[Investigation of the arsenic levels in ecosystem aspect in water type of endemic arsenicosis area in Datong City].

OBJECTIVE: To investigate the arsenic levels in endemic arsenism in Datong City, Shanxi Province. METHODS: A total of 85 inhabitants from one village in endemic arsenism area in Datong City, Shanxi Province were collected as research subjects. The People's Republic of China health industry standard for endemic arsenism was used to identify and diagnosis the patients. Daily drinking water and soil were collected and detected by atomic fluorescence spectrometry. The content of vegetables were detected by inductively coupled plasma mass spectrometry (ICP-MS). RESULTS: In the study, 85 samples were collected. Arsenic concentration in the daily drinking water were 14.41 - 90.34 µg/L, and the median value was 43.88 µg/L. The arsenic concentration of vegetables were 0.001 - 0.771 mg/kg, and 43.04% of samples, were higher than the maximal permissible limit of As in food. The results that the arsenic concentration of vegetables constant changes in the leaf vegetables > tubers > fruit vegetables. The health risk of intaking arsenic pollution in vegetables up to 71.77%. The arsenic levels in village of four directions were not exceeded the Chinese standards. CONCLUSIONS: Arsenic concentration in drinking water and vegetables are high in waterborn endemic arsenicosis area of Shanxi province. Arsenic in drinking water has been considered as a primary cause of arsenism, but direct intake of arsenic from vegetables can not be ignored.

Oxidative DNA damage of peripheral blood polymorphonuclear leukocytes, selectively induced by chronic arsenic exposure, is associated with extent of arsenic-related skin lesions.

There is increasing evidence that oxidative stress is an important risk factor for arsenic-related diseases. Peripheral blood leukocytes constitute an important defense against microorganisms or pathogens, while the research on the impact of chronic arsenic exposure on peripheral blood leukocytes is much more limited, especially at low level arsenic exposure. The purpose of the present study was to explore whether chronic arsenic exposure affects oxidative stress of peripheral blood leukocytes and possible linkages between oxidative stress and arsenic-induced skin lesions. 75 male inhabitants recruited from an As-endemic region of China were investigated in the present study. The classification of arsenicosis was based on the degree of skin lesions. Arsenic levels were measured in drinking water and urine by Atomic Fluorescence Spectroscopy. Urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) was tested by Enzyme-Linked Immunosorbent Assay. 8-OHdG of peripheral blood leukocytes was evaluated using immunocytochemical staining. 8-OHdG-positive reactions were only present in polymorphonuclear leukocytes (PMNs), but not in monocytes (MNs). The 8-OHdG staining of PMN cytoplasm was observed in all investigated populations, while the 8-OHdG staining of PMN nuclei was frequently found along with the elevated amounts of cell debris in individuals with skin lesion. Urinary arsenic levels were increased in the severe skin lesion group compared with the normal group. No relationship was observed between drinking water arsenic or urine 8-OHdG and the degree of skin lesions. These findings indicated that the target and persistent oxidative stress in peripheral blood PMNs may be employed as a sensitive biomarker directly to assess adverse health effects caused by chronic exposure to lower levels of arsenic.

Heat-shock protein 70 (HSP70) polymorphisms affect the risk of coke-oven emission-induced neurobehavioral damage.

OBJECTIVES: Epidemiology studies indicated that coke-oven workers with long-term exposure to polycyclic aromatic hydrocarbons (PAHs) often have some neurobehavioral abnormalities especially impairment for cognitive function, while the underlying mechanisms are not fully understood. Numerous studies have indicated the antioxidant and anti-apoptosis roles of heat shock protein 70 (Hsp70). The genetic polymorphisms in HSP70 genes are associated with multiple diseases including neurotoxicity. However, it is unclear whether HSP70 polymorphisms are related to the neurotoxicity of PAH. We, therefore, investigate the possible association between HSP70 polymorphisms and neurobehavioral abnormalities. METHODS: 188 coke-oven workers and 137 control workers were recruited in this study. Emotional and cognitive function was assessed using the WHO/NCTB. HSP70 polymorphisms (HSP70-1 G190C, HSP70-2 G1267 A and HSP70-hom T2437C) were checked by PCR-RFLP. RESULTS: The results indicated that HSP70-1 CC genotypes in coke-oven workers were associated with poor neurobehavioral performance such as the attention /response speed and visual perception/memory, while the HSP70-2 AA genotypes were associated with lower short-term auditory memory. CONCLUSIONS: HSP70-1 CC and HSP70-2 AA genotypes in coke-oven workers may increase the risk for neurobehavioral damage, especially attention, learning and memory.

