Mechanism and optimization of supramolecular complexation-enhanced fluorescence spectroscopy for the determination of SN-38 in plasma and cells.

Quantitative detection of two different forms of SN-38 in biological samples is, currently, cumbersome and difficult. A revisit to the mechanism of supramolecular complexation-enhanced fluorescence spectroscopy helps to optimize the determination of SN-38 in plasma and the cellular pharmacokinetics in A549 cells based on the supramolecular complexation. Firstly, the inclusion mechanism dominated by thermodynamic constants was determined by measuring kinetic/thermodynamic parameters (kon , koff , ΔG, ΔH, ΔS). On this basis, the best effect of fluorescence sensitization was optimized through screening the interaction conditions (cyclodextrin species and concentrations, drug levels, temperature, pH of the buffer, and reaction time). Furthermore, the proportional relationship between the concentration of the inclusion complex and the fluorescence intensity was confirmed. Finally, a highly sensitive, selective spectrofluorimetric method was established and validated for quantitative analysis of the lactone and carboxylate molecular states of SN-38 plasma levels in rats and cell membrane transfer kinetics in A549 cell lines. The limits of detection for the lactone and carboxylate forms in plasma were found to be 0.44 ng·ml-1 and 0.28 ng·ml-1 , respectively. Precision and accuracy met the requirements of biological samples analysis. The proposed detection method provided a reference for elucidating the biodistribution of SN-38.

Periconceptional use of cod liver oil, a vitamin D source, could decrease the risk of CHD in offspring.

OBJECTIVE: To explore if there is association between vitamin D supplementation through cod liver oil ingestion around the periconceptional period and the risk of developing severe CHD in offspring. Furthermore, we would examine the interaction between vitamin D and folic acid supplementation in the association. METHODS: A case-control study was conducted in Shanghai Children's Medical Center, in which, a total of 262 severe CHD cases versus 262 controls were recruited through June 2016 to December 2017. All children were younger than 2 years. To reduce potential selection bias and to minimise confounding effects, propensity score matching was applied. RESULTS: After propensity score matching, vitamin D supplementation seemed to be associated with decreased odds ratio of severe CHD (odds ratio = 0.666; 95% confidence intervals: 0.449-0.990) in the multivariable conditional logistic analysis. Furthermore, we found an additive interaction between vitamin D and folic acid supplementation (relative excess risk due to interaction = 0.810, 95% confidence intervals: 0.386-1.235) in the association. CONCLUSION: The results suggested that maternal vitamin D supplementation could decrease the risk of offspring severe CHD; moreover, it could strengthen the protective effect of folic acid. The significance of this study lies in providing epidemiological evidence that vitamin D supplementation around the periconceptional period could be a potential nutritional intervention strategy to meet the challenge of increasing CHD.

Associations between maternal social support and stressful life event with ventricular septal defect in offspring: a case-control study.

BACKGROUND: Previous studies suggested that maternal subjective feeling of stress seemed to be involved in the incidence of congenial heart disease in offspring. To better understand the findings, our study would discuss the relationships of maternal exposure to stressful life event and social support, which are more objective and comprehensive indicators of stress, around periconceptional period with the risk of ventricular septal defect (VSD), the most popular subtype of congenital heart disease. METHODS: A hospital-based case-control study was conducted through June, 2016 to December, 2017. We collected maternal self-reports of 8 social support questions in 3 aspects and 8 stressful life events among mothers of 202 VSD cases and 262 controls. Social support was categorized into low, medium high, and high (higher is better), and stressful life event was indexed into low, medium low, and high (higher is worse). Logistic regression models were applied to estimate adjusted odds ratios and 95% confidence intervals (95% CI). RESULTS: The adjusted odds ratio of high stressful life event was 2.342 (95% CI: 1.348, 4.819) compared with low stressful life event. After crossover analysis, compared with low event & high support, the adjusted odds ratio of low event & low support, high event & high support, and high event & low support were 2.059 (95% CI: 1.104, 3.841), 2.699 (95% CI: 1.042, 6.988) and 2.781 (95% CI: 1.033, 7.489), respectively. CONCLUSIONS: In summary, we observed an increased risk of VSD when pregnant women exposed to stressful life events, however, social support could, to some extent, reduce the risk of stressful life event.

Sleep Habits, Sleep Problems, Sleep Hygiene, and Their Associations With Mental Health Problems Among Adolescents.

