes can induce excessive release of inflammation factors like interleukin-1, interleukin-6, interleukin-12, tumor necrosis factor-α, interferon-α, interferon-β, interferon-γ, Monocyte Chemoattractant Protein-1 and interleukin-8 thereby leading to a disease that appears similar to sepsis [30,31]. Importantly, excessive inflammation reactions can also induce acute oxidative stress [2]. Together, both cytokine storm and oxidative stress promote each other and induce MODS, result in high mortality rate [15].

Until now, numerous studies have indicated cytokine storm and oxidative stress are highly associated with the pathological process when getting infected with these viruses [32- 34]. Although cytokine storm and oxidative stress probably try to eliminate theses pathogens, they seem to generate multi-organ damage resulting in lethal clinical symptoms such as extensive pulmonary oedema, alveolar and other tissue haemorrhage, and acute respiratory distress syndrome, etc [6,7,33]. Moreover, when inflammation and oxidative stress damage tissue and organs, healing occurs with fibrosis, aggravating persistent multiple organ dysfunction [30]. Therefore, timely elimination of these mass of cytokines and oxidative stress would presumably protect normal organs from the damaging effects of pathogen infection.

At present, there are some therapies which include vaccines and drugs such as Oseltamivir, Amantadine, Curcumin and Ribavirin, as well as S1P1R for influenza and other viral infections diseases [35-39]. However, due to the highly variable nature of these pathogens, no ideal therapy comprehensively conforms to the criteria of effective, selective, non-toxic, and tolerance-inducing anti-influenza and other viral therapy. Furthermore, studies have showen that oseltamivir can alleviate clinical diseases symptoms and reduce morbidity and mortality [40,41]. However, there are still controversies over the prevention, treatment, and tolerance effects of oseltamivir on influenza virus [22]. Importantly, recent research showed that epistatic interactions between neuraminidase mutations promote the number of oseltamivir-resistant influenza virus populations [42]. Moreover, the clinical application revealed oseltamivir had many adverse effects such as nausea, vomiting, and an increased risk of headaches as well as renal and psychiatric syndromes [22]. Therefore, more attention should be paid to the trade-off between benefits and drawbacks when deciding to choose oseltamivir for a therapy.

Although anti-influenza and other viral therapies have been widely studied in the past decades, no therapy can achieve the desired standards. Medical researchers have been striving hard to identify effective, novel, non-toxic, and convenient compounds to protect patients against influenza and other viral infections.

Our hypothesis is that hydrogen-rich solution therapy may be a safe, reliable, effective, and specific treatment for MODS induced by influenza and other viral infectious diseases. Given the theory that molecular hydrogen can both significantly down regulate expressions of inflammation-related genes and selectively reduce hydroxyl radical and Peroxynitrite, we have reasons to consider that cytokine storm and oxidative stress can be suppressed when getting infected with avian influenza and other severe viruses [1,11,12,43-45]. Our theory is unique because it not only puts forward a new kind of non-toxic antiviral therapy but also makes hydrogen-based medicine able to heal disease in to the whole body.

Proinflammatory cytokines including interleukin-1β , interleukin-6, Interferon-γ, intercellular cell adhesion molecule-1, inducible nitric oxide synthase, monocyte chemotactic protein 1, chemokine ligand 2 and tumor necrosis factor -α as well as Proliferating Cell Nuclear Antigen are the main contributors for cytokine storm[30,46-48]. Numerous studies have consistently shown that the contributors were significantly down regulated after applying hydrogen medicine therapy [10,14,26,27,41]. Besides, with the deepening biological mechanism of hydrogen research being developed, scientists gradually found that hydrogen therapy can significantly suppress many pathological signal transduction channels such as NF-κβ, MAPK, Lyn-P, and MEK-1 as well as ERK1/2 pathways and ultimately achieve the goal of recovery from many diseases [44,45,49-53]. In addition, it is worth noting that as H₂ is moderate enough, it can selectively react with only hydroxyl radicals (•OH) and peroxynitrite(ONOO-), the main contributors of oxidative stress in vitro and in vivo without disturbing metabolic redox reactions [1]. Last but not least, as H₂ is an endogenous substance, the goal of better tissue compatibility than other anti-viral drugs can be achieved [54-56].

Since persistent tissue damage, MODS and high mortality rate are highly associated with oxidative stress and cytokine storm induced by influenza and other severe viral infections hydrogen can significantly reduce oxidative stress and restrain excessive production of cytokines [9,57-60]. We hypothesize that hydrogen can be potentially effective for MODS induced by influenza and other viral infectious diseases. That is to say, hydrogen may be a promising novel anti-influenza and other severe viral infections protectant. We believe work on hydrogen-based medicine for anti-viral therapy in vitro and in vivo should commence as soon as possible. In view of the outbreak, transmission, and widespread nature of these viruses, and the global issues caused by new variation pandemics threats, hydrogen medicine may give us more hope for greater survival and fewer human morbidity and mortality (Figure 1).

It’s interesting to note that H₂ therapy can be administered through inhalation, oral intake of hydrogen-rich water, injection of hydrogen-rich saline, direct diffusion of hydrogen: bath, eye drops and immersion, as well as increase hydrogen in intestine [11,61-69]. Although each delivery way has its own characteristic and advantages, injection of hydrogen-rich saline allows enough amount of hydrogen to have its own antioxidant, anti-inflammation, anti-apoptosis effect at the shortest time [11,70]. Moreover, it is emergency to cure patients with influenza and other severe viral infectious diseases. Therefore, it would be most suitable to choose injection hydrogen-rich saline method as the primary hydrogen therapy for influenza and other severe viral infectious diseases.

This work was supported by National Nature Science Foundation of China (No. 81400274), National Student’s Platform for Innovation and Entrepreneurship Training Program(Program Number :201510439107).

1. Ohsawa I, Ishikawa M, Takahashi K, Watanabe M, Nishimaki K, Yamagata K, et al. Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med. 2007;13(6):688-694.
2. Radi R. Peroxynitrite, a stealthy biological oxidant. J Biol Chem. 2013;288(37):26464-26472. doi: 10.1074/jbc.R113.472936
3. Mishra A, Vijayakumar P, Raut AA. Emerging avian influenza infections: Current understanding of innate immune response and molecular pathogenesis. Int Rev Immunol. 2017;36(2):89-107. doi: 10.1080/08830185.2017
4. Oliveira JH, Talyuli OA, Goncalves RL, Paiva-Silva GO, Sorgine MH, Alvarenga PH, et al. Catalase protects Aedes aegypti from oxidative stress and increases midgut infection prevalence of Dengue but not Zika. PLoS neglected tropical diseases. 2017;11(4):e0005525.
5. Behrens EM, Koretzky GA. Cytokine Storm Syndrome: Looking Toward the Precision Medicine Era. Arthritis Rheumatol. 2017. doi: 10.1002/art.40071
6. Zhu HY, Huang H, Shi XL, Zhou W, Zhou P, Yan QL, et al. Qiangzhi decoction protects mice from influenza A pneumonia through inhibition of inflammatory cytokine storm. Chin J Integr Med. 2015;21(5):376-383. doi: 10.1007/s11655-014-2020-2
7. Sordillo PP, Helson L. Curcumin suppression of cytokine release and cytokine storm. A potential therapy for patients with Ebola and other severe viral infections. In Vivo. 2015;29(1):1-4.
8. Qian L, Shen J. Hydrogen therapy may be an effective and specific novel treatment for acute graft-versus-host disease (GVHD). Journal of cellular and molecular medicine. J Cell Mol Med. 2013;17(8):1059-1063. doi: 10.1111/jcmm.12081
9. Zhao S, Mei K, Qian L, Yang Y, Liu W, Huang Y, et al. Therapeutic effects of hydrogen-rich solution on aplastic anemia in vivo. Cell Physiol Biochem. 2013;32(3):549-560. doi: 10.1159/000354459
10. Xia C, Liu W, Zeng D, Zhu L, Sun X. Effect of hydrogen-rich water on oxidative stress, liver function, and viral load in patients with chronic hepatitis B. Clin Transl Sci. 2013;6(5):372-375. doi: 10.1111/cts.12076
11. Ohta S. Molecular hydrogen as a preventive and therapeutic medical gas: initiation, development and potential of hydrogen medicine. Pharmacol Ther. 2014;144(1):1-11. doi: 10.1016/j.pharmthera.2014.04.006
12. Ohno K, Ito M, Ichihara M. Molecular hydrogen as an emerging therapeutic medical gas for neurodegenerative and other diseases. Oxidative medicine and cellular longevity. 2012;2012:353152.
13. Zhang J, Wu Q, Song S, Wan Y, Zhang R, Tai M, et al. Effect of hydrogen-rich water on acute peritonitis of rat models. Int Immunopharmacol. 2014;21(1):94-101. doi: 10.1016/j.intimp.2014.04.011
14. Li GM, Ji MH, Sun XJ, Zeng QT, Tian M, Fan YX, et al. Effects of hydrogen-rich saline treatment on polymicrobial sepsis. J Surg Res. 2013;181(2):279-286. doi: 10.1016/j.jss.2012.06.058
15. Liang SY, Kumar A. Empiric antimicrobial therapy in severe sepsis and septic shock: optimizing pathogen clearance. Curr Infect Dis Rep. 2015;17(7):493. doi: 10.1007/s11908-015-0493-6
16. Bhatia PK, Biyani G. Fluid resuscitation in severe sepsis and septic shock: Shifting goalposts. Indian J Anaesth. 2015;59(5):269-271. doi: 10.4103/0019-5049.156863
17. Chen H, Sun YP, Li Y, Liu WW, Xiang HG, Fan LY, et al. Hydrogen-rich saline ameliorates the severity of l-arginine-induced acute pancreatitis in rats. Biochem Biophys Res Commun. 2010;393(2):308-313. doi: 10.1016/j.bbrc.2010.02.005
18. Rha B, Rudd J, Feikin D, Watson J, Curns AT, Swerdlow DL, et al. Update on the epidemiology of Middle East respiratory syndrome coronavirus (MERS-CoV) infection, and guidance for the public, clinicians, and public health authorities - January 2015. MMWR Morbidity and mortality weekly report. 2015;64(3):61-62.
19. Tong YG, Shi WF, Liu D, Qian J, Liang L, Bo XC, et al. Genetic diversity and evolutionary dynamics of Ebola virus in Sierra Leone. Nature. 2015;524(7563):93-96. doi: 10.1038/nature14490
20. Ware LB, Matthay MA. The acute respiratory distress syndrome. N Engl J Med. 2000;342(18):1334-1349.
21. Teijaro JR, Walsh KB, Rice S, Rosen H, Oldstone MB. Mapping the innate signaling cascade essential for cytokine storm during influenza virus infection. Proc Natl Acad Sci U S A. 2014;111(10):3799-3804. doi: 10.1073/pnas.1400593111
22. Jefferson T, Jones M, Doshi P, Spencer EA, Onakpoya I, Heneghan CJ. Oseltamivir for influenza in adults and children: systematic review of clinical study reports and summary of regulatory comments. BMJ. 2014;348:g2545.
23. Dole M, Wilson FR, Fife WP. Hyperbaric hydrogen therapy: a possible treatment for cancer. Science. 1975;190(4210):152-154.
24. Gralinski LE, Bankhead A 3rd, Jeng S, Menachery VD, Proll S, Belisle SE, et al. Mechanisms of severe acute respiratory syndrome coronavirus-induced acute lung injury. MBio. 2013;4(4). pii: e00271-13. doi: 10.1128/mBio.00271-13
25. Nieto-Torres JL, DeDiego ML, Verdia-Baguena C, Jimenez-Guardeno JM, Regla-Nava JA, Fernandez-Delgado R, et al. Severe acute respiratory syndrome coronavirus envelope protein ion channel activity promotes virus fitness and pathogenesis. PLoS Pathog. 2014;10(5):e1004077. doi: 10.1371/journal.ppat.1004077
26. Barrios-Payan J, Revuelta A, Mata-Espinosa D, Marquina-Castillo B, Villanueva EB, Gutierrez ME, et al. The contribution of the sympathetic nervous system to the immunopathology of experimental pulmonary tuberculosis. J Neuroimmunol. 2016;298:98-105. doi: 10.1016/j.jneuroim.2016.07.012
27. Roggero E, Perez AR, Pollachini N, Villar SR, Wildmann J, Besedovsky H, et al. The sympathetic nervous system affects the susceptibility and course of Trypanosoma cruzi infection. Brain Behav Immun. 2016;58:228-236. doi: 10.1016/j.bbi.2016.07.163
28. Tappia PS, Hata T, Hozaima L, Sandhu MS, Panagia V, Dhalla NS. Role of oxidative stress in catecholamine-induced changes in cardiac sarcolemmal Ca2+ transport. Arch Biochem Biophys. 2001;387(1):85-92.