Low selenium status affects arsenic metabolites in an arsenic exposed population with skin lesions.

BACKGROUND: The antagonistic effects between selenium (Se) and arsenic (As) suggest that low selenium status plays important roles in arsenism development. However, no study has been reported for humans suffering from chronic arsenic exposure with low selenium status. METHODS: Sixty-three subjects were divided into 2 experimental groups by skin lesions (including hyperkeratosis, depigmentation, and hyperpigmentation). Total urine and serum concentrations of arsenic and selenium were determined by ICP-MS with collision/reaction cell. Arsenic species were analysed by ICP-MS coupled with HPLC. RESULTS: The mean concentration of As in the drinking waters was 41.5 microg/l. The selenium dietary intake for the studied population was 31.7 microg Se/d, and which for the cases and controls were 25.9 and 36.3 microg Se/d, respectively. Compared with the controls, the skin lesions cases had lower selenium concentrations in serum and urine (41.4 vs 49.6 microg/l and 71.0 vs 78.8 microg/l, respectively), higher inorganic arsenic (iAs) in serum (5.2 vs 3.4 microg/l, P<0.01), higher percentages of iAs in serum and urine (20.2) vs 16.9% and 18.3 vs 14.5%, respectively, P<0.01) but lower percentages of monomethylarsonate (MMA) in serum (15.5 vs 18.8%, P<0.01) ans dimethylarsinate acid (DMA) in urine (65.1 vs 69.8%, P<0.01). Subjects with lower selenium concentrations in serum (<50 microg/l) had a stronger tendency to the risk of skin lesions than individual having higher selenium concentrations [odd ratio (OR), 7.3; 95% confidence interval (95% CI), 1.5-35.7; P=0.014]. This OR estimation was confirmed in those subjects having higher ratios of As/Se in urine and serum, with OR as high as 10.3 and 3.8 respectively. CONCLUSIONS: Lower serum selenium status (<50 microg/l) is significantly correlated to the arsenic-associated skin lesions in the arsenic exposed population. The accumulation of iAs and its inhibition to be biotransformed to DMA occurred in human due to chronic exposure of low selenium status.

A novel transdermal patch incorporating isosorbide dinitrate with bisoprolol: in vitro and in vivo characterization.

The combination therapy of nitrate and selective beta-adrenoceptor antagonist has shown benefits for treatment of hypertension and heart disease than either drug alone. The objectives of the present study were to define effects on the anti-hypertension activity and pharmacokinetics of a novel transdermal patch incorporating isosorbide dinitrate (ISDN) with bisoprolol (BP). The 3:2 ratio of ISDN to BP (mg/mg) in the transdermal patches exhibited better anti-hypertension effect synergistically with a similar inhibiting heart rates effect to that of BP alone in renovascular hypertensive rats, and was therefore selected as a final formulation. The in vitro transdermal penetration of both ISDN and BP from the patches displayed a zero-order process, and the penetration rate constants were 7.4 microg/(cm(2)h) for ISDN, and 5.9 microg/(cm(2)h) for BP, respectively. After transdermal administration at single dose or multiple doses, the synergistic anti-hypertension effect was confirmed in spontaneously hypertensive rats also. The effect of each patch lasts for 3 days, and increased with the total dose of two drugs (2mg/cm(2), ISDN:BP=3:2, mg/mg), showing a dose dependant manner. After transdermal administration to rabbits, the absolute bioavailabilities were 33.6% for ISDN, and 31.3% for BP, respectively. The maximal concentrations (C(max)) of both drugs were significantly reduced while the areas under the plasma concentration-time curve (AUC), and mean residence times (MRT) were evidently increased and extended, respectively. As a patient-friendly, convenient, and multi-day dosing therapeutic system, the transdermal patches incorporating ISDN and BP could be promising for prevention and treatment of hypertension.

Protective effects of hepatocellular canalicular conjugate export pump (Mrp2) on sodium arsenite-induced hepatic dysfunction in rats.

