Progression prediction in idiopathic inflammatory myopathy-associated interstitial lung disease: a combination of initial high attenuation areas and anti-melanoma differentiation-associated gene 5-positive.

OBJECTIVES: To analyse quantitative lung densitometry and clinical baseline data of individuals with idiopathic inflammatory myopathy (IIM) and identify risk factors capable of predicting the progression of interstitial lung disease (ILD). METHODS: We utilised quantitative lung densitometry and clinical baseline data as explanatory variables. Univariate and multivariate Cox regression analyses were employed to pinpoint effective risk factors for predicting ILD progression in IIM patients. RESULTS: The findings from the Cox univariate regression analysis indicate that elevated carcinoembryonic antigen levels (HR=1.036, 95% CI 1.004-1.069) are connected to an elevated risk of ILD progression in patients with IIM (P=0.027), while PO2 (HR=0.980, 95% CI 0.962-0.997) , forced vital capacity (HR=0.551, 95% CI 0.320-0.946) are protective factors for ILD progression in patients with IIM (p=0.025, p=0.031, respectively), anti-EJ positivity (HR=0.399, 95% CI 0.175-0.912) and anti-Ro52 positivity (HR=0.437, 95% CI 0.199-0.960) are risk factors for ILD progression in patients with IIM (p=0.029, p=0.039, respectively). Furthermore, the results of Cox multivariate regression analysis reveal that high attenuation areas (HAA) (>465.745 cm3) (HR=5.007, 95% CI 1.773-14.144) and anti-melanoma differentiation-associated gene 5 (Anti-MDA5) positivity (HR=0.127, 95% CI 0.041-0.396) are autonomous prognostic risk factors for ILD progression in individuals with IIM (p=0.002, p<0.001, respectively). CONCLUSIONS: Among IIM patients, those who are anti-MDA5-positive, and exhibit HAA (>465.745cm3) are more likely to experience ILD progression.

Pyrazolone compounds could inhibit CES1 and ameliorates fat accumulation during adipocyte differentiation.

Carboxylesterase 1 (CES1), a member of the serine hydrolase superfamily, is involved in a wide range of xenobiotic and endogenous substances metabolic reactions in mammals. The inhibition of CES1 could not only alter the metabolism and disposition of related drugs, but also be benefit for treatment of metabolic disorders, such as obesity and fatty liver disease. In the present study, we aim to develop potential inhibitors of CES1 and reveal the preferred inhibitor structure from a series of synthetic pyrazolones (compounds 1-27). By in vitro high-throughput screening method, we found compounds 25 and 27 had non-competitive inhibition on CES1-mediated N-alkylated d-luciferin methyl ester (NLMe) hydrolysis, while compound 26 competitively inhibited CES1-mediated NLMe hydrolysis. Additionally, Compounds 25, 26 and 27 can inhibit CES1-mediated fluorescent probe hydrolysis in live HepG2 cells with effect. Besides, compounds 25, 26 and 27 could effectively inhibit the accumulation of lipid droplets in mouse adipocytes cells. These data not only provided study basis for the design of newly CES1 inhibitors. The present study not only provided the basis for the development of lead compounds for novel CES1 inhibitors with better performance, but also offered a new direction for the explore of candidate compounds for the treatment of hyperlipidemia and related diseases.

Discovery of natural anthraquinones as potent inhibitors against pancreatic lipase: structure-activity relationships and inhibitory mechanism.

Obesity is acknowledged as a significant risk factor for various metabolic diseases, and the inhibition of human pancreatic lipase (hPL) can impede lipid digestion and absorption, thereby offering potential benefits for obesity treatment. Anthraquinones is a kind of natural and synthetic compounds with wide application. In this study, the inhibitory effects of 31 anthraquinones on hPL were evaluated. The data shows that AQ7, AQ26, and AQ27 demonstrated significant inhibitory activity against hPL, and exhibited selectivity towards other known serine hydrolases. Then the structure-activity relationship between anthraquinones and hPL was further analysed. AQ7 was found to be a mixed inhibition of hPL through inhibition kinetics, while AQ26 and AQ27 were effective non-competitive inhibition of hPL. Molecular docking data revealed that AQ7, AQ26, and AQ27 all could associate with the site of hPL. Developing hPL inhibitors for obesity prevention and treatment could be simplified with this novel and promising lead compound.

