UHPLC-qTOF-MS-Based Nontargeted Metabolomics to Characterize the Effects of Capsaicin on Plasma and Skin Metabolic Profiles of C57BL/6 Mice-An In vivo Experimental Study.

Background: Capsaicin is the main compound found in chili pepper and has complex pharmacologic effects. This study aimed to elucidate the mechanism of the effect of capsaicin on physiological processes by analyzing changes in metabolites and metabolic pathways. Methods: Female C57BL/6 mice were divided into two groups(n = 10/group) and fed with capsaicin-soybean oil solution(group T) or soybean oil(group C) for 6 weeks. Ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC-qTOF-MS) based metabolomics was undertaken to assess plasma and skin metabolic profile changes and identify differential metabolites through multivariate analysis. Results: According to the OPLS-DA score plots, the plasma and skin metabolic profiles in the group T and group C were significantly separated. In plasma, 38 significant differential metabolites were identified. KEGG pathway enrichment analysis revealed that the most significant plasma metabolic pathways included pyruvate metabolism and ABC transporters. In skin, seven significant differential metabolites were found. Four metabolic pathways with p values < 0.05 were detected, including sphingolipid metabolism, sphingolipid signaling pathway, apoptosis, and necroptosis. Conclusion: These findings will provide metabolomic insights to assess the physiological functions of capsaicin and contribute to a better understanding of the potential effects of a capsaicin-rich diet on health.

Plant growth and heavy meal accumulation characteristics of Spathiphyllum kochii cultured in three soil extractions with and without silicate supplementation.

A hydroponic method was conducted to test whether Spathiphyllum kochii is tolerant to multiple HMs as well as to evaluate whether sodium silicate promotes plant growth and alleviates HM stress mainly by assessing biomass, HM accumulation characteristics and antioxidant enzyme activities (AEAs). Three soil extractions from an uncontaminated soil, a comparable lightly HM-contaminated soil (EnSE), and a comparable heavily HM-contaminated soil (ExSE) with or without 1 mM sodium silicate supplementation were used. S. kochii showed no obvious symptoms when cultured in EnSE and ExSE, indicating that it was a multi-HM-tolerant species. The biomass and photosynthesis followed the order: UnSE > EnSE > ExSE, but the opposite order was found for HM concentration, AEAs, and malondialdehyde content. Silicate had no effects on the growth and HM bioaccumulation characteristics of S. kochii cultured in UnSE but exhibited a novel role in decreasing HM uptake by 13.61-41.51% in EnSE and ExSE, respectively, corresponding upregulated AEAs, and reduced malondialdehyde contents, resulting in increased biomass and alleviating HM stress. The activities of peroxidase and superoxide dismutase were upregulated by an increase in soil extraction HM concentration and further upregulated by silicate supplementation, indicating that they were important mechanisms alleviating HM stress in S. kochii.

Phytoremediation is an economical and environmentally friendly technology for the alleviation of heavy metal (HM)-contaminated soil. Improving bioremediation efficiency is crucial for this kind of technology. Many studies have shown that silicon plays a novel role in plant growth and adversity responses, but studies in the field of phytoremediation are limited. In addition, phytoremediation plant species are usually hyperaccumulators or may be tolerant crops, commercial crops, or wild species from mining areas, and the use of landscape species in phytoremediation is limited. This is the first report on the effects of silicate on the multi-HM bioaccumulation characteristics of a garden plant (Spathiphyllum kochii) cultured in uncontaminated and HM-contaminated soil extractions. This study will broaden phytoremediation species screening and enrich our understanding of the mechanisms by which Si supports the bioremediation of HM-contaminated environments.HIGHLIGHTSS. kochii was a multi-heavy metal-tolerant species.Silicon played a novel role in reducing heavy metal concentrations by 14­40% and 14­42% in shoots and roots, respectively.Silicon upregulated antioxidant enzyme activities to alleviate heavy metal stress in plants.

Emodin regulates the autophagy via the miR-371a-5p/PTEN axis to inhibit hepatic malignancy.

