Research progress on ethnobotany, phytochemistry, pharmacological action, and applications of Engelhardia roxburghiana Wall: a review.

OBJECTIVES: Engelhardia roxburghiana Wall is a plant of the Juglandaceae family, and its leaves is the main part used as a medicine. It is used to relieve heat and pain, gasification, and dampness. The purpose of this review is to provide a systematic review about the botany, traditional uses, phytochemistry, pharmacology, and toxicology of this plant. KEY FINDINGS: Many compounds have been isolated and identified from the plant, including flavonoids, triterpenoids, steroids, quinones, essential oils, and other types of chemical constituents. Extensive pharmacological activities of the extracts or compounds of E. roxburghiana Wall in vivo and in vitro were mainly confirmed, including anti-cancer, anti-diabetic, anti-inflammatory, and anti-allergic effects. SUMMARY: In this paper, the botany, traditional uses, phytochemistry, and pharmacology of E. roxburghiana Wall were reviewed. In the future, E. roxburghiana Wall needs further study, such as paying more attention to quality control and the utilization on agriculture. In addition, discussing the medicinal components of decoction as well as the toxicity will also contribute to the progress of clinical trial studies.

Quality assessment and Q-markers discovery in Citri Sarcodactylis Fructus by integrating serum pharmacochemistry and network pharmacology.

INTRODUCTION: Citri Sarcodactylis Fructus (CSF), a common fruit and traditional Chinese medicine (TCM), has been hindered in its further development and research owing to the lack of comprehensive and specific quality evaluation standards. OBJECTIVE: This study aimed to establish clear TCM quality standards related to the therapeutic mechanisms of CSF and to provide a basis for subsequent research and development. METHODS: Ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry (UPLC-Q-orbitrap HRMS) technology was used to comprehensively identify CSF components and explore their absorbance levels in rat serum. Network pharmacology research methods were employed to investigate the potential mechanisms of action of the identified components in the treatment of major clinical diseases. Subsequently, a combination of HPLC chromatographic fingerprinting for qualitative analysis and multi-index content determination was used to evaluate the detectability of the identified quality markers (Q-markers). RESULTS: Twenty-six prototype components were tentatively characterized in rat serum. Network pharmacology analysis showed six effective components, namely 7-hydroxycoumarin, isoscopoletin, diosmin, hesperidin, 5,7-dimethoxycoumarin, and bergapten, which played important roles in the treatment of chronic gastritis, functional dyspepsia, peptic ulcer, and depression and were preliminarily identified as Q-markers. The results of content determination in 15 batches of CSF indicated significant differences in the content of medicinal materials from different origins. However, compared with the preliminarily determined Q-markers, all six components could be measured and were determined as Q-markers of CSF. CONCLUSION: The chemical Q-markers obtained in this study could be used for effective quality control of CSF.

Tai Chi improves non-motor symptoms of Parkinson's disease: One-year randomized controlled study with the investigation of mechanisms.

BACKGROUND: Tai Chi was found to improve motor symptoms in Parkinson's disease (PD). Whether long-term Tai Chi training could improve non-motor symptoms (NMS) and the related mechanisms were unknown. OBJECTIVE: To investigate Tai Chi's impact on non-motor symptoms in PD and related mechanisms. METHODS: 95 early-stage PD patients were recruited and randomly divided into Tai Chi (N = 32), brisk walking (N = 31), and no-exercise groups (N = 32). All subjects were evaluated at baseline, 6 months, and 12 months within one-year intervention. Non-motor symptoms (including cognition, sleep, autonomic symptoms, anxiety/depression, and quality of life) were investigated by rating scales. fMRI, plasma cytokines and metabolomics, and blood Huntingtin interaction protein 2 (HIP2) mRNA levels were detected to observe changes in brain networks and plasma biomarkers. RESULTS: Sixty-six patients completed the study. Non-motor functions assessed by rating scales, e.g. PD cognitive rating scale (PDCRS) and Epworth Sleepiness scale (ESS), were significantly improved in the Tai Chi group than the control group. Besides, Tai Chi had advantages in improving NMS-Quest and ESS than brisk walking. Improved brain function was seen in the somatomotor network, correlating with improved PDCRS (p = 0.003, respectively). Downregulation of eotaxin and upregulation of BDNF demonstrated a positive correlation with improvement of PDCRS and PDCRS-frontal lobe scores (p ≤ 0.037). Improvement of energy and immune-related metabolomics (p ≤ 0.043), and elevation of HIP2 mRNA levels (p = 0.003) were also found associated with the improvement of PDCRS. CONCLUSIONS: Tai Chi improved non-motor symptoms in PD, especially in cognition and sleep. Enhanced brain network function, downregulation of inflammation, and enhanced energy metabolism were observed after Tai Chi training.