Added Value of Frequency of Imaging Markers for Prediction of Outcome After Intracerebral Hemorrhage: A Secondary Analysis of Existing Data.

BACKGROUND: Frequency of imaging markers (FIM) has been identified as an independent predictor of hematoma expansion in patients with intracerebral hemorrhage (ICH), but its impact on clinical outcome of ICH is yet to be determined. The aim of the present study was to investigate this association. METHODS: This study was a secondary analysis of our prior research. The data for this study were derived from six retrospective cohorts of ICH from January 2018 to August 2022. All consecutive study participants were examined within 6 h of stroke onset on neuroimaging. FIM was defined as the ratio of the number of imaging markers on noncontrast head tomography (i.e., hypodensities, blend sign, and island sign) to onset-to-neuroimaging time. The primary poor outcome was defined as a modified Rankin Scale score of 3-6 at 3 months. RESULTS: A total of 1253 patients with ICH were included for final analysis. Among those with available follow-up results, 713 (56.90%) exhibited a poor neurologic outcome at 3 months. In a univariate analysis, FIM was associated with poor prognosis (odds ratio 4.36; 95% confidence interval 3.31-5.74; p < 0.001). After adjustment for age, Glasgow Coma Scale score, systolic blood pressure, hematoma volume, and intraventricular hemorrhage, FIM was still an independent predictor of worse prognosis (odds ratio 3.26; 95% confidence interval 2.37-4.48; p < 0.001). Based on receiver operating characteristic curve analysis, a cutoff value of 0.28 for FIM was associated with 0.69 sensitivity, 0.66 specificity, 0.73 positive predictive value, 0.62 negative predictive value, and 0.71 area under the curve for the diagnosis of poor outcome. CONCLUSIONS: The metric of FIM is associated with 3-month poor outcome after ICH. The novel indicator that helps identify patients who are likely within the 6-h time window at risk for worse outcome would be a valuable addition to the clinical management of ICH.

Insomnia trajectories predict chronic inflammation over 2 years at the transition to adulthood.

Insomnia in adolescents is an important public health concern, as its impacts on both their current and future physical and mental health has been discussed. However, few longitudinal studies have examined insomnia and chronic inflammation at the transition from adolescence to adulthood. This study aimed to examine the predictive effects of insomnia and insomnia trajectories on inflammation in college students by using a prospective design. Using data from the College Student Behaviour and Health Cohort Study, which was conducted between April 2019 and April 2021, with an interval of 6 months. We investigated the associations between insomnia trajectories from Year 1 to Year 3 and five inflammatory biomarkers (C-reactive protein [CRP], tumour necrosis factor [TNF]-α, interleukin [IL]-6, IL-1ß, IL-10) at Year 3. The association of insomnia symptoms at baseline, Wave 1 or Wave 2 with inflammatory biomarkers at Wave 4 were also assessed. A total of 312 college students (males: 51.6%) aged 16-26 years (mean [SD] 18.82 [1.22] years) were analysed. We identified two insomnia trajectory classes: increasing insomnia (n = 63 [20.2%]) and decreasing insomnia (n = 249 [79.8%]). Generalised linear model analysis revealed that insomnia symptoms at Wave 1 were associated with significantly elevated CRP and TNF-α levels at Wave 4. Increasing insomnia trajectories predicted consistently higher levels of CRP, TNF-α and IL-10. However, after adjusting for potential confounders, these associations were significantly attenuated. Overall, the findings suggest that insomnia symptoms affect chronic inflammation at the transition to adulthood. Our study needs to be replicated in larger cohorts to further explore how inflammation interacts with insomnia to increase the susceptibility to adverse health conditions.

Licochalcone A Derivatives as Selective Dipeptidyl Peptidase 4 Inhibitors with Anti-Inflammatory Effects.