Emodin has been reported to fulfill an important function in suppressing the vicious outcome of liver cancer. We aimed to elucidate the partial underlying molecular mechanism of emodin in inhibiting liver cancer, and we applied miRNA-sequence analysis and corresponding molecular functional experiments to find that the inhibitory effect of emodin on liver cancer was partly mediated by cellular autophagy through the miR-371a-5p/PTEN axis. The expression level of miR-371a-5p was down-regulated after emodin treatment in liver cancer cell lines (LCCLs). Restoring the expression level of miR-371a-5p attenuated the suppression of emodin on LCCLs. Additionally, we performed the prediction in relevant online databases and found that PTEN might functioned as a downstream target of miR-371a-5p to participate in the regulation on the above process. What's more, the detection of autophagy-related protein markers showed that LC3II was elevated accompanied by the decreased P62. The above results revealed that PTEN functioned as a key target to regulate the autophagy in the process where emodin inhibited the malignant outcome of LCCLs via miR-371a-5p, which further provided a theoretical basis for the application of traditional Chinese medicine (TCM) on clinical tumors.

Association between serum vitamin A levels and premature ovarian insufficiency: a case-control, cross-sectional survey study.

BACKGROUND: Although vitamin A is known to play an important role in ovarian function, its association with ovarian insufficiency has not been reported yet. Therefore, the aim of the study was to explore the association between serum vitamin A levels and premature ovarian insufficiency (POI). METHODS: This cross-sectional survey included women with POI (n = 47) and normo-ovulatory controls (n = 67) who were enrolled between December 2016 and May 2018 in Zhejiang, China. The serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), anti-Müllerian hormone (AMH), vitamin A, and total cholesterol (TC) were measured for each participant. The association of TC-adjusted vitamin A levels with the risk of POI was assessed using binary logistic regression analysis. RESULTS: Serum vitamin A levels appeared to be slightly higher in the POI group than in the control group, but there was no evidence of a statistically significant difference (728.00 ± 176.00 µg/L vs. 503.93 ± 145.64 µg/L, p = 0.13). After adjustment for serum lipid levels, the serum vitamin A/TC ratio was significantly lower in the POI group than in the control group (143.14 ± 35.86 vs. 157.56 ± 35.21 µg/mmol, p = 0.04). Further, the serum vitamin A/TC ratio was significantly and inversely associated with POI risk (unadjusted odds ratio [OR] = 0.988, 95% confidence interval [CI]: 0.977-0.999, p = 0.04). The association remained after adjusting for confounding factors (age, BMI, annual household income, and education) (OR = 0.986, 95% CI: 0.972-0.999, p = 0.04). CONCLUSIONS: Serum vitamin A/TC ratio was inversely associated with POI risk. Therefore, the serum vitamin A/TC ratio may serve as a predictive factor for POI, and vitamin A supplementation may play help prevent or treat POI.

Heavy metal pollution in coastal wetlands: A systematic review of studies globally over the past three decades.

Coastal wetlands are ecosystems lying between land and ocean and are subject to inputs of heavy metals (HMs) from terrestrial, oceanic and atmospheric sources. Although the study on HM pollution in coastal wetlands has been rapidly developing over the past three decades, systematic reviews are still unavailable. Here, by analyzing 3343 articles published between 1990 and 2019, we provided the first holistic systematic review of studies on HM pollution in coastal wetlands globally. The results showed a trend of rapid increases in publications in this field globally, especially over the past ten years. Trends varied greatly among coastal countries, and global trends were primarily driven by the US before 2000, and in China after 2010. We also found that mercury (Hg), cadmium (Cd), and copper (Cu) were the most widely studied HM elements globally, but patterns differed geographically, with Hg being most widely examined in the Americas, Cd in China and India, and lead (Pb) in the western Europe and Australia, respectively. Among different types of coastal wetlands, salt marshes, mangrove forests, and estuaries were the most widely studied, in contrast to seagrass beds and tidal flats. As for ecosystem components, soils/sediments and plants were most extensively investigated, while algae, microbes, and animals were much less examined. Our analysis further revealed rapid emergence of topics on anthropogenic sources, interactions with other anthropogenic environmental changes (climate change in particular), and control and remediation methodology in the literature in the recent ten years. Moving forward, we highlight that future studies are needed to i) better understand the impacts of HM pollution in less studied coastal wetland systems and species, ii) deepen current understanding of the biogeochemical behaviors of HMs under anthropogenic activities, iii) examine interactions with other anthropogenic environmental changes, iv) conceive ecological remediation (i.e., "ecoremediation" as compared to traditional physiochemical remediation and bioremediation) strategies, and v) develop advanced analysis instruments and methods. The perspectives we brought forward can help stimulate many new advances in this field.

