Frizzled 6 mutation regulates reserpine-induced depression-like behavior and Wnt signaling pathway in mice.

BACKGROUND: Frizzled 6 (Fzd6) is involved in the development of various disorders; however, its role in the etiology of depression remains unclear. We aimed to determine the potential regulatory mechanisms of Fzd6 as a Wnt receptor in depression. METHODS: Mice were divided into four groups: wild-type control (Fzd6WT-control), Fzd6 mutant control (Fzd6Q152E-control), wild-type reserpine (Fzd6WT-reserpine), and Fzd6 mutant reserpine (Fzd6Q152E-reserpine). Reserpine (0.5 mg/kg) was injected intraperitoneally for 10 days. Four behavioral experiments were performed to assess the effects of Fzd6Q152E on depression-like behaviors in the reserpine-treated mice. Blood samples were collected for an enzyme-linked immunosorbent assay (ELISA). Gene expression in the hippocampus was quantified using quantitative real-time polymerase chain reaction (qRT-PCR), and protein expression levels in the hippocampus were identified using western blotting. RESULTS: The Fzd6 mutation affected reserpine-induced depression-like behavioral changes in mice. ELISA revealed significantly reduced serum levels of 5-hydroxytryptamine (5-HT), brain-derived neurotrophic factor (BDNF), and norepinephrine in both Fzd6Q152E-reserpine and Fzd6WT-reserpine mice, with a more pronounced decrease in Fzd6Q152E-reserpine mice, especially in norepinephrine expression. The qRT-PCR results showed significantly decreased Fzd6 expression in Fzd6Q152E-reserpine mice and altered expression of Dkk2, Gsk-3ß, Lrp6, Wnt2, Wnt3, and Wnt3a in the Wnt pathway. Western blotting revealed decreased Fzd6 protein expression in Fzd6Q152E-control mice compared to Fzd6WT-control mice, whereas Fzd6 protein expression was restored in Fzd6Q152E-reserpine mice, and Gsk-3ß expression was significantly changed. CONCLUSION: Fzd6 potentially influences reserpine-induced depressive behavioral changes and serum depressive factor alterations and modulates the expression of the Wnt signaling pathway in the hippocampus of depressed mice.

In vitro transdifferentiated signatures of goat preadipocytes into mammary epithelial cells revealed by DNA methylation and transcriptome profiling.

During mammary development, the transdifferentiation of mammary preadipocytes is one of the important sources for lactating mammary epithelial cells (MECs). However, there is limited knowledge about the mechanisms of dynamic regulation of transcriptome and genome-wide DNA methylation in the preadipocyte transdifferentiation process. Here, to gain more insight into these mechanisms, preadipocytes were isolated from adipose tissues from around the goat mammary gland (GM-preadipocytes). The GM-preadipocytes were cultured on Matrigel in conditioned media made from goat MECs to induce GM-preadipocyte-to-MEC transdifferentiation. The transdifferentiated GM-preadipocytes showed high abundance of keratin 18, which is a marker protein of MECs, and formed mammary acinar-like structures after 8 days of induction. Then, we performed transcriptome and DNA methylome profiling of the GM-preadipocytes and transdifferentiated GM-preadipocytes, respectively, and the differentially expressed genes and differentially methylated genes that play underlying roles in the process of transdifferentiation were obtained. Subsequently, we identified the candidate transcription factors in regulating the GM-preadipocyte-to-MEC transdifferentiation by transcription factor-binding motif enrichment analysis of differentially expressed genes and differentially methylated genes. Meanwhile, the secretory proteome of GM-preadipocytes cultured in conditioned media was also detected. By integrating the transcriptome, DNA methylome, and proteome, three candidate genes, four proteins, and several epigenetic regulatory axes were further identified, which are involved in regulation of the cell cycle, cell polarity establishment, cell adhesion, cell reprogramming, and adipocyte plasticity. These findings provide novel insights into the molecular mechanism of preadipocyte transdifferentiation and mammary development.

Construction of Fzd6Q152E mice through CRISPR/Cas9 technology and their reproduction and identification.

BACKGROUND: The CRISPR/Cas9 system is widely used for genome editing in human, rat and mouse cells. In this study, we established Fzd6 mutant mice using CRISPR/Cas9 technology, and obtained Fzd6 homozygous mutant (Fzd6Q152E) mice through breeding. Fzd6 plays a role in depression, but there are few related reports. We used this model to investigate the mechanism of Fzd6 involved in depression, and build a solid foundation for subsequent in-depth studies. METHODS AND RESULTS: The target of Fzd6 mutation was obtained by CRISPR/Cas9 technology and hippocampal tissue was collected for Nissl staining and histological analysis. Blood was collected for enzyme linked immunosorbent assay (ELISA); The gene expression of Fzd6 and the related genes expression in wnt pathway was quantified by quantitative real-time PCR (qRT-PCR), and then expression of Fzd6 and proteins in the Wnt pathway were identified by western blotting. ELISA results showed that the expression levels of brain derived neurotrophic factor (BDNF), 5-hydroxytryptamine (5-HT), and Noradrenaline (NE) in serum were significantly decreased in Fzd6Q152E mice, whereas the mRNA expression of Lrp5, Lrp6, and Dkk2 is increased. The western blotting revealed that the expression of Fzd6 and Lrp6 is decreased, although the expression of Dkk2 and Gsk-3ß increased. CONCLUSION: Our study successfully established homozygous Fzd6 mutant mice model. The relationship between Fzd6-Wnt and depression was preliminarily clarified, which provides an ideal animal model for subsequent research on diseases induced by the Fzd6 mutation.

5-HTP decreases goat mammