OBJECTIVE: To confirm whether formaldehyde disturb detecting carbon monoxide in blood. To give an evidence that can be used for detecting carboxyhemoglobin more accurately in carbon monoxide posioning appraises. METHODS: Blood samples came from carbon monoxide poisoning and the health were collected. Regular methods for detecting carboxyhemoglobin were used. Observing and comparing the detection results between which were spiked with methanal and no spiked one were performed. RESULTS: Methanal will affect the result of following experiments such as heating, adding NaOH, absorbed by PdCl2 and spectrophotometry. CONCLUSION The samples which contaminated by formaldehyde couldn't be used for detecting carboxyhemoglobin.
Carbon Monoxide/blood* ; Carbon Monoxide Poisoning/diagnosis* ; Carboxyhemoglobin/analysis* ; Forensic Medicine ; Formaldehyde/pharmacology* ; Humans ; Spectrophotometry/methods* ; Temperature

OBJECTIVETo explore the effects of exogenous carbon monoxide-releasing molecule 2 (CORM-2) on formation of human neutrophil extracellular traps (NETs) stimulated by endotoxin/lipopolysaccharide (LPS) and its relevant mechanism.

METHODSVenous blood samples were collected from a healthy adult volunteer to isolate neutrophils. The neutrophils were divided into normal control (NC) group, LPS group, LPS+ 10 μmol/L CORM-2 group, LPS+ 50 μmol/L CORM-2 group, and LPS+ inactive CORM-2 (iCORM-2) group according to the random number table. No treatment was given to the neutrophils in NC group. The neutrophils in LPS group underwent LPS stimulation (1 μL, 1 μg/mL). The neutrophils in LPS+ 10 μmol/L CORM-2 group, LPS+ 50 μmol/L CORM-2 group, and LPS+ iCORM-2 group underwent the same LPS stimulation as that in LPS group and treatment of 10 μmol/L CORM-2, 50 μmol/L CORM-2, and 50 μmol/L iCORM-2, respectively, with the volune of 1 μL. After conventional culture for 1 h, the number of NETs was determined with propidium iodide staining method; the early cell apoptosis rate was determined with flow cytometer; the generation level of reactive oxygen species (ROS) was assessed with dihydrogenrhodamine 123 fluorescent probe staining method (denoted as mean fluorescence intensity); the expression level of phosphorylated extracellular regulated kinase 1/2 (p-ERK1/2) was determined by Western blotting. The sample numbers of each group in the 4 experiments were all 5. Data were processed with one-way analysis of variance and SNK test.

RESULTS(1) The numbers of NETs per 400-time visual field in cells of LPS and LPS+ iCORM-2 groups were close to the number in NC group (with P values above 0.05). The number of NETs per 400-time visual field was significantly larger in cells of LPS+ 10 μmol/L CORM-2 and LPS+ 50 μmol/L CORM-2 groups than in NC and LPS groups (with P values below 0.05). The number of NETs per 400-time visual field in cells of LPS+ iCORM-2 group was close to that of LPS group (P>0.05). (2) The early cell apoptosis rate was significantly increased in LPS, LPS+ 10 μmol/L CORM-2, LPS+ 50 μmol/L CORM-2, and LPS+ iCORM-2 groups than in NC group (with P values below 0.05). The early cell apoptosis rates in LPS+ 10 μmol/L CORM-2, LPS+ 50 μmol/L CORM-2, and LPS+ iCORM-2 groups were close to the rate in LPS group (with P values above 0.05). (3) The generation level of ROS was significantly higher in cells of LPS, LPS+ 10 μmol/L CORM-2, and LPS+ iCORM-2 groups than in NC group (with P values below 0.05). The generation level of ROS in cells of LPS+ 50 μmol/L CORM-2 group was close to that of NC group (P>0.05). The generation level of ROS was lower in cells of LPS+ 10 μmol/L CORM-2 and LPS+ 50 μmol/L CORM-2 groups than in LPS group (with P values below 0.05), while the generation level of ROS in cells of LPS+ iCORM-2 group was close to that of LPS group (P>0.05). (4) The expression levels of p-ERK1/2 in cells of LPS and LPS+ iCORM-2 groups (respectively 0.0311±0.001 and 0.0309±0.0018) were close to the level in NC group (0.0304±0.0046, with P values above 0.05). The expression level of p-ERK1/2 was significantly higher in cells of LPS+ 10 μmol/L CORM-2 and LPS+ 50 μmol/L CORM-2 groups (respectively 0.7891±0.0201 and 1.2970±0.0056) than in NC group (with P values below 0.05). The expression level of p-ERK1/2 was significantly higher in cells of LPS+ 10 μmol/L CORM-2 and LPS+ 50 μmol/L CORM-2 groups than in LPS group (with P values below 0.05). The expression level of p-ERK1/2 in cells of LPS+ iCORM-2 group was close to that of LPS group (P>0.05).

