Five-layer-funnel filtering mode discovers effective components of Chinese medicine formulas: Zhishi-Xiebai-Guizhi decoction as a case study.

BACKGROUND: How to screen and identify the effective components in the complex substance system is one of the core issues in achieving the modernization of traditional Chinese medicine (TCM) formulas. However, it is still challenging to systematically screen out the effective components from the hundreds or thousands of components in a TCM formula. PURPOSE: An innovative five-layer-funnel filtering mode stepwise integrating chemical profile, quantitative analysis, xenobiotic profile, network pharmacology and bioactivity evaluation was successfully presented to discover the effective components and implemented on a case study of Zhishi-Xiebai-Guizhi decoction (ZXG), a well-known TCM formula for coronary heart disease (CHD). METHODS: Initially, the chemical profile of ZXG was systemically characterized. Subsequently, the representative constituents were quantitatively analyzed. In the third step, the multi-component xenobiotics profile of ZXG was systemically delineated, and the prototypes absorbed into the blood were identified and designated as the primary bioavailable components. Next, an integrated network of "bioavailable components-CHD targets-pathways-therapeutic effects" was constructed, and the crucial bioavailable components of ZXG against CHD were screened out. Lastly, the bioactivities of crucial bioavailable components were further evaluated to pinpoint effective components. RESULTS: First of all, the chemical profile of ZXG was systemically characterized with the detection of 201 components. Secondly, 37 representative components were quantified to comprehensively describe its content distribution characteristics. Thirdly, among the quantified components, 24 bioavailable components of ZXG were identified based on the multi-component xenobiotic profile. Fourthly, an integrated network led to the identification of 11 crucial bioavailable components against CHD. Ultimately, 9 components (honokiol, magnolol, naringenin, magnoflorine, hesperidin, hesperetin, naringin, neohesperidin and narirutin) exhibiting myocardial protection in vitro were identified as effective components of ZXG for the first time. CONCLUSION: Overall, this innovative strategy successfully identified the effective components of ZXG for the first time. It could not only significantly contribute to elucidating the therapeutic mechanism of ZXG in the treatment of CHD, but also serve as a helpful reference for the systematic discovery of effective components as well as ideal quality markers in the quality assessment of TCM formulas.

Correlation Between Chronotypes and Depressive Symptoms Mediated by Sleep Quality Among Chinese College Students During the COVID-19 Pandemic.

Purpose: The COVID-19 pandemic has adversely impacted the mental health of the population. The current study aimed to determine the prevalence of depressive symptoms and sleep disturbances among Chinese college students during the COVID-19 pandemic and investigate the correlations between chronotypes, sleep quality, and depressive symptoms. Participants and Methods: In the current study, 2526 college students responded anonymously to an online questionnaire survey from 26 May 2020 to 20 July 2020. The participants' chronotypes, sleep quality, and depressive symptoms were evaluated using the Chinese version of the Morning and Evening Questionnaire-5 (MEQ-5), Pittsburgh Sleep Quality Index (PSQI), and Patient Health Questionnaire-9 (PHQ-9). Sociodemographic information of the participants was also acquired. Statistical analyses were performed using Statistical Package for Social Sciences (SPSS) 19.0 software, with the mediating effect assessed by Hayes' PROCESS Macro. Results: During the COVID-19 pandemic, the prevalence of depressive symptoms and sleep disturbances among Chinese college students surveyed was 54.95% and 48.18%, respectively. From absolute evening chronotype to absolute morning chronotype, the surveyed college students' chronotypes were negatively correlated with their depressive symptoms. Moreover, the mediation analysis showed that the correlation between chronotypes and depressive symptoms was fully mediated by sleep quality. Eveningness college students with poorer sleep quality were more likely to report higher levels of depressive symptoms. Conclusion: Our findings suggest that during the COVID-19 pandemic, delayed circadian preference (ie, eveningness) may be linked to worse depressive symptoms among Chinese college students, and call for more attention to the sleep quality of Chinese college students as sleep quality fully mediated the correlation between chronotypes and depressive symptoms among them. Reasonable adjustment in bedtime/circadian preference and improvement in sleep quality may help to reduce the prevalence and severity of depressive symptoms among Chinese college students.

