Recent advances in understanding the regulatory mechanism of plasma membrane H+-ATPase through the brassinosteroid signaling pathway.

The polyhydroxylated steroid phytohormone brassinosteroids (BRs) control many aspects of plant growth, development and responses to environmental changes. Plasma membrane (PM) H+-ATPase, the well-known PM proton pump, is a central regulator in plant physiology, which mediates not only plant growth and development, but also adaptation to stresses. Recent studies highlight that PM H+-ATPase is at least partly regulated via the BR signaling. Firstly, the BR cell surface receptor BRASSINOSTEROID-INSENSITIVE 1 (BRI1) and multiple key components of BR signaling directly or indirectly influence PM H+-ATPase activity. Secondly, the SMALL AUXIN UP RNA (SAUR) gene family physically interacts with BRI1 to enhance organ development of Arabidopsis by activating PM H+-ATPase. Thirdly, RNA-sequencing (RNA-seq) assays showed that the expression of some SAUR genes is upregulated under the light or sucrose conditions, which is related to the phosphorylation state of the penultimate residue of PM H+-ATPase in a time-course manner. In this review, we describe the structural and functional features of PM H+-ATPase, and summarize recent progress toward understanding the regulatory mechanism of PM H+-ATPase by BRs, and briefly introduce how PM H+-ATPase activity is modulated by its own biterminal regions and the post-translational modifications.

The Universally Conserved Unconventional G Protein YchF Is Critical for Growth and Stress Response.

The ancient guanine nucleotide-binding (G) proteins are a group of critical regulatory and signal transduction proteins, widely involved in diverse cellular processes of all kingdoms of life. YchF is a kind of universally conserved novel unconventional G protein that appears to be crucial for growth and stress response in eukaryotes and bacteria. YchF is able to bind and hydrolyze both adenine nucleoside triphosphate (ATP) and guanosine nucleoside triphosphate (GTP), unlike other members of the P-loop GTPases. Hence, it can transduce signals and mediate multiple biological functions by using either ATP or GTP. YchF is not only a nucleotide-dependent translational factor associated with the ribosomal particles and proteasomal subunits, potentially bridging protein biosynthesis and degradation, but also sensitive to reactive oxygen species (ROS), probably recruiting many partner proteins in response to environmental stress. In this review, we summarize the latest insights into how YchF is associated with protein translation and ubiquitin-dependent protein degradation to regulate growth and maintain proteostasis under stress conditions.

Development and validation of a survival prediction model for patients received mechanical ventilation in the intensive care unit: a large sample size cohort from the MIMIC database.

BACKGROUND: Mechanical ventilation remains one of the primary management measures for critically ill patients in intensive care units (ICUs). However, previous studies on the prognosis prediction of ICU patients received mechanical ventilation were limited. This study was to develop and validate a nomogram for predicting short- and long-term survival among patients who received mechanical ventilation in the ICU. METHODS: This was a retrospective cohort study with a 3-year follow-up. Demographic, laboratory, clinical data of 16,775 participants aged ≥18 years who received mechanical ventilation in the ICU were extracted from the Medical Information Mart for Intensive Care III (MIMIC-III) database. The outcomes of this study were 1-month, 3-month, 1-year, and 3-year survival. All eligible patients were randomly classified into the training and testing groups with a ratio of 7:3. A multivariate Cox regression in the training group was used to explore the predictors and develop the predictive nomogram. Internal and subgroup validations were performed, and the C-index was calculated to estimate the predictive performance of the nomogram. The time-dependent receiver operating characteristic curves were drawn, and corresponding areas under the curve (AUC) were calculated. RESULTS: Totally 6,291 patients died during the follow-up duration. Age, gender, ethnicity, ICU type, comorbidity, days of mechanical ventilation, white blood cell count, blood urea nitrogen, the fraction of inspiration O2, Sequential Organ Failure Assessment scores, and the Glasgow coma score were predictors of the survival of ICU patients who received mechanical ventilation (P<0.05). The C-index of the nomogram was 0.819 and was validated in the testing group at 0.816. The AUCs for the prognostic nomogram for 1-month, 3-month, 1-year, and 3-year survival were 0.889, 0.892, 0.882, and 0.866, respectively. CONCLUSIONS: This nomogram showed good predictive performance for short- and long-term survival in ICU patients treated with mechanical ventilation, which may be a useful tool for clinicians to assess the prognosis of patients and to adjust treatment strategies in time.