Epiberberine ameliorated diabetic nephropathy by inactivating the angiotensinogen (Agt) to repress TGFß/Smad2 pathway.

BACKGROUND: Diabetic nephropathy (DN) is a severe microvascular complication of diabetes with prominent morbidity and mortality. At present, there are hardly any effective drugs to treat DN. Epiberberine (EPI), an isoquinoline alkaloid, has attracted considerable attention due to its anti-hyperglycemic, anti-hyperlipidemic, and anti-inflammatory functions. However, whether there is a protective effect of EPI on DN has not been reported. PURPOSE: The research was aimed to investigate the activities of EPI alleviating kidney damage in db/db mice and to explore its possible mechanisms. STUDY DESIGN: The db/db mice and high-glucose (HG) induced glomerular mesangial cells (GMCs) were used to explore the protective effect of EPI on DN in vivo and in vitro. METHODS: The changes in fasting blood glucose, metabolic index, renal function, and histopathological morphology in db/db mice were detected to evaluate the therapeutic effect of EPI. Then, renal transcriptome and molecular docking were used to screen the key targets. Subsequently, HG-induced GMCs through mimicing the pathological changes in DN were utilized to study the renal protective effects of EPI and its potential mechanism. RESULTS: The results in vivo showed that EPI administration for 8 weeks significantly alleviated diabetes-related metabolic disorders, improved renal functions, and relieved the histopathological abnormalities of renal tissue, especially renal fibrosis in db/db mice. The results in vitro showed that EPI inhibited the proliferation and induced the G2/M phase arrest of HG-induced GMCs. Moreover, a key gene Angiotensinogen (Agt) was screen out by the RNA-seq of kidney and molecular docking, and EPI reduced Agt, TGFß1, and Smad2 expression in vitro and in vivo. Noteworthy, Agt knockdown by siRNA significantly attenuated these beneficial efficacies exerted by EPI, indicating that Agt played a crucial role in the process of EPI improving DN. CONCLUSION: These findings suggested that EPI might be a potential drug for the treatment of DN dependent on the Agt-TGFß/Smad2 pathway.

An integrated network pharmacology and transcriptomic method to explore the mechanism of the total Rhizoma Coptidis alkaloids in improving diabetic nephropathy.