BACKGROUND: Studies in adults suggested that sleep could be a significant contributor to mental health. However, little is known about their relationship in adolescents. OBJECTIVE: The present study aimed to examine the overall associations of full-spectrum sleep behaviors, including sleep habits, sleep problems, and sleep hygiene, with mental health problems among adolescents in Shanghai, China. DESIGN: A stratified, cluster random sample of 4,823 adolescents aged 11 to 20 years participated in a cross-sectional survey. The Adolescent Sleep Disturbance Questionnaire and the modified Adolescent Sleep Hygiene Scale were used to examine sleep behaviors. The Strengths and Difficulties Questionnaire was used to evaluate mental health problems. RESULTS: Five sleep variables were found to be associated with adolescents' mental health. The five factors covered three sleep domains: sleep habits (later bedtime during weekdays), sleep problems (maintaining sleep difficulties, disorders of arousal), and sleep hygiene (poor emotion at bedtime, unstable sleep schedule). CONCLUSIONS: The clinical significance of the findings lies in the emphasis of comprehensive screening of sleep in the predicting, diagnosis, nursing, and intervention of adolescents' mental health problems.

TCS2 Increases Olaquindox-Induced Apoptosis by Upregulation of ROS Production and Downregulation of Autophagy in HEK293 Cells.

Olaquindox, a feed additive, has drawn public attention due to its potential mutagenicity, genotoxicity, hepatoxicity and nephrotoxicity. The purpose of this study was to investigate the role of tuberous sclerosis complex (TSC2) pathways in olaquindox-induced autophagy in human embryonic kidney 293 (HEK293) cells. The results revealed that olaquindox treatment reduced the cell viability of HEK293 cells and downregulated the expression of TSC2 in a dose- and time-dependent manner. Meanwhile, olaquindox treatment markedly induced the production of reactive oxygen species (ROS), cascaded to autophagy, oxidative stress, and apoptotic cell death, which was effectively eliminated by the antioxidant N-acetylcysteine (NAC). Furthermore, overexpression of TSC2 attenuated olaquindox-induced autophagy in contrast to inducing the production of ROS, oxidative stress and apoptosis. Consistently, knockdown of TSC2 upregulated autophagy, and decreased olaquindox-induced cell apoptosis. In conclusion, our findings indicate that TSC2 partly participates in olaquindox-induced autophagy, oxidative stress and apoptosis, and demonstrate that TSC2 has a negative regulation role in olaquindox-induced autophagy in HEK293 cells.

Quinocetone triggered ER stress-induced autophagy via ATF6/DAPK1-modulated mAtg9a trafficking.

The present study is undertaken to explore quinocetone-induced autophagy and its possible mechanism. Western blotting and green fluorescence protein (GFP)-LC3 vector transfection were performed to determine the ratio of LC3 conversion and its subcellular localization. Results revealed that the quinocetone induced autophagy in time- and dose-dependent manners. Besides, we tested the expressions of immunoglobulin heavy chain binding protein (BiP) and C/EBP homologous protein (CHOP) and the transcription of BiP, HerpUD, and sec24D by western blotting and RT-PCR, respectively. Results showed that quinocetone also induced endoplasmic reticulum (ER) stress during quinocetone-induced autophagy. Furthermore, we observed the cleavage of ATF6, the phosphorylation of MRLC, and the expression of death-associated protein kinase (DAPK1) by western blotting; the transcription of DAPK1 by RT-PCR; and the subcellular localization of ATF6 and mAtg9 by immunofluorescence. These results suggest that quinocetone stimulates the MRLC-mediated mAtg9 trafficking, which is critical for autophagosome formation, via the ATF6 upregulated expression of DAPK1. Last, we generated ATF6 and DAPK1 stable knockdown HepG2 cell lines and found that the conversion ratios of LC3 were decreased upon the treatment of quinocetone. Together, we propose that quinocetone induces autophagy through ER stress signaling pathway-induced cytoskeleton activation.

ML-7 amplifies the quinocetone-induced cell death through akt and MAPK-mediated apoptosis on HepG2 cell line.

