The purpose of this study was to investigate the anti-injury effect and protective mechanism of hydrogen-enriched water in a rat model of acute liver injury induced by aflatoxin B (AFB). Healthy male Sprague-Dawley (SD) rats were randomly divided into control group, model group (AFB group) and hydrogen-enriched water treatment group (AFB+H group). The rat model of acute liver injury induced by AFB was established by single intragastric administration of AFB (2.0 mg/kg), and then the rats were treated with hydrogen-enriched water intragastrically. HE staining was used to observe the pathological changes of liver tissue. Blood samples were taken from vena cava to measure serum liver function indexes. Live tissue was sampled to detect malondialdehyde (MDA) and reduced glutathione (GSH) contents. Western blot was used to detect phosphorylation levels of MAPK signaling pathway proteins (ERK, JNK and p38 MAPK). The results showed that, compared with the AFB group, the AFB+H group exhibited increased body weights, alleviated acute liver injury, decreased activities of serum glutamic-pyruvic transaminase and glutamic oxaloacetic transaminase, as well as total bilirubin level in the serum. Meanwhile, hydrogen-enriched water decreased MDA content and increased GSH content in liver tissue. AFB-increased phosphorylation levels of ERK, JNK and p38 MAPK in liver tissue were down-regulated significantly by hydrogen-enriched water treatment. These results suggest that hydrogen-enriched water can alleviate liver injury induced by AFB, and its mechanism may be related to the reduction of oxidative stress and the inhibition of MAPK signal transduction pathway activation.
Aflatoxin B1 ; Animals ; Chemical and Drug Induced Liver Injury ; pathology ; prevention & control ; Deuterium Oxide ; therapeutic use ; Liver ; pathology ; MAP Kinase Signaling System ; Male ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley

The aim of the present study was to measure the kinetic parameters of skeletal muscle protein synthesis in rats by deuterated water (HO). Twenty Sprague-Dawley (SD) rats were labeled byHO through intraperitoneal injection and drinking. At the each end of the 1st, 3rd, 5th, 6th and 10th week after the firstHO labeling, four rats were sacrificed by cardiac puncture for blood plasma and quadriceps femoris sampling. Skeletal muscle protein and free amino acids in plasma were purified, hydrolyzed by hydrochloric acid and derived. The deuterium enrichments ofH-labeled alanyl in skeletal muscle protein and plasma protein-boundH-labeled alanine were determined by gas chromatography-mass spectrometry (GC-MS). The fractional synthesis rate of skeletal muscle protein and synthetic dynamic equation were calculated. The fractional synthetic rate of skeletal muscle protein was 12.8%/week, and synthetic dynamic equation was f= 0.158 × (1 - e). The results suggest that the kinetic parameters of skeletal muscle protein synthesis can be measured byHO labeling, and the method can be applied in long-term labeling experiment.
Alanine ; Amino Acids ; blood ; Animals ; Deuterium ; Gas Chromatography-Mass Spectrometry ; Kinetics ; Male ; Muscle Proteins ; biosynthesis ; Muscle, Skeletal ; metabolism ; Protein Biosynthesis ; Rats ; Rats, Sprague-Dawley ; Water

In silico acquirement of the accurate residue details of protein on chromatographic media is a bottleneck in protein chromatography separation and purification. Here we developed a novel approach by coupling with H/D exchange and nuclear magnetic resonance to observe hen egg white lysozyme (HEWL) unfolding behavior adsorbed on cation exchange media (SP Sepharose FF). Analysis of 1D 1H-NMR shows that protein unfolding accelerated H/D exchange rate, leading to more loss of signal of amide hydrogen owing to exposure of residues and the more unfolding of protein. Analysis of two-dimensional hydrogen-hydrogen total correlation spectroscopy shows that lysozyme lost more signals and experienced great unfolding during its adsorption on media surface. However, for several distinct fragments, the protection degrees varied, the adsorbed lysozyme lost more signal intensity and was less protected at disorder structures (coil, bend, and turn), but was comparatively more protected against exchange at secondary structure domains (α-helix, β-sheet). Finally, the binding site was determined by electrostatic calculations using computer simulation methods in conjunction with hydrogen deuterium labeled protein and NMR. This study would help deeply understand the microscopic mechanism of protein chromatography and guide the purposely design of chromatographic process and media. Moreover, it also provide an effective tool to study the protein and biomaterials interaction in other applications.
Adsorption ; Amides ; Cations ; Computer Simulation ; Deuterium ; Hydrogen ; Magnetic Resonance Spectroscopy ; Muramidase ; chemistry ; Protein Structure, Secondary ; Protein Unfolding ; Proteins ; chemistry

OBJECTIVEUsing the stable isotopes as the internal standard of microdialysis technology to establish a new method to study the whole and local brain dynamics of nicotine percutaneous preparations.

METHODUsing th healthy rats as experimental animals, administrating nicotine in abdominal transdermal way, then sample in the blood and brain simultaneously by microdialysis which use deuterium nicotine (DL-nicotine) as internal standard. Detecting the samples by LC-MS/MS method.

RESULTThe configuration process in blood and brain both conforms to 2 compartments model, t1/2 is 29.38 min, t1/2beta is 208.51 min, AUC(0-infinity) is 152 127.10 microg x min x L(-1) in the blood t1/2 is 86.64 min, t1/2beta is 386.00 min, AUC(0-infinity) is 152 820.90 microg x min x L(-1) in the brain.

CONCLUSIONDl-nicotine can be used as internal standard of nicotine to correcte the recovery; Stable isotopes internal standard microdialysis technology can be used for studing the whole and the local pharmacokinetic of nicotine and also provide new ideas and methods to studing the process of new drug delivery system.

Animals ; Brain ; metabolism ; Brain Chemistry ; Deuterium ; chemistry ; Isotope Labeling ; methods ; Male ; Microdialysis ; methods ; Nicotine ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

Deuterium is an important predisposing factor for cancer. Deuterium-depleted Water, also known as low deuterium water, ultra-light water or no deuterium water, can be obtained by removing deuterium from natural water. Studies have shown that water with a low deuterium concentration (<65% percent of volume) can inhibit cancer growth. Clinical trials demonstrated that drinking DDW (10-20 ppm) caused growth arrest of malignant cells in cancer patients and significantly prolonged the patient survival with also improved quality of life. A wide range of anti-cancer drugs in current use are associated with severe adverse effects, while deuterium-depleted water appears to have virtually no pharmacological side effects and is convenient to administer. The authors review the advances in the researches of anti-cancer effects and the underlying mechanisms of deuterium-depleted water.
Animals ; Antineoplastic Agents ; Deuterium ; therapeutic use ; Humans ; Neoplasms ; therapy ; Water ; chemistry

OBJECTIVETo evaluate the inhibitory effect of deuterium-depleted water (DDW) on the proliferation of nasopharyngeal carcinoma (NPC) cells in vitro and explore the possible mechanism.

METHODSThe growth inhibition of NPC cells and preosteoblast MC3T3-E1 cells following DDW treatment was measured by MTT assay and plate colony formation assay. The changes in migration and invasion of NPC cells were evaluated using Transwell and boyden chamber assays. The protein expression of proliferating cell nuclear antigen (PCNA) was determined using Western blotting. Flow cytometry was employed to evaluate the changes in cell cycle distribution after DDW treatment.

RESULTSDDW with deuterium concentrations of 100, 75 and 50 ppm significantly suppressed the cell proliferation (P<0.05) and lowered colony formation capacity and invasiveness of the NPC cells (P<0.01). Western blotting demonstrated a down-regulated expression of PCNA in the cells by DDW. DDW also caused obvious cell cycle arrest in the NPC cells with reduced cells in S phase and significantly increased cells in G(1) phase (P<0.05). Rather than causing growth inhibition, DDW promoted the growth of normal control MC3T3-E1 cells.

CONCLUSIONDDW possesses selective biological effects to inhibit the proliferation of NPC cells in vitro, suggesting the potential of DDW as a novel nontoxic adjuvant therapeutic agent in antitumor therapy.

