Identification of the Metabolites of Both Formononetin in Rat Hepatic S9 and Ononin in Rat Urine Samples and Preliminary Network Pharmacology Evaluation of Their Main Metabolites.

Astragalus membranaceus is a traditional Chinese medicine derived from the roots of Astragalus membranaceus (Fisch.) Bge., which has the same medicinal and edible uses in China. It is also widely used in daily food, and its pharmacological effects mainly include antioxidant effects, vascular softening effects, etc. Currently, it is increasingly widely used in the prevention of hypertension, cerebral ischemia, and stroke in China. Formononetin and its glucopyranoside (ononin) are both important components of Astragalus membranaceuss and may play important roles in the treatment of cardiovascular diseases (CVDs). This study conducted metabolic studies using formononectin and its glucopyranoside (ononin), including a combination of the in vitro metabolism of Formonetin using rat liver S9 and the in vivo metabolism of ononin administered orally to rats. Five metabolites (Sm2, 7, 9, 10, and 12) were obtained from the solution incubated with formononetin and rat hepatic S9 fraction using chromatographic methods. The structures of the five metabolites were elucidated as (Sm2)6,7,4'-trihydroxy-isoflavonoid; (Sm7)7,4'-dihydroxy-isoflavonoid; (Sm9)7,8,4'-trihydroxy-isoflavonoid; (Sm10)7,8,-dihydroxy-4'-methoxy-isoflavonoid; and (Sm12)6,7-dihydroxy-4'-methoxy- isoflavonoid on the basis of UV, NMR, and MS data. Totally, 14 metabolites were identified via HPLC-DAD-ESI-IT-TOF-MSn analysis, from which the formononetin was incubated with rat hepatic S9 fraction, and the main metabolic pathways were hydroxylation, demethylation, and glycosylation. Then, 21 metabolites were identified via HPLC-DAD-ESI-IT-TOF-MSn analysis from the urine samples from SD rats to which ononin was orally administered, and the main metabolic pathways were glucuronidation, hydroxylation, demethylation, and sulfonation. The main difference between the in vitro metabolism of formononetin and the in vivo metabolism of ononin is that ononin undergoes deglycemic transformation into Formonetin in the rat intestine, while Formonetin is absorbed into the bloodstream for metabolism, and the metabolic products also produce combined metabolites during in vivo metabolism. The six metabolites obtained from the aforementioned separation indicate the primary forms of formononetin metabolism, and due to their higher contents of similar isoflavone metabolites, they are considered the main active compounds that are responsible for pharmacological effects. To investigate the metabolites of the active ingredients of formononetin in the rat liver S9 system, network pharmacology was used to evaluate the cardiovascular disease (CVD) activities of the six primary metabolites that were structurally identified. Additionally, the macromolecular docking results of six main components and two core targets (HSP90AA1 and SRC) related to CVD showed that formononetin and its main metabolites, Sm10 and Sm12, may have roles in CVD treatment due to their strong binding activities with the HSP90AA1 receptor, while the Sm7 metabolite may have a role in CVD treatment due to its strong binding activity with the SRC receptor.

Fibroblast growth factor 21 associating with serotonin and dopamine in the cerebrospinal fluid predicts impulsivity in healthy subjects.

BACKGROUND: Impulsivity is more commonly reported in subjects with mental disorders compared to healthy subjects, suggesting a potential application of impulsivity in predicting impulsivity-related mental disorders. However, no biomarker of impulsivity available so far. This study explored the association between cerebrospinal fluid (CSF) fibroblast growth factor 21 (FGF21), a key hormonal mediator of the stress response, and impulsivity in healthy subjects. METHODS: A total of 126 healthy persons subjected to surgery of anterior cruciate ligament were recruited in the present study. The impulsiveness of the subjects was evaluated by the Chinese version of the Barratt Impulsiveness Scale (BIS)-11 before surgery. CSF and blood samples of the subjects were collected before spinal anesthesia for surgery. The levels of FGF21, serotonin and dopamine in CSF and the level of FGF21 in blood of the subjects were measured by ELISA using commercial kits. RESULTS: Negative correlations were found between BIS-11 total score and either FGF21, serotonin or dopamine in CSF. However, BIS-11 total score was not correlated with FGF21 in blood. In addition, FGF21 was positively correlated with serotonin and dopamine in CSF, respectively. Multivariable linear regression models indicated that the decrease of FGF21 level associating with the decrease of serotonin and dopamine level in CSF contributed to the higher impulsivity. Furthermore, receiver operating characteristic curve (ROC) analysis indicated an important role of CSF FGF21 predicting high impulsivity. CONCLUSIONS: FGF21, serotonin and dopamine in CSF associate with impulsivity in opposite directions. The decrease of CSF FGF21 is related to higher impulsivity, and indicate that CSF FGF21 may predict impulsivity in healthy subjects.