Anthriscus sylvestris: An overview on Bioactive Compounds and Anticancer Mechanisms from a Traditional Medicinal Plant to Modern Investigation.

Anthriscus sylvestris (L.) Hoffm. Gen. is a biennial or perennial herb commonly found in China. It has a long history of use in traditional Chinese medicine to treat various ailments such as cough, gastric disorders, spleen deficiency, and limb weakness. Recently, its potential as an anticancer agent has gained considerable attention and has been the subject of extensive research focusing on extract efficacy, identification of active compounds, and proposed molecular mechanisms. Nevertheless, further high-quality research is still required to fully evaluate its potential as an anticancer drug. This review aims to comprehensively summarize the anticancer properties exhibited by the active components found in Anthriscus sylvestris. We conducted a comprehensive search, collation, and analysis of published articles on anticancer activity and active compounds of A. sylvestris using various databases that include, but are not limited to, PubMed, Web of Science, Science Direct and Google Scholar. The primary chemical composition of A. sylvestris consists of phenylpropanoids, flavonoids, steroids, fatty acids, and organic acids, showcasing an array of pharmacological activities like anticancer, antioxidant, anti-aging, and immunoregulatory properties. Thus, this review highlights the active compounds isolated from A. sylvestris extracts, which provide potential leads for the development of novel anticancer drugs and a better understanding of the plant's pharmacological effects, particularly its anticancer mechanism of action.

Trichoderma asperellum boosts nitrogen accumulation and photosynthetic capacity of wolfberry (Lycium chinense) under saline soil stress.

Trichoderma can promote plant growth under saline stress, but the mechanisms remain to be revealed. In this study, we investigate photosynthetic gas exchange, photosystem II (PSII) performance, nitrogen absorption and accumulation in a medicinal plant wolfberry (Lycium chinense) in saline soil supplemented with Trichoderma biofertilizer (TF). Larger nitrogen and biomass accumulation were found in plants supplemented with TF than with organic fertilizer (OF), suggesting that Trichoderma asperellum promoted plant growth and nitrogen accumulation under saline stress. T. asperellum strengthened root nitrogen (N) absorption according to greater increased root NH4+ and NO3- influxes under supplement with TF than OF, while nitrogen assimilative enzymes such as nitrate reductase, nitrite reductase and glutamine synthetase activities in roots and leaves were also stimulated. Thus, the elevated N accumulation derived from the induction of T. asperellum on nitrogen absorption and assimilation. Greater increased photosynthetic rate (Pn) and photosynthetic N-use efficiency under supplement with TF than OF illustrated that T. asperellum enhanced photosynthetic capacity and N utilization under saline stress. Although increased leaf stomatal conductance contributed to carbon (C) isotope fractionation under TF supplement, leaf 13C abundance was significantly increased by supplement with TF rather than OF, indicating that T. asperellum raised CO2 assimilation to a greater extent, reducing C isotope preference. Trichoderma asperellum optimized electron transport at PSII donor and acceptor sides under saline stress because of lower K and J steps in chlorophyll fluorescence transients under supplement with TF than OF. The amount of PSII active reaction centers was also increased by T. asperellum. Thus, PSII performance was upgraded, consistent with greater heightened delayed chlorophyll fluorescence transients and I1 peak under supplement with TF than OF. In summary, TF acted to increase N nutrient acquisition and photosynthetic C fixation resulting in enhanced wolfberry growth under saline soil stress.

Isolation, phytochemistry, characterization, biological activity, and application of Morinda officinalis How oligosaccharide: a review.

