Quantitative Analysis of Bioactive Compounds in Commercial Teas: Profiling Catechin Alkaloids, Phenolic Acids, and Flavonols Using Targeted Statistical Approaches.

Tea, an extensively consumed and globally popular beverage, has diverse chemical compositions that ascertain its quality and categorization. In this investigation, we formulated an analytical and quantification approach employing reversed-phase ultra-high-performance liquid chromatography (UHPLC) methodology coupled with diode-array detection (DAD) to precisely quantify 20 principal constituents within 121 tea samples spanning 6 distinct variants. The constituents include alkaloids, catechins, flavonols, and phenolic acids. Our findings delineate that the variances in chemical constitution across dissimilar tea types predominantly hinge upon the intricacies of their processing protocols. Notably, green and yellow teas evinced elevated concentrations of total chemical moieties vis à vis other tea classifications. Remarkably divergent levels of alkaloids, catechins, flavonols, and phenolic acids were ascertained among the disparate tea classifications. By leveraging random forest analysis, we ascertained gallocatechin, epigallocatechin gallate, and epicatechin gallate as pivotal biomarkers for effective tea classification within the principal cadre of tea catechins. Our outcomes distinctly underscore substantial dissimilarities in the specific compounds inherent to varying tea categories, as ascertained via the devised and duly validated approach. The implications of this compositional elucidation serve as a pertinent benchmark for the comprehensive assessment and classification of tea specimens.

Condition optimization of iron-air fuel cell to treat phosphate-containing wastewater regarding sustainable development.

Increasing use of phosphorus products and excessive exploitation of phosphorus resources become two major problems in perspective of phosphorus sustainable development. Phosphorus recovery is the shortcut to solve this dilemma. Combining electrochemistry, an iron-air fuel cell was adopted to recover phosphate and electricity from phosphate-containing wastewater in our previous studies. The present study focused on investigating the effects of catholyte/anolyte conductivity, external resistance, and anolyte pH on the performance of iron-air fuel cell, and obtaining the optimized conditions. Furthermore, the electrochemical methods of phosphate recovery were compared and assessed, and it is concluded that iron-air fuel cell has great potential for energy recovery. The phosphate removal efficiencies and vivianite yield roughly positively correlated with the catholyte conductivity and the anolyte pH, but negatively correlated with the external resistance and the anolyte conductivity. The electricity generation roughly positively correlated with the catholyte conductivity and anolyte conductivity, but showed limitations in the test range of anolyte pH and external resistance. To pursue high phosphate removal efficiencies and vivianite yield, the catholyte conductivity, external resistance, anolyte pH and anolyte conductivity were suggested to be 35 g-NaCl/L, 10 Ω, 8 and 0 g-NaCl/L. While if electricity generation was the primary goal, these parameters should be 35 g-NaCl/L, 220 Ω, 5 and 70 g-NaCl/L. The optimized conditions will help to improve the phosphate removal efficiency, vivianite yield and electricity generation, and to promote the development of iron-air fuel cell technology.

Recovering phosphate and energy from anaerobic sludge digested wastewater with iron-air fuel cells: Two-chamber cell versus one-chamber cell.

Anaerobic sludge digested (ASD) wastewater is widespread in wastewater treatment plants. Recovering phosphate from ASD wastewater not only removes pollutants but also solves the phosphorus deficiency problem. Iron-air fuel cells were chosen to recover phosphate and generate electricity from ASD wastewater. To optimize cell configuration, a two-chamber and a one-chamber iron-air fuel cell were set up. The phosphate removal efficiency, the vivianite yield and the electricity generation efficiency of the two fuel cells were evaluated. It turned out that the volumetric removal rate (VRR) of phosphate of the two-chamber cell was 11.60 mg P·L-1·h-1, which was about five times of that in the one-chamber cell. The phosphate recovery product vivianite was detected on the surface of the iron anodes and the calculated purities of the two-chamber fuel cell and one-chamber fuel cell were 90.6% and 58.7%, respectively. Considering the content and purity, the iron anode surface in the two-chamber fuel cell was the best point to recover phosphate. The proton exchange membrane (PEM) in the two-chamber fuel cell provided low pH conditions suitable for vivianite formation. Moreover, under the low pH condition, metal ions of Fe2+, Ca2+, Al3+ and so on were kept soluble, leading to a high conductivity. The high conductivity caused low internal resistance, which benefited the electricity generation. The total output electric power of the two-chamber fuel cell was 2.4 times that of the one-chamber fuel cell when treating 25 mL ASD wastewater (0.62 vs. 0.26 mW·h). Overall, the two-chamber fuel cell was the better choice for phosphate recovery and electricity generation from ASD wastewater. Further studies on the long-term operation of two-chamber fuel cells should be carried out.

N6 -methyladenosine modification of circular RNA circ-ARL3 facilitates Hepatitis B virus-associated hepatocellular carcinoma via sponging miR-1305.