Pharmacokinetics integrated with network pharmacology to clarify effective components and mechanism of Wendan decoction for the intervention of coronary heart disease.

ETHNOPHARMACOLOGICAL RELEVANCE: Coronary heart disease (CHD), one of the leading causes of mortality in the world among chronic non-infectious diseases, is closely associated with atherosclerosis, which ultimately leads to myocardial injury. Wendan decoction (WDD), a classical famous formula, exerted an intervention effect on CHD according to numerous reports. However, the effective components and underlying mechanisms for the treatment of CHD have not been fully elucidated. AIM OF THE STUDY: An in-depth investigation of the effective components and mechanisms of WDD for the intervention of CHD was further explored. MATERIALS AND METHODS: Firstly, based on our previous metabolic profile results, a quantification method for absorbed components was established by ultra-performance liquid chromatography triple quadrupole-mass spectrometry (UPLC-TQ-MS) and applied to the pharmacokinetics study of WDD. Then the network pharmacology analysis for considerable exposure components in rat plasma was employed to screen key components of WDD. Gene ontology and KEGG pathway enrichment analysis were further performed to obtain putative action pathways. The effective components and mechanism of WDD were confirmed by in vitro experiments. RESULTS: A rapid and sensitive quantification method was successfully applied to the pharmacokinetic study of 16 high-exposure components of WDD at three different doses. A total of 235 putative CHD targets were obtained for these 16 components. Then, 44 core targets and 10 key components with high degree values were successively screened out by the investigation of protein-protein interaction and the network of "herbal medicine-key components-core targets". Enrichment analysis suggested that the PI3K-Akt signaling pathway was closely related to this formula's therapeutic mechanism. Furthermore, pharmacological experiments demonstrated that 5 of 10 key components (liquiritigenin, narigenin, hesperetin, 3,5,6,7,8,3',4'-heptamethoxyflavone, and isoliquiritigenin) significantly enhanced DOX-induced H9c2 cell viability. The cardioprotective effects of WDD against DOX-induced cell death through the PI3K-Akt signaling pathway were verified by western blot experiments. CONCLUSION: The integration of pharmacokinetics and network pharmacology approaches successfully clarified 5 effective components and therapeutic mechanism of WDD for the intervention of CHD.

Effects and dose-response relationships of exercise intervention on weight loss in overweight and obese children: a meta-regression and system review.

The aim of this study was to determine the effect of different exercise doses on weight loss in obese/overweight children. PubMed, Embase, SPORTDiscus, and the Cochrane library were searched from inception to November 2020 for randomized controlled trials. Fourty six trials involving 2,599 obese/overweight children were finally included. Different exercise dose interventions had different impacts. Exercise intervention reduce body weight (BW) by 1.46 kg (95% CI, -2.35 to -0.56, p=0.001), body fat percentage (BF%) by 2.24 (95% CI, -2.63 to -1.84, p<0.001) and body mass index (BMI) by 1.09 kg/m2 (95% CI, -1.45 to -0.73, p<0.001). Each MET-h/week was association with 0.147 kg (95% CI, -0.287 to -0.007, p=0.039) decrease in BW, 0.060 (95% CI, -0.118 to -0.002, p=0.042) decrease in BF%, and 0.069 kg/m2 (95% CI, -0.125 to -0.014, p=0.015) decrease in BMI. The findings suggest that there is a positive liner between exercise dose and weight loss, each MET-h/week associated with 0.147 kg, 0.060 and 0.069 kg/m2 decrease in body weight, BF%, BMI, respectively.

The mRNA expression levels of GABAA receptor α1 and α2 subunits in patients with major depressive disorder during onset and remission.