Arsenic is a double-edged sword to human health. The excretion of various organic anions into bile is mediated by an adenosine triphosphate-dependent conjugate export pump, which has been identified as the canalicular isoform of the multidrug resistance protein 2 (Mrp2). It has been proved that Mrp2 can transport arsenite in vitro, but its effects in vivo are not clear. The aim of this study was to investigate whether Mrp2 plays a role in exportation of arsenic in vivo and its protective effects on liver function. Mrp2 protein level in rat liver was determined by Western blot analysis. Total arsenic concentrations in whole blood and bile were measured using hydride generation atomic absorption spectrometry. Alanine aminotransferase (ALT) activity, aspartate aminotransferase activity (AST), glutathione peroxidase (GSH-PX) activity, malon dialdehyde (MDA) and total bilirubin were measured by biochemical assays. The morphological changes were observed by electron microscopy. Total arsenic levels in blood and bile of arsenite-treated rats were significantly higher than those of control rats (P<0.05) at all three different time points. The overexpression of Mrp2 was 36.61%, 32.36% and 12.73% at 2, 4 and 6 weeks, respectively (percentage of controls, P<0.05), which was significantly higher than controls. A positive correlation between Mrp2 expression level and total arsenic concentration in bile indicated that Mrp2 accelerated the transport of arsenic. Electron microscopy showed that microvilli of bile canaliculi became swollen and sparse. ALT and AST activities in serum were markedly raised at 6 weeks. MDA level in serum increased (P<0.05) and GSH-PX activity in serum decreased except for 2 weeks. Damage of liver function became worse following decreased expression of Mrp2. In conclusion, overexpression of Mrp2 may explain increased biliary excretion of arsenic and it may protect liver function.

[Effects of MRP2-GSH cotransport system on hepatic arsenic metabolism in rats].

OBJECTIVE: To investigate the role of multidrug resistant protein 2 (MRP2) and glutathione (GSH) cotransport system in hepatic arsenic metabolism in rats. METHODS: Thirty healthy Wistar rats were divided randomizedly into five groups. The first group was the control group and the rats in this group were administered with normal saline. In the second, third and fourth group the rats were administered with 4, 10 and 20 mg As(+)3/kg BW of sodium arsenite respectively every other day for two weeks. The fifth group was the benzene-soluble organics (BSO) intervention group and in this group the rats were administered with 2 mmol/kg BW BSO intraperitoneally every day three days before the end of the experiment. The other treatment was the same as in other groups. All rats were sacrificed two weeks after the treatments. Arsenic contents in bile, liver and blood were detected by atomic absorption spectroscopy (AAS), and the expression of MRP2 in the membrane of hepatocyte was determined by Western-blot analysis. RESULTS: The level of total arsenic (including organic arsenic and inorganic arsenic) in bile, liver and blood in all three different dose groups was higher than those in the control groups (P < 0.05). Arsenic levels of bile and liver were increased with intragastric arsenic dose. Blood arsenic levels were not significantly different in three different dose groups. Expression of hepatic MRP2 was increased with intragastric arsenic concentration. A positive correlation between biliary arsenic concentration and MRP2 levels was found in liver (r = 0.986, P < 0.05). For the rats pretreated with BSO, the biliary arsenic was significantly higher than that in the control group but lower than that in the high dose group; the liver and blood arsenic was higher than that in the control group and in the high dose group. Expression of MRP2 pretreated with BSO was decreased. CONCLUSION: Sodium arsenite can induce expression of MRP2 and the up-regulation of MRP2 may play an important role in the bile secretion of arsenite and its metabolites. The function of MRP2 for transportation of arsenic and its metabolites is associated with the intracellular GSH level. BSO inhibits the synthesis of GSH, which weakens the function of the MRP2-GSH cotransport system and makes the liver arsenic increased.

[Toxic effect of arsenite on the expression of liver multidrug resistance-associated protein 2 in rat].

OBJECTIVE: To investigate the role of multidrug resistance-associated protein 2 (MRP2) in the hepatic cell membrane of rats. METHODS: Thirty healthy Wistar rats were divided randomly into six groups based on time of administration (2 w, 4 w, 6 w) of 20 mg/kg of sodium arsenite, and their corresponding control groups. Animals were administered every other day. Arsenic content in blood and bile were detected by atomic absorption spectroscopy (AAS), and the expression of MRP2 in the membrane of hepatocyte by Western blotting was determined. RESULTS: Total arsenic levels (including organic arsenic and inorganic arsenic) in blood and bile were significantly higher than control groups (P < 0.05) at all three different time points, especially in 2 w and 4 w group (16.8 and 13.8 fold greater than that in control). The expression of MRP2 increased 36.61%, 32.36%, 12.73% more respectively in 2 w, 4 w, 6 w groups than those in control groups (P < 0.05). The expression of MRP2 was correlated with total arsenic content in bile (r = 0.713, P < 0.05). CONCLUSIONS: Bile is one of the major routes for the excretion of arsenite and its metabolites, and the overexpression of MRP2 may play an important role in the bile excretion of them at early stage.