29. Ingoglia G, Martin Sag C, Rex N, De Franceschi L, Vinchi F, Cimino J, et al. Hemopexin counteracts systolic dysfunction induced by heme-driven oxidative stress. Free Radic Biol Med. 2017;108:452-464. doi: 10.1016/j.freeradbiomed.2017.04.003
30. Tisoncik JR, Korth MJ, Simmons CP, Farrar J, Martin TR, Katze MG. Into the eye of the cytokine storm. Microbiol Mol Biol Rev. 2012;76(1):16-32. doi: 10.1128/MMBR.05015-11
31. Canna SW, Behrens EM. Making sense of the cytokine storm: a conceptual framework for understanding, diagnosing, and treating hemophagocytic syndromes. Pediatr Clin North Am. 2012;59(2):329-44. doi: 10.1016/j.pcl.2012.03.002
32. Us D. Cytokine storm in avian influenza. Mikrobiyoloji bulteni. 2008;42(2):365-380.
33. Martines RB, Ng DL, Greer PW, Rollin PE, Zaki SR. Tissue and cellular tropism, pathology and pathogenesis of Ebola and Marburg viruses. J Pathol. 2015;235(2):153-174. doi: 10.1002/path.4456
34. Hong Y, Shao A, Wang J, Chen S, Wu H, McBride DW, et al. Neuroprotective effect of hydrogen-rich saline against neurologic damage and apoptosis in early brain injury following subarachnoid hemorrhage: possible role of the Akt/GSK3beta signaling pathway. PLoS One. 2014;9(4):e96212. doi: 10.1371/journal.pone.0096212
35. McQuade B, Blair M. Influenza treatment with oseltamivir outside of labeled recommendations. Am J Health Syst Pharm. 2015;72(2):112-116. doi: 10.2146/ajhp140390
36. Spritzer SD, Kinney CL, Condie J, Wellik KE, Hoffman-Snyder CR, Wingerchuk DM, et al. Amantadine for patients with severe traumatic brain injury: a critically appraised topic. Neurologist. 2015;19(2):61-64. doi: 10.1097/NRL.0000000000000001
37. Rivas-Aravena A, Guajardo S, Valenzuela B, Cartagena J, Imarai MI, Spencer E, et al. Ribavirin stimulates the immune response of Atlantic salmon. Vet Immunol Immunopathol. 2015;164(1-2):93-100. doi: 10.1016/j.vetimm.2015.01.001
38. Dilnawaz F, Sahoo SK. Enhanced accumulation of curcumin and temozolomide loaded magnetic nanoparticles executes profound cytotoxic effect in glioblastoma spheroid model. Eur J Pharm Biopharm. 2013;85(3 Pt A):452-462. doi: 10.1016/j.ejpb.2013.07.013
39. Woo JH, Kim YH, Choi YJ, Kim DG, Lee KS, Bae JH, et al. Molecular mechanisms of curcumin-induced cytotoxicity: induction of apoptosis through generation of reactive oxygen species, down-regulation of Bcl-XL and IAP, the release of cytochrome c and inhibition of Akt. Carcinogenesis. 2003;24(7):1199-1208.
40. Marty FM, Vidal-Puigserver J, Clark C, Gupta SK, Merino E, Garot D, et al. Intravenous zanamivir or oral oseltamivir for hospitalised patients with influenza: an international, randomised, double-blind, double-dummy, phase 3 trial. Lancet Respir Med. 2017;5(2):135-146. doi: 10.1016/S2213-2600(16)30435-0
41. Ye M, Jacobs A, Khan MN, Jaipaul J, Oda J, Johnson M, et al. Evaluation of the use of oseltamivir prophylaxis in the control of influenza outbreaks in long-term care facilities in Alberta, Canada: a retrospective provincial database analysis. BMJ open. 2016;6(7):e011686.
42. Duan S, Govorkova EA, Bahl J, Zaraket H, Baranovich T, Seiler P, et al. Epistatic interactions between neuraminidase mutations facilitated the emergence of the oseltamivir-resistant H1N1 influenza viruses. Nat Commun. 2014;5:5029. doi: 10.1038/ncomms6029
43. Jazwa A, Cuadrado A. Targeting heme oxygenase-1 for neuroprotection and neuroinflammation in neurodegenerative diseases. Current drug targets. Curr Drug Targets. 2010;11(12):1517-1531.