Morinda officinalis How oligosaccharide (MOO) stands as one of the principal active constituents of M. officinalis How, widely employed in traditional Chinese medicine. The methods for MOO extraction predominantly encompass hot water extraction, ethanol extraction, ultrasonic-assisted extraction, and microwave-assisted extraction. Distinct extraction techniques yield varying MOO quantities. MOO encompasses a diversity of oligosaccharides, including bajijiasu, sucrose, 1-kestose, nystose, mannose, 1F-fructofuranosylnystose, 1,1,1,1-kestohexose, fructoheptasaccharide, inulin-type hexasaccharide, inulin-type heptasaccharide, inulotriose, inulotetraose, inulopentaose, and mannose. MOO exhibits a wide spectrum of biological activities, exerting specific effects on the nervous system, cardiovascular system, motor system, reproductive system, and immune system. It demonstrates antidepressant properties, offers potential in mitigating Alzheimer's disease, stimulates angiogenesis, and possesses anti-osteoporotic and other pharmacological effects. Clinically, when combined with various antidepressants, MOO exhibits specific therapeutic efficacy across multiple forms of depression. As a naturally occurring plant oligosaccharide, MOO holds diverse pharmaceutical applications. This article conducts a review of the latest extraction and purification methodologies, structural characterization analysis, biological activity assessment, and clinical applications of MOO. Such a comprehensive analysis yields innovative insights for advancing the research and application of MOO in the future.

Development and validation of risk prediction model for premenstrual syndrome in nurses: results from the nurses-based the TARGET cohort study.

Objective: Premenstrual syndrome (PMS) stands as a significant concern within the realm gynecological disorders, profoundly impacting women of childbearing age in China. However, the elusive nature of its risk factors necessitates investigation. This study, therefore, is dedicated to unraveling the intricacies of PMS by focusing on nurses, a cohort with unique occupational stressors, to develop and validate a predictive model for assessing the risk of PMS. Methods: This investigation employed a multi-center cross-sectional analysis drawing upon data from the TARGET Nurses' health cohort. Utilizing online survey versions of the Premenstrual Syndrome Scale (PMSS), a comprehensive dataset encompassing physiological, social, psychological, occupational, and behavioral variables was collected from 18,645 participants. A stepwise multivariate logistic regression analysis was conducted to identify independent risk factors for PMS. Furthermore, a refined variable selection process was executed, combining the Least Absolute Shrinkage and Selection Operator (LASSO) method with 10-fold cross-validation. The visualization of the risk prediction model was achieved through a nomogram, and its performance was evaluated using the C index, receiver operating characteristic (ROC) curves, and the calibration curves. Results: Among the diverse variables explored, this study identified several noteworthy predictors of PMS in nurses, including tea or coffee consumption, sleep quality, menstrual cycle regularity, intermenstrual bleeding episodes, dysmenorrhea severity, experiences of workplace bullying, trait coping style, anxiety, depression and perceived stress levels. The prediction model exhibited robust discriminatory power, with an area under the curve of 0.765 for the training set and 0.769 for the test set. Furthermore, the calibration curve underscored the model's high degree of alignment with observed outcomes. Conclusion: The developed model showcases exceptional accuracy in identifying nurses at risk of PMS. This early alert system holds potential to significantly enhance nurses' well-being and underscore the importance of professional support.

Schisandrin B Inhibits Cell Viability and Malignant Progression of Melanoma Cells via Wnt/ß-catenin Signaling Pathway.

BACKGROUND: Melanoma is of great interest due to its aggressive behavior and less favorable prognosis. The need for the development of novel drugs for the treatment of melanoma is urgent. Considerable evidence indicated that Schisandrin B (Sch B), a bioactive compound extracted from Schisandra chinensis, has numerous anti-tumor properties in multiple malignant tumors. A few studies have reported the effect of Sch B on melanogenesis in the melanoma B16F10 cell line; however, the specific anti-tumor effects and mechanisms need to be further explored. OBJECTIVE: This study aimed to investigate the effects of Sch B on the cell viability, migration, invasion, and cell cycleblocking of melanoma cells and explore its potential anti-tumor mechanism in vitro and in vivo. METHODS: Melanoma cells (A375 and B16) were treated with different concentrations of Sch B (0, 20, 40, 60, or 80 µM), with dimethyl sulfoxide (DMSO) as control. The inhibitory effect of Sch B on A375 and B16 melanoma cells was verified by crystal violet assay and CCK8 assay. The flow cytometry was performed to observe cell cycle blocking. The effect of Sch B on the migration and invasion of melanoma cells was detected by wound healing assay and transwell assay, respectively. Western blot analysis was used to determine protein expression levels. The growth of the A375 melanoma xenograft-treated groups and immunohistochemical staining were conducted to assess the anti-tumor effect of Sch B in vivo. RESULTS: The crystal violet assay and CCK8 assay showed that Sch B significantly inhibited melanoma cell viability in a dose-dependent manner. Meanwhile, the flow cytometry analysis revealed that Sch B induced melanoma cell cycleblocking at the G1/S phase. In addition, the wound healing assay and transwell assay showed that Sch B inhibited the migration and invasion of melanoma cells. Furthermore, by establishing an animal model, we found that Sch B significantly inhibited the growth of melanoma in vivo. The potential mechanism could be that Sch B inhibited the activity of the Wnt/ß-catenin signaling pathway. CONCLUSION: These findings indicated that Sch B inhibits the cell viability and malignant progression of melanoma cells via the Wnt/ß-catenin pathway and induces cell cycle arrest. Our study suggests that Sch B has potential as a bioactive compound for the development of new drugs for melanoma.

