Atractylodin alleviates nonalcoholic fatty liver disease by regulating Nrf2-mediated ferroptosis.

Nonalcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease worldwide. Oxidative stress is one of the main inducers of NAFLD. Atractylodin (ART), a major active ingredient of Atractylodes lancea, possesses potential antioxidant and anti-inflammatory activity in many types of disease. In the current study, the underlying mechanism by which ART alleviates the progression of NAFLD was explored. The function of ART in facilitating NAFLD was investigated in vitro and in vivo. Functionally, ART attenuated high-fat diet (HFD)-induced NAFLD in mice and palmitic acid (PA)-induced oxidative stress in HepG2 cells. Furthermore, our data verified that ART attenuated HFD-induced NAFLD by inhibiting ferroptosis of hepatocyte cells, as evidenced by decreased Fe2+ concentration, reactive oxygen species (ROS) level, malondialdehyde (MDA) content, and increased glutathione (GSH) content. The protective effect of ART on the cell viability of hepatocytes was blocked by a specific ferroptosis inhibitor (ferrostatin-1). Mechanistically, ART treatment promoted the translocation of nuclear factor erythroid 2-related Factor 2 (NFE2L2/NRF2) and thus increased glutathione peroxidase 4 (GPX4), ferritin heavy chain 1 (FTH1), and solute carrier family 7 member 11 (SLC7A11) expression. Taken together, ART alleviates NAFLD by regulating Nrf2-mediated ferroptosis.

Depression and anxiety of medical students at Kunming Medical University during COVID-19: A cross-sectional survey.

An isolation strategy was used to control the transmission and rapid spread of COVID-19 in Yunnan. As a result, students were supposed to stay at home and disrupted their outside activities. It led to a detrimental influence on students' mental health. The purpose of this study was to investigate the prevalence and risk factors of depression and anxiety among medical students and to provide ideas for the prevention of depression and anxiety in medical students. A cross-sectional survey was conducted among 2,116 medical students at Kunming Medical University from July 8 to July 16, 2020. Participants' demographic and living conditions were collected. Depression and anxiety were measured using the Patient Health Questionnaire 9 and General Anxiety Disorder-7, respectively. Uni- and multivariate logistic regression analyses were performed to detect risk factors associated with depression and anxiety. The prevalence rates of depression and anxiety among medical students were 52.5 and 29.6%, respectively. Depression was more likely to be caused by low grades, lack of physical exercise, drug use, irregular diet, extensive screen time on mobile phones, being greatly affected by the COVID-19 pandemic, and inadaptability to offline courses. Anxiety was more likely to be caused by lack of physical exercise, drug use, irregular diet, and inadaptability to offline courses. Depression and anxiety are highly comorbid. Our study showed predictive factors for depression and anxiety and identified a major mental health burden on medical students during the COVID-19 outbreak. More targeted measures should be taken to improve the mental state of students to reduce the incidence of depression and anxiety.

Neuroprotection of Oral Edaravone on Middle Cerebral Artery Occlusion in Rats.

Edaravone has been widely used in the treatment of acute ischemic stroke. However, there has been no oral preparation of edaravone in the clinic. In this study, we assessed the effect and possible mechanisms of oral edaravone on the middle cerebral artery occlusion (MCAO) model in rats. Highly bioavailable form of novel edaravone formulation developed using self-nanomicellizing solid dispersion strategy which showed up to 16.1-fold improved oral bioavailability was considered oral edaravone. The male rats (n = 84) were randomly divided into sham; model; oral edaravone in low dose (10 mg/kg), medium dose (20 mg/kg), and high dose (30 mg/kg); and edaravone by intraperitoneal administration group (IP group, 10 mg/kg). Rats were treated with different drugs 5 h after the operation, twice a day for 7 days. The behavioral data were dose-dependently improved by oral edaravone and sensorimotor functions of the high dose group were similar to those of the edaravone by IP route group. Furthermore, oral edaravone significantly reduced cerebral infarction area and downregulated the levels of caspase-3, GFAP, Iba1, 3-NT, and 4-HNE, whereas upregulated those of Vamp-2 and Map-2 in a dose-dependent manner. Especially effect of the high dose on these molecules was equal to that of edaravone by IP administration. Taken together, our data suggest that the improvement of sensorimotor deficits by oral edaravone in high doses after ischemia is similar to that in edaravone by IP administration. Neuroprotection of oral edaravone is at least partial by minimizing oxidative stress, the overactivation of glial cells, and the levels of the apoptosis-associated proteins, and alleviating synaptic damage in a dose-dependent manner.