Hepatitis B virus (HBV) infection is a major risk factor for hepatocellular carcinoma (HCC), whether circular RNA (circRNA) is involved in this process remains unknown. In this study, we performed circRNA microarray profile and found an HBV-related circRNA, circ-ARL3 (hsa_circ_0092493). Stable knockdown of circ-ARL3 inhibited the proliferation and invasion of HBV+ HCC cells. High circ-ARL3 was positively correlated with malignant clinical features and poor prognosis. In terms of mechanism, HBx protein upregulated N6 -methyladenosine (m6 A) methyltransferases METTL3 expression, increasing the m6 A modification of circ-ARL3; then, m6 A reader YTHDC1 bound to m6 A-modified of circ-ARL3 and favored its reverse splicing and biogenesis. Furthermore, circ-ARL3 was able to sponge miR-1305, antagonizing the inhibitory effects of miR-1305 on a cohort of target oncogenes, thereby promoting HBV+ HCC progression. Importantly, depletion of circ-ARL3 significantly retarded HBV+ HCC cell growth in vivo, whereas this effect was evidently blocked after silencing of miR-1305. Collectively, our data suggest that circ-ARL3 is a critical regulator in HBV-related HCC, targeting the axis of circ-ARL3/miR-1305 may be a promising treatment for HBV+ HCC patients.

HMGB1 Inhibits HNF1A to Modulate Liver Fibrogenesis via p65/miR-146b Signaling.

High mobility group box 1 (HMGB1) is essential for the pathogenesis of liver injury and liver fibrosis. We previously revealed that miR-146b promotes hepatic stellate cells (HSCs) activation and proliferation. Nevertheless, the potential mechanisms are still unknown. Herein, HMGB1 increased HSCs proliferation and COL1A1 and α-SMA protein levels. However, the knockdown of miR-146b inhibited HSCs proliferation and COL1A1 and α-SMA protein levels induced via HMGB1 treatment. miR-146b was upregulated by HMGB1 and miR-146b targeted hepatocyte nuclear factor 1A (HNF1A) 3'-untranslated region (3'UTR) to modulate its expression negatively. Further, we confirmed that HMGB1 might elicit miR-146b expression via p65 within HSCs. Knockdown or block of HMGB1 relieved the CCl4-induced liver fibrosis. In fibrotic liver tissues, miR-146b expression was positively correlated with p65 mRNA, but HNF1A mRNA was inversely correlated with p65, and miR-146b expression. In summary, our findings suggest that HMGB1/p65/miR-146b/HNF1A signaling exerts a crucial effect on liver fibrogenesis via the regulation of HSC function.

miR-206 inhibits cell proliferation, invasion, and migration by down-regulating PTP1B in hepatocellular carcinoma.

Protein tyrosine phosphatase 1B (PTP1B) has been reported as an oncogene in hepatocellular carcinoma (HCC). However, how PTP1B is regulated in HCC remains unclear. MicroRNAs (miRNAs) are a class of small non-coding RNAs involved many biological processes including tumorigenesis. In this study, we investigated whether miRNA participated in the regulation of PTP1B in HCC. We found that miR-206, which was down-regulated during tumorigenesis, inhibited HCC cell proliferation and invasion. Overexpression of miR-206 inhibited proliferation, invasion, and migration of HCC cell lines HepG2 and Huh7. Mechanistically, we demonstrated that miR-206 directly targeted PTP1B by binding to the 3'-UTR of PTP1B mRNA as demonstrated by the luciferase reporter assay. Overexpression miR-206 inhibited PTP1B expression while miR-206 inhibition enhanced PTP1B expression in HepG2 and Huh7 cells. Functionally, the regulatory effect on cell proliferation/migration/invasion of miR-206 was reversed by PTP1B overexpression. Furthermore, tumor inoculation nude mice model was used to explore the function of miR-206 in vivo Our results showed that overexpression of miR-206 drastically inhibited tumor development. In summary, our data suggest that miR-206 inhibits HCC development by targeting PTP1B.

Role of growth factor receptor-bound 2 in CCl4-induced hepatic fibrosis.

BACKGROUND: Growth Factor Receptor-bound 2 (GRB2) plays a crucial role in regulation of cellular function including proliferation and differentiation, and we previously identified GRB2 as promoting HSCs (HSCs) proliferation. However, the underlying mechanisms that are involving in the regulation of GRB2 in hepatic fibrogenesis remain unknown. METHODS: In the present study, we tested the function of GRB2 in hepatic fibrosis. Hepatic fibrosis was induced by subcutaneous CCl4 administration at a dose of 3mL/kg in rats. The rat HSC cell line HSC-T6 were cultured for proliferation investigation by CCK-8 and BrdU incorporation method. The levels of GRB2, HMGB1, PI3K/AKT, COL1A1 and α-SMA were analyzed by western blot or real-time PCR. RESULTS: showed that the expression of GRB2 and HMGB1 was obviously increased in liver tissues of hepatic fibrosis rats accompanied by up-regulation of COL1A1 and α-SMA. In cultured HSCs, application of exogenous HMGB1 induced cell proliferation and cell proliferation rate concomitantly with up-regulation of GRB2 expression and PI3K/AKT phosphorylation. The effects of HMGB1-induced proliferation of HSCs and up-regulation of COL1A1 and α-SMA were abolished by GRB2 siRNA. HMGB1-induced proliferation of HSCs and up-regulation of COL1A1 and α-SMA was reversed in the presence of LY294002, an inhibitor of PI3K inhibitor. CONCLUSIONS: These findings suggest that GRB2 plays an important role in CCl4-induced hepatic fibrosis by regulating HSCs' function, and up-regulation of GRB2 induced by HMGB1 is mediated via the PI3K/AKT pathway.