A set of 22 analogs of licochalcone A was designed and synthesized to explore their potentials as dipeptidyl peptidase 4 (DPP4) inhibitors with anti-inflammatory effects. The anti-DPP4 effects of these analogs were evaluated using the fluorescent substrate Gly-Pro-N-butyl-4-amino-1,8-naphthalimide (GP-BAN). The nitro-substituted analogue 27 exhibited the most potent activity (Ki = 0.96 µM). A structure-activity relationship investigation revealed that 4-hydroxyl and 5-chloro substituents are essential for DPP4 inhibition, while the 3'-nitro substituent improved both DPP4 inhibition and microsomal stability. Furthermore, compound 27 demonstrated good selectivity for DPP4 over other proteases, including dipeptidyl peptidase 9 (DPP9), thrombin, prolyl endopeptidase (PREP), and fibroblast activation protein (FAP). The cytotoxic effect of 27 was evaluated in cancer cell lines HepG-2 and Caco-2 and in somatic RAW264.7 cells and RPTECs. Compound 27 showed no toxicity to normal cells and weak toxicity to cancer cells. In a living cell imaging assay, 27 blocked the dipeptidase activity of DPP4 in both Caco-2 and HepG-2 cells. This compound also dose-dependently suppressed the expression levels of the chemokines tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1ß (IL-1ß).

Association between healthy sleep patterns and depressive trajectories among college students: a prospective cohort study.

BACKGROUND: The purpose of this study was to identify different develpment trajectories of depression symptoms during college period, and prospectively investigate the associations healthy sleep patterns with trajectories of depression symptoms among college students from freshman through junior year. METHODS: A total of 999 participants from the College Student Behavior and Health Cohort Study were included between April 2019 and June 2021. Healthy sleep patterns were defined by chronotype, sleep duration, insomnia, snoring, and daytime sleepiness. Latent growth curve model was used to identify trajectories of depression symptoms. Then binary logistic regression was used to examine association of the healthy sleep patterns with these trajectories. RESULTS: In baseline survey, we found that a total of 100 (10.0%) participants had healthy sleep patterns' score equal to 5. Then, we used 5 surveys' data to identify 2 distinct trajectories of depression symptoms during college (decreasing: 82.5%; increasing: 17.5%). The healthy sleep patterns were associated with these trajectories, the better healthy sleep patterns significantly decrease the risk of increasing trajectories of depression symptoms in males (OR: 0.72, 95%CI: 0.54 ~ 0.97, P = 0.031). Moreover, we found out that the healthy sleep patterns of college students can predict the future depressive symptoms in this study (all P < 0.001). CONCLUSION: Our findings indicate that the better healthy sleep patterns may significantly decrease the risk of increasing trajectory of depression symptoms only in male college students. The results speak to a need for college student with depression symptoms to identify and address sleep problems when present, which could prevent or reduce depression detriments in later life.

Prospective study of the association between chronotype and cardiometabolic risk among Chinese young adults.

BACKGROUND: The association of evening chronotype with cardiometabolic disease has been well established. However, the extent to which circadian rhythm disturbances independently result in risk remains unclear. This study aimed to investigate the cross-sectional and prospective longitudinal associations between chronotype and cardiometabolic risk among Chinese young adults. METHODS: From April to May 2019, a total of 1 135 young adults were selected to complete the self-administered questionnaire, and 744 fasting blood samples were collected to quantify cardiometabolic parameters. From April to May 2021, 340 fasting blood samples were collected to quantify cardiometabolic parameters. The Morning and Evening Questionnaire 5 (MEQ-5) was used to assess chronotype. The cardiometabolic (CM)-risk score was the sum of standardized Z scores based on gender for the 5 indicators: waist circumference (WC), mean arterial pressure (MAP), triglyceride (TG), homeostasis model assessment for insulin resistance (HOMA-IR), and high-density lipoprotein cholesterol (HDL-C), where the HDL-C is multiplied by-1. The generalized linear model was used to determine the cross-sectional and prospective longitudinal associations between chronotype and each cardiometabolic parameter. RESULTS: Cross-sectional association analysis showed that lower MEQ-5 scores were correlated with higher fasting insulin (ß=-1.420, 95%CI: -2.386~-0.453), higher HOMA-IR (ß=-0.301, 95%CI: -0.507~-0.095), and higher CM risk score (ß=-0.063, 95%CI: -0.122~-0.003), even after adjustment for covariates. Prospective longitudinal association analysis also showed that lower MEQ-5 scores were associated with 2 years later higher fasting glucose (ß=-0.018, 95%CI: -0.034~-0.003), higher fasting insulin (ß=-0.384, 95%CI: -0.766~-0.003), higher HOMA-IR (ß=-0.089, 95%CI: -0.176~-0.002), and higher CM-risk score (ß=-0.109, 95%CI: -0.214~-0.003) after adjustment for covariates. CONCLUSIONS: Evening chronotype was significantly correlated with higher CM risk among young adults. Our findings suggest that biologically and socially affected sleep timing misalignment is a contributing factor to cardiovascular disease risk.