The combination of living Bifidobacterium, Lactobacillus, and Streptococcus improves social ranking and relieves anxiety-like behaviors in competitive mice in a social dominance tube test.

INTRODUCTION: Social rank has a profound influence on the behavior and health of humans and animals. METHODS: To explore the effect of a combination of living Bifidobacterium, Lactobacillus and Streptococcus (CLB) on anxiety- and depression-like behaviors and social rank, mice were subjected to a social dominance tube test (SDTT). The behaviors, rank, gut microbiota, and expression of inflammatory cytokines and brain-derived neurotrophic factor (BDNF) in the hippocampus were measured. RESULTS: The results indicated that CLB improved the SDTT ranking score of the losers and alleviated anxiety-like behaviors of the winners. CLB decreased the level of Desulfovibrio and augmented the level of Mollicutes in the feces, increased BDNF content, and reduced the level of tumor necrosis factor-α in the hippocampus. CONCLUSION: These findings indicated that CLB may be used for the treatment of anxiety and improvement of the rank score via regulation of gut microbiota and anti-inflammatory effects.

Protective effect of pogostone on murine norovirus infected-RAW264.7 macrophages through inhibition of NF-κB/NLRP3-dependent pyroptosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Pogostemon cablin, the dry overground parts of Pogostemon cablin (Blanco) Benth, has been widely used in the treatment of gastrointestinal dysfunction, such as nausea, diarrhea, headaches and fever. Pogostone (PO) is a major component of Pogostemon cablin which has a variety of pharmacological properties, including antiinflammatory, and immunosuppressive activities, and antioxidant. However, the effect of PO on norovirus gastroenteritis and the underlying molecular mechanism remain unclear. AIM OF THE STUDY: The purpose of our study is to investigate the effects of PO against MNV infection using RAW264.7 cells and to elucidate its active mechanisms. MATERIALS AND METHODS: The cell viability was assessed using Cell Counting Kit-8 (CCK-8) assay and Fluorescein diacetate (FDA) staining. The activation of nuclear factor kappa B (NF-κB) signaling and NOD-like receptor 3 (NLRP3) inflammasome was evaluated by assessing the level of phospho-NF-κB p65, interleukin (IL)-6, TNF-α, NLRP3, cleaved caspase-1, IL-18, IL-1ß using Western blot and quantitative real-time PCR (qPCR), respectively. The number of infected cells were determined by immunofluorescence microscopic assay. RESULTS: PO did not possess a cytotoxic effect toward RAW264.7 cells. The cytotoxic damage caused by MNV infection in RAW264.7 cells decreased significantly in the presence of PO. Cell viability assays showed that pyroptosis is the major mechanism of death in MNV-infected RAW264.7 cells. PO could decreased the expression levels of p-p65, IL-6, TNF-α, NLRP3, cleaved caspase-1, IL-1ß, and IL-18. CONCLUSIONS: These results demonstrate that PO decreases MNV-induced RAW264.7 macrophages death and MNV replication through repressing NF-κB/NLRP3-dependent pyroptosis. Therefore PO may be considered as a potential therapeutic agent for preventing and treating norovirus gastroenteritis.

[Frontometaphyseal dysplasia 1 caused by variant of FLNA gene in a case].