Clinical Characteristics of Patients with Drug-induced Liver Injury.

BACKGROUND: Drug is an important cause of liver injury and accounts for up to 40% of instances of fulminant hepatic failure. Drug-induced liver injury (DILI) is increasing while the diagnosis becomes more difficult. Though many drugs may cause DILI, Chinese herbal medicines have recently emerged as a major cause due to their extensive use in China. We aimed to provide drug safety information to patients and health carers by analyzing the clinical and pathological characteristics of the DILI and the associated drug types. METHODS: A retrospective analysis was conducted in 287 patients diagnosed with DILI enrolled in our hospital from January 2011 to December 2015. The categories of causative drugs, clinical and pathological characteristics were reviewed. RESULTS: Western medicines ranked as the top cause of DILI, accounting for 163 out of the 287 DILI patients (56.79%) in our study. Among the Western medicine, antituberculosis drugs were the highest cause (18.47%, 53 patients) of DILI.   Antibiotics (18 patients, 6.27%) and antithyroid (18 patients, 6.27%) drugs also ranked among the major causes of DILI. Chinese herbal medicines are another major cause of DILI, accounting for 36.59% of cases (105 patients). Most of the causative Chinese herbal medicines were those used to treat osteopathy, arthropathy, dermatosis, gastropathy, leukotrichia, alopecia, and gynecologic diseases. Hepatocellular hepatitis was prevalent in DILI, regardless of Chinese herbal medicine or Western medicine-induced DILI. CONCLUSIONS: Risks and the rational use of medicines should be made clear to reduce the occurrence of DILI. For patients with liver injury of unknown origin, liver tissue pathological examination is recommended for further diagnosis.

Acute liver failure caused by pembrolizumab in a patient with pulmonary metastatic liver cancer: A case report.

RATIONALE: Keytruda (pembrolizumab) is an inhibitor of programmed cell death receptor-1 (PD-1), which was approved to treat advanced melanoma and nonsmall cell lung cancer patients who do not respond to other treatment. However, its efficacy and security in the treatment of advanced liver cancer is still under investigation. PATIENT CONCERNS: A 60-year-old man was diagnosed with pulmonary metastatic liver cancer who accepted pembrolizumab treatment after the failure of sorafenib. When injected pembrolizumab, in spite of pulmonary metastatic lesion shrink, the patient experienced severe liver dysfunction. DIAGNOSES: Based on the features of the clinical signs and laboratory examination,the patient was diagnosed with pembrolizumab-induced immune-related hepatitis by excluding other etiologies and drug-induced side effects. INTERVENTIONS: The patient received glucocorticoid and artificial liver (plasma exchange) therapy after failed conservative liver protection treatment. OUTCOMES: The patient's liver dysfunction continuously progressed and he finally died of liver failure and its complications during his hospitalization. LESSONS: Pembrolizumab showed efficacy in an advanced hepatocellular carcinoma patient with lung metastases. However, it can generate immune-related adverse events such as immune-related hepatitis which can be lethal.

Muscarinic cholinergic regulation of L-type calcium channel in heart of embryonic mice at different developmental stages.

AIM: To investigate the muscarinic regulation of L-type calcium current (I(Ca-L)) during development. METHODS: The whole cell patch-clamp technique was used to record II(Ca-L) in mice embryonic cardiomyocytes at different stages (the early developmental stage, EDS; the intermediate developmental stage, IDS; and the late developmental stage, LDS). Carbachol (CCh) was used to stimulate M-receptor in the embryonic cardiomyocytes of mice. RESULTS: The expression of I(Ca-L) density did not change in different developmental stages (P>0.05). There was no difference in the sensitivity of I(Ca-L) to CCh during development (P>0.05). This inhibitory action of CCh was mediated by inhibition of cyclic AMP since 8-bromo-cAMP completely reversed the muscarinic inhibitory action. IBMX, a non-selective inhibitor of phosphodiesterase (PDE), reversed the inhibitory action of M-receptor on I(Ca-L) current by 71.2 %+/-9.2 % (n=8) and 11.3 %+/-2.5 % (n=9) in EDS and LDS respectively. However forskolin, an agonist of adenylyl cyclase (AC), reversed the action of CCh by 14.5 %+/-3.5 % (n=5) and 82.7 %+/-10.4 % (n=7) in EDS and LDS respectively. CONCLUSION: The inhibitory action of CCh on I(Ca-L) current was mediated in different pathways: in EDS, the inhibitory action of M-receptor on I(Ca-L) channel mainly depended on the stimulation of PDE. However, in LDS, the regulation by M-receptor on I(Ca-L) channel mainly depended on the inactivation of AC.