Wogonin alleviates BaP-induced DNA damage and oxidative stress in human airway epithelial cells by dual inhibiting CYP1A1 activity and expression.

Benzo[a]pyrene (BaP) is a common air pollutant that has been reported to cause oxidative stress and carcinogenesis. Wogonin, a flavonoid compound extracted from the roots of Scutellaria baicalensis, has been found to possess a variety of pharmacological activities, including anti-inflammatory and anti-cancer effects. The purpose of this study was to examine the ability of wogonin to alleviate the cytotoxicity induced by BaP in human airway epithelial cells and explore the corresponding mechanism. Our study found that wogonin treatment inhibited DNA damage and reactive oxygen species overproduction induced by BaP in human airway epithelial cells. In vitro enzyme assays showed that wogonin significantly inhibited the enzymatic activity of CYP1A1. In addition, wogonin decreased the basal level of CYP1A1 and inhibited the CYP1A1 overexpression induced by BaP, whereas overexpression of CYP1A1 partially reversed the effect of wogonin on BaP-induced DNA damage. Meanwhile, a CYP1A1 inhibitor and CYP1A1 knockdown also showed these same effects. Further studies showed that wogonin regulates CYP1A1 expression by inhibiting CDK7 and CDK9 activity. The use of CDK7 or CDK9 inhibitors decreased BaP-induced cytotoxicity and CYP1A1 expression. Finally, we found that the methoxy group of wogonin was crucial for its inhibitory activity. In conclusion, our data indicated that wogonin could effectively relieve BaP induced cytotoxicity, and its mechanism was related to the dual inhibition of CYP1A1 activity and expression.

[Association between changes in physical activity and comorbid symptoms of anxiety and depression in college students].

OBJECTIVE: To describe the prevalence of physical activity and comorbid symptoms of anxiety and depression in college students, and to explore the correlation strength between changes in physical activity and comorbid symptoms of anxiety and depression, so as to provide a reference for promoting college students' mental health. METHODS: From April to May 2019, 1179 freshmen majoring in public health, nursing, chemistry and physical education were randomly sampled from one university in Hefei City, Anhui Province, and Shangrao City, Jiangxi Province, respectively. A baseline questionnaire survey was conducted. A follow-up survey was conducted in May 2021, and a total of 1046 subjects were included, including 647 female and 399 male. The International Physical Activity Questionnaire-Short Form was used to evaluate the physical activity level of college students, and the Patient Health Questionnaire and Generalized Anxiety Disorder Scale were used to evaluate the anxiety and depression symptoms of college students during follow-up. Determining the coexistence of anxiety and depression symptoms in college students as anxiety-depression comorbid symptoms. RESULTS: In the follow-up survey, the detection rate of anxiety and depression comorbid symptoms of college students was 16.9%(n=177), and the detection rates of sufficient, decreased, increased, and insufficient physical activity changes were 72.5%(n=758), 13.8%(n=144), 9.2%(n=96), and 4.6%(n=48), respectively. The result of multiple Logistic regression model showed that, after controlling for confounding factors, compared with those with sustained high level of physical activity, i. e. , adequate physical activity, increased physical activity(OR=1.89, 95%CI 1.10-3.25), decreased physical activity(OR =2.80, 95% CI 1.72-4.57), and insufficient physical activity(OR = 3.66, 95% CI 1.85-7.23) increased the risk of anxiety-depression comorbidity symptoms of college students(P<0.05). However, there was no significant increase in the risk of anxiety or depressive symptoms in those who increased, decreased, or insufficient physical activity compared with those who were sufficient physical activity(P>0.05). CONCLUSION: The level of physical activity and its changes are related to mental health of college students. The continuous low level of physical activity is associated with the increased risk of comorbidity of anxiety and depression in college students.