Association of FIB-4 index and clinical outcomes in critically ill patients with acute kidney injury: a cohort study.

BACKGROUND: The relationship between fibrosis-4 (FIB-4) index and clinical outcomes in patients with acute kidney injury (AKI) is unclear. We aimed to investigate the association between FIB-4 index and all-cause mortality in critically ill patients with AKI. METHODS: We used data from the Multiparameter Intelligent Monitoring in Intensive Care III (MIMIC-III) database (v1.4). The FIB-4 score was calculated using the existing formulas. logistic regression model, and Cox proportional hazards model were used to assessed the relationship between the FIB-4 index and in-hospital,28-day and 90-day mortality, respectively. RESULTS: A total of 3592 patients with AKI included in the data analysis. 395 (10.99%) patients died during hospitalization and 458 (12.74%) patients died in 28-day. During the 90-day follow-up, 893 (22.54%) patients were dead. An elevated FIB-4 value was significantly associated with increased in-hospital mortality when used as a continuous variable (odds ratio [OR] 1.183, 95% confidence interval [CI] 1.072-1.305, P = 0.002) and as a quartile variable (OR of Q2 to Q4 1.216-1.744, with Q1 as reference). FIB-4 was positively associated with 28-day mortality of AKI patients with hazard ratio (HR) of 1.097 (95% CI 1.008, 1.194) and 1.098 (95% 1.032, 1.167) for 90-day mortality, respectively. CONCLUSION: This study demonstrated the FIB-4 index is associated with clinical outcomes in critically ill patients with acute kidney injury.

The association between triglyceride glucose index and depression: data from NHANES 2005-2018.

BACKGROUND: The association between triglyceride glucose (TyG) index and depression is unclear. We conducted this analysis to explore whether higher TyG index is associated with a higher odd of depression. METHODS: This was an observational study using data from the National Health and Nutrition Examination Survey (2005-2018), a cross-sectional and nationally representative database. Depression was assessed using the Patient Health Questionnaire-9 (PHQ-9). TyG index was calculated based on the equation as follows: ln [triglyceride (mg/dL) × fasting blood glucose (mg/dL)/2], and participants were divided into quartiles based on TyG index. Weighted multivariable logistic regression models were used to explore the relationship between the TyG index and depression. RESULTS: A total of 13,350 patients were included, involving 1001 (7.50%) individuals with depression. Higher TyG index is significantly associated with elevated depressive symptoms in U.S. adults. Multivariate-adjusted HRs for patients in the TyG index 4th quartile were higher for depression (OR = 1.46; 95% confidence interval (CI) 1.30, 1.64) compared with the 1st quartile of TyG index. Similar results were seen in men and women, across age groups, and baseline comorbidities. CONCLUSION: In this large cross-sectional study, our result suggests that population with higher TyG index are significantly more likely to have depressive symptoms in U.S. adults.

Isatin dimers and their biological activities.

Dimerization is a promising strategy to develop novel drug candidates that could extend the biological spectrum, enhance the activity, overcome drug resistance, as well as improve pharmacological, pharmacokinetic, and physicochemical profiles. Isatin dimers possess a broad spectrum of biological properties and the isatin dimer indirubin has already been used in the clinic, revealing the potential of isatin dimers as putative drugs. This review covers the recent advances of isatin dimers as pharmacologically significant scaffolds and the structure-activity relationship to set up the direction for the design and development of isatin dimers with higher efficiency and lower toxicity.

A Meta-Analysis on the Effect of Agricultural Conservation Practices on Nutrient Loss.