ETHNOPHARMACOLOGICAL RELEVANCE: Rhizoma Coptidis (RC) is a traditional Chinese medicine (TCM) used for treating diabetes (Xiao Ke Zheng), which is firstly recorded in Shennong Bencao Jing. Modern pharmacological studies have confirmed that RC has beneficial effects on diabetes and its complications. Alkaloids are the main active pharmacological component of RC. However, the effect and molecular mechanism of total Rhizoma Coptidis alkaloids (TRCA) in improving diabetic nephropathy (DN) are still unclear. AIM OF THE STUDY: To verify the effect of TRCA in the treatment of DN and clarify the molecular mechanism by combining network pharmacology and transcriptomic. MATERIALS AND METHODS: Eight-week-old db/db mice were orally administered with normal saline, 100 mg/kg TRCA, and 100 mg/kg berberine (BBR) for 8 weeks. Serum, urine, and kidney samples were collected to measure biological indicators and observe renal pathological changes. Then, the molecular mechanism of TRCA improving DN was predicted by the network pharmacology. Briefly, the main active alkaloids components of TRCA and their targets were collected from the database, as well as the potential targets of DN. Using the Cytoscape software to visualize the interactive network diagram of "ingredient-target". The GO and KEGG pathways enrichment analysis of the core targets were executed by Metascape. Furthermore, RNA-seq was used to get whole transcriptomes from the kidneys of db/m mice, db/db mice, and db/db mice treated with TRCA. The key differentially expressed genes (DEGs) were gathered to conduct the GO and KEGG pathways enrichment analysis. Finally, the potential pathways were validated by western blotting. RESULTS: The administration of BBR or TRCA for 8 weeks significantly reduced the fasting blood glucose (FBG) and body weight of db/db mice, and improved their renal function and lipid disorders. According to H&E, PAS, and Masson staining, both the BBR and TRCA could alleviate renal damage and fibrosis. The Venn diagram had shown that seven alkaloids ingredients collected from TRCA regulated 85 common targets merged in the TRCA and DN. The results of RNA-seq indicated that there are 121 potential targets for TRCA treatment on DN. Intriguingly, both the AGE-RAGE signaling pathway and the PI3k-Akt signaling pathway were included in the KEGG pathways enrichment results of network pharmacology and RNA-seq. Moreover, we verified that TRCA down-regulated the expression of related proteins in the AGEs-RAGE-TGFß/Smad2 and PI3K-Akt pathways in the kidney tissues. CONCLUSIONS: In summary, the renal protection of TRCA on DN may be related to activation of the AGEs-RAGE-TGFß/Smad2 and PI3K-Akt signaling pathways.

Hydrocotyle vulgaris L.: a new cadmium-tolerant landscape species and its physiological responses to cadmium exposure.

Landscape plants have both ecological and aesthetic value and may also represent ideal candidates for phytoremediation. In the present study, one round of hydroponic culture for 14 days with different cadmium (Cd) concentrations (0, 0.5, 1, and 2 mg L-1 Cd) was carried out to test whether Hydrocotyle vulgaris L. is a Cd-tolerant plant. Furthermore, physiological parameters, including pigment concentrations, photosynthesis, antioxidant enzyme activities (AEAs), and nutrient uptake, were also examined to determine the tolerance of H. vulgaris to Cd exposure. The results showed that H. vulgaris could grow normally under all Cd supply levels. The Cd removal efficiency reached 100% at Cd concentrations ≤1.0 mg L-1. The concentrations of Cd in roots and shoots increased (P < 0.05) with Cd supplementation. The maximum concentrations of Cd reached 26.4 and 118 mg kg-1 in shoots and roots, respectively. The translocation factor values were similar under all Cd treatments. The highest mean daily increase in biomass (MDIB) was obtained under 1 mg L-1 Cd exposure, which increased by 69.86% compared to that in the control, which may be due to the increased photosynthetic pigments, photosynthetic rate, and the consistent nutrient concentrations under this Cd level, as there were positive relationships between these parameters and MDIB. Moreover, the activities of AEA also generally explicated highest among all Cd levels. All these results indicate that the above physiological parameters play a positive role in promoting plant growth and alleviating Cd stress. In summary, H. vulgaris was verified as a potential Cd-tolerant plant, providing new information for Cd phytoremediation. Furthermore, given its extensive habitat distribution, this species might be tested for phytoremediation of contaminated soils in future work.

Effect of epiberberine from Coptis chinensis Franch on inhibition of tumor growth in MKN-45 xenograft mice.