Inhibition of drug-metabolizing enzymes by Jingyin granules: implications of herb-drug interactions in antiviral therapy.

Jingyin granules, a marketed antiviral herbal medicine, have been recommended for treating H1N1 influenza A virus infection and Coronavirus disease 2019 (COVID-19) in China. To fight viral diseases in a more efficient way, Jingyin granules are frequently co-administered in clinical settings with a variety of therapeutic agents, including antiviral drugs, anti-inflammatory drugs, and other Western medicines. However, it is unclear whether Jingyin granules modulate the pharmacokinetics of Western drugs or trigger clinically significant herb-drug interactions. This study aims to assess the inhibitory potency of the herbal extract of Jingyin granules (HEJG) against human drug-metabolizing enzymes and to clarify whether HEJG can modulate the pharmacokinetic profiles of Western drug(s) in vivo. The results clearly demonstrated that HEJG dose-dependently inhibited human CES1A, CES2A, CYPs1A, 2A6, 2C8, 2C9, 2D6, and 2E1; this herbal medicine also time- and NADPH-dependently inhibited human CYP2C19 and CYP3A. In vivo tests showed that HEJG significantly increased the plasma exposure of lopinavir (a CYP3A-substrate drug) by 2.43-fold and strongly prolonged its half-life by 1.91-fold when HEJG (3 g/kg) was co-administered with lopinavir to rats. Further investigation revealed licochalcone A, licochalcone B, licochalcone C and echinatin in Radix Glycyrrhizae, as well as quercetin and kaempferol in Folium Llicis Purpureae, to be time-dependent CYP3A inhibitors. Collectively, our findings reveal that HEJG modulates the pharmacokinetics of CYP substrate-drug(s) by inactivating CYP3A, providing key information for both clinicians and patients to use herb-drug combinations for antiviral therapy in a scientific and reasonable way.

Interaction between physical activity and outdoor time on allostatic load in Chinese college students.

BACKGROUND: Physical activity (PA) deficiency, outdoor time reduction during college have been associated with higher cumulative physiological burden as measured by allostatic load (AL). Therefore, the present research sought to analyze the independent and interaction effects of PA and outdoor time on AL in college students. METHODS: A cross-sectional survey was conducted in two universities from April to May 2019. Self-assessment questionnaire and International Physical Activity Questionnaire Short Version (IPAQ-SF) were used in the investigation, AL level was assessed according to the results of biochemical examination, blood pressure and human body morphological measurements. Binary Logistic Analysis was used to analyze the relationships between PA, outdoor time and AL. RESULTS: The prevalence of low PA, low outdoor time and high AL were 16.3%, 71.1% and 47.6%, respectively. Low PA (OR=1.83, 95%CI: 1.20~2.78) and low outdoor time (OR=1.90, 95%CI: 1.35~2.67) are independently associated with high AL (P<0.05, for each). Interaction analysis indicated that low PA and low outdoor time were interactively associated with high AL (OR=2.93, 95%CI: 1.73~4.94, P<0.05). CONCLUSIONS: There were the significant independent and interaction effects between PA and outdoor time on AL. In the future, college students' physical education can be arranged reasonably to reduce the health risks.

Discovery of triterpenoids as potent dual inhibitors of pancreatic lipase and human carboxylesterase 1.