Conservation practices are widely used to reduce N and P loads from agricultural fields and minimize their impact on water quality, but research using field-scale data to model the national average impact of conservation practices for different forms of N and P is needed. Thus, we quantified the effects of conservation practices (grassed waterways, terraces, contour farming, filter strips, and riparian buffers) on total, particulate, and dissolved N and P runoff from farmlands. Specifically, we conducted a meta-analysis of the Measured Annual Nutrient loads from AGricultural Environments (MANAGE) database using propensity score matching and multilevel modeling to remove the influence of confounding factors. There is no best method for addressing this influence, so we applied two alternative methods because similar results increase confidence in our findings. Propensity score matching found that conservation practices reduced total P, particulate P, and particulate N loading by an average of 67, 83, and 67%, respectively. Multilevel modeling estimated reductions of 58, 76, and 64% for the same nutrients. Although the propensity score method only yields a mean rate of reduction, multilevel modeling further estimates the reduction for different subgroups (i.e., different crops and fertilizer application methods) when such groupings are feasible. The multilevel models indicated that conservation practices affected row crops the most (e.g., corn [ L.] and soybean [ (L.) Merr.]) and fields with injected or surface-applied fertilizers. Our analysis used field-scale data to estimate the average effectiveness of conservation practices in reducing N and P runoff, providing valuable insight for regional and national decision making.

On Abandoning Hypothesis Testing in Environmental Standard Compliance Assessment.

We use basic characteristics of statistical significance test to argue the abandonment of hypothesis testing in environmental standard (or criterion) compliance assessment. The typical sample size used for environmental assessment is small, and the natural variation of many water quality constituent concentrations is high. These conditions lead to low statistical power of the hypothesis tests used in the assessment process. As a result, using hypothesis testing is often inefficient in detecting noncompliance. When a noncompliance is detected, it is frequently due to sampling or other types of error. We illustrate the problems using two examples, through which we argue that these problems cannot be resolved under the current practice of assessing compliance one water at a time. We recommend that the hypothesis testing framework be replaced by a statistical estimation approach, which can more effectively leverage information from assessments on similar waters using a probabilistic assessment approach.

The multiple-comparison trap and the Raven's paradox-perils of using null hypothesis testing in environmental assessment.

Detecting and quantifying environmental thresholds is frequently an important step in understanding ecological responses to environmental stressors. We discuss two statistical issues often encountered in threshold detection and quantification when statistical null hypothesis testing is used as a main analytical tool. The hidden multiple-comparison trap (leading to a much higher risk of a false detection) and Raven's paradox (rendering a "detection" meaningless) are often obscured when statistical hypothesis testing is used as part of a more elaborate model, especially models based on computer-intensive methods. Using two examples, we show that the hidden multiple-comparison trap can be exposed using computer simulation to estimate the probability of making a false detection; Raven's paradox can be avoided by clearly stating the null and alternative hypotheses using scientific terms to substantiate that the rejection of the null is equivalent to proving that the alternative of interest is true. The hidden multiple-comparison trap implies that a null hypothesis testing based on a computer-intensive method should be used with caution. The implication of Raven's paradox requires that we focus on providing evidence supporting the proposed hypothesis or model, rather than seeking evidence against the frequently irrelevant null hypothesis. These two problems, and many others related to null hypothesis testing, suggest that statistical hypothesis testing should be used only as a component of the body of evidence, perhaps, as the devil's advocate.

Response to CO2 enrichment of understory vegetation in the shade of forests.

Responses of forest ecosystems to increased atmospheric CO2 concentration have been studied in few free-air CO2 enrichment (FACE) experiments during last two decades. Most studies focused principally on the overstory trees with little attention given to understory vegetation. Despite its small contribution to total productivity of an ecosystem, understory vegetation plays an important role in predicting successional dynamics and future plant community composition. Thus, the response of understory vegetation in Pinus taeda plantation at the Duke Forest FACE site after 15-17 years of exposure to elevated CO2 , 6-13 of which with nitrogen (N) amendment, was examined. Aboveground biomass and density of the understory decreased across all treatments with increasing overstory leaf area index (LAI). However, the CO2 and N treatments had no effect on aboveground biomass, tree density, community composition, and the fraction of shade-tolerant species. The increases of overstory LAI (~28%) under elevated CO2 resulted in a reduction of light available to the understory (~18%) sufficient to nullify the expected growth-enhancing effect of elevated CO2 on understory vegetation.