Research Trends of the Research and Development of Acupuncture and Moxibustion Therapy on Lumbar Disc Herniation: A Bibliometric Analysis.

Background: Acupuncture and moxibustion has been applied worldwide in the treatment of various pain diseases including lumbar disc herniation (LDH) and other pain, However, there has been no bibliometric analysis on this aspect in the past five years. Therefore, this study was carried out for finding research trends and fronts in this field using Citespace and VOSviewer. Methods: Publications about acupuncture therapy for LDH were extracted from the Web of Science database and PubMed with an unlimited time frame. A bibliometric analysis and visualization of results was conducted using CiteSpace 6.1.R3 and VOSviewer 1.6.18 on the information of the annual publication, countries, journals, institutions, authors, references, and keywords. Results: A total of 127 publications were included, and the number of publications had increased noticeably over the past 30 years and reached a peak in the past 3 years. The most productive country with the most publications was China, whose Medical University was the institution with the highest volume of publications. The most productive author was Chen Rixin, while the most-cited author was Kreiner DS. Chinese Acupuncture and Moxibustion was the journal with the most publications, and Spine Journal was the most frequently cited journal. In cited references, an article published in The New England Journal of Medicine by Deyo RA had the most citations and the highest centrality. Of the keywords, the five most frequently used keywords include lumbar disc herniation, acupuncture, low back pain, intervertebral disc displacement, and management. Conclusion: Acupuncture and moxibustion can help to relieve symptoms in patients. However, this field is in the early stages of development and requires more high-quality research studies and international collaborations. In addition, exploring the effectiveness and mechanism of acupuncture for LDH is the hot trend in the future.

Structure-based virtual screening for discovery of paederosidic acid from Paederia scandens as novel P2Y14R antagonist.

BACKGROUND: The activation of P2Y14 receptor (P2Y14R) promotes osteoclast formation and causes neuropathic pain, exhibiting possible link to osteoarthritis (OA). Given lack of P2Y14R antagonist, the present study aims to search a novel P2Y14R antagonist with low toxicity and high activity from natural products as a possible drug candidate in treatment of OA. METHODS: The role of P2Y14R on OA was verified using P2Y14R knockout (KO) rats. Molecular docking virtual screening strategy and activity test in P2Y14R stably-expressed HEK293 cells were used to screen target compound from natural product library. The MM/GBSA free energy calculation/decomposition technique was used to determine the principal interaction mechanism. Next, the binding of target compound to P2Y14R was examined using cellular thermal shift assay and drug affinity responsive target stability test. Finally, the therapeutic effect of target compound was performed in monosodium iodoacetate (MIA)-induced OA mouse model. To verify whether the effect of target compound was attributed to P2Y14R, we establish the osteoarthritis model in P2Y14R KO mice to perform pharmacodynamic evaluation. Importantly, to investigate the potential mechanism by which target compound attenuate OA, expressions of the major transcription factors involved in osteoclast differentiation were detected by western blot, while markers of nerve damage in dorsal root ganglion (DRG) were evaluated by RT-qPCR and immunofluorescence techniques. RESULTS: Deficiency of P2Y14R alleviated pain behavior and cartilage destruction in MIA-induced OA rats. 14 natural compounds were screened by Glide docking-based virtual screening, among which paederosidic acid exhibited the highest antagonistic activity to P2Y14R with IC50 of 8.287 µM. As a bioactive component extracted from Paederia scandens, paederosidic acid directly interacted with P2Y14R to enhance the thermostability and decrease the protease sensitivity of target protein, which significantly inhibited receptor activator for nuclear factor-κB ligand (RANKL)-mediated osteoclastogenesis. More importantly, paederosidic acid suppressed osteoclast formation by downregulating expressions of NFAT2 and ATP6V0D2, as well as relieved neuropathic pain by decreasing expressions of CGRP, CSF1 and galanin in DRG. CONCLUSIONS: Paederosidic acid targeted P2Y14R to improve OA through alleviating osteoclast formation and neuropathic pain, which provided an available strategy for developing novel drug leads for treatment of OA.