CONCLUSIONSCORM-2 can obviously increase the production of NETs in LPS-induced neutrophils, and it might be attributable to the promotion of inhibition of ROS generation and phosphorylation of ERK1/2.

Apoptosis ; Carbon Monoxide ; metabolism ; Extracellular Traps ; Humans ; Lipopolysaccharides ; pharmacology ; Organometallic Compounds ; pharmacology ; Phosphorylation ; drug effects

The aim of the present study was to investigate the effect of endogenous carbon monoxide (CO) on respiratory rhythm. The experiments were carried out on the medullary slices of newborn Sprague-Dawley rats. The rhythmic discharge frequency (DF) of hypoglossal rootlets was taken as an index of rhythmic respiratory activity. The slices of medulla oblongata were superfused with ZnPP-9 (inhibitor of heme oxygenase), CO and hemin (substrate of heme oxygenase), respectively, to observe their effects on respiratory rhythm. The preparations were divided into 5 groups: control group of artificial cerebrospinal fluid (ACSF), group of ZnPP-9, group of exogenous CO, group of hemin and group of ZnPP-9 + hemin. The results obtained were as follows. In ZnPP-9 group, the rhythmic DF of the hypoglossal rootlets was increased (P<0.05); while in exogenous CO group, it was decreased (P<0.05). In the groups of hemin and ZnPP-9 + hemin, the rhythmic DF of the hypoglossal rootlets was increased (P<0.05). It is suggested that endogenous CO may play an important role in the regulation of respiratory rhythm.
Animals ; Animals, Newborn ; Carbon Monoxide ; physiology ; Female ; Hemin ; pharmacology ; Hypoglossal Nerve ; drug effects ; physiology ; Male ; Protoporphyrins ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Respiration

The organosulfur compound sulforaphane (SFN; C₆H₁₁NOS₂) is a potent cytoprotective agent promoting antioxidant, anti-inflammatory, antiglycative, and antimicrobial effects in in vitro and in vivo experimental models. Mitochondria are the major site of adenosine triphosphate (ATP) production due to the work of the oxidative phosphorylation (OXPHOS) system. They are also the main site of reactive oxygen species (ROS) production in nucleated human cells. Mitochondrial impairment is central in several human diseases, including neurodegeneration and metabolic disorders. In this paper, we describe and discuss the effects and mechanisms of action by which SFN modulates mitochondrial function and dynamics in mammalian cells. Mitochondria-related pro-apoptotic effects promoted by SFN in tumor cells are also discussed. SFN may be considered a cytoprotective agent, at least in part, because of the effects this organosulfur agent induces in mitochondria. Nonetheless, there are certain points that should be addressed in further experiments, indicated here as future directions, which may help researchers in this field of research.
Animals ; Antioxidants/pharmacology* ; Apoptosis/drug effects* ; Brain/ultrastructure* ; Carbon Monoxide Poisoning/metabolism* ; Cytoprotection ; Humans ; Isothiocyanates/pharmacology* ; Membrane Potential, Mitochondrial/drug effects* ; Mitochondria/metabolism* ; Sulfoxides

AIMTo explore the effects of heme- heme oxygenase-1 (HO-1)-carbon monoxide(CO)-cyclic GMP (cGMP)on aortic vascular reactivity in endotoxemic rats and its molecular mechanism.

METHODSBy using isolated vascular ring tension detecting technique, cumulative responses of thoracic aortic rings (TARs)to phenylephrine (PE) were measured at 6 h after lipopolysaccharide administration. Effects on contractile responses to PE were measured under which the TARs were incubated with hemin (He, donor of CO), zinc-protoporphyrin-IX(ZnPP-IX, selective inhibitor of HO-1), or methylene blue (MB, inhibitor of guanylyl cyclase), respectively. The content of CO and the activity of HO-1 were measured. The protein and the gene expression of HO-1 were examined by Western blot and RT-PCR.