OBJECTIVE: To explore the clinical and genetic characteristics of a child with frontometaphyseal dysplasia 1 (FMD1) due to variant of FLNA gene. METHODS: Clinical phenotype of the patient was analyzed. Whole exome sequencing (WES) was carried out to detect pathogenic genetic variants. Sanger sequencing was used to verify the result in his parents. RESULTS: The 2-year-and-9-month-old boy presented with facial dysmorphism (supraorbital hyperostosis, down-slanting palpebral fissure and ocular hypertelorism), skeletal deformities (bowed lower limbs, right genu valgum, left genu varus, slight deformity of index and middle fingers, and flexion contracture of little fingers). He also had limited left elbow movement. High-throughput sequencing revealed that he has carried a de novo heterogeneous c.3527G>A (p.Gly1176Glu) missense variant of the FLNA gene. The same variant was found in neither parent. CONCLUSION: The clinical manifestations of FMD1 such as joint contracture and bone dysplasia can occur in infancy and deteriorate with age, and require long-term follow-up and treatment. Above finding has expanded the spectrum of FLNA gene variants.

Baicalin Is Curative Against Rotavirus Damp Heat Diarrhea by Tuning Colonic Mucosal Barrier and Lung Immune Function.

BACKGROUND: Previous studies have indicated that rotavirus (RV) is a causative factor for diarrhea and gastroenteritis in pediatric and neonatal settings. Baicalin has many functions, including antibacterial, antiinflammatory, and antihypertensive activities. However, the immunological mechanism of RV-induced diarrhea with heat-dampness syndrome (RV-DH) remains unclear. AIMS: The aim of this study is to explore the role of baicalin in RV-DH diarrhea and its underlying mechanism. METHODS: A mouse model of pediatric RV-DH diarrhea was established and treated with baicalin. The concentrations of cytokines were detected by enzyme-linked immunosorbent assay. Messenger RNA (mRNA) expression levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR), while protein expression levels were determined by Western blotting and immunohistochemistry. Flow cytometry was used to detect the frequency of lymphocytes. RESULTS: The concentrations of interleukin-1ß (IL-1ß), IL-2, IL-6, IL-8, RVvb, and secretory immunoglobulin A (SIgA) in bronchoalveolar lavage fluid (BALF) and colonic mucosa were significantly increased in the RV-DH group. Decreased expression of occludin, claudin-1, and zonula occludens-1 (ZO-1) indicated loss of tight junction function and disturbances in intestinal mucosal permeability in the RV-DH group. Flow cytometry analysis showed a high rate of CD8+ lymphocytes and low amount of CD4+ lymphocytes in the RV-DH group. Treatment of RV-DH mice with baicalin significantly reduced the duration of diarrhea and ameliorated the symptoms and pathological and immunological changes. Furthermore, baicalin inhibited STAT1 and activated STAT3 signaling pathways. CONCLUSIONS: These findings indicate the curative and immunoregulatory properties of baicalin and have direct practical and clinical relevance for the treatment of RV-DH enteritis in humans.

Interference of miR-943-3p with secreted frizzled-related proteins4 (SFRP4) in an asthma mouse model.