BACKGROUND AND PURPOSE: Gastric cancer is one of the major malignancies worldwide. Epiberberine (EPI) is a major alkaloid from Coptis chinensis Franch and the antitumor property of EPI remains poorly understood. METHOD: The inhibition on gastric cancer cells was observed by MTT assays and colony formation experiments. The apoptosis, cell cycle, and reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) in gastric cancer cells were analyzed by Flow cytometry. The anti-tumor effect of EPI was evaluated with the MKN-45-beraring nude mice, and the potential mechanisms were explored by RNA-seq, qPCR, siRNA silencing and western blotting. RESULTS: EPI inhibited the proliferation of human gastric cancer cell lines MKN-45 (harboring wild-type p53) and HGC-27 (harboring mutant p53) in a dose dependent manner. EPI induced the apoptosis and cell cycle arrest in these two cell lines, of which MKN-45 cells are more sensitive to EPI than HGC-27 cells. Further experiments indicated that EPI induced the accumulation of ROS and decreased of ΔΨm in MKN-45 cells. The significant differentially expressed genes obtained by RNA-seq were distinctly enriched in the p53 signaling pathway. The apoptosis induced by EPI in MKN-45 cells would be effectively inhibited with the treatment of p53 siRNA and p53 inhibitor PFT-α. Western blotting demonstrated that EPI diminished the expression of Bcl-2 and XIAP, and increased those of p53, Bax, p21, p27, Cytochrome C and Cleaved-caspase 3. Animal experiments confirmed that EPI significantly alleviated tumor growth in MKN-45 xenograft mice via p53/Bax pathway. CONCLUSIONS: These data indicated that EPI could be a novel anti-tumor candidate against MKN-45-related gastric cancer via targeting p53-dependent mitochondria-associated pathway.

Comparative analysis of the seed germination of pakchoi and its phytoremediation efficacy combined with chemical amendment in four polluted soils.

The seed germination plant growth parameters and level of heavy metal accumulation were investigated in pakchoi cultured in four contaminated soils with different levels of heavy metals supplemented with citric acid (CA) or calcium phosphate (CP). Results showed that the seed germination energy, germination percentage and germination index parameters were similar, while the seed vigor (SV) significantly (p < 0.05) decreased as the soil pollution level increased. The lengths of the shoots and roots presented the same trend as SV. All the seedlings grew in heavily polluted soil without any amendments before harvesting; therefore, no plant material was available for subsequent analyses. The photosynthesis parameters of pakchoi cultured in lightly polluted soil without amendment (LPS), lightly polluted soil with CA (LPSA) and moderately polluted soil with CP (MPSP) were similar. The concentrations of Pb, Zn, Mn, Cu and Cd in the shoots, roots and whole plants were in the order of MPSP > LPSA > LPS. Pakchoi cultured in MPSP showed the most promising results in terms of plant height, biomass and heavy metal accumulation. Pakchoi presented the highest translocation and bioaccumulation factors for Cd and the lowest for Pb.HighlightsSoil pollution and the type of chemical amendment had no effect on the seed germination of pakchoi.Citric acid addition in lightly polluted soil improved pakchoi growth and heavy metal extraction.Pakchoi cultured in moderately polluted soil with calcium phosphate amendment presented the highest biomass and heavy metal concentration.

A54 peptide-mediated functionalized gold nanocages for targeted delivery of DOX as a combinational photothermal-chemotherapy for liver cancer.

The combination of photothermal therapy and chemotherapy (photothermal-chemotherapy) is a promising strategy for cancer therapy. Gold nanocages (AuNCs), with hollow and porous structures and unique optical properties, have become a rising star in the field of drug delivery. Here, we designed a novel targeted drug delivery system based on functionalized AuNCs and evaluated their therapeutic effects in vitro and in vivo. We then loaded doxorubicin into this promising system, designated as DHTPAuNCs consisting of hyaluronic acid-grafted and A54 peptide-targeted PEGylated AuNCs. Its formation was corroborated by ultraviolet-visible spectroscopy, transmission electron microscopy and dynamic light scattering. This delivery platform needed hyaluronidase to release encapsulated drugs, meanwhile the acidic pH and near-infrared irradiation could accelerate the release. In addition, the results of cellular uptake demonstrate that this system could bind specifically with BEL-7402 cells. In vitro, we evaluated therapeutic effects of the DHTPAuNCs in BEL-7402 cells by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide assay. Moreover, in BEL-7402 tumor-bearing nude mice, its therapy effect in vivo was also evaluated. As expected, DHTPAuNCs exhibited excellent therapeutic effect by photothermal-chemotherapy, both in vitro and in vivo. In short, DHTPAuNCs with low toxicity showed great potential as a drug delivery system for cancer therapy.