RESULTSContractile responses of TARs to cumulative doses of PE were depressed by pretreated with LPS. The hyporesponsiveness was partly reversed by incubation with ZnPP-IX and was restored to normal by incubation with MB in endotoxemic rats. Incubation with He could contribute to the vascular hyporeactivity. The content of CO, the activity and the protein and the gene expression of HO-1 were significantly increased in aorta of endotoxemic rats.

CONCLUSIONLPS could induce the HO-1 mRNA and the protein expression, the activity of HO-1 increase in aorta, lead to active the pathway of heme-HO-1-CO-cGMP, which is one of the important mechanisms of the vascular hyporeactivity in endotoxemic rats.

Animals ; Aorta ; drug effects ; metabolism ; Carbon Monoxide ; metabolism ; Cyclic GMP ; metabolism ; Heme Oxygenase (Decyclizing) ; metabolism ; Lipopolysaccharides ; adverse effects ; Male ; Phenylephrine ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

AIMTo investigate the expression of heme oxygenase-1 gene and production of endogenous carbon monoxide in the rat lung tissue at different time points of chronic hypoxic pulmonary hypertension and the effect of hemin, an inducer of heme oxygenase, on the expression of HO-1 gene and production of endogenous carbon monoxide and pulmonary hypertension.

METHODSWe recreated a rat model of hypoxic pulmonary hypertension by intermittent normal pressure hypoxia (10% O2). The following assays were carried out: Reverse transcriptase polymerase chain reaction (RT-PCR) were performed to determine the level of HO-1 mRNA in rat lung tissue, double wave length spectrophotometry was used to evaluate the quantity of COHb in arterial blood, cardiac catheterization was used to measure the right ventricular systolic pressure (RVSP) and HE staining was performed in dissected lung tissue to observe the pathologic changes of the intra-acinar pulmonary arteries(IAPA).

RESULTS(DT here was low level of HO-1 mRNA in normal rat lung tissue, but the level of HO-1 mRNA increased by 2-4 times in the lung tissue of hypoxic rats (P < 0.01). The quantity of COHb was 2-3 times as those of control group (P < 0.01 or P < 0.05). These were accompanied by the increase of RVSP and the thickness of IAPA. (2) Hemin could maintain the HO-1 mRNA and COHb in the hypoxic rat lung tissue at a high level, and partially suppressed the increase of rat RVSP, ameliorated the pathologic changes of IAPA.

CONCLUSIONThe upregulation of the expression of HO-1 gene and production of CO in the rat lung of hypoxic pulmonary hypertension plays a role of inhibition in the development of hypoxic pulmonary hypertension. Hemin has a therapeutic effect on hypoxic pulmonary hypertension.

Animals ; Carbon Monoxide ; metabolism ; Heme Oxygenase (Decyclizing) ; metabolism ; Hemin ; pharmacology ; Hypertension, Pulmonary ; metabolism ; pathology ; physiopathology ; Hypoxia ; metabolism ; pathology ; Male ; Pulmonary Artery ; metabolism ; physiopathology ; Rats ; Rats, Wistar

AIMTo investigate the protective effect of exogenous carbon monoxide (CO) on the liver injury induced by ischemia/reperfusion (I/R) of hind limbs in rats.

METHODS100 SD rats were divided randomly into sham operated group (S), S+ CO group (SC), I/R group (I/R), I/ R+ CO group (RC). A rat model of ischemia in hind limbs and the reperfusion liver injury was established with the occlusion of the femoral arteries for 4 h and re-opening for 6 - 72 h, 10 d. The rats in SC and RC groups were exposed to air containing CO (the volume traction of CO: 0.05%) for 2 h before and after reperfusion or the corresponding control time point, while the other two groups were exposed to the routine air. The pathologic changes of liver tissue were morphologically observed by HE stain. Serum GPT activity was tested by Automatic Biochemical Analyzer. The percentage of apoptosis, expression levels of bax and bcl-2 protein in liver tissue were detected by Flow Cytometry.

