Acute Effects of Individual Exposure to Fine Particulate Matter on Pulmonary Function in Schoolchildren.

OBJECTIVE: This panel study aimed to determine the acute effects of exposure to fine particulate matter (PM 2.5) on schoolchildren's pulmonary function. METHODS: We selected 51 schoolchildren aged 9-12 years attending a full-time boarding school in Beijing, China, measured the indoor and outdoor PM 2.5 concentrations for five consecutive days, calculated the PM 2.5 time-weighted individual exposure levels based on the school micro-environmental concentrations and the time activity pattern recorded by schoolchildren, measured schoolchildren's pulmonary function on the fifth day. The survey was performed three times from December 2018 to April 2019. We used a linear mixed-effects model to evaluate the associations between PM 2.5 and pulmonary function. RESULTS: During the three surveys, the median PM 2.5 time-weighted individual exposure concentrations were 15.30 µg/m 3, 48.92 µg/m 3, and 42.89 µg/m 3, respectively. There was a significant difference between the three surveys in vital capacity (VC), forced vital capacity (FVC), forced expiratory volume in one second (FEV 1) and forced expiratory volume in one second/forced vital capacity (FEV 1/FVC) ( P < 0.05). The relevance analysis found that PM 2.5 had lag effect on schoolchildren's pulmonary function, each 10 µg/m 3increase in PM 2.5could cause largest decreases in FEF 25%-75%, FEV 1/FVC, FEF 75%, and FEV 1 on lag 0-1 d (80.44 mL/s, 35.85%, 78.58 mL/s, and 61.34 mL, respectively), and largest decreases in FEF 25% on lag 1 d (83.68 mL/s), in VC on lag 4 d (32.34 mL), and in FVC on lag 0-4 d (37.76 mL). Gender subgroup analysis revealed that the increase in PM 2.5 caused a decrease in FEV 1/FVC and VC on the day of physical examination only in boys, and on lag days it caused changes in different pulmonary function indicators, both for boys and girls, but most of the pulmonary function indicators decreased more in boys than in girls. CONCLUSION: Our findings show that acute PM 2.5 exposure has significant effects on pulmonary function within 0-4 d, on both small airway indicators and large airway indicators. Boys' pulmonary function is more sensitive to PM 2.5 than girls.

Verification on the Developmental Toxicity of Short-term Exposure to Phenol in Rats.

OBJECTIVE: To verify the health advisory for short-term exposure to phenol. METHODS: The method of this validation experiment was the same as the US Environmental Protection Agency (EPA) methodology for toxicology experiments used to determine phenol drinking water equivalent level (DWEL). Pregnant female Sprague-Dawley rats were administered phenol in distilled water by gavage at daily doses of 15, 30, 60, 120, and 240 mg/kg body weight (b.w.) from implantation (the 6th day post-mating) to the day prior to the scheduled caesarean section (the 20th day of pregnancy). The following information was recorded: general behavior; body weight; number of corpus luteum, live birth, fetus, stillbirth, and implantation; fetal gender; body weight; body length; tail length; and abnormalities and pathomorphological changes in the dams. RESULTS: In the 60 mg/kg b.w. dose group, the mortality of pregnant rats increased with increasing doses, suggesting maternal toxicity. Fetal and placental weights decreased as phenol dose increased from 30 mg/kg b.w., and were significantly different compared those in the vehicle control group, which suggested developmental toxicity in the fetuses. However, the phenol-exposed groups showed no significant change in other parameters compared with the vehicle control group ( P > 0.05). CONCLUSION: Despite using the same method as the US EPA, a different NOEAL of 15 mg/(kg·d) was obtained in this study.

FOXC1 up-regulates the expression of toll-like receptors in myocardial ischaemia.

Myocardial ischaemia (MI) remains a major cause of death and disability worldwide. Accumulating evidence suggests a significant role for innate immunity, in which the family of toll-like receptors (TLRs) acts as an essential player. We previously reported and reviewed the changes of Tlr expression in models of MI. However, the underlying mechanisms regulating Tlr expression in MI remain unclear. The present study first screened transcription factors (TFs) that potentially regulate Tlr gene transcription based on in silico analyses followed by experimental verification, using both in vivo and in vitro models. Forkhead box C1 (FOXC1) was identified as a putative TF, which was highly responsive to MI. Next, by focusing on two representative TLR subtypes, an intracellular subtype TLR3 and a cell-surface subtype TLR4, the regulation of FOXC1 on Tlr expression was investigated. The overexpression or knockdown of FoxC1 was observed to up- or down-regulate Tlr3/4 mRNA and protein levels, respectively. A dual-luciferase assay showed that FOXC1 trans-activated Tlr3/4 promoter, and a ChIP assay showed direct binding of FOXC1 to Tlr3/4 promoter. Last, a functional study of FOXC1 was performed, which revealed the pro-inflammatory effects of FOXC1 and its destructive effects on infarct size and heart function in a mouse model of MI. The present study for the first time identified FOXC1 as a novel regulator of Tlr expression and described its function in MI.