Characterization of Dendrobium catenatum CBL-CIPK signaling networks and their response to abiotic stress.

Dendrobium catenatum is a traditional Chinese medicine listing as rare and endangered due to environmental impacts. But little is known about its stress resistance mechanism. The CBL-CIPK signaling pathway played vital roles in various stress responses. In this study, we identified 9 calcineurin B-like (CBL) genes and 28 CBL-interacting protein kinase (CIPK) genes from D. catenatum. Phylogenetic analysis showed that DcCBL and DcCIPK families could be divided into four and six subgroups, respectively. Members in each subgroup had similar gene structures. Cis-acting element analyses showed that these genes were involved in stress responses and hormone signaling. Spatial expression profiles showed that they were tissue-specific, and expressed lower in vegetative organs than reproductive organs. Gene expression analyses revealed that these genes were involved in drought, heat, cold, and salt responses and depended on abscisic acid (ABA) and salicylic acid (SA) signaling pathways. Furthermore, we cloned 19 DcCIPK genes and 9 DcCBL genes and detected ten interacting CBL-CIPK combinations using yeast two-hybrid system. Finally, we constructed 20 CBL-CIPK signaling pathways based on their expression patterns and interaction relationships. These results established CBL-CIPK signaling pathway responding to abiotic stress and provided a molecular basis for improving D. catenatum stress resistance in the future.

[Impounding Impacts of the Three Gorges Reservoir on Phytoplankton Function Groups and Its Relationship with Resource Use Efficiency].

In order to investigate the relationship between phytoplankton community functional group compositions and resource use efficiency in important tributaries of the Three Gorges Reservoir, phytoplankton and environment parameters were sampled from five tributaries, the Xiangxi River, Daning River, Meixi River, Pengxi River, and Huangjin River, in August and November, 2020. There were 119 species (variants) belonging to 62 genera and 7 phyla identified in summer, whereas 118 species (variants) belonging to 7 divisions of 58 genera were found in winter. According to Padisak's theory, all phytoplankton were divided into 25 functional groups, of which there were six important functional groups in both summer and winter:L0, H1, D, Y, MP, and P in summer and L0, H1, A, M, MP, and Y in winter. The α-diversity of the phytoplankton functional group in summer was higher than that in winter. Moreover, a higher α-diversity was also found in downstream samples relative to that in upstream samples, indicating that the community structure was more complex, and the community stability was relatively better in downstream regions of the rivers. Redundancy analysis (RDA) showed that the environment factors, i.e., ν, pH, permanganate index, WT, and RUETN, significantly affected phytoplankton functional groups (P<0.05). Variance partitioning analysis (VPA) indicated that environmental factors had a higher explanatory degree for the change in functional group composition in summer (45.23%); on the contrary, resource use efficiency had a higher explanatory degree in winter (42.33%). The linear fitted model showed that functional groups L0, H1, D, and Y showed a significant positive correlation relationship with RUETN and RUETP in summer, whereas only four functional groups (M, MP, Y, and A) had a linear relationship with RUETP, and all function groups had a good linear relationship with RUETN in winter. These results indicated that the functional groups belonging to cyanobacteria, dinoflagellates, and cryptophyta were more efficient at using limited resources in summer, whereas the diatoms had a good linear relationship with resource use efficiency and formed a dominant group in the low temperature environment of winter. These results suggest that the impounding of the Three Gorges Reservoir area can significantly change the resource use efficiency of phytoplankton, resulting in changes in the phytoplankton functional group composition and community structure.

Efficacy Evaluation of the Mahuang-Fuzi-Xixin Decoction in Combination with Shenmai Injection for Bradyarrhythmia Treatment: A Systematic Review and Meta-Analysis.