RESULTSThere was no difference between S and SC groups. Compared with SC group: (1) Pathological changes in liver tissue were significant in I/R and RC groups. (2) The serum GPT activity of I/R and RC groups was obviously increased. (3) In IR and RC groups, the percentage of apoptosis in liver tissue was all significantly increased. (4) The bax expression level was significantly increased. Compared RC group with I/R group: (1) Pathological change was slight. (2) The serum GPT activity was depressed. (3) The percentage of apoptosis and expression level of bax protein in liver tissue were depressed. (4) The expression level of bcl-2 protein in liver tissue was increased.

CONCLUSIONExogenous CO could attenuate liver tissue injury induced by limbs I/R in rats.

Animals ; Carbon Monoxide ; pharmacology ; Extremities ; blood supply ; Female ; Liver ; blood supply ; pathology ; Liver Diseases ; etiology ; pathology ; prevention & control ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; prevention & control

OBJECTIVETo study the effect of endogenous carbon monoxide (CO) on the smooth muscle function of the dog penile corpus cavernosum in vitro.

METHODSTissue bioassay was used to measure the corpus cavernosum muscle contraction and relaxation. The production of CO was induced in the corpus cavernosum smooth muscle, and the effect of CO on the penile corpus cavernosum smooth muscle pre-contracted by phenylephrine (PE) was determined.

RESULTSChlorinous hemoglobin could relax the smooth muscle stripes pre-contracted by 10 micromol/L PE. A dose-dependent relaxation was observed. The relaxation responses by 10 -100 micromol/L chlorinous hemoglobin were significant compared with the control group (P < 0. 01). The pretreatment of the muscle stripes with ZnPP-IX or methylthioninium significantly reduced the relaxing effect of chlorinous hemoglobin (P < 0.01).

CONCLUSIONThe relaxing effect of endogenous CO on the smooth muscle of the penile corpus cavernosum depends on the concentration of endogenous CO. The underlying mechanism may involve the pathway from CO to cGMP production.

Animals ; Carbon Monoxide ; physiology ; Dogs ; Dose-Response Relationship, Drug ; Hemin ; pharmacology ; In Vitro Techniques ; Male ; Muscle, Smooth ; drug effects ; physiology ; Penile Erection ; drug effects ; physiology ; Penis ; drug effects ; physiology

The discovery of nitric oxide (NO) initiated a novel research field, gaseous signal molecules. Other two gaseous signal molecules, carbon monoxide (CO) and hydrogen sulfide (H2S), have been approved subsequently. All of these three gaseous signal molecules are generated endogenously and exert extensive biological effects. This article reviews the significance of gaseous signal molecules in the pathogenesis of cardiovascular diseases. In the cardiovascular system, the endogenous NO, CO and H2S form three specific and relevant pathways with their corresponding enzymes including nitric oxide synthase (NOS), heme oxygenase (HO), and cystathionine-gamma-lyase (CSE). The gaseous signal molecules not only participate in the maintenance of physiological function and structure of the cardiovascular system, but also exert important pathophysiological effects in the pathogenesis of cardiovascular diseases, such as hypertension, pulmonary hypertension, septic shock and atherosclerosis. Further studies on gaseous signal molecules will remarkably promote the basic research in the cardiovascular fields, and provide a novel direction for the clinical research as well.
Animals ; Carbon Monoxide ; physiology ; Cardiovascular Diseases ; etiology ; Humans ; Hydrogen Sulfide ; metabolism ; pharmacology ; Hypertension ; etiology ; Hypertension, Pulmonary ; etiology ; Nitric Oxide ; physiology ; Shock, Septic ; etiology ; Signal Transduction

The use of exogenous carbon monoxide releasing molecules (CORMs) provides promise for clinical application; however, the hazard potential of CORMs in vivo remains poorly understood. The developmental toxicity of CORM-3 was investigated by exposure to concentrations ranging from 6.25 to 400 μmol/L during 4-144 h post fertilization. Toxicity endpoints of mortality, spontaneous movement, heart rate, hatching rate, malformation, body length, and larval behavior were measured. CORM-3 disrupted the progression of zebrafish larval development at concentrations exceeding 50 μmol/L, resulting in embryonic developmental toxicity.
Animals ; Carbon Monoxide ; pharmacology ; Cardiotonic Agents ; toxicity ; Dose-Response Relationship, Drug ; Embryo, Nonmammalian ; drug effects ; Embryonic Development ; drug effects ; Organometallic Compounds ; toxicity ; Zebrafish ; embryology ; metabolism