The study aims at evaluating the combination of the quinocetone and the ML-7 in preclinical hepatocellular carcinoma models. To this end, the effect of quinocetone and ML-7 on apoptosis induction and signaling pathways was analyzed on HepG2 cell lines. Here, we report that ML-7, in a nontoxic concentration, sensitized the HepG2 cells to quinocetone-induced cytotoxicity. Also, ML-7 profoundly enhances quinocetone-induced apoptosis in HepG2 cell line. Mechanistic investigations revealed that ML-7 and quinocetone act in concert to trigger the cleavage of caspase-8 as well as Bax/Bcl-2 ratio up-regulation and subsequent cleavage of Bid, capsases-9 and -3. Importantly, ML-7 weakened the quinocetone-induced Akt pathway activation, but strengthened the phosphorylation of p-38, ERK and JNK. Further treatment of Akt activator and p-38 inhibitor almost completely abolished the ML-7/quinocetone-induced apoptosis. In contrast, the ERK and JNK inhibitor aggravated the ML-7/quinocetone-induced apoptosis, indicating that the synergism critically depended on p-38 phosphorylation and HepG2 cells provoke Akt, ERK and JNK signaling pathways to against apoptosis. In conclusion, the rational combination of quinocetone and ML-7 presents a promising approach to trigger apoptosis in hepatocellular carcinoma, which warrants further investigation.

Curcumin attenuates quinocetone-induced oxidative stress and genotoxicity in human hepatocyte L02 cells.

Quinocetone (QCT), a new quinoxaline 1,4-dioxides, has been used as antimicrobial feed additive in China. Potential genotoxicity of QCT was concerned as a public health problem. This study aimed to investigate the protective effect of curcumin on QCT-induced oxidative stress and genotoxicity in human hepatocyte L02 cells. Cell viability and intracellular reactive oxygen species (ROS), biomarkers of oxidative stress including superoxide dismutase (SOD) activity and glutathione (GSH) level were measured. Meanwhile, comet assay and micronucleus assay were carried out to evaluate genotoxicity. The results showed that, compared to the control group, QCT at the concentration ranges of 2-16 µg/mL significantly decreased L02 cell viability, which was significantly attenuated with curcumin pretreatment (2.5 and 5 µM). In addition, QCT significantly increased cell oxidative stress, characterized by increases of intracellular ROS level, while decreased endogenous antioxidant biomarkers GSH level and SOD activity (all p < 0.05 or 0.01). Curcumin pretreatment significantly attenuated ROS formation, inhibited the decreases of SOD activity and GSH level. Furthermore, curcumin significantly reduced QCT-induced DNA fragments and micronuclei formation. These data suggest that curcumin could attenuate QCT-induced cytotoxicity and genotoxicity in L02 cells, which may be attributed to ROS scavenging and anti-oxidative ability of curcumin. Importantly, consumption of curcumin may be a plausible way to prevent quinoxaline 1,4-dioxides-mediated oxidative stress and genotoxicity in human or animals.

Melatonin inhibits embryonic rat H9c2 cells growth through induction of apoptosis and cell cycle arrest via PI3K-AKT signaling pathway.

BACKGROUND: Our recent epidemiological study revealed that maternal sleep during the periconceptional period should be involved in the risk of congenital heart disease (CHD) in offspring. Melatonin, a sleep related hormone, has been suggested to play a crucial role in embryonic development based on the emerging evidence. In this study, we set out to assess the effect of melatonin on the embryonic cardiac cell growth and to explore the underlying mechanisms. METHODS: We observed the effect of different gradient doses of melatonin as 10, 100, or 1,000 µM on cell proliferation in H9c2 embryonic rat cardiac cells. Furthermore, flow cytometry was applied to evaluate the impact on apoptosis and cell cycle. RNA-seq was conducted to screen the changes in expression of mRNA and signaling pathways. Quantitative Real-Time-PCR (qRT-PCR) was then conducted to validate the results. RESULTS: It was observed that melatonin could inhibit H9c2 cell growth, at the doses of 100 and 1,000 µM, but not at 10 µM. Moreover, melatonin ranged from 100 to 1,000 µM could instigate cell cycle arrest at G1 phase and simulate apoptosis, in a dose-dependent manner. In addition, melatonin was found to down-regulate the expression of a number of genes, which are related to heart development (SPARC, IFITM3, TNNT2, LOX), and PI3K-Akt signaling pathway activation (FN1, HSP90B1, THBS1, MFGE8, and CLU). CONCLUSIONS: Our findings suggested that high level of melatonin could be capable of inhibiting growth through the induction of apoptosis and cell cycle arrest via PI3K-AKT signaling pathway, thereby interfering with embryonic heart development. Considering this study is based on H9c2 embryonic rat cardiac cells, future additional studies using human embryonic cardiac cell are warranted.

Associations of sleep characteristics with atopic disease: a cross-sectional study among Chinese adolescents.

