A novel controllable hydrogen sulfide-releasing molecule protects human skin keratinocytes against methylglyoxal-induced injury and dysfunction.

BACKGROUND/AIM: Delayed wound healing is a common skin complication of diabetes, which is associated with keratinocyte injury and dysfunction. Levels of methylglyoxal (MGO), an α-dicarbonyl compound, are elevated in diabetic skin tissue and plasma, while levels of hydrogen sulfide (H2S), a critical gaseous signaling molecule, are reduced. Interestingly, the gas has shown dermal protection in our previous study. To date, there is no evidence demonstrating whether MGO affects keratinocyte viability and function or H2S donation abolishes these effects and improves MGO-related impairment of wound healing. The current study was conducted to examine the effects of MGO on the injury and function in human skin keratinocytes and then to evaluate the protective action of a novel H2S-releasing molecule. METHODS: An N-mercapto-based H2S donor (NSHD)-1 was synthesized and its ability to release H2S was observed in cell medium and cells, respectively. HaCaT cells, a cell line of human skin keratinocyte, were exposed to MGO to establish an in vitro diabetic wound healing model. NSHD-1 was added to the cells before MGO exposure and the improvement of cell function was observed in respect of cellular viability, apoptosis, oxidative stress, mitochondrial membrane potential (MMP) and behavioral function. RESULTS: Treatment with MGO decreased cell viability, induced cellular apoptosis, increased intracellular reactive oxygen species (ROS) content and depressed MMP in HaCaT cells. The treatment also damaged cell behavioral function, characterized by decreased cellular adhesion and migration. The synthesized H2S-releasing molecule, NSHD-1, was able to increase H2S levels in both cell medium and cells. Importantly, pretreatment with NSHD-1 inhibited MGO-induced decreases in cell viability and MMP, increases in apoptosis and ROS accumulation in HaCaT cells. The pretreatment was also able to improve adhesion and migration function. CONCLUSION: These results demonstrate that the novel synthesized H2S donor is able to protect human skin keratinocytes against MGO-induced injury and behavior dysfunction. We believe that more reasonable H2S-releasing molecules will bring relief to patients suffering from delayed wound healing in diabetes mellitus in the future.

[Key parameters and formulas for approximate average height and weight of normal Chinese children under 7 years of age].

OBJECTIVE: In order to understand the normal physical growth of Chinese children, data of children aged from 0 to 7 years from urban and rural areas of nine Chinese cities in 2005 were analyzed. METHODS: The original data of height and weight were drawn into growth curves charts by Graphpad Prism 5.0 software according to the different age groups. The children were classified into five age groups: 0-3 months, 4-6 months, 7-12 months, 13-24 months, and 2-7 years. RESULTS: The average birth weight was 3.3 kg and the height averaged 50 cm. The average monthly weight gain was 1.0-1.2 kg and the average monthly increase of height was 4 cm in the 0-3 months group. By 3 months of age, the weight and height averaged 6.6 kg and 62 cm, respectively. In the 4-6 months group, the growth rate was reduced to a half of the 0-3 months group, with an average monthly weight and height gain was 0.5-0.6 kg and 2 cm respectively. The growth rate in the 7-2 months group was a half of the 4-6 months group. By 12 months of age, the weight and height average 9.9 kg and 75 cm, respectively. The average monthly weight and height gain in the 13-24 months group was 0.2 kg and 1 cm respectively, with an average weight and height of 12 kg and 87 cm respectively by 24 months of age. A steady growth was found in the 2-7 years group, with a yearly average weight and height increment of about 2 kg and 7 cm respectively. The formulas for approximate average weight and height in children between 2 and 7 years were as follows: age (yr)*2+8 (kg) (weight); age (yr)*7+75 (cm) (height). CONCLUSIONS: The approximate weight and height of normal Chinese children under 7 years of age can be evaluated by the key parameters and formulas above mentioned.

Calcitriol prevents peripheral RSC96 Schwann neural cells from high glucose & methylglyoxal-induced injury through restoration of CBS/H2S expression.