Effectiveness of Secundum Atrial Septal Defect Occlusion with the Septal Occluder through Right-chest Small Incision: Clinical Analysis of 140 Cases.

Objective To evaluate the feasibility and effectiveness of secundum atrial septal defect(ASD)occlusion with the septal occluder through right-chest small incision. Methods The clinical data of 140 secundum ASD patients (47 males and 93 females) aged 3-63 years who were treated in our center from August 2004 to July 2014 were retrospectively analyzed. The diameter of ASD was 6 to 36 mm. Under general anesthesia, all patients underwent intraoperative transtsophageal echocardiography (TEE), during which no associated cardiac deformity was found. All patients received ASD occlusion via a small incision (3-4 cm) at the right anterior chest. The occluders were released with the help of TEE. Results The atrial septal defect closure was successfully completed in 134 cases. Six cases received surgical closure of ASD after the failure of occlusion. The reasons of conversion included postoperative dislodgement of occlusion device (n=2, both were central type with large size) and technically unsuitable for occlusion (n=4, in whom residual shunt was found in 2 case, sieve pore type in 1 case, and intraoperative dislodgement in 1 case). All of these 6 patients were treated surgically under cardiopulmonary bypass. No dislocation of the device or atrial shunt was found within 3 to 48 months after the operation. Conclusion Occlusion via small chest incision of ASD under TEE guidance without cardiopulmonary bypass is a safe, minimally invasive, effective, and convenient treatment and worth clinical application.

Implications of Stein's Paradox for Environmental Standard Compliance Assessment.

The implications of Stein's paradox stirred considerable debate in statistical circles when the concept was first introduced in the 1950s. The paradox arises when we are interested in estimating the means of several variables simultaneously. In this situation, the best estimator for an individual mean, the sample average, is no longer the best. Rather, a shrinkage estimator, which shrinks individual sample averages toward the overall average is shown to have improved overall accuracy. Although controversial at the time, the concept of shrinking toward overall average is now widely accepted as a good practice for improving statistical stability and reducing error, not only in simple estimation problems, but also in complicated modeling problems. However, the utility of Stein's insights are not widely recognized in the environmental management community, where mean pollutant concentrations of multiple waters are routinely estimated for management decision-making. In this essay, we introduce Stein's paradox and its modern generalization, the Bayesian hierarchical model, in the context of environmental standard compliance assessment. Using simulated data and nutrient monitoring data from wadeable streams around the Great Lakes, we show that a Bayesian hierarchical model can improve overall estimation accuracy, thereby improving our confidence in the assessment results, especially for standard compliance assessment of waters with small sample sizes.

Quantifying and Reducing Uncertainty in Estimated Microcystin Concentrations from the ELISA Method.

We discuss the uncertainty associated with a commonly used method for measuring the concentration of microcystin, a group of toxins associated with cyanobacterial blooms. Such uncertainty is rarely reported and accounted for in important drinking water management decisions. Using monitoring data from Ohio Environmental Protection Agency and from City of Toledo, we document the sources of measurement uncertainty and recommend a Bayesian hierarchical modeling approach for reducing the measurement uncertainty. Our analysis suggests that (1) much of the uncertainty is a result of the highly uncertain "standard curve" developed during each test and (2) the uncertainty can be reduced by pooling raw test data from multiple tests. Based on these results, we suggest that estimation uncertainty can be effectively reduced through the effort of either (1) regional regulatory agencies by sharing and combining raw test data from regularly scheduled microcystin monitoring program or (2) the manufacturer of the testing kit by conducting additional tests as part of an effort to improve the testing kit.

Photoperiodic regulation of the seasonal pattern of photosynthetic capacity and the implications for carbon cycling.