Tea and tea drinking: China's outstanding contributions to the mankind.

BACKGROUND: Tea trees originated in southwest China 60 million or 70 million years ago. Written records show that Chinese ancestors had begun drinking tea over 3000 years ago. Nowadays, with the aging of populations worldwide and more people suffering from non-communicable diseases or poor health, tea beverages have become an inexpensive and fine complementary and alternative medicine (CAM) therapy. At present, there are 3 billion people who like to drink tea in the world, but few of them actually understand tea, especially on its development process and the spiritual and cultural connotations. METHODS: We searched PubMed, Google Scholar, Web of Science, CNKI, and other relevant platforms with the key word "tea", and reviewed and analyzed tea-related literatures and pictures in the past 40 years about tea's history, culture, customs, experimental studies, and markets. RESULTS: China is the hometown of tea, tea trees, tea drinking, and tea culture. China has the oldest wild and planted tea trees in the world, fossil of a tea leaf from 35,400,000 years ago, and abundant tea-related literatures and art works. Moreover, tea may be the first Chinese herbal medicine (CHM) used by Chinese people in ancient times. Tea drinking has many benefits to our physical health via its antioxidant, anti-inflammatory, immuno-regulatory, anticancer, cardiovascular-protective, anti-diabetic, and anti-obesity activities. At the moment, COVID-19 is wreaking havoc across the globe and causing severe damages to people's health and lives. Tea has anti-COVID-19 functions via the enhancement of the innate immune response and inhibition of viral growth. Besides, drinking tea can allow people to acquire a peaceful, relaxed, refreshed and cheerful enjoyment, and even longevity. According to the meridian theory of traditional Chinese medicine, different kinds of tea can activate different meridian systems in the human body. At present, black tea (fermented tea) and green tea (non-fermented tea) are the most popular in the world. Black tea accounts for over 90% of all teas sold in western countries. The world's top-grade black teas include Qi Men black in China, Darjeeling and Assam black tea in India, and Uva black tea in Sri Lanka. However, all top ten famous green teas in the world are produced in China, and Xi Hu Long Jing tea is the most famous among all green teas. More than 700 different kinds of components and 27 mineral elements can be found in tea. Tea polyphenols and theaflavin/thearubigins are considered to be the major bioactive components of black tea and green tea, respectively. Overly strong or overheated tea liquid should be avoided when drinking tea. CONCLUSIONS: Today, CAM provides an array of treatment modalities for the health promotion in both developed and developing countries all over the world. Tea drinking, a simple herb-based CAM therapy, has become a popular man-made non-alcoholic beverage widely consumed worldwide, and it can improve the growth of economy as well. Tea can improve our physical and mental health and promote the harmonious development of society through its chemical and cultural elements.

Cardiomyocyte Stim1 Deficiency Exacerbates Doxorubicin Cardiotoxicity by Magnification of Endoplasmic Reticulum Stress.