44. Kawamura T, Wakabayashi N, Shigemura N, Huang CS, Masutani K, Tanaka Y, et al. Hydrogen gas reduces hyperoxic lung injury via the Nrf2 pathway in vivo. American journal of physiology-Lung cellular and molecular physiology. 2013;304(10):L646-656.
45. Zhai X, Chen X, Shi J, Shi D, Ye Z, Liu W, et al. Lactulose ameliorates cerebral ischemia-reperfusion injury in rats by inducing hydrogen by activating Nrf2 expression. Free Radic Biol Med. 2013;65:731-741. doi: 10.1016/j.freeradbiomed.2013.08.004
46. Hale BG, Jackson D, Chen YH, Lamb RA, Randall RE. Influenza A virus NS1 protein binds p85beta and activates phosphatidylinositol-3-kinase signaling. Proc Natl Acad Sci U S A. 2006;103(38):14194-14199.
47. Phung TT, Luong ST, Kawachi S, Nunoi H, Nguyen LT, Nakayama T, et al. Interleukin 12 and myeloperoxidase (MPO) in Vietnamese children with acute respiratory distress syndrome due to Avian influenza (H5N1) infection. J Infect. 2011;62(1):104-106. doi: 10.1016/j.jinf.2010.11.012
48. Grattendick K, Stuart R, Roberts E, Lincoln J, Lefkowitz SS, Bollen A, et al. Alveolar macrophage activation by myeloperoxidase: a model for exacerbation of lung inflammation. Am J Respir Cell Mol Biol. 2002;26(6):716-722.
49. Hu W, Wang G, Li P, Wang Y, Si CL, He J, et al. Neuroprotective effects of macranthoin G from Eucommia ulmoides against hydrogen peroxide-induced apoptosis in PC12 cells via inhibiting NF-kappaB activation. Chem Biol Interact. 2014;224:108-116. doi: 10.1016/j.cbi.2014.10.011
50. Wang W, Zheng JP, Zhu SX, Guan WJ, Chen M, Zhong NS. Carbocisteine attenuates hydrogen peroxide-induced inflammatory injury in A549 cells via NF-kappaB and ERK1/2 MAPK pathways. Int Immunopharmacol. 2015;24(2):306-313. doi: 10.1016/j.intimp.2014.12.018
51. Itoh T, Fujita Y, Ito M, Masuda A, Ohno K, Ichihara M, et al. Molecular hydrogen suppresses FcepsilonRI-mediated signal transduction and prevents degranulation of mast cells. Biochemical and biophysical research communications. 2009;389(4):651-656.
52. Cardinal JS, Zhan J, Wang Y, Sugimoto R, Tsung A, McCurry KR, et al. Oral hydrogen water prevents chronic allograft nephropathy in rats. Kidney Int. 2010;77(2):101-109. doi: 10.1038/ki.2009.421
53. Chen Y, Jiang J, Miao H, Chen X, Sun X, Li Y. Hydrogen-rich saline attenuates vascular smooth muscle cell proliferation and neointimal hyperplasia by inhibiting reactive oxygen species production and inactivating the Ras-ERK1/2-MEK1/2 and Akt pathways. Int J Mol Med. 2013;31(3):597-606. doi: 10.3892/ijmm.2013.1256
54. Azuma T, Yamane M, Ekuni D, Kawabata Y, Kataoka K, Kasuyama K, et al. Drinking Hydrogen-Rich Water Has Additive Effects on Non-Surgical Periodontal Treatment of Improving Periodontitis: A Pilot Study. Antioxidants (Basel). 2015;4(3):513-522. doi: 10.3390/antiox4030513
55. Bai X, Liu S, Yuan L, Xie Y, Li T, Wang L, et al. Hydrogen-rich saline mediates neuroprotection through the regulation of endoplasmic reticulum stress and autophagy under hypoxia-ischemia neonatal brain injury in mice. Brain Res. 2016;1646:410-417. doi: 10.1016/j.brainres.2016.06.020
56. Beheshti SM, Ghassemi H, Shahsavan-Markadeh R, Fremaux S. Hydrogen-rich gas production via CaO sorption-enhanced steam gasification of rice husk: a modelling study. Environ Technol. 2015;36(9-12):1327-1333. doi: 10.1080/09593330.2014.988185
57. Tong YG, Shi WF, Liu D, Qian J, Liang L, Bo XC, et al. Erratum: Genetic diversity and evolutionary dynamics of Ebola virus in Sierra Leone. Nature. 2015;526:595.