Sub-chronic Toxicity of Defoamer Used in Seawater Desalination.

OBJECTIVE: To explore the possible long-term health effects of the defoamer used in seawater desalination by sub-chronic toxicity testing. METHODS: Blood analysis, internal organ assessment, and histopathological examination were carried out in rats exposed to low, medium, and high (0.5, 1.0, and 2.0 g/kg BW, respectively) doses of defoamer for 90 days through oral administration. RESULTS: The high dose group showed decreased blood alanine aminotransferase and aspartate aminotransferase (P < 0.05). All doses resulted in a significant increase in albumin and decrease in globulin (P < 0.05). The direct bilirubin and indirect bilirubin were decreased in the medium and high dose groups (P < 0.05). All dose groups showed significant induction of alkaline phosphatase (P < 0.05). Pathological examination revealed a case of liver mononuclear cell infiltration in the medium dose group and three cases of liver congestion, steatosis of hepatic cells around the central vein, and punctate necrosis with multiple focal mononuclear cell infiltration in male rats administered the high dose. The No Observed Adverse Effect Level was 0.5 g/kg BW in rats, with albumin and total bilirubin as health effect indices. CONCLUSION: Long-term defoamer exposure may cause liver injury but has no significant impact on renal function in rats. The effect on blood cells in female rats was more prominent than that in male rats.

TLR3 contributes to persistent autophagy and heart failure in mice after myocardial infarction.

Toll-like receptors (TLRs) are essential immunoreceptors involved in host defence against invading microbes. Recent studies indicate that certain TLRs activate immunological autophagy to eliminate microbes. It remains unknown whether TLRs regulate autophagy to play a role in the heart. This study examined this question. The activation of TLR3 in cultured cardiomyocytes was observed to increase protein levels of autophagic components, including LC3-II, a specific marker for autophagy induction, and p62/SQSTM1, an autophagy receptor normally degraded in the final step of autophagy. The results of transfection with a tandem mRFP-GFP-LC3 adenovirus and use of an autophagic flux inhibitor chloroquine both suggested that TLR3 in cardiomyocytes promotes autophagy induction without affecting autophagic flux. Gene-knockdown experiments showed that the TRIF-dependent pathway mediated the autophagic effect of TLR3. In the mouse model of chronic myocardial infarction, persistent autophagy was observed, concomitant with up-regulated TLR3 expression and increased TLR3-Trif signalling. Germline knockout (KO) of TLR3 inhibited autophagy, reduced infarct size, attenuated heart failure and improved survival. These protective effects were abolished by in vivo administration of an autophagy inducer rapamycin. Similar to the results obtained in cultured cardiomyocytes, TLR3-KO did not prevent autophagic flux in mouse heart. Additionally, this study failed to detect the involvement of inflammation in TLR3-KO-derived protection, as wild-type and TLR3-KO hearts were comparable in inflammatory activity. It is concluded that up-regulated TLR3 expression and signalling contributes to persistent autophagy following MI, which promotes heart failure and lethality.

Dimethylation of Histone 3 Lysine 9 is sensitive to the epileptic activity, and affects the transcriptional regulation of the potassium channel Kcnj10 gene in epileptic rats.

Potassium channels can be affected by epileptic seizures and serve a crucial role in the pathophysiology of epilepsy. Dimethylation of histone 3 lysine 9 (H3K9me2) and its enzyme euchromatic histone­lysine N­methyltransferase 2 (G9a) are the major epigenetic modulators and are associated with gene silencing. Insight into whether H3K9me2 and G9a can respond to epileptic seizures and regulate expression of genes encoding potassium channels is the main purpose of the present study. A total of 16 subtypes of potassium channel genes in pilocarpine­modelled epileptic rats were screened by reverse transcription­quantitative polymerase chain reaction, and it was determined that the expression ATP­sensitive inward rectifier potassium channel 10 (Kcnj10) increased in hippocampus and insular cortex, while the expression of most of the other subtypes decreased. The total level of H3K9me2 decreased in the model group compared with the control. The Kcnj10 gene encoding the Kir4.1 channel was selected to analyse changes in H3K9me2 in the promoter region by the chromatin immuno­precipitation method. Anti­H3K9me2 and anti­G9a antibodies were used to identify the modified DNAs. Five primers were designed across the promoter region of the Kcnj10 gene. In epileptic hippocampi, the relative abundance of H3K9me2 and G9a in the promoter region of Kcnj10 decreased markedly. Removal of the H3K9me2 repressive mark resulted in decreased transcriptional inhibition of the Kcnj10 gene and therefore increased its expression. In the cultured C6 cells, specific inhibition of the enzymatic activity of G9a by 2­(Hexahydro­4­methyl­1H­1,4­diazepin­1­yl)­6,7­di­ methoxy­N­(1­(phenyl­methyl)­4­piperidinyl)­4­quinazolinamine tri­hydrochloride hydrate (bix01294) resulted in upregulation of the expression of Kir4.1 proteins. The present study demonstrated that H3K9me2 and G9a are sensitive to epileptic seizure activity during the acute phase of epilepsy and can affect the transcriptional regulation of the Kcnj10 channel.