BACKGROUND: Adolescence, as a transition between childhood and adulthood, is a critical stage for the long-term control of atopic diseases. We aim to determine if sleep characteristics are involved in the increased risk of atopic disease among adolescents. METHODS: Adopting the stratified cluster random sampling method, this cross-sectional survey included 4932 participants aged 12-18 years. The Chinese version of adolescent sleep disturbance questionnaire and the adolescent sleep hygiene scale were used to collect information on sleep problems and sleep hygiene, respectively. Logistic regression models were implemented to examine the associations of sleep with atopic diseases. RESULTS: Sleep duration was not found to be related with allergic diseases. By contrast, sleep-disordered breathing was associated with an increased risk of asthma (adjusted OR = 1.79, 95% CI 1.25-2.55), allergic rhinitis (adjusted OR = 1.95, 95% CI 1.52-2.49), and eczema (adjusted OR = 1.63, 95% CI 1.23-2.16); poor sleep physiology was correspondent to increased odds of asthma (adjusted OR = 1.69, 95% CI 1.24-2.29), allergic rhinitis (adjusted OR = 1.40, 95% CI 1.13-1.73) and eczema (adjusted OR = 1.66, 95% CI 1.32-2.09); non-optimal sleep environment was associated with an increased prevalence of asthma (adjusted OR = 1.52, 95% CI 1.08-2.12), allergic rhinitis (adjusted OR = 1.32, 95% CI 1.04-1.69) and eczema (adjusted OR = 1.53, 95% CI 1.19-1.96). CONCLUSIONS: As sleep-disordered breathing, poor sleep physiology and non-optimal sleep environment were associated with a higher risk of allergic diseases, the results of this study provide a new concept for the adjuvant treatment of allergic diseases in adolescents. Management strategies of allergic diseases should take regular screening and targeted treatment of sleep issues into account.

Glycoside scutellarin enhanced CD-MOF anchoring for laryngeal delivery.

It is essential to develop new carriers for laryngeal drug delivery in light of the lack of therapy in laryngeal related diseases. When the inhalable micron-sized crystals of γ-cyclodextrin metal-organic framework (CD-MOF) was utilized as dry powder inhalers (DPIs) carrier with high fine particle fraction (FPF), it was found in this research that the encapsulation of a glycoside compound, namely, scutellarin (SCU) in CD-MOF could significantly enhance its laryngeal deposition. Firstly, SCU loading into CD-MOF was optimized by incubation. Then, a series of characterizations were carried out to elucidate the mechanisms of drug loading. Finally, the laryngeal deposition rate of CD-MOF was 57.72 ± 2.19% improved by SCU, about two times higher than that of CD-MOF, when it was determined by Next Generation Impactor (NGI) at 65 L/min. As a proof of concept, pharyngolaryngitis therapeutic agent dexamethasone (DEX) had improved laryngeal deposition after being co-encapsulated with SCU in CD-MOF. The molecular simulation demonstrated the configuration of SCU in CD-MOF and its contribution to the free energy of the SCU@CD-MOF, which defined the enhanced laryngeal anchoring. In conclusion, the glycosides-like SCU could effectively enhance the anchoring of CD-MOF particles to the larynx to facilitate the treatment of laryngeal diseases.

Exploring associations of maternal sleep during periconceptional period with congenital heart disease in offspring.

BACKGROUND: In general, the existing evidence points to a role for maternal sleep in pregnancy complications and fetal growth, however, little has been focused on birth defects. We aimed to explore the association between periconceptional poor sleep and the risk of congenital heart disease (CHD), and to examine if daytime napping could to some extent change the association. METHODS: A case-control study was conducted in Shanghai Children's Medical Center, in which, a total of 524 cases (262 simple CHD vs. 262 severe CHD), along with 262 controls. RESULTS: In the multivariable logistic analysis, poor sleep could increase the risk of both simple CHD (OR = 2.486, 95% CI = 1.619-3.818) and severe CHD (OR = 1.950, 95% CI = 1.269-2.997), while routine daytime nap could decrease risk of simple CHD (OR = 0.634, 95% CI = 0.435-0.923). In the stratified analysis, the concurrence with routine daytime nap could weaken the risk of simple CHD caused by poor sleep (OR = 3.183, 95% CI: 1.830-5.537 decreased to OR = 2.236, 95% CI: 1.200-4.165). The examinations were repeated in ventricular septal defect and tetralogy of Fallot, and the established associations can be verified. Moreover, all these findings were also similarly observed in both propensity-score-adjusted and propensity-score-matched analyses. CONCLUSIONS: Poor maternal sleep around periconceptional period seems to be an independent risk factor for CHD. The concurrence with daytime nap could to some extent reduce the risk in simple CHD. The results individually and collectively put forward the importance of maternal sleep in embryonic heart development.