Pancreatic lipase (PL) is a well-known key target for the prevention and treatment of obesity. Human carboxylesterase 1A (hCES1A) has become an important target for the treatment of hyperlipidaemia. Thus, the discovery of potent dual-target inhibitors based on PL and hCES1A hold great potential for the development of remedies for treating related metabolic diseases. In this study, a series of natural triterpenoids were collected and the inhibitory effects of these triterpenoids on PL and hCES1A were determined using fluorescence-based biochemical assays. It was found that oleanolic acid (OA) and ursolic acid (UA) have the excellent inhibitory effects against PL and hCES1A, and highly selectivity over hCES2A. Subsequently, a number of compounds based on the OA and UA skeletons were synthesised and evaluated. Structure-activity relationship (SAR) analysis of these compounds revealed that the acetyl group at the C-3 site of UA (compound 41) was very essential for both PL and hCES1A inhibition, with IC50 of 0.75 µM and 0.014 µM, respectively. In addition, compound 39 with 2-enol and 3-ketal moiety of OA also has strong inhibitory effects against both PL and hCES1A, with IC50 of 2.13 µM and 0.055 µM, respectively. Furthermore, compound 39 and 41 exhibited good selectivity over other human serine hydrolases including hCES2A, butyrylcholinesterase (BChE) and dipeptidyl peptidase IV (DPP-IV). Inhibitory kinetics and molecular docking studies demonstrated that both compounds 39 and 41 were effective mixed inhibitors of PL, while competitive inhibitors of hCES1A. Further investigations demonstrated that both compounds 39 and 41 could inhibit adipocyte adipogenesis induced by mouse preadipocytes. Collectively, we found two triterpenoid derivatives with strong inhibitory ability on both PL and hCES1A, which can be served as promising lead compounds for the development of more potent dual-target inhibitors targeting on PL and hCES1A.

[Mediating role of IL-10 in the association between health-risk behaviors and depressive symptoms of college students].

OBJECTIVE: To examine the relationship between health-risk behaviors and depressive symptoms among college students, and explore the mediating role of plasma IL-10 level in the relationship between the two. METHODS: Freshman students in two universities in Hefei City, Anhui Province and Shangrao City, Jiangxi Province were recruited between April and May 2019, and follow-up investigation was conducted 6 months later. Health risk behaviors were measured based on the Young Risk Behavior Surveillance System(YRBSS) questionnaire, and depressive symptoms was evaluated by using the Depression Anxiety Stress Scale(DASS-21) among college students at baseline and 6 months follow-up survey. Plasma interleukin-10(IL-10) level was measured at baseline. Univariate analysis was used to compare the correlation between health risk behaviors and depressive symptoms among college students. Binary Logistic regression analyzed the relationship between health risk behaviors, IL-10 and depressive symptoms. The mediation model was used to explore the mediating role of IL-10 levels in the association between health risk behaviors and depressive symptoms. RESULTS: At baseline, boys reported a higher rate of depressive symptoms than that of girls(χ~2=6.33, P=0.01); higher rates of depressive symptoms were observed in students who were from a family with a low perceived economic status(χ~2=7.31, P=0.03)or in poor health(χ~2=6.71, P=0.04). Participants who reported low physical activity(χ~2=19.09, P<0.01), smoking(χ~2=7.03, P<0.01), and poor sleep quality(χ~2=68.78, P<0.01)at baseline were more likely to experience depressive symptoms. Multiple health-risk behaviors at baseline were positively correlated with depressive symptoms among college students. After adjusting gender, self-reported family economy and self-rated health, the regression model showed that plasma IL-10 at baseline was negatively associated with the prevalence of depressive symptoms(OR=0.36, 95% CI 0.18-0.72) and the incidence of depressive symptoms after 6 months(OR=0.20, 95% CI 0.08-0.49). Structural equation model showed that health-risk behaviors was negatively correlated to IL-10 level(ß=-0.13, SE=0.04), IL-10 negatively predicted depressive symptoms at follow-up(ß=-0.09, SE=0.04), and IL-10 play a mediating role between health risk behavior and depressive symptoms. CONCLUSION: Health risk behaviors are positively correlated with depressive symptoms among college students. Plasma IL-10 level at baseline was negatively associated with the incidence of depressive symptoms after 6 months, and IL-10 level at baseline has a partial mediating effect between baseline health risk behavior clustering and depressive symptoms at follow-up.

Discovery of pyrazolones as novel carboxylesterase 2 inhibitors that potently inhibit the adipogenesis in cells.