Carcinoma ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Deuterium ; administration & dosage ; pharmacology ; Humans ; Nasopharyngeal Neoplasms ; pathology ; Proliferating Cell Nuclear Antigen ; metabolism ; Water ; chemistry

The paper is to report the study of pharmacokinetics of transdermal administered nicotine in the brain of freely moving rat by using microdialysis with stable labeled isotope as internal standard. The pharmacokinetic behavior of nicotine in Sprague Dawley rat brain was investigated after intranasal administration (3.75 mg). Brain fluid samples were collected by intracerebral microdialysis with DL-nicotine as internal standard. Concentrations of nicotine and DL-nicotine in the sample were measured by HPLC-MS/MS. Main pharmacokinetic parameters were calculated and analyzed by Das 2.0 pharmacokinetic software. The recovery of nicotine and the delivery of DL-nicotine were the same. The fate of absorption and distribution was two compartment model and the values of t1/2alpha was 170.31 min, t1/2beta was 263.30 min and the AUC(0-infinity) was 2.75 x 10(5) microg x L(-1) min separately. DL-nicotine can be used to calibrate the recovery of nicotine, and the new method of stable isotope microdialysis can be used to study the pharmacokinetics of freely moving rat. It will make sense for the treatment of addiction of tobacco and provide a new thought for the research of pharmacokinetics-pharmacodynamic combination.
Administration, Cutaneous ; Administration, Intranasal ; Animals ; Area Under Curve ; Brain ; metabolism ; Chromatography, High Pressure Liquid ; Deuterium ; Female ; Isotope Labeling ; methods ; Male ; Microdialysis ; methods ; Nicotine ; administration & dosage ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

PURPOSE: To evaluate the antiproliferative activity of deuterium oxide (D2O) on urological cancer cells for the application of D2O in the treatment of urological cancer. MATERIALS AND METHODS: Urological cancer cell A-498 (kidney), T-24 (bladder) and DU 145 (prostate) were used in this study. The changes in cellular proliferation and the expressions of the bcl-2 and bax genes, according to changes in the D2O concentrationand exposure time were measured. The changes in cellular proliferation were measured using a hemocytometer and the MTT assay, and the changes in gene expression by Western hybridization. RESULTS: D2O had antiproliferative effects, DU-145 was most resistant and T-24 was most sensitive to D2O. The proliferation of cells in T-24, as measured by the MTT assay, showed a reduced growth rate, which was the inverse of the increased D2O concentration and exposure time. The expression of bcl-2 was reduced with increasing exposure time and D2O concentration, and that of bax was increased with increasing exposure time and D2O concentration. CONCLUSIONS: From this study, the authors believe D2O has antiproliferative effects on urological cancers, and the effect on bladder cancer cells suggests that D2O shows potential as an agent for the treatment of early small bladder cancer or the prevention of superficial bladder cancer recurrence following transurethral resection.
Cell Proliferation ; Deuterium Oxide* ; Deuterium* ; Gene Expression ; Recurrence ; Urinary Bladder Neoplasms ; Urologic Neoplasms*

AIMTo study the NMR phenomena of cetirizine hydrochloride and assign all proton and carbon signals in NMR spectra.

METHODSTo record the 1D and 2D NMR spectra of cetirizine hydrochloride while changing the experimental temperature and adding D2O into the solution.

RESULTSMore than one NMR signal or broad peak resulting from piperazine and the attached groups with N atom were given in DMSO-d6 solution at room temperature. "Coalescence" or narrowing had occurred for the proton and carbon signals when the experimental temperature was increased or D2O was added into the solution.

CONCLUSIONCompared with the NMR "time scale", there are more than one conformation of cetirizine hydrochloride in DMSO-d6 solution at room temperature. The different conformation will be exchanged fast while temperature rise and the stable conformation will be existed while D2O was added into the solution.

Cetirizine ; chemistry ; Deuterium ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Protein Conformation ; Temperature

Hydrogen sulfide poisoning is frequently encountered in the workplace. Two workers lost their consciousness in an underground tank at a factory producing paper. The tank contained liquid mixture of used paper, sodium oxygenate chloride(NaOC1), and sodium thiosulfate pentahydrate(NaSO3 5H90). A worker(worker A; 36-year-old man) entered tank to remove sludge. When worker A lost his consciousness, worker B entered the tank to rescue worker A, however he lost consciousness inside the tank. We discuss in detail the clinical features of this condition. Hydrogen sulfide poisonings have occurred in industries involving petroleum refining, the manufacture of heavy water, tanning of hides, vulcanization of rubber, and the manufacture of rayon. And it is necessary to stress the health education for workers and managers in these industries.
Adult ; Consciousness ; Deuterium Oxide ; Health Education ; Humans ; Hydrogen Sulfide* ; Hydrogen* ; Oxygen ; Petroleum ; Poisoning* ; Rescue Work ; Rubber ; Sewage ; Sodium ; Tanning ; Triacetoneamine-N-Oxyl