A meta-analysis has suggested that vitamin D deficiency is involved in diabetic peripheral neuropathy (DPN) and the levels of hydrogen sulfide (H2S) are also decreased in type 2 diabetes. The injection of vitamin D induces cystathionine-ß-synthase (CBS) expression and H2S generation. However, it remains unclear whether the supplementation of vitamin D prevents DPN through improvement of CBS/H2S expression. In the present study, RSC96 cells, a rat Schwann cell line, were exposed to high glucose and methylglyoxal (HG&MG) to simulate diabetic peripheral nerve injury in vivo. Before the exposure to HG&MG, the cells were preconditioned with calcitriol (CCT), an active form of vitamin D, and then CCT-mediated neuroprotection was investigated in respect of cellular viability, superoxide anion (O2(-)) generation, inducible nitric oxide (NO) synthase (iNOS)/NO expression, mitochondrial membrane potential (MMP), as well as CBS expression and activity. It was found that both high glucose and MGO decreased cell viability and co-treatment with the two induced a more serious injury in RSC96 cells. Therefore, the exposure to HG&MG was used in the present study. The exposure to HG&MG markedly induced iNOS expression, NO and O2(-) generation, as well as MMP loss. In addition, the exposure to HG&MG depressed CBS expression and activity in RSC96 cells. However, the preconditioning with CCT significantly antagonized HG&MG-induced cell injury including the decreased viability, iNOS overexpression, NO and O2(-) accumulation, as well as MMP loss. CCT also partially restored the decreased CBS expression and activity triggered by HG&MG, while the inhibition of CBS with hydroxylamine attenuated CCT-mediated neuroprotection. Moreover, the exogenous donation of H2S produced similar cellular protective effects to CCT. The data indicate that the supplementation of vitamin D prevents HG&MG-induced peripheral nerve injury involving the restoration of endogenous H2S system, which may provide a basal support for the treatment of DPN with vitamin D clinically.

[Comparison of RR-formoterol with rac-formoterol in bronchodilating effect on human bronchus].

OBJECTIVE: To compare the bronchodilating effect of RR-formoterol (RR-FMT) with that of racemic formoterol (rac-FMT) on human bronchus. METHODS: Human bronchial spiral strips (2 - 4 mm internal diameter,15 mm length) were suspended in tissue baths under resting tension of 1.0 g. The changes of tension induced by RR-FMT and rac-FMT(10 pmol x L(-1) - 3.2 micromol x L(-1)) in a cumulative concentration manner were studied under resting tension conditions or precontraction with carbamylcholine (10 micromol x L(-1)) or histamine(100 micromol x L(-1)) in human bronchus. RESULT: The bronchodilating effect of RR-FMT was more potent than that of rac-FMT under resting condition(P<0.05). RR-FMT and rac-FMT reversed histamine or carbamylcholine-induced contraction, and the bronchodilating effect of RR-FMT was more potent than that of rac-FMT (P<0.05). CONCLUSION: The bronchodilating effect of RR-FMT is more potent than that of rac-FMT in both the resting condition and carbamylcholine or histamine-induced contraction in human bronchus in vitro.

[The protective effects of ischemic preconditioning on reperfusion injury of rat lung].

AIM: To investigate the effects of ischemic preconditioning on reperfusion injury of rat lung. METHODS: Rat isolated lungs (n=8 in each group) were stabilized and perfused by Krebs-Henseleit solution on a modified langendorff perfusion apparatus. 36 wistar rats were divided into three groups: ischemic preconditioning group, ischemia/reperfusion group and control group. Mean pulmonary artery pressure, wet/dry ratio, pulmonary surfactant phospholipid and alveolar surface tension in bronchoalveolar lavage fluid and electron microscope were detected. RESULTS: The morphological changes of lung injury were alleviated in the preconditioning group under electron microscope. Wet/dry ratio, mean pulmonary artery pressure and SA/LA ratio were significantly lower in the preconditioning group after ischemic/reperfusion (P < 0.01). Total phospholipid and large aggregate in the BALF were significantly increased in the preconditioning group (P < 0.01). Small aggregate showed no change in three groups. Surfactant activity test showed that surface tension markedly decreased in IPC group (P < 0.01). CONCLUSION: These results indicates that ischemic preconditioning may have a protective effect in ischemic/reperfusion injuries lung by ameliorating the content and function of surfactant phospholipid.