Carboxylesterase 2 (CES2) is one of the most important Phase I drug metabolizing enzymes in the carboxylesterase family. It plays crucial roles in the bioavailability of oral ester prodrugs and the therapeutic effect of some anticancer drugs such as irinotecan (CPT11) and capecitabine. In addition to the well-known roles of CES2 in xenobiotic metabolism, the enzyme also participates in endogenous metabolism and the production of lipids. In this study, we synthesized a series of pyrazolones and assayed their inhibitory effects against CES2 in vitro. Structure-activity relationship analysis of these pyrazolones reveals that the introduction of 4-methylphenyl unit (R1), 4-methylbenzyl (R2) and cyclohexyl (R3) moieties are beneficial for CES2 inhibition. Guided by these SARs results, 1-cyclohexyl-4-(4-methylbenzyl)-3-p-tolyl-1H- pyrazol-5(4H)-one (27) was designed and synthesized. Further investigations demonstrated that the compound 27 exhibited stronger CES2 inhibition activity with a lower IC50 value (0.13 µM). The inhibition kinetic study demonstrated that compound 27 inhibited the hydrolysis of CES2-fluorescein diacetate (FD) through non-competitive inhibition. In addition, the molecular docking showed that the core of pyrazolone, the cyclohexane moiety, 4-methylbenzyl and 4-methylphenyl groups in compound 27 all played important roles with the amino acid residues of CSE2. Also, compound 27 could inhibit adipocyte adipogenesis induced by mouse preadipocytes. In brief, we designed and synthesized a novel pyrazolone compound with a strong inhibitory ability on CES2 and could inhibit the adipogenesis induced by mouse preadipocytes, which can be served as a promising lead compound for the development of more potent pyrazolone-type CES2 inhibitors, and also used as a potential tool for exploring the biological functions of CES2 in human being.

Differences in susceptibility of HT-29 and A549 cells to statin-induced toxicity: An investigation using high content screening.

Statins are a group of hydroxymethylglutaryl coenzyme A reductase inhibitors that are used in the treatment of cardiovascular diseases. However, statins have been found to be cytotoxic, and many unexpected side effects have been reported in clinical applications. The susceptibilities of different cell lines toward statins are diverse, and the mechanisms of cytotoxicity remain unknown. Therefore, the present study aimed to investigate differences in the susceptibility to and mechanisms of statin-induced cytotoxicity in two cell lines, HT-29 and A549, using a high content screening-based multiparametric toxicity assay panel. We found that the two cell types exhibited differing susceptibilities to the cytotoxic effects of the different statins. Additionally, the cytotoxicity was inconsistent between different statins in the two cell lines. Four statins with strong cytotoxicity decreased the viability of HT-29 cells via the mitochondrial pathway, as evidenced by decreased mitochondrial membrane potential, and elevated mitochondrial mass, calcium release and cell apoptosis, and reactive oxygen species. In contrast, these four statins only induced a decrease in the mitochondrial membrane potential in A549 cells. The above results provide an objective reason for future evaluations of cytotoxic differences in cell types and the underlying mechanisms of cytotoxicity in different statins, and provide a good scientific basis for further research on countermeasures against statin-induced cell injuries.

Inhibition of catechol-O-methyltransferase by natural pentacyclic triterpenes: structure-activity relationships and kinetic mechanism.

Inhibitors of COMT are clinically used for the treatment of Parkinson's disease. Here, we report the first natural pentacyclic triterpenoid-type COMT inhibitors and their structure-activity relationships and inhibition mechanism. The most potent compounds were found to be oleanic acid, betulinic acid and celastrol with IC50 values of 3.89-5.07 µM, that acted as mixed (uncompetitive plus non-competitive) inhibitors of COMT, representing a new skeleton of COMT inhibitor. Molecular docking suggested that they can specifically recognise and bind with the unique hydrophobic residues surrounding the catechol pocket. Furthermore, oleanic acid and betulinic acid proved to be less disruptive of mitochondrial membrane potential (MMP) compared to tolcapone, thus reducing the risk of liver toxicity. These findings could be used to produce an ideal lead compound and to guide synthetic efforts in generating related derivatives for further preclinical testing.

Theophylline Acetaldehyde as the Initial Product in Doxophylline Metabolism in Human Liver.

