Network pharmacology analysis of the regulatory effects and mechanisms of ALAE on sow reproduction in vivo and in vitro.

ETHNOPHARMACOLOGICAL RELEVANCE: Reproductive ability of sows is a primary element influencing the development of pig farming. Herbal extracts of Angelica sinensis (Oliv.) Diels, Astragalus mongholicus Bunge, Eucommia ulmoides Oliv., and Polypodium glycyrrhiza D.C.Eaton showed effects on improvement of reproduction in sows. AIMS OF THE STUDY: To investigate the mechanism of the treatment effects by a compound of these four Chinese herbs in a 1:1:1:1 ratio (ALAE) on endometriosis, endometritis, uterine adhesion, intrauterine growth retardation, pre-eclampsia, and its enhancement of reproductive efficiency in sows. MATERIALS AND METHODS: Active components of ALAE were identified by using ultra-performance liquid chromatography-mass spectrometry analysis and network pharmacology. Then we used the results to construct a visualization network. Key targets and pathways of ALAE involved in sow reproduction improvement were validated in sow animals and porcine endometrial epithelial cells (PEECs). RESULTS: A total of 62 active compounds were found in ALAE (41 in Polypodium glycyrrhiza D.C.Eaton, 5 in Astragalus mongholicus Bunge, 11 in Eucommia ulmoides Oliv., 5 in Angelica sinensis (Oliv.) Diels) with 563 disease-related targets (e.g. caspase-3, EGFR, IL-6) involved in EGFR tyrosine kinase inhibitor resistance, PI3K-AKT, and other signaling pathways. Molecular docking results indicated GC41 (glabridin), GC18 (medicarpin), EGFR and CCND1 are possible key components and target proteins related to reproductive improvement in sows. In PEECs, EGFR expression decreased at the mRNA and protein levels by three doses (160, 320, and 640 µg/mL) of ALAE. The phosphorylation of downstream pathway PI3K-AKT1 was enhanced. The expression of inflammatory factors (IL-6, IL-1ß), ESR1 and caspase-3 decreased through multiple pathways. Additionally, the expression levels of an anti-inflammatory factor (IL-10), angiogenesis-related factors (MMP9, PIGF, PPARγ, IgG), and placental junction-related factors (CTNNB1, occludin, and claudin1) increased. Furthermore, the total born number of piglets, the number of live and healthy litters were significantly increased. The number of stillbirths decreased by ALAE treatment in sow animals. CONCLUSIONS: Dministration of ALAE significantly increased the total number of piglets born, the numbers of live and healthy litters and decreased the number of stillbirths through improving placental structure, attenuating inflammatory response, modulating placental angiogenesis and growth factor receptors in sows. The improvement of reproductive ability may be related to activation of the EGFR-PI3K-AKT1 pathway in PEECs. Moreover, ALAE maybe involved in modulation of estrogen receptors, apoptotic factors, and cell cycle proteins.

PLA inhibits TNF-α-induced PANoptosis of prostate cancer cells through metabolic reprogramming.

Previous studies have shown that phenyllactic acid (alpha-Hydroxyhydrocinnamic acid, 2-Hydroxy-3-phenylpropionic acid, PLA), a type of organic acid metabolite, has excellent diagnostic efficacy when used to differentiate between prostate cancer, benign prostatic hyperplasia, and prostatitis. This research aims to explore the molecular mechanism by which PLA influences the PANoptosis of prostate cancer (PCa) cell lines. First, we found that PLA was detected in all prostate cancer cell lines (PC-3, PC-3 M, DU145, LNCAP). Further experiments showed that the addition of PLA to prostate cancer cells could promote ATP generation, enhance cysteine desulfurase (NFS1) expression, and reduce tumor necrosis factor alpha (TNF-α) levels, thereby inhibiting apoptosis in prostate cancer cells. Notably, overexpression of NFS1 can inhibit the binding of TNF-α to serpin mRNA binding protein 1 (SERBP1), suggesting that NFS1 competes with TNF-α for binding to SERBP1. Knockdown of SERBP1 significantly reduced the level of small ubiquity-related modifier (SUMO) modification of TNF-α. This suggests that NFS1 reduces the SUMO modification of TNF-α by competing with SERBP1, thereby reducing the expression and stability of TNF-α and ultimately inhibiting apoptosis in prostate cancer cell lines. In conclusion, PLA inhibits TNF-α induced panapoptosis of prostate cancer cells through metabolic reprogramming, providing a new idea for targeted treatment of prostate cancer.

New Alkaloids and Steroids from Hydranth of Goniopora columna Corals and Their Inhibiting Lung Cancer Cell Activities.