INTRODUCTION: Doxorubicin (Dox) is an effective anticancer agent; however, its cardiotoxicity remains a challenge. Dysfunction of intracellular calcium ion (Ca2+) is implicated in the process of Dox-induced cardiomyocyte apoptosis. Although store-operated Ca2+ entry (SOCE) is suggested to be responsible for Ca2+ entry in cardiomyocytes, the direct role of store-operated Ca2+ channels in Dox-related cardiomyocyte apoptosis is unknown. MATERIALS AND METHODS: Cardiomyocyte Stim1-specific knockout or overexpression mice were treated with Dox. Cardiomyocytes were pretreated with Stim1 adenovirus or siRNA followed by Dox incubation in vitro. Cardiac function and underlying mechanisms echocardiography were assessed via immunofluorescence, flow cytometry, real-time PCR, Western blotting and immunoprecipitation. RESULTS: We observed the inhibition of Stim1 expression, association of Stim1 to Orai1 or Trpc1, and SOCE in Dox-treated mouse myocardium and cardiomyocytes. Orai1 and Trpc1 expression remained unchanged. Cardiomyocyte-specific deficiency of Stim1 exacerbated Dox-induced cardiac dysfunction and myocardial apoptosis. However, specific overexpression of Stim1 in the myocardium was associated with amelioration of cardiac dysfunction and myocardial apoptosis. In vitro, STIM1 knockdown potentiated Dox-induced AC16 human cardiomyocyte apoptosis. This apoptosis was attenuated by STIM1 upregulation. Moreover, STIM1 downregulation enhanced Dox-induced endoplasmic reticulum (ER) stress in cardiomyocytes. In contrast, STIM1 overexpression inhibited the activation of the above molecular markers of ER stress. Immunoprecipitation assay showed that STIM1 interacted with GRP78 in cardiomyocytes. This interaction was attenuated in response to Dox treatment. CONCLUSION: Our data demonstrate that cardiomyocyte STIM1 binding to GRP78 ameliorates Dox cardiotoxicity by inhibiting pro-apoptotic ER stress.

Pyrroloquinoline quinone (PQQ) alleviated sepsis-induced acute liver injury, inflammation, oxidative stress and cell apoptosis by downregulating CUL3 expression.

PQQ has anti-inflammatory and anti-oxidant effects. PQQ can relieve high glucose-induced renal cell damage by suppressing Keap1 expression. Keap1 can interact with CUL3. Upregulation of CUL3 facilitates the apoptosis of LPS-induced podocytes. Based on knowledge above, this current work was designed to explore the role of PQQ in sepsis and determine the molecular function of CUL3 in the pathogenesis of sepsis. Rats received CLP surgery to establish sepsis models in vivo. Kupffer cells were pretreated with PQQ (10, 50 and 100 nmol/L) for 2 h and then treated with 100 ng/mL LPS for 24 h, simulating sepsis-induced acute liver injury in vitro. H&E staining was performed to evaluate liver injury of SD rats. Levels of inflammatory factors and oxidative stress markers were detected to assess inflammatory response and oxidative stress. Moreover, TUNEL staining, flow cytometric analysis and western blot were applied to determine cell apoptosis. It was confirmed that PQQ treatment relieved acute liver injury, inflammatory and oxidative stress damage and apoptosis of liver tissue cells in sepsis rats. In addition, PQQ therapy could alleviate inflammation, oxidative stress and apoptosis in LPS-induced Kupffer cells. Notably, LPS stimulation enhanced CUL3 expression and PQQ repressed CUL3 expression in Kupffer cells suffered from LPS. Overall, CUL3 overexpression weakened the remission effects of PQQ on LPS-induced inflammatory and oxidative damage and apoptosis of Kupffer cells. Mechanistically, PQQ treatment may mitigate sepsis-induced acute liver injury through downregulating CUL3 expression.

Evaluation of Beneficial and Adverse Effects of a Diet Supplemented with Schisandrae Fructus Seed Ethanol Extract on Lipid and Glucose Metabolism in Normal and Hypercholesterolemic/Hyperglycemic Mice.