Background: Bradyarrhythmia treatment is often not timely enough, posing a potential threat to health. It is necessary to find a strategy with stable curative effects and high safety. Mahuang-Fuzi-Xixin (MFX) decoction and Shenmai injection (SMI) are compound Chinese Patent Medicines. Recent evidence has shown that the combined use of these two drugs can effectively treat bradyarrhythmia. Purpose: To evaluate the effect of MFX decoction combined with SMI on bradyarrhythmia. Methods: PRISMA was followed as the guideline for this systematic review. RevMan 5.4 software was applied for meta-analysis. Results: Eight studies were included in the analysis, with a total of 340 patients in the intervention and control groups. The results of the meta-analysis showed that the effective rate of MFX combined with SMI treatment was higher than that of SMI alone (OR = 3.27, 95% CI (2.18, 4.89), P < 0.01); after treatment, MFX combined with SMI treatment and SMI alone had no significant difference in heart rate. In the subgroup of patients with an age less than 60, the effect of MFX combined with SMI treatment on the 24-hour mean heart rate was better than that of SMI alone (MD = 3.68, 95% CI (3.14, 4.22), P < 0.01), as did the 24-hour minimal heart rate (MD = 3.48, 95% CI (3.03, 3.93), P < 0.01). In addition, the effect of MFX combined with SMI treatment on the Traditional Chinese Medicine syndrome score (TCMSS) was significantly better than that of SMI alone (MD = -2.69, 95% CI (-3.10, -2.28), P < 0.01). In terms of safety, two adverse events were reported in the SMI combined with MFX group compared to 12 in the SMI alone group. Conclusions: MFX combined with SMI treatment is effective in treating bradyarrhythmia. However, the results were heterogeneous. The safety of MFX combined with SMI treatment should be verified and treated with caution.

Investigating the effect of Shenmai injection on cardiac electrophysiology and calcium signaling using human-induced pluripotent stem cell-derived cardiomyocytes.

Traditional Chinese medicine injection (TCMI) refers to the use of modern technology to make Chinese patent medicines in injectable forms, which shorten the onset time of the traditional Chinese medicine (TCM). Although there have been clinical cases in which Shenmai injection (SMI) was used to treat cardiovascular diseases (CVDs), there are no pharmacological experiments that investigate the efficacy of the drug in vitro or the underlying mechanisms. Aim of the study: We aimed to systemically evaluate the efficacy and investigate the mechanisms of SMI in modulating electrophysiology and calcium (Ca2+) signaling using a microelectrode array (MEA) and a genetically encoded Ca2+ indicator, GCaMP6s, respectively, in human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Materials and methods: A MEA system was employed to record field potentials (FPs) in hiPSC-CMs. The QT interval is corrected by the RR interval, the reciprocal of the beating rate. GCaMP6s was used to measure Ca2+ signaling in hiPSC-CMs. Meanwhile, the transcriptome changes in hiPSC-CMs treated with 2% SMI were examined using RNAseq. In addition, the ingredients of SMI were investigated using liquid chromatography-mass spectrometry (LC-MS). Results: It was found that 0.5%, 1%, and 2% (v/v) SMIs could increase corrected QT (QTc) but did not change other FP parameters. GCaMP6s was successfully applied to measure the chronic function of SMI. The full width at half maximum (FWHM), rise time, and decay time significantly decreased after treatment with SMI for 1 h and 24 h, whereas an increased Ca2+ transient frequency was observed. Conclusions: We first used the Ca2+ indicator to measure the chronic effects of TCM. We found that SMI treatment can modulate electrophysiology and calcium signaling and regulate oxidative phosphorylation, cardiac muscle contraction, and the cell cycle pathway in hiPSC-CMs.

Alantolactone Inhibits Melanoma Cell Culture Viability and Migration and Promotes Apoptosis by Inhibiting Wnt/ß-Catenin Signaling.