Doxophylline (DOXO) and theophylline are widely used as bronchodilators for treating asthma and chronic obstructive pulmonary disease, and DOXO has a better safety profile than theophylline. How DOXO's metabolism and disposition affect its antiasthmatic efficacy and safety remains to be explored. In this study, the metabolites of DOXO were characterized. A total of nine metabolites of DOXO were identified in vitro using liver microsomes from human and four other animal species. Among them, six metabolites were reported for the first time. The top three metabolites were theophylline acetaldehyde (M1), theophylline-7-acetic acid (M2), and etophylline (M4). A comparative analysis of DOXO metabolism in human using liver microsomes, S9 fraction, and plasma samples demonstrated the following: 1) The metabolism of DOXO began with a cytochrome P450 (P450)-mediated, rate-limiting step at the C ring and produced M1, the most abundant metabolite in human liver microsomes. However, in human plasma, the M1 production was rather low. 2) M1 was further converted to M2 and M4, the end products of DOXO metabolism in vivo, by non-P450 dismutase in the cytosol. This dismutation process also relied on the ratio of NADP+/NADPH in the cell. These findings for the first time elucidated the metabolic sites and routes of DOXO metabolism in human. SIGNIFICANCE STATEMENT: We systematically characterized doxophylline metabolism using in vitro and in vivo assays. Our findings evolved the understandings of metabolic sites and pathways for methylxanthine derivatives with the aldehyde functional group.

Discovery of natural alkaloids as potent and selective inhibitors against human carboxylesterase 2.

Human Carboxylesterase 2A (hCES2A), one of the most important serine hydrolases, plays crucial roles in the hydrolysis and the metabolic activation of a wide range of esters and amides. Increasing evidence has indicated that potent inhibition on intestinal hCES2A may reduce the excessive accumulation of SN-38 (the hydrolytic metabolite of irinotecan with potent cytotoxicity) in the intestinal tract and thereby alleviate the intestinal toxicity triggered by irinotecan. In this study, more than sixty natural alkaloids have been collected and their inhibitory effects against hCES2A are assayed using a fluorescence-based biochemical assay. Following preliminary screening, seventeen alkaloids are found with strong to moderate hCES2A inhibition activity. Primary structure-activity relationships (SAR) analysis of natural isoquinoline alkaloids reveal that the benzo-1,3-dioxole group and the aromatic pyridine structure are beneficial for hCES2A inhibition. Further investigations demonstrate that a steroidal alkaloid reserpine exhibits strong hCES2A inhibition activity (IC50 = 0.94 µM) and high selectivity over other human serine hydrolases including hCES1A, dipeptidyl peptidase IV (DPP-IV), butyrylcholinesterase (BChE) and thrombin. Inhibition kinetic analyses demonstrated that reserpine acts as a non-competitive inhibitor against hCES2A-mediated FD hydrolysis. Molecular docking simulations demonstrated that the potent inhibition of hCES2A by reserpine could partially be attributed to its strong σ-π and S-π interactions between reserpine and hCES2A. Collectively, our findings suggest that reserpine is a potent and highly selective inhibitor of hCES2A, which can be served as a promising lead compound for the development of more efficacious and selective alkaloids-type hCES2A inhibitors for biomedical applications.

Interaction between physical activity and problematic mobile phone use on suicidality in Chinese college students.

BACKGROUND: Previous research has found a relationship between problematic mobile phone use (PMPU) and suicidality. However, few studies have examined the interaction effects between low physical activity (PA) and PMPU on suicidality among college students. This study aimed to examine the interactions of PA and PMPU and their impact on suicidality in a school-based sample among Chinese college students. METHODS: Analysis is based on date from two university in China, which recruited 4787 participants. Binomial logistic regression models were used to explore the associations of PA, PMPU with suicidal ideation and suicide attempt, as well as the interaction of PA and PMPU with suicidality. RESULTS: The prevalence of suicide attempt and suicidal ideation were 3.5 and 7.2%, respectively. Low PA was significantly associated with suicide attempt (OR = 3.48, 95%CI: 2.52-4.81) and suicidal ideation (OR = 1.90, 95%CI: 1.46-2.46). PMPU was significantly associated with suicide attempt (OR = 3.65, 95%CI: 2.66-5.01) and suicidal ideation (OR = 2.83, 95%CI: 2.25-3.54). Interaction analysis indicated that low PA and PMPU were interactively associated with suicide attempt (OR = 9.51, 95%CI: 6.15-14.73, P < 0.001), RERI = 4.85(1.20-8.50), AP = 0.51(0.29-0.73), SI = 2.32(1.34-4.04). There was no additive interaction effects between PA and PMPU on suicidal ideation. CONCLUSIONS: The findings reveals that the intervention programs of suicide attempt should consider the students PA levels and PMPU.