A new alkaloids, aplysingoniopora A (1), and new configuration pregnane type steroid compound, 9,17-α-pregn-1,4,20-en-3-one (2), and two known pregnane type steroid compounds (3 and 4) were isolated from hydranth of Goniopora columna corals. The compounds structures and absolute configurations were determined by extensive spectroscopic analysis, MS data, single-crystal X-ray diffraction analysis and quantum chemical calculation. The anticancer effect of the compounds were explored in human non-small-cell lung cancer (NSCLC) A549 cell lines. As the results, the compound 3 and 4 induces toxicity and has proliferation inhibitory effects on A549 cells (IC50=58.99 µM and 58.77 µM, respectively) in vitro.

Validity across four common street-crossing distraction indicators to predict pedestrian safety.

BACKGROUND: Multiple distraction indicators have been applied to measure street-crossing distraction but their validities in predicting pedestrian safety are poorly understood. METHODS: Based on a video-based observational study, we compared the validity of four commonly used distraction indicators (total duration of distraction while crossing a street, proportion of distracted time over total street-crossing time, duration of the longest distraction time, and total number of distractions) in predicting three pedestrian safety outcomes (near-crash incidence, frequency of looking left and right, and speed crossing the street) across three types of distraction (mobile phone use, talking to other pedestrians, eating/drinking/smoking). Change in Harrell's C statistic was calculated to assess the validity of each distraction indicator based on multivariable regression models including only covariates and including both covariates and the distraction indicator. RESULTS: Heterogeneous capacities in predicting the three safety outcomes across the four distraction indicators were observed: 1) duration of the longest distraction time was most predictive for the occurrence of near-crashes and looks left and right among pedestrians with all three types of distraction combined and talking with other pedestrians (Harrell's C statistic changes ranged from 0.0310 to 0.0335, P < 0.05), and for the occurrence of near-crashes for pedestrians involving mobile phone use (Harrell's C statistic change: 0.0053); 2) total duration of distraction was most predictive for speed crossing the street among pedestrians with the combination and each of the three types of distraction (Harrell's C statistic changes ranged from 0.0037 to 0.0111, P < 0.05), frequency of looking left and right among pedestrians distracted by mobile phone use (Harrell's C statistic change: 0.0115), and the occurrence of near-crash among pedestrians eating, drinking, or smoking (Harrell's C statistic change: 0.0119); and 3) the total number of distractions was the most predictive indicator of frequency of looking left and right among pedestrians eating, drinking, or smoking (Harrell's C statistic change: 0.0013). Sensitivity analyses showed the results were robust to change in grouping criteria of the four distraction indicators. CONCLUSIONS: Future research should consider the pedestrian safety outcomes and type of distractions to select the best distraction indicator.

The therapeutic role and potential mechanism of EGCG in obesity-related precocious puberty as determined by integrated metabolomics and network pharmacology.

Objective: (-)-Epigallocatechin-3-gallate (EGCG) has preventive effects on obesity-related precocious puberty, but its underlying mechanism remains unclear. The aim of this study was to integrate metabolomics and network pharmacology to reveal the mechanism of EGCG in the prevention of obesity-related precocious puberty. Materials and methods: A high-performance liquid chromatography-electrospray ionization ion-trap tandem mass spectrometry (LC-ESI-MS/MS) was used to analyze the impact of EGCG on serum metabolomics and associated metabolic pathways in a randomized controlled trial. Twelve weeks of EGCG capsules were given to obese girls in this trail. Additionally, the targets and pathways of EGCG in preventing obesity-related precocious puberty network pharmacology were predicted using network pharmacology. Finally, the mechanism of EGCG prevention of obesity-related precocious puberty was elucidated through integrated metabolomics and network pharmacology. Results: Serum metabolomics screened 234 endogenous differential metabolites, and network pharmacology identified a total of 153 common targets. These metabolites and targets mainly enrichment pathways involving endocrine-related pathways (estrogen signaling pathway, insulin resistance, and insulin secretion), and signal transduction (PI3K-Akt, MAPK, and Jak-STAT signaling pathways). The integrated metabolomics and network pharmacology indicated that AKT1, EGFR, ESR1, STAT3, IGF1, and MAPK1 may be key targets for EGCG in preventing obesity-related precocious puberty. Conclusion: EGCG may contribute to preventing obesity-related precocious puberty through targets such as AKT1, EGFR, ESR1, STAT3, IGF1, and MAPK1 and multiple signaling pathways, including the estrogen, PI3K-Akt, MAPK, and Jak-STAT pathways. This study provided a theoretical foundation for future research.

Cyanidin-3-glucoside induces mesenchymal to epithelial transition via activating Sirt1 expression in triple negative breast cancer cells.