BACKGROUND: Melanoma is a highly invasive and metastatic malignant tumor originating from melanocytes and is associated with a poor prognosis. Surgical resection and chemotherapy are currently the main therapeutic options for malignant melanoma; however, their efficacy is poor, highlighting the need for the development of new, safe, and effective drugs for the treatment of this cancer. OBJECTIVE: To investigate the effects of alantolactone (ALT) on the proliferative, migratory, invasive, and apoptotic ability of malignant melanoma cells and explore its potential anticancer mechanism. METHODS: Melanoma cells (A375 and B16) were treated with different concentrations (4, 6, 8, and 10 µmol/L) of ALT, with DMSO and no treatment serving as controls. The effects of the different concentrations of the drug on cell proliferation were assessed by crystal violet staining and CCK-8 assay. The effects on cell migration and invasion were detected by wound healing and Transwell assays, respectively. Flow cytometry was used to evaluate the effects of the drug on apoptosis and the cell cycle. ALT target genes in melanoma were screened using network pharmacology. Western blotting was used to measure the expression levels of the proliferation-related protein PCNA; the apoptosisrelated proteins Bax, Bcl-2, and caspase-3; the invasion and metastasis-related proteins MMP-2, MMP-7, MMP-9, vimentin, E-cadherin, and N-cadherin; and the canonical Wnt signaling pathway-related proteins ß-catenin, c-Myc, and p-GSK3ß. In addition, an l model of melanoma was established by the subcutaneous injection of A375 melanoma cells into nude mice, following which the effects of ALT treatment on malignant melanoma were determined in vivo. RESULTS: Compared with the controls, the proliferative, migratory, and invasive capacity of ALT-treated melanoma cells was significantly inhibited, whereas apoptosis was enhanced (P<0.01), showing effects that were exerted in a dose-dependent manner. The expression levels of the pro-apoptotic proteins Bax and caspase-3, as well as those of the interstitial marker E-cadherin, were upregulated in melanoma cells irrespective of the ALT concentration (P<0.05). In contrast, the expression levels of the anti-apoptotic protein Bcl-2, the proliferation-related protein PCNA, and the invasion and metastasis-related proteins MMP-2, MMP-7, MMP-9, N-cadherin, and vimentin were downregulated (P<0.05). The network pharmacology results indicated that GSK3ß may be a key ALT target in melanoma. Meanwhile, western blotting assays showed that ALT treatment markedly suppressed the expression of ß-catenin as well as that of its downstream effector c-Myc, and could also inhibit GSK3ß phosphorylation. CONCLUSION: ALT can effectively inhibit the culture viability, migration, and invasion of A375 and B16 melanoma cells while also promoting their apoptosis. ALT may exert its anti-melanoma effects by inhibiting the Wnt/ß-catenin signaling pathway. Combined, our data indicate that ALT has the potential as an effective and safe therapeutic drug for the treatment of melanoma.

Iron quantitative analysis of motor combined with bulbar region in M1 cortex may improve diagnosis performance in ALS.

OBJECTIVES: To explore whether the combined analysis of motor and bulbar region of M1 on susceptibility-weighted imaging (SWI) can be a valid biomarker for amyotrophic lateral sclerosis (ALS). METHODS: Thirty-two non-demented ALS patients and 35 age- and gender-matched healthy controls (HC) were retrospectively recruited. SWI and 3D-T1-MPRAGE images were obtained from all individuals using a 3.0-T MRI scan. The bilateral posterior band of M1 was manually delineated by three neuroradiologists on phase images and subdivided into the motor and bulbar regions. We compared the phase values in two groups and performed a stratification analysis (ALSFRS-R score, duration, disease progression rate, and onset). Receiver operating characteristic (ROC) curves were also constructed. RESULTS: ALS group showed significantly increased phase values in M1 and the two subregions than the HC group, on the all and elderly level (p < 0.001, respectively). On all-age level comparison, negative correlations were found between phase values of M1 and clinical score and duration (p < 0.05, respectively). Similar associations were found in the motor region (p < 0.05, respectively). On both the total (p < 0.01) and elderly (p < 0.05) levels, there were positive relationships between disease progression rate and M1 phase values. In comparing ROC curves, the entire M1 showed the best diagnostic performance. CONCLUSIONS: Combining motor and bulbar analyses as an integral M1 region on SWI can improve ALS diagnosis performance, especially in the elderly. The phase value could be a valuable biomarker for ALS evaluation. KEY POINTS: • Integrated analysis of the motor and bulbar as an entire M1 region on SWI can improve the diagnosis performance in ALS. • Quantitative analysis of iron deposition by SWI measurement helps the clinical evaluation, especially for the elderly patients. • Phase value, when combined with the disease progression rate, could be a valuable biomarker for ALS.

Qiliqiangxin Capsule Modulates Calcium Transients and Calcium Sparks in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.

Background: The therapeutic effects of Qiliqiangxin capsule (QLQX), a Chinese patent medicine, in patients with chronic heart failure are well established. However, whether QLQX modulates cardiac calcium (Ca2+) signals, which are crucial for the heart function, remains unclear. Aim of the Study. This study aimed to evaluate the role of QLQX in modulating Ca2+ signals in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Materials and Methods: Fluorescence imaging was used to monitor Ca2+ signals in the cytosol and nuclei of hiPSC-CMs. For Ca2+ spark measurements, the line-scan mode of a confocal microscope was used. Results: The QLQX treatment substantially decreased the frequency of spontaneous Ca2+ transients, whereas the amplitude of Ca2+ transients elicited by electrical stimulation did not change. QLQX increased the Ca2+ spark frequency in both the cytosol and nuclei without changing the sarcoplasmic reticulum Ca2+ content. Interestingly, QLQX ameliorated abnormal Ca2+ transients in CMs differentiated from hiPSCs derived from patients with long-QT syndrome. Conclusions: Our findings provide the first line of evidence that QLQX directly modulates cardiac Ca2+ signals in a human cardiomyocyte model.