The aim of this study is to investigate the potential roles of miR-943-3p and its target gene secreted frizzled-related proteins4 (SFRP4) in allergic asthma and elucidate its underlying mechanism, which may prompt a new clue about developing novel treatments of this disease. An allergic asthma mouse model was generated by challenging with ovalbumin (OVA); lung pathological features of mice were viewed using H&E staining; thickness of subepithelial fibrosis and smooth muscle was measured using Masson's trichrome staining. Inflammatory cells from bronchoalveolar lavage fluid (BALF) were counted based on Diff-Quik staining and morphometric analysis. Expressions of miR-943-3p, SFRP4 and Wnt signal pathway-associated proteins were detected using RT-PCR or immunoblotting, respectively. SFRP4 was downregulated in the bronchial biopsies of allergic asthma patients and represented a unique intersection between differentially expressed genes (DEGs) and genes in the Wnt signal pathway. Both miR-943-3p upregulation and SFRP4 downregulation were detected in allergic asthma patients and OVA-induced mice. Besides, OVA-induced mice possessed more inflammatory cells in BALF including macrophage (mac), eosinophil (eos), lymphocyte (lym) and neutrophil (neu), higher expression of collagen, ß-catenin and c-Myc as well as thicker subepithelial fibrosis and smooth muscle in lung than control mice. In vivo delivery of miR-943-3p agomir worsened these symptoms, while both miR-943-3p antagomir and Ad-SFRP4 administration effectively alleviated this disease. Taken together, miR-943-3p accelerated the progression of airway inflammation and remodeling in allergic asthma via suppressing the activity of SFRP4 through Wnt signaling pathway in asthma patients and OVA-induced mice.

Regulating effect of baicalin on IKK/IKB/NF-kB signaling pathway and apoptosis-related proteins in rats with ulcerative colitis.

OBJECTIVE: This study was aimed to explore effect of baicalin on IKK/IKB/NF-kB signaling pathway and apoptosis-related proteins in rats with ulcerative colitis (UC). METHODS: Histopathological observation and scores of colon tissue were performed in the UC rat model. IKK/IKB/NF-kB signaling pathway and apoptosis-related proteins were measured by Western blotting. RESULTS: Baicalin significantly increased the activity of SOD, CAT and GSH-Px in colon tissue of rats with UC, but significantly decreased the content of MDA, IL-1ß, MPO, PEG2 and TNF-α in colon tissue of rats with UC. In the molecular mechanism, baicalin significantly decreased the expression of cleaved-caspase3, cleaved-caspase9, Bcl-2/Bax, cyt-c, NF-kB p-65, p-IKKß/IKKß and p-IKBα/IKBα. Baicalin could significantly inhibit p-IKBα/IKBα content change, but had no significant effect on p-IKKß/IKKß. CONCLUSION: Baicalin may have a regulating effect on IKK/IKB/NF-kB signaling pathway and apoptosis-related proteins in UC rats.

[Pathogenic role of leukotriene B4 in pulmonary microvascular endothelial cell hyper- permeability induced by one lung ventilation in rabbits].

OBJECTIVE: To elucidate the pathogenic role of leukotriene B4 (LTB4) in increased pulmonary microvascular endothelial cell permeability induced by one lung ventilation (OLV) in rabbits. METHODS: Forty-eight healthy Japanese white rabbits were randomly divided into control group (group C), saline pretreatment group (group S), bestatin (a leukotriene A4 hydrolase (LTA4H) inhibitor) plus saline pretreatment group (group B), OLV group (group O), saline pretreatment plus OLV group (group SO) and bestatin plus saline pretreatment with OLV group (group BO). ELISA was used to detect LTB4 content in the lung tissues, and LTA4H and phospholipase Cεl (PLCEl) expressions were examined by Western blotting and quantitative PCR. The wet/dry weight (W/D) ratio of the lung, lung permeability index and the expressions of myosin light chain kinase (MLCK) protein and mRNA in the lung tissues were determined to evaluate the permeability of the pulmonary microvascular endothelial cells (PMVECs). The severities of lung injury were evaluated by lung histomorphological scores. RESULTS: No significant differences were found among groups C, S and B except that LTA4H expressions was significantly lower in group B than in groups C and S (P<0.05). OLV significantly increased the expressions of LTA4H (P<0.05) and resulted in LTB4 overproduction in the lungs (P<0.05) accompanied by significantly enhanced PLCE1 expression and PMVEC permeability (P<0.05). Pretreatment with bestatin, significantly reduced the expression of LTA4H and LTB4 production (P<0.05) and down-regulated the expression of PLCE1 in the lungs of the rabbits receiving OLV (P<0.05). CONCLUSION: Bestatin plays a protective role in OLV-induced rabbit lung injury by downregulating LTA4H to reduce the production of LTB4 in the lungs. LTB4 can increase PMVEC permeability by up-regulating PLCE1 expression in rabbits with OLV-induced lung injury.