Schisandrae Fructus (SF), the fruit of Schisandra chinensis (Turcz.) Baillon, has been used for the treatment of liver injury and metabolism-related disorders in China. The objective of this study was to investigate the effects of supplementation with ethanol extract of SF seed (EtSF-S) on serum/hepatic lipid and glucose levels as well as fecal total cholesterol (TC) contents in mice fed a normal diet (ND) or high-fat/fructose diet (HFFD) containing 15% lard oil and 15% fructose. Female ICR mice (18-20 g in body weight) were fed with ND or HFFD for 3 months, and then EtSF-S was added to both chow diets at increasing concentrations of 1, 5, and 10% (w/w). Thirty days later, serum and hepatic lipids, including TC, triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), and glucose, were measured. Dietary supplementation with EtSF-S reduced hepatic TC (36 and 18%) and TG levels (38 and 28%) and increased serum HDL/LDL ratio (16 and 26%) in both ND- and HFFD-fed mice, respectively. Moreover, supplementation with EtSF-S elevated serum HDL (31%) in HFFD-fed mice and reduced serum LDL (27%) in ND-fed mice. EtSF-S treatment reduced fat mass (40%) in ND-fed mice and increased fecal TC contents (33%) in HFFD-fed mice. EtSF-S supplementation decreased hepatic glucose contents (29%) in both ND- and HFFD-fed mice. However, diet supplemented with EtSF-S elevated serum TG levels (up to 123%) and hepatic size (28%), but more importantly, suppressed the body weight gain (approximately 130%) in mice fed with HFFD. These findings suggested that dietary supplementation with EtSF-S as natural herbal function food may be a useful strategy for the treatment of patients with fatty liver disease or overweight without a high intake of sugar and fat.

Thal protects against paraquat-induced lung injury through a microRNA-141/HDAC6/IκBα-NF-κB axis in rat and cell models.

The protective functions of thalidomide in paraquat (PQ)-induced injury have been reported. But the mechanisms remain largely unknown. In this research, a PQ-treated rat model was established and further treated with thalidomide. Oedema and pathological changes, oxidative stress, inflammation, fibrosis and cell apoptosis in rat lungs were detected. A PQ-treated RLE-6TN cell model was constructed, and the viability and apoptosis rate of cells were measured. Differentially expressed microRNAs (miRNAs) after thalidomide administration were screened out. Binding relationship between miR-141 and histone deacetylase 6 (HDAC6) was validated. Altered expression of miR-141 and HDAC6 was introduced to identify their involvements in thalidomide-mediated events. Consequently, thalidomide administration alone exerted no damage to rat lungs; in addition it reduced PQ-induced oedema. The oxidative stress, inflammation and cell apoptosis in rat lungs were reduced by thalidomide. In RLE-6TN cells, thalidomide increased cell viability and decreased apoptosis. miR-141 was responsible for thalidomide-mediated protective events by targeting HDAC6. Overexpression of HDAC6 blocked the protection of thalidomide against PQ-induced injury via activating the IkBα-NF-κB signalling pathway. Collectively, this study evidenced that thalidomide protects lung tissues from PQ-induced injury through a miR-141/HDAC6/IkBα-NF-κB axis.

FoxF1 protects rats from paraquat-evoked lung injury following HDAC2 inhibition via the microRNA-342/KLF5/IκB/NF-κB p65 axis.

PURPOSE: Forkhead box f1 (FoxF1), a transcription factor, was implicated in lung development. However, the molecular mechanism of FoxF1 in lung injury, specifically in injury caused by paraquat (PQ), one of the most frequently used herbicides, is unknown. Accordingly, we performed this study to investigate whether FoxF1 attenuates PQ-induced lung injury and to determine the possible mechanism. METHODS: We used PQ-treated Beas-2B cells to measure the expression of FoxF1. Later, ChIP-qPCR was applied to detect the levels of histone acetylation in cells, followed by the validation of the relationship between histone deacetylase-2 (HDAC2) and FoxF1. Subsequently, the correlation between FoxF1 and microRNA (miR)-342 and the downstream mechanism of miR-342 were evaluated by bioinformatics analysis. The apoptosis and the content of reactive oxygen species (ROS) in PQ-treated cells were detected to evaluate the roles of HDAC2, FoxF1 and miR-342 in vitro. Finally, a rat model was developed to evaluate the effects of HDAC2, miR-342 and Krüppel-like factor 5 (KLF5) on PQ-induced lung injury in vivo. RESULTS: PQ treatment significantly enhanced FoxF1 promoter deacetylation, thereby inhibiting FoxF1 expression. After inhibition of HDAC2 activity, apoptosis and oxidative stress induced by PQ were significantly reversed. Nevertheless, further inhibition of miR-342 or overexpression of KLF5 promoted apoptosis and oxidative stress induced by PQ, and IκB/NF-κB p65 signaling was significantly activated after PQ treatment. CONCLUSION: PQ treatment inhibited miR-342 expression by promoting HDAC2-induced deacetylation of the FoxF1 promoter, thereby promoting KLF5 expression and the IκB/NF-κB p65 signaling activation, and finally exacerbating PQ-induced lung injury in rats.