The relationship between insomnia symptoms and school performance among 4966 adolescents in Shanghai, China.

PURPOSE: Insomnia symptoms are common in adolescents. This study examined the associations of insomnia symptoms with school performance among adolescents in Shanghai, China. METHODS: A total of 4966 adolescents aged 11-20 years participated in a cross-sectional survey during November of 2009. The Adolescent Sleep Disturbance Questionnaire was used to examine insomnia symptoms, and the Teacher School Achievement Form was applied to evaluate adolescents' school performance. RESULTS: The results demonstrated that, after adjusting for possible confounders, all 3 dimensions of insomnia symptoms were associated with adolescents' school performance. Difficulty falling asleep problem was associated with poor attention and concentration (odds ratio [OR] = 1.30, 95% confidence interval [CI] 1.11-1.52, P = .001), academic frustration (OR = 1.24, 95% CI: 1.06-1.45, P = .007), and poor school relationships (OR = 1.26, 95% CI: 1.08-1.48, P = .003); maintaining sleep problem was associated with poor attention and concentration (OR = 1.25, 95% CI: 1.06-1.46, P = .007) and poor interest and motivation (OR = 1.22, 95% CI: 1.04-1.44, P = .017); and reinitiating sleep problem was related to poor interest and motivation (OR = 1.20, 95% CI: 1.00-1.45, P = .048). Coexisting insomnia symptoms were associated with poorer school performance, especially in maintaining attention and concentration. CONCLUSIONS: The findings of this study highlight the importance of sleep in the social and cognitive development of adolescents.

Association between Sleep-Disordered Breathing during Pregnancy and Maternal and Fetal Outcomes: An Updated Systematic Review and Meta-Analysis.

BACKGROUND: Due to the high prevalence in pregnant women and potential association with pregnancy complications or perinatal outcomes, sleep-disordered breathing (SDB) has become an increasing concern. METHODS: Pubmed and Embase were retrieved from inception until 2017 to conduct a meta-analysis to explore the association of SDB and several outcomes during gestation. A stratified analysis differentiated by the type of SDB [snoring alone/obstructive sleep apnea (OSA)] was also performed. Pooled odds ratios were produced for binary outcomes. Weighted mean differences were also produced for continuous outcomes. Sensitivity analysis was performed to identify the impact of individual studies on summary results and estimation of publication bias was performed by funnel plot. RESULTS: 35 studies with a total of 56,751,837 subjects were included. SDB during pregnancy was associated with a significantly increased risk of gestational diabetes mellitus (GDM), pregnancy-induced hypertension (PIH), and preeclampsia (PEC), but not significantly associated with fetal maternal outcomes, namely APGAR score and birth weight. Moreover, OSA was linked with an increasing risk of GDM, PIH, PEC and preterm birth while snoring appeared to increase the risk of GDM, PIH, and PEC. CONCLUSION: The finding provided potential evidence for association between SDB and adverse perinatal outcomes. SDB increased the risk of some pregnancy complications while its influence to fetal outcomes was not clear.

Effect of GADD45a on olaquindox-induced apoptosis in human hepatoma G2 cells: Involvement of mitochondrial dysfunction.

Olaquindox, a quinoxaline 1, 4-dioxide derivative, has been widely used as a feed additive for promoting animal growth in China. The aim of present study was to investigate the effect of grow arrest and DNA damage 45 alpha (GADD45a) on olaquindox-induced apoptosis in HepG2 cells. The result showed that olaquindox induced the decrease of cell viability in a dose dependent manner. Compared to the control group, olaquindox treatment at 400 and 800µg/mL increased the expression level of GADD45a protein and reactive oxygen species (ROS) production, decreased mitochondrial membrane potential (MMP), and subsequently increased the expression of Bax while decreased the expression of Bcl-2, leading to the release of cytochrome c (Cyt c). However, knockdown of GADD45a enhanced olaquindox-induced ROS production, disrupted MMP and subsequently caused Cyt c release, then further increased olaquindox- induced cell apoptosis by increasing the activities of caspase-9, caspase-3, and poly (ADP-ribose) polymerase (PARP). In conclusion, the results revealed that GADD45a played a critical role in olaquindox-induced apoptosis in HepG2 cells, which may embrace the regulatory ability on the mitochondrial apoptosis pathway.