58. Sieghart D, Liszt M, Wanivenhaus A, Broll H, Kiener H, Klosch B, et al. Hydrogen sulphide decreases IL-1beta-induced activation of fibroblast-like synoviocytes from patients with osteoarthritis. J Cell Mol Med. 2015;19(1):187-197. doi: 10.1111/jcmm.12405
59. Qian L, Mei K, Shen J, Cai J. Administration of hydrogen-rich saline protects mice from lethal acute graft-versus-host disease (aGVHD). Transplantation. 2013;95(5):658-662. doi: 10.1097/TP.0b013e31827e6b23
60. Cai WW, Zhang MH, Yu YS, Cai JH. Treatment with hydrogen molecule alleviates TNFα-induced cell injury in osteoblast. Mol Cell Biochem. 2013;373(1-2):1-9. doi: 10.1007/s11010-012-1450-4
61. Watanabe S, Fujita M, Ishihara M, Tachibana S, Yamamoto Y, Kaji T, et al. Protective effect of inhalation of hydrogen gas on radiation-induced dermatitis and skin injury in rats. J J Radiat Res. 2014;55(6):1107-1113. doi: 10.1093/jrr/rru067
62. Koyama Y, Taura K, Hatano E, Tanabe K, Yamamoto G, Nakamura K, et al. Effects of oral intake of hydrogen water on liver fibrogenesis in mice. Hepatol Res. 2014;44(6):663-677. doi: 10.1111/hepr.12165
63. Qian L, Shen J, Chuai Y, Cai J. Hydrogen as a new class of radioprotective agent. Int J Biol Sci. 2013;9(9):887-894. doi: 10.7150/ijbs.7220
64. Shao A, Wu H, Hong Y, Tu S, Sun X, Wu Q, et al. Hydrogen-Rich Saline Attenuated Subarachnoid Hemorrhage-Induced Early Brain Injury in Rats by Suppressing Inflammatory Response: Possible Involvement of NF-kappaB Pathway and NLRP3 Inflammasome. Mol Neurobiol. 2016;53(5):3462-3476. doi: 10.1007/s12035-015-9242-y
65. Ge Y, Wu F, Sun X, Xiang Z, Yang L, Huang S, et al. Intrathecal infusion of hydrogen-rich normal saline attenuates neuropathic pain via inhibition of activation of spinal astrocytes and microglia in rats. PLoS One. 2014;9(5):e97436. doi: 10.1371/journal.pone.0097436
66. Kato S, Saitoh Y, Iwai K, Miwa N. Hydrogen-rich electrolyzed warm water represses wrinkle formation against UVA ray together with type-I collagen production and oxidative-stress diminishment in fibroblasts and cell-injury prevention in keratinocytes. J Photochem Photobiol B. 2012;106:24-33. doi: 10.1016/j.jphotobiol.2011.09.006
67. Buchholz BM, Masutani K, Kawamura T, Peng X, Toyoda Y, Billiar TR, et al. Hydrogen-enriched preservation protects the isogeneic intestinal graft and amends recipient gastric function during transplantation. Transplantation. 2011;92(9):985-992. doi: 10.1097/TP.0b013e318230159d
68. Chen X, Zhai X, Shi J, Liu WW, Tao H, Sun X, et al. Lactulose mediates suppression of dextran sodium sulfate-induced colon inflammation by increasing hydrogen production. Dig Dis Sci. 2013;58(6):1560-1568. doi: 10.1007/s10620-013-2563-7
69. Suzuki Y, Sano M, Hayashida K, Ohsawa I, Ohta S, Fukuda K. Are the effects of alpha-glucosidase inhibitors on cardiovascular events related to elevated levels of hydrogen gas in the gastrointestinal tract? FEBS Lett. 2009;583(13):2157-2159. doi: 10.1016/j.febslet.2009.05.052
70. Ohta S. Initiation, development and potential of hydrogen medicine: Toward therapeutic and preventive applications of molecular hydrogen against a variety of diseases. Seikagaku. 2015;87(1):82-90.