Selenium Nanoparticles Improved Intestinal Health Through Modulation of the NLRP3 Signaling Pathway.

Selenium nanoparticles (SeNPs) play important roles in promoting animal health, however, their impact on intestinal health remains elusive. This study was intended to evaluate the effects of different doses of SeNPs on the intestinal health, especially the development of goblet cells in the broiler jejunum. A total of 480 1-day-old Arbor Acres broilers were randomly allotted to 5 treatments with 6 replications of 16 chicks each. Birds were fed with low selenium corn-soybean meal-based diets supplemented with 0.1, 0.2, 0.3, or 0.4 mg/kg of SeNPs. On d 21, dietary supplementation of SeNPs effectively reduced the mortality of broilers. The villus height and the villus height/crypt depth ratio of the jejunum showed significant quadratic effects with the increasing concentration of SeNPs (P < 0.05). The mRNA expression of zonula occluden-1 (ZO-1), ZO-2, claudin-3, and claudin-5 in the jejunum decreased linearly with the increasing dose of SeNPs (P < 0.05). The mRNA expression levels of interleukin 1 beta (IL-1ß), IL-18, and the concentration of reactive oxygen species (ROS) in the jejunum decreased linearly with the increase of SeNPs concentration (P < 0.05). Compared with the control group, the number of goblet cells in the jejunum was significantly increased by adding 0.1 and 0.4 mg/kg SeNPs(P < 0.05). In addition, the mRNA expression of Mucin2 (Muc2) showed a significant quadratic relationship that increased after adding 0.1 mg/kg SeNPs (P < 0.05). Dietary SeNPs also linearly reduced the expression of v-myc avian myelocytomatosis viral oncogene homolog (c-myc) (P < 0.05). The mean density of TUNEL positive cells in the 0.2 and 0.4 mg/kg SeNPs groups were lower than the control group (P < 0.05). Similarly, the mRNA expression levels of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), NLR family pyrin domain containing 3 (NLRP3), cysteinyl aspartate specific proteinase-1 (Caspase-1), toll-like receptor-2 (TLR-2), and myeloid differentiation factor 88 (MyD88) in the jejunum decreased linearly with the increase of SeNPs concentration (P < 0.05). Results show that supplementation with 0.2 mg/kg SeNPs may decrease intestinal oxidative stress and inflammation by modifying the activation of NLRP3 signaling pathway, which can effectively promote intestinal goblet cells of 21-day-old broilers.

Mechanism of Danhong Injection in the Treatment of Arrhythmia Based on Network Pharmacology, Molecular Docking, and In Vitro Experiments.

Background: Danhong injection (DHI) is widely used in the treatment of cardiovascular and cerebrovascular diseases, and its safety and effectiveness have been widely recognized and applied in China. However, the potential molecular mechanism of action for the treatment of arrhythmia is not fully understood. Aim: In this study, through network pharmacology and in vitro cell experiments, we explored the active compounds of DHI for the treatment of arrhythmia and predicted the potential targets of the drug to investigate its mechanism of action. Materials and Methods: First, the potential therapeutic effect of DHI on arrhythmia was investigated in an in vitro arrhythmia model using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), in which calcium transients were recorded to evaluate the status of arrhythmia. Next, the active compounds and key targets in the treatment of arrhythmia were identified through network pharmacology and molecular docking, and the key signaling pathways related to the treatment of arrhythmia were analyzed. Furthermore, we used real-time quantitative reverse transcription PCR (qRT-PCR) to verify the expression levels of key genes. Results: Early afterdepolarizations (EADs) were observed during aconitine treatment in hiPSC-CMs, and the proarrhythmic effect of aconitine was partially rescued by DHI, indicating that the antiarrhythmic role of DHI was verified in an in vitro human cardiomyocyte model. To further dissect the underlying molecular basis of this observation, network pharmacology analysis was performed, and the results showed that there were 108 crosstargets between DHI and arrhythmia. Moreover, 30 of these targets, such as AKT1 and HMOX1, were key genes. In addition, the mRNA expression of AKT1 and HMOX1 could be regulated by DHI. Conclusion: DHI can alleviate aconitine-induced arrhythmia in an in vitro model, presumably because of its multitarget regulatory mechanism. Key genes, such as AKT1 and HMOX1, may contribute to the antiarrhythmic role of DHI in the heart.