The effects of estradiol on inflammatory and endothelial dysfunction in rats with preeclampsia.

Preeclampsia (PE), a hypertensive disorder during pregnancy, has adverse effects to both the mother and the fetus. Maternal inflammatory and vascular endothelial dysfunction are important factors in the pathogenesis of PE. The present study aimed to investigate the effects of estradiol (E2) on inflammatory and endothelial dysfunction in an N (omega)­nitro­L­arginine methyl ester (L­NAME)­induced rat model of PE. Adult pregnant female Sprague­Dawley rats were divided into four equal groups between days 7 and 11 of gestation and treated as follows: i) Pregnant rats receiving daily intraperitoneal (i.p.) injections of equal volume of 0.9% normal saline (NS) (Control group, n=12); ii) pregnant rats receiving daily i.p. injections of L­NAME at 50 mg/kg (L­NAME group, n=12); iii) pregnant rats receiving a daily i.p. injection of 50 mg/kg L­NAME and NS from day 11 (L­NAME + NS group, n=12); and iv) pregnant rats receiving daily i.p. injections of 50 mg/kg L­NAME and 100 µg/kg/day E2 from day 11 (L­NAME + E2 group, n=12). On day 21, blood pressure (BP) and the level of 24­h urine protein in the maternal rats, fetal weight and percentage of stillbirths following a cesarean section were recorded. The activities of nitric oxide (NO) and inducible NO synthase (iNOS), the levels of inflammatory cytokines [interleukin (IL)­1ß, IL­6, interferon­Î³ and monocyte chemoattractant protein­1], adherence factors (CD49d, intracellular adhesion molecule 1 and lymphocyte function­associated antigen­1) and uterine angiogenic status (Fms­like tyrosine kinase­1, vascular cell adhesion molecule and matrix metalloproteinase 2/9) were also assessed. In addition, the histopathology of the placenta, the expression of estrogen receptor α 36 (ERα36), ERα, ERß and G protein­coupled ER, as well as the activation of the toll­like receptor 4 (TLR4) signaling pathway (TLR4, myeloid differentiation primary response 88, IL­1 receptor­associated kinase 4 and tumor necrosis factor receptor­associated factor 6) were evaluated by H&E staining, immunofluorescence and western blot assays. Treatment with L­NAME increased the BP, urine protein and rate of stillbirths and suppressed fetal weight compared with those in the control group. The L­NAME­induced effects were attenuated by the administration of E2. In addition, the administration of E2 decreased inflammation and NO levels and altered the uterine angiogenic status. The histological analysis of PE rat placenta in the E2­treated group confirmed the effects on biochemical parameters. Of note, E2 treatment significantly suppressed the TLR4 signaling pathway. In the rat model of PE, adverse outcomes including BP, fetal rat weight and proteinuria, high neonatal death rate, inflammatory response, oxidative stress and endothelial dysfunction were attenuated by exogenous E2 administration, which may present a novel approach for the clinical treatment of PE.

Fish oils protects against cecal ligation and puncture­induced septic acute kidney injury via the regulation of inflammation, oxidative stress and apoptosis.