[Distribution of Syndrome Types of Chinese Medicine in Acute Infectious Diarrhea].

OBJECTIVE: To observe syndrome types of Chinese medicine (CM) and distribution features of acute infectious diarrhea patients. METHODS: A retrospective study was performed in 465 acute infectious diarrhea patients. The distribution of CM syndrome and syndrome types in different seasons and genders were analyzed. RESULTS: Lack of appetite [381 (81.94%)], fatigue [350 (75.27%)], abdominal pain [338 (72.69%)], tenesmus [325 (69.89%)], anal scorching hot [276 (59.35%)], nausea [25 (55.48%)], diarrhea [249 (53.55%)], short yellow-urine [240 (51.61%)], thirsty [210 (45.16%)], and abdominal distention [206 (44.30%)] were most often seen. The syndrome distribution were sequenced as intestinal damp heat syndrome [268 (57.63%)], dyspeptic retention in intestine and stomach syndrome [106 (22.80%)], cold-damp invading exterior syndrome [47 (10.11%)], Pi-Wei qi deficiency syndrome [23 (4.95%)], cold-damp disturbing Pi syndrome [21 (4.52%)]. The incidence ratio of intestinal damp heat syndrome was the highest in autumn (P < 0.01), while that of cold-damp invading exterior syndrome was the highest in winter (P < 0.01). Cold-damp disturbing Pi syndrome was more often seen in females than in males (P < 0.01), and its incidence ratio was the highest in autumn (P < 0.05). The incidence ratio of dyspeptic retention in intestine and stomach syndrome was the highest in winter (P < 0.01). CONCLUSIONS: Intestinal damp heat syndrome was the most often seen in acute infectious diarrhea. Incidence ratios of in- testinal damp heat syndrome and cold-damp disturbing Pi syndrome were higher in autumn, while those of cold-damp invading exterior syndrome and dyspeptic retention in intestine and stomach syndrome were higher in winter. Cold-damp disturbing Pi syndrome was more often seen in females.

Baseline factors associated with treatment response in patients infected with hepatitis C virus 1b by stratification of IL28B polymorphisms.

Although the single-nucleotide polymorphism (SNP) rs12979860 in the IL28B gene is a better predictor of sustained virological response to treatment of chronic hepatitis C (CHC) than other baseline factors, some CHC patients with the favorable C allele cannot achieve a sustained virological response when treated with peginterferon plus ribavirin. The aim of this study was to examine baseline factors as predictors of rapid virological response (RVR) and complete early virological response (cEVR) to peginterferon alfa-2a plus ribavirin treatment in Chinese CHC patients with hepatitis C virus (HCV) genotype 1b, with emphasis on the difference between the rs129860 CC and CT/TT genotypes. A total of 337 treatment-naïve patients participated in this study. All patients were treated with peginterferon alfa-2a plus ribavirin at standard dosage. Serum samples from all patients were collected at baseline, week 4, and week 12 for testing of laboratory parameters, and IL28B genotypes were determined. Multivariate analysis showed that among rs12979860 CC genotype patients, glucose level and aspartate amino transferase (AST) activity were inversely associated with RVR, while abnormal platelet count and allergy inversely associated with cEVR. Among rs12979860 CT genotype patients, age below 40 years and short infection duration were associated with RVR, while age below 40 years, short infection duration, high body mass index (BMI), and no history of allergies were associated with cEVR. The baseline factors associated with the response to CHC treatment may depend on the IL28B genotype. Refinement of the baseline predictors based on IL28B genotypes may be useful for management of HCV infection.