Electroacupuncture Ameliorates Cerebral Ischemia-Reperfusion Injury by Regulation of Autophagy and Apoptosis.

Background. The therapeutic mechanisms of cerebral ischemia treatment by acupuncture are yet not well addressed. Objective. We investigated the effects of electroacupuncture (EA) at GV26 observing the expression of autophagy-related proteins Beclin-1 and LC3B and proportion of apoptotic cells and Bcl-2 positive cells in MCAO/R model rats. Methods. Sprague-Dawley (SD) male rats were randomly assigned to 7 groups: model groups (M6h, M24h, and M72h), EA treatment groups (T6h, T24h, and T72h), and sham operation group (S). Neurological deficit and cerebral infarction volume were measured to assess the improvement effect, while the expression of Beclin-1 and LC3B and proportion of Tunel-positive and Bcl-2 positive cells were examined to explore EA effect on autophagy and apoptosis. Results. EA significantly decreased neurological deficit scores and the volume of cerebral infarction. Beclin-1 was significantly decreased in T24h, while LC3B-II/LC3B-I ratio markedly reduced in 6th hour. EA groups markedly reduced the number of Tunel positive cells, especially in T24h. Meanwhile, the number of Bcl-2 positive cells obviously increased after EA treatment, especially in T6h and T24h. Conclusions. The alleviation of inadequate autophagy and apoptosis may be a key mechanism involved in the reflex regulation of EA at GV26 to treat cerebral ischemia.

Wrist-ankle acupuncture (WAA) for precompetition nervous syndrome: study protocol for a randomized controlled trial.

BACKGROUND: Precompetition nervous syndrome comprises an excessive nervous and anxiety response to the high-pressure environment preceding a sporting competition. The use of acupuncture as a treatment option for anxiety, and wrist-ankle acupuncture (WAA) specifically in this instance, has been identified as a growing trend within the Western world. In our previous study, we have confirmed the efficacy of WAA for pre-examination anxiety. In this paper, we present a randomized controlled single-blind trial evaluating the use of WAA for precompetition nervous syndrome, comparing it with the intervention of sham acupuncture. METHODS/DESIGN: The study was designed as a randomized controlled single-blind trial to evaluate the effects of WAA for precompetition anxiety. The trial will be conducted in annual track and field events of Shanghai University of Sport. A total of 100 participants who meet inclusion criteria are randomly assigned by computerized randomization to receive WAA therapy or sham acupuncture. The group allocations and interventions are concealed to participants and statisticians. The Competition State Anxiety Scale (CSAI-2) is used as the primary outcome measure, while heart rate, blood pressure, respiratory frequency, tension syndrome curative effect evaluation and participants' feeling of acupuncture questionnaire are applied as secondary outcome measures. DISCUSSION: The results of this trial will confirm whether WAA is effective to treat precompetition anxiety in annual track and field events. TRIAL REGISTRATION: Chinese Clinical Trial Registry (identifier: ChiCTR-TRC-13003931; registration date: 22 October 2013).

A level set method for cupping artifact correction in cone-beam CT.

PURPOSE: To reduce cupping artifacts and improve the contrast-to-noise ratio in cone-beam computed tomography (CBCT). METHODS: A level set method is proposed to reduce cupping artifacts in the reconstructed image of CBCT. The authors derive a local intensity clustering property of the CBCT image and define a local clustering criterion function of the image intensities in a neighborhood of each point. This criterion function defines an energy in terms of the level set functions, which represent a segmentation result and the cupping artifacts. The cupping artifacts are estimated as a result of minimizing this energy. RESULTS: The cupping artifacts in CBCT are reduced by an average of 90%. The results indicate that the level set-based algorithm is practical and effective for reducing the cupping artifacts and preserving the quality of the reconstructed image. CONCLUSIONS: The proposed method focuses on the reconstructed image without requiring any additional physical equipment, is easily implemented, and provides cupping correction through a single-scan acquisition. The experimental results demonstrate that the proposed method successfully reduces the cupping artifacts.