Potential Effects of Desalinated Seawater on Arteriosclerosis in Rats.

To evaluate the potential risk of arteriosclerosis caused by desalinated seawater, Wistar rats were provided desalinated seawater over a 1-year period, and blood samples were collected at 0, 90, 180, and 360 days. Blood calcium, magnesium, and arteriosclerosis-related indicators were investigated. Female rats treated with desalinated seawater for 180 days showed lower magnesium levels than the control rats (P < 0.05). The calcium and magnesium levels in female rats and the magnesium level in male rats were lower than the levels in the controls, following treatment with desalinated seawater for 360 days (P < 0.05). Blood levels of arteriosclerosis-related lipid peroxidation indicators and C-reactive protein (CRP) in the treatment group did not differ from those in the controls. The levels of lipid peroxidation indicators and CRP in rats were not significantly affected by drinking desalinated seawater, and no increase in risk of arteriosclerosis was observed.

MiR-126 Suppresses the Glucose-Stimulated Proliferation via IRS-2 in INS-1 ß Cells.

BACKGROUND: Increasing evidence suggests that miR-126 participates in the glucose homeostasis through its target molecules. Although bioinformatics analysis predicts that miR-126 can bind with the insulin receptor substrate-2(IRS-2) mRNA at the "seed sequence", but there are still no definitely reports to support it. In this study, we provided evidences that IRS-2 was one of the target genes of miR-126. And miR-126 has a proliferation inhibiting effects in INS-1 ß cells, mainly through the suppression of IRS-2. METHODS: The 3'-UTR of IRS-2 regulated by miR-126 was analyzed by the luciferase assay and western blot. Furthermore, proliferation of INS-1 ß cells stimulated by glucose was tested, and the association between IRS-2 and miR-126 were analyzed. RESULTS: We found that mutation of only three of the 6 "seed sequences" can eliminate the inhibition effect of miR-126. In INS-1 ß cells, administration of miR-126 suppresses the proliferation, together with the unbalanced down-regulation of IRS-2 and IRS-1. Over-expression of IRS-2 can reverse the proliferation effect of miR-126, while not of IRS-1. These results suggested that miR-126 inhibited the ß-cell proliferation via the inhibition of IRS-2 instead of IRS-1.Additionally, we also found that high glucose and insulin could stimulate the rapid production of endogenous miR-126 within 6 hours, together with the short term suppression of IRS-1 and IRS-2 expression, and intensify the unbalanced expression of IRS-1 and IRS-2. CONCLUSIONS: IRS-2 was one of the targets of miR-126. MiR-126 inhibited the ß-cell proliferation through IRS-2 instead of IRS-1. MiR-126 may take part in the glucose homeostasis both through its target IRS-2 and IRS-1. The unbalance between IRS-1 and IRS-2 caused by miR-126 may play an important role in type 2 diabetes.

[Heavy metals in the surface sediment of the dumping ground outside Jiaozhou Bay and their potential ecological risk].

Based on the monitoring data of heavy metals (Cr, Hg, Cd, Pb, Zn, Cu) in the surface sediment of the dumping ground outside Jiaozhou Bay from 2003 to 2008, the distribution patterns, factors controlling the distribution, and the potential ecological risks of heavy metals were studied with the data in 2007-08, and the fluctuation trends of heavy metals in the surface sediment over the 6 years were also discussed. The average concentrations of heavy metals Cr, Hg, Cd, Pb, Zn, Cu in the surface sediment were 29.47, 0.065, 0.105, 1.145, 9.63, 3.355 microg/g, respectively. Except for Cr, the concentration of heavy metals was high in the central dumping area while low outside the dumping ground, suggesting that the dredged material dumped was the main source of heavy metals. Organic carbon content in the surface sediment had a significant positive correlation with heavy metals except for Cr. Based on the results of ecological risk assessment, Hg had a medium potential ecological risk, while the other heavy metals had low potential ecological risk. The overall risk index (RI) of the heavy metals was 100.50, which was considered as a level of low potential ecological risk. The average concentration of heavy metals showed a decreasing trend over the 6 years, except Hg. In conclusion, the quality of surface sediment in term of heavy metals in the dumping ground outside Jiaozhou Bay is relatively good.