Septic acute kidney injury (AKI) is usually caused by sepsis. ω3 fatty acid has been reported to suppress sepsis­induced organ dysfunction to a certain degree. The present study aimed to investigate the effects of ω3 fatty acid in septic renal injury. Sprague Dawley rats were used to establish a cecal ligation and puncture (CLP) model in order to mimic the development of septic injury. The rats were treated with dexamethasone and fish oils (FOs) for 4 days prior to CLP. Alterations in the morphology of the tissues, the renal function and the induction of inflammation, oxidative stress and apoptosis were evaluated. The effects of FOs on nuclear factor­κB (NF­κB), JAK2/STAT3 and p38­MAPK were determined. The rats of the CLP model group exhibited low survival rates and increased expression of serum creatine, blood urea nitrogen, neutrophil gelatinase­associated lipocalin, kidney injury molecule­1 and of proinflammatory cytokines. In addition, the levels of the markers of oxidative injury and apoptosis were increased. The induction of renal injury was notably reversed by administration of dexamethasone and FOs. The expression levels of the protein markers involved in inflammation and apoptosis were measured and the results indicated that FOs inhibited JAK/STAT3 and p­38MAPK signaling, while they concomitantly increased the expression of NF­κB. The present study highlighted that FOs improve CLP­induced mortality and renal injury by inhibiting inflammation, oxidative stress and apoptosis.

Angiopoietin-Like 7 Contributes to Angiotensin II-Induced Proliferation, Inflammation and Apoptosis in Vascular Smooth Muscle Cells.

INTRODUCTION: Angiotensin II (AngII) induces hypertension and pathophysiological vascular thickening and atherosclerosis. This study aims to validate the effects of Angiopoietin-like 7 (ANGPTL7) in AngII-induced hypertension. METHODS: ANGPTL7 in blood samples were determined by quantitative real-time polymerase chain reaction. AngII-induced cell growth were detected by CCK-8. Cell cycle arrest and cell apoptosis by downregulation of ANGPTL7 were detected by flow cytometric assay. AngII-induced inflammation was evaluated by Western blotting and ELISA. RESULTS: ANGPTL7 was highly expressed in patients with hypertension. AngII promoted cell viability and the expression level of ANGPTL7 in vascular smooth muscle cells (VSMC). Downregulation of ANGPTL7 inhibited AngII-induced cell proliferation and cell inflammation. Moreover, ANGPTL7 expression decreases also promoted cell apoptosis. CONCLUSIONS: Downregulation of ANGPTL7 reversed AngII-induced cell proliferation and cell inflammation and promoted apoptosis in AngII-induced VSMC cells. Therefore, ANGPTL7 can be a potential target in AngII-induced hypertension.

Netrin-1 prevents the attachment of monocytes to endothelial cells via an anti-inflammatory effect.

Netrin-1 is best known for its function guiding axon growth and migration. Netrin-1 has been shown to be involved in regulating cardiovascular function. In this study, we aimed to understand the biological role of Netrin-1 and its receptor Unc5b in endothelial cells. Our results demonstrate that Unc5b was moderately expressed in human aortic endothelial cells (HAECs) and TNF-α had a dose-dependent inhibitory effect on Unc5b level. Netrin-1 potently suppressed TNF-α-induced vascular adhesion molecules VCAM-1, ICAM-1, E-selectin and blocked the adhesion of monocytes to endothelial cells. Netrin-1 also suppressed TNF-α-induced production of cytokines including MCP-1, IL-1ß, and IL-6. Importantly, Netrin-1 suppressed toll like receptor 4 (TLR4) expression and prevented NF-κB activation. Mechanistically, Netrin-1 reduced TNF-α-induced IKK and IκBα activation and prevented degradation of IκBα. Netrin-1 reduced nuclear accumulation of p65 and strongly suppressed NF-κB promoter activation. Collectively, our data demonstrated that signaling of Netrin-1 and its receptor Unc5b had an anti-inflammatory effect in endothelial cells. Netrin-1 signaling could be imperative for normal endothelial function.