Although temperature is an important driver of seasonal changes in photosynthetic physiology, photoperiod also regulates leaf activity. Climate change will extend growing seasons if temperature cues predominate, but photoperiod-controlled species will show limited responsiveness to warming. We show that photoperiod explains more seasonal variation in photosynthetic activity across 23 tree species than temperature. Although leaves remain green, photosynthetic capacity peaks just after summer solstice and declines with decreasing photoperiod, before air temperatures peak. In support of these findings, saplings grown at constant temperature but exposed to an extended photoperiod maintained high photosynthetic capacity, but photosynthetic activity declined in saplings experiencing a naturally shortening photoperiod; leaves remained equally green in both treatments. Incorporating a photoperiodic correction of photosynthetic physiology into a global-scale terrestrial carbon-cycle model significantly improves predictions of seasonal atmospheric CO(2) cycling, demonstrating the benefit of such a function in coupled climate system models. Accounting for photoperiod-induced seasonality in photosynthetic parameters reduces modeled global gross primary production 2.5% (∼4 PgC y(-1)), resulting in a >3% (∼2 PgC y(-1)) decrease of net primary production. Such a correction is also needed in models estimating current carbon uptake based on remotely sensed greenness. Photoperiod-associated declines in photosynthetic capacity could limit autumn carbon gain in forests, even if warming delays leaf senescence.

The Frequency Component of Water Quality Criterion Compliance Assessment Should be Data Driven.

A numerical water quality criterion in the U.S. consists of three components representing magnitude, duration, and frequency. While magnitude and duration are well defined and conceptually unambiguous, the meaning of the frequency component is often debatable. We interpret the frequency component as a tool for accounting for uncertainty in estimating the mean concentration of a water quality constituent, after revisiting early works on environmental standards and criteria. Based on this interpretation, we illustrate management-related issues when using the default frequency of one exceedance in 3 years in compliance assessment. We propose a data-driven approach for estimating an appropriate frequency to ensure a consistent level of confidence in a water's compliance of a water quality criterion. The data-driven frequency is determined by water quality constituent concentration distribution characteristics and sample size. The method is illustrated using two examples.

Depolymerization and Re/Upcycling of Biodegradable PLA Plastics.

With the escalating utilization of plastic products, global attention has been increasingly drawn to environmental pollution and recycling challenges stemming from plastic waste. Against this backdrop, biodegradable plastics have emerged as viable alternatives owing to their sustainability and capacity for biodegradation. Polylactic acid (PLA) presently commands the largest market share among biodegradable plastics, finding extensive application in products such as thin films, medical materials, and biodegradable straws. However, the widespread adoption of PLA is hindered by challenges such as high cost, low recycling rates, and complete degradation to H2O and CO2 in natural conditions. Therefore, it is imperative and time-sensitive to explore solutions for the depolymerization and re/upcycling of PLA waste plastics. This review comprehensively outlines the current landscape of PLA recycling methods, emphasizing the advantages and significance of chemical re/upcycling. The subsequent exploration encompasses recent breakthroughs and technical obstacles inherent in diverse chemical depolymerization methods. Ultimately, this review accentuates the impediments and forthcoming possibilities in the realm of PLA plastics, emphasizing the pursuit of closed-loop recycling and upcycling.

Enhancing Quantitative Analysis of Xenobiotics in Blood Plasma through Cross-Matrix Calibration and Bayesian Hierarchical Modeling.

This study addresses the challenges of matrix effects and interspecies plasma protein binding (PPB) on measurement variability during method validation across diverse plasma types (human, rat, rabbit, and bovine). Accurate measurements of small molecules in plasma samples often require matrix-matched calibration approaches with the use of specific plasma types, which may have limited availability or affordability. To mitigate the costs associated with human plasma measurements, we explore in this work the potential of cross-matrix-matched calibration using Bayesian hierarchical modeling (BHM) to correct for matrix effects associated with PPB. We initially developed a targeted quantitative approach utilizing biocompatible solid-phase microextraction coupled with liquid chromatography-mass spectrometry for xenobiotic analysis in plasma. The method was evaluated for absolute matrix effects across human, bovine, rat, and rabbit plasma comparing pre- and postmatrix extraction standards. Absolute matrix effects from 96 to 108% for most analytes across plasma sources indicate that the biocompatibility of the extraction phase minimizes interference coextraction. However, the extent of PPB in different media can still affect the accuracy of the measurement when the extraction of small molecules is carried out via free concentration, as in the case of microextraction techniques. In fact, while matrix-matched calibration revealed high accuracy, cross-matrix calibration (e.g., using a calibration curve generated from bovine plasma) proved inadequate for precise measurements in human plasma. A BHM was used to calculate correction factors for each analyte within each plasma type, successfully mitigating the measurement bias resulting from diverse calibration curve types used to quantify human plasma samples. This work contributes to the development of cost-effective, efficient calibration strategies for biofluids. Leveraging easily accessible plasma sources, like bovine plasma, for method optimization and validation prior to analyzing costly plasma (e.g., human plasma) holds substantial advantages applicable to biomonitoring and pharmacokinetic studies.