Triple-negative breast cancer (TNBC) is a heterogeneous group of breast cancer with one common feature: distinctly metastatic nature with higher rate of relapse and shorter survival compared with other subtypes of breast cancer. The epithelial to mesenchymal transition (EMT) is highly associated with cancer metastasis. Cyanidin-3-glucoside (C3G), the most abundant anthocyanin pigment enriched in fresh fruits and vegetables, showed ideal anti-oxidant property. C3G could also inhibit certain malignant behaviors of cancer cells, however, whether repression of EMT was involved in its anti-cancer especially TNBC effect remains unknown. Herein, we report that C3G decreases the migratory and invasive nature of TNBC lines MDA-MB-231 and BT-549. Mechanistically, C3G induces reversion of EMT characterized by phenotype modulation with increased epithelial marker E-ca and ZO-1, decreased mesenchymal marker Vimentin, N-ca and EMT-associated transcription factors Snail1, Snail2. NF-κB is pivotal for EMT and Sirt1 is a NF-κB inhibitor. We show that NF-κB is attenuated and Sirt1 is induced by C3G in TNBC, respectively. And later evidence demonstrates that abrogation of Sirt1 with small interfering RNA transfection abolished NF-κB inhibition and EMT reversion by C3G. Subsequently, we show that microRNA-138(miR-138) represses Sirt1 via mRNA translation inhibition and is inhibited by C3G. Moreover, miR-138 repression is involved in Sirt1 re-activation and migratory and invasive inhibition of TNBC by C3G. Taken together, we supplied more evidence to the anti-breast cancer mechanisms of C3G.

Heavy Metal Contamination in Soil and Brown Rice and Human Health Risk Assessment near Three Mining Areas in Central China.

BACKGROUND: Metal mining and waste discharge lead to regional heavy metal contamination and attract major concern because of the potential risk to local residents. METHODS: This research was conducted to determine lead (Pb), cadmium (Cd), arsenic (As), manganese (Mn), and antimony (Sb) concentrations in soil and brown rice samples from three heavy metal mining areas in Hunan Province, central China, and to assess the potential health risks to local inhabitants. RESULTS: Local soil contamination was observed, with mean concentrations of Cd, Pb, Sb, and As of 0.472, 193.133, 36.793, and 89.029 mg/kg, respectively. Mean concentrations of Cd, Pb, Sb, Mn, and As in brown rice were 0.103, 0.131, 5.175, 6.007, and 0.524 mg/kg, respectively. Daily intakes of Cd, As, Sb, Pb, and Mn through brown rice consumption were estimated to be 0.011, 0.0002, 0.004, 0.0001, and 0.0003 mg/(kg/day), respectively. The combined hazard index for the five heavy metals was 22.5917, and the total cancer risk was 0.1773. Cd contributed most significantly to cancer risk, accounting for approximately 99.77% of this risk. CONCLUSIONS: The results show that potential noncarcinogenic and carcinogenic health risks exist for local inhabitants and that regular monitoring of pollution to protect human health is urgently required.

A novel damage mechanism: Contribution of the interaction between necroptosis and ROS to high glucose-induced injury and inflammation in H9c2 cardiac cells.

Recently, a novel mechanism known as 'programmed necrosis' or necroptosis has been shown to be another important mechanism of cell death in the heart. In this study, we investigated the role of necroptosis in high glucose (HG)-induced injury and inflammation, as well as the underlying mechanisms. In particular, we focused on the interaction between necroptosis and reactive oxygen species (ROS) in H9c2 cardiac cells. Our results demonstrated that the exposure of H9c2 cardiac cells to 35 mM glucose (HG) markedly enhanced the expression level of receptor-interacting protein 3 (RIP3), a kinase which promotes necroptosis. Importantly, co-treatment of the cells with 100 µM necrostatin-1 (a specific inhibitor of necroptosis) and HG for 24 h attenuated not only the increased expression level of RIP3, but also the HG-induced injury and inflammation, as evidenced by an increase in cell viability, a decrease in ROS generation, the attenuation of the dissipation of mitochondrial membrane potential and a decrese in the secretion levels of inflammatory cytokines, i.e., interleukin (IL)-1ß and tumor necrosis factor (TNF)-α. Furthermore, treatment of the cells with 1 mM N-acetyl­L­cysteine (a scavenger of ROS) for 60 min prior to exposure to HG significantly reduced the HG-induced increase in the RIP3 expression level, as well as the injury and inflammatory response described above. Taken together, the findings of this study clearly demonstrate a novel damage mechanism involving the positive interaction between necroptosis and ROS attributing to HG-induced injury and inflammation in H9c2 cardiac cells.