OBJECTIVE: We aimed to investigate the expression levels of GABA and GABAA receptor α1 and α2 subunits in patients with major depressive disorder (MDD) during onset and remission. MATERIALS AND METHODS: 48 patients with MDD during onset and 45 patients with MDD during remission who were treated in our university were selected. Moreover, the control group included 46 healthy volunteers recruited in the community. The depression and anxiety symptoms were assessed by Hamilton Depression (HAMD) Scale and Hamilton Anxiety (HAMA) Scale. ELISA was used to determine the serum GABA levels. The mRNA expression of GABAA receptor α1 and α2 subunits in peripheral blood were detected by RT-PCR. RESULTS: The expression levels of serum GABA and of GABAA receptor α1 and α2 subunits in MDD depression attack group were notably decreased in comparison with those in MDD remission group and control group ((4.10 ± 0.73) v.s. (5.91 ± 1.25) and (5.83 ± 1.17) umol/L, F = 5.61, p < 0.001; (0.53 ± 0.32) v.s. (0.91 ± 0.18) and (0.93 ± 0.21), F = 8.37, p < 0.001; (1.45 ± 0.86) v.s. (2.33 ± 1.49) and (2.28 ± 1.50), F = 8.23, p < 0.001). However, there were no marked difference in the levels of these three indices between the MDD remission group and the control group (p > 0.05). Serum GABA levels were negatively correlated with HAMA total score (r = -0.34, p = 0.02), HAMD total score (r = -0.46, p = 0.01) and depression core symptom score (r = -0.32, p = 0.03). CONCLUSIONS: During the onset of MDD, there may be GABA neuronal dysfunction and abnormal expression of GABAA receptor subunits, and those changes showed a state change, which gradually returned to normal during remission.

High-dose omega-3 polyunsaturated fatty acid supplementation might be more superior than low-dose for major depressive disorder in early therapy period: a network meta-analysis.

BACKGROUND: The application of n-3 Polyunsaturated Fatty Acids (n-3 PUFAs) supplementation for major depressive disorder (MDD) has been widely discussed in recent years, but its efficacy and application are still controversial. This network meta-analysis was conducted to compare the efficacy of different dosages of n-3 PUFAs on MDD patients in the early period of treatment. METHODS: Randomized controlled trials (RCTs) exploring the efficacy of n-3 PUFA supplementation for patients with MDD were retrieved from the databases of Pubmed, Embase and the Cochrane Library. RCTs comparing the efficacy of n-3 PUFA for adult (≥18 years) MDD patients without comorbidity were eligible for our study. The score of depressive symptoms in early therapy period of the treatment (≤9 weeks) was extracted. Standardized mean deviations (SMDs) of all the sores from the eligible RCTs were synthesized in a pairwise meta-analysis in frequentist framework and a random-effects network meta-analysis in Bayesian framework for the overall and subgroups (high- and low-dose) efficacy of n-3 PUFAs. RESULTS: A total of 910 MDD patients in 10 trials with 3 adjuvant therapy strategies (high-dose n-3 PUFAs, low-dose n-3 PUFAs and placebo) were included. Results of pairwise meta-analysis showed that n-3 PUFAs were superior to placebo (SMD: 1.243 ± 0.596; 95% CI: 0.060 ~ 2.414). Results of the network meta-analysis showed that both the high (SMD: 0.908 ± 0.331; 95% CI: 0.262 ~ 1.581) and the low-dose (SMD: 0.601 ± 0.286; 95% CI: 0.034 ~ 1.18) n-3 PUFAs were superior to placebo, and the efficacy of high-dose n-3 PUFAs is superior to that of low-dose. CONCLUSIONS: High-dose n-3 PUFAs supplementation might be more superior than low-dose in the early therapy period for MDD. More head-to-head clinical trials need to be carried out to provide more direct comparison and enhance the evidence of the efficacy of n-3PUFAs for MDD.

Salvianolic acid B abolished chronic mild stress-induced depression through suppressing oxidative stress and neuro-inflammation via regulating NLRP3 inflammasome activation.

This study was framed to investigate the molecular mechanism behind the anti-depressant effect of salvianolic acid B (SB) against unpredictable chronic mild stress (CMS) induced depression rat model. Control rats received only saline without CMS exposure, whereas CMS model rats were induced to several stress (CMS) for 6 weeks. Treatment group rats were induced with CMS for 6 weeks but received either 20 or 40 mg/kg of SB or 20 mg/kg imipramine (CMS+IMP) from the 4th week to 6th week. Treatment with SB or IMP significantly ameliorated body weight, sucrose consumption rate with shorter immobility time than the control group. Also, administration with SB or IMP could reverse the hyperactivity of hypothalamic-pituitary-adrenal axis as well as decreased inflammatory cytokines with improved antioxidant status. Furthermore, the protein expression of NLRP3 (inflammasome) was markedly downregulated upon treatment with SB (both 20 and 40 mg) or IMP and thereby confirming its potent anti-depressant activity. PRACTICAL APPLICATIONS: Salvianolic acid B (SB) is a phenolic acid extracted from Salvia militiorrhiza Bunge, a popular Chinese herb, which has been prescribed for various pathological conditions. SB has been previously reported with anti-depressant activity but, the in-depth mechanism behind the anti-depressant effect of SB against CMS is still elusive. Hence, the current study was plotted to explore the in-depth mechanism behind the anti-depressant effect of SB against CMS model of depression in rats. The outcome of the current study has confirmed the anti-depressant activity by abolishing oxidative stress, and neuroinflammatory response in the hippocampus through inhibiting NLRP3 inflammasome activation. Hence, SB can be prescribed to major depression patients with standard anti-depressant agents to abolish oxidative stress, neuro-inflammatory response, and related neurological changes.