Human umbilical cord mesenchymal stem cells combined with porcine small intestinal submucosa promote the healing of full-thickness skin injury in SD rats.

Aim: To assess the therapeutic potential of human umbilical cord mesenchymal stem cells (hUCMSCs) combined with porcine small intestinal submucosa (SIS) on full-thickness skin injuries in rats. Methods: We established full-thickness skin injury models in Sprague-Dawley rats, dividing them into blank control, SIS, hUCMSCs and hUCMSCs combined with SIS. We monitored wound healing, scores and area, and analyzed inflammatory cells, microvessel density and collagen fibers after 12 days. Results: The blank group showed no healing, forming a scar of 0.6 × 0.5 cm2, while SIS and hUCMSCs groups exhibited incomplete healing with 0.4 × 0.5 cm2 scabs. Wound healing was significantly better in the hUCMSCs combined with the SIS group. Conclusion: Local application of hUCMSCs combined with SIS enhances full-thickness skin injury wound healing in rats.

Our skin protects us from infections and injuries, but severe damage can lead to health problems. In this study, we explored a promising new treatment to enhance skin healing. We used mesenchymal stem cells derived from umbilical cords in combination with a biological material called porcine small intestinal submucosa (SIS) to conduct experiemnts on rats with skin wounds. This treatment led to much better healing in rats with deep skin wounds compared with standard approaches. This approach is promising for treating severe skin injuries, offering hope for quicker recovery and better outcome, including faster recovery, reduced pain and inflammation and less scarring.

Amorphous curcumin-based hydrogels to reduce the incidence of post-surgical intrauterine adhesions.

The incidence of intrauterine adhesions (IUA) has increased with the rising utilization of intrauterine surgery. The postoperative physical barrier methods commonly used, such as balloons and other fillers, have limited effectiveness and may even cause further damage to the remaining endometrial tissue. Herein, we developed an injectable thermosensitive hydrogel using Pluronic F127/F68 as pharmaceutical excipients and curcumin as a natural active molecule. The hydrogel effectively addresses solubility and low bioavailability issues associated with curcumin. In vitro, drug release assays revealed that the amorphous curcumin hydrogel promotes dissolution and sustained release of curcumin. In vitro experiments reveal high biocompatibility of the hydrogel and its ability to enhance vascular formation while inhibiting the expression of fibrotic factor TGF-ß1. To assess the effectiveness of preventing IUAs, in vivo experiments were conducted using IUA rats and compared with a class III medical device, a new-crosslinked hyaluronic acid (NCHA) gel. According to the study, curcumin hydrogel is more effective than the NCHA group in improving the regeneration of the endometrium, increasing the blood supply to the endometrium and reducing the abnormal deposition of fibrin, thus preventing IUA more effectively. This study provides a promising strategy for treating and preventing IUA.

Phlebosclerosis: An overlooked complication of varicose veins that affects clinical outcome: A case report.

BACKGROUND: Phlebosclerosis is a common age-related fibrotic degeneration of the venous wall. It is a disorder rather than a disease, which may cause venous dysfunction and even venous thrombosis. It is rarely reported in patients with varicose veins. CASE SUMMARY: The present report describes the case of a 70-year-old man with varicose veins, vitiligo, and phlebosclerosis. Venous angiography revealed blood reflux in the superficial and deep veins. The patient underwent surgery to remove the saphenous veins. During the operation, a calcified vein resembling a wooden stick was found, which was surprisingly extracted from the thickened venous wall. A cross-section of this wooden stick-like vein revealed venous fibrosis and calcification, obvious thickening of the venous wall, extensive collagen deposition on the venous wall, hyaline degeneration, and venous sclerosis causing closure of the venous lumen. CONCLUSION: This is probably the first report of a wooden stick-like structure being found in the venous wall in patients with varicose veins and venous ulcers. Phlebosclerosis can be observed in the late stage of varicose veins complicated by frequent infections and worse clinical outcomes. Therefore, it is important to be aware of this condition and address it rather than overlook it.