[Analyses of exercise-induced muscle damage-specific microRNA expression and molecular target of sarcolemmal damage in rats].

In the present study, we were to screen the specific microRNA (miRNA) of exercise-induced muscle damage (EIMD) and assess the EIMD-specific miRNAs-regulated target of sarcolemmal damage in rats. Twenty-four male Sprague-Dawley (SD) rats were randomly divided into 3 groups, which included sedentary (C), 24 h post-exercise (E24) and 48 h post-exercise (E48) groups. Rat EIMD model was established by an acute eccentric exercise, i.e., a downhill running treatment at -16º gradient. EIMD characteristics were verified by Evans blue dye staining, differentially expressed miRNAs were detected by microarray assay, EIMD-specific miRNAs expressions were further validated by real-time quantitative RT-PCR (RT-qPCR), and targets of the miRNAs were predicted based on mRNA expressions of associated proteins and related pathway core molecules of sarcolemmal damage. Two EIMD-specific expressed miRNAs, including miR-206-3p and miR-139-3p, were found in the study. There was a significantly negative correlation (P < 0.05) between miR-206-3p expression and dystrophin (r = -0.68), utrophin (r = -0.64), JNK (r = -0.62) or ERK1 (r = -0.68) respectively, but no correlation was found between miR-139-3p and these biomolecules. The results suggest that: i) the expression profile of miRNAs in rat is significantly affected by EIMD, ii) miR-206-3p and miR-139-3p are the EIMD-specific miRNAs, and iii) miR-206-3p may control sarcolemmal damage by regulating dystrophin, utrophin, JNK and ERK1.

Plasma neuropeptide Y levels in Chinese patients with primary insomnia.

PURPOSE: The present study aimed to explore the possible difference in plasma neuropeptide Y (NPY) level between patients with primary insomnia and healthy normal sleepers. METHODS: The sample comprised 42 patients with primary insomnia and 38 age- and sex-matched healthy controls. Clinical measures, including the Pittsburgh Sleep Quality Index (PSQI), State-Trait Anxiety Inventory (STAI), and Beck Depression Inventory (revised edition, BDI-R), were recorded, respectively. Morning fasting plasma NPY levels of all participants were determined by using enzyme-linked immunosorbent assay (ELISA). Student's t-test or chi-square test was used to compare the differences in demographic and clinical factors and scores of psychometric assessments between groups. Bivariate correlation test was used to analyze the relationship between NPY level and factors, such as age, body mass index (BMI), PSQI, STAI, and BDI-R score. Analysis of covariance (ANCOVA) was used to study the difference of plasma NPY level between two groups with adjustment for age, sex, BMI, STAI, and BDI-R total score. RESULTS: We found that morning plasma NPY levels in patients with primary insomnia were significantly lower than those in the normal controls (5.11 ± 2.87 vs. 7.01 ± 3.44 ng/ml, p = 0.009). The difference in plasma NPY level persisted even after adjustment for age, sex, BMI, STAI, and BDI-R total score (p = 0.026). For all subjects, plasma NPY level was found decreasing significantly with age (r = -0.232, p = 0.038). In addition, there was a trend that plasma NPY level was negatively associated with PSQI total score (r = -0.209, p = 0.063). CONCLUSIONS: Our findings suggest that NPY system may involve in the pathophysiological process of primary insomnia. Further studies are warranted to determine the causal relationship between low plasma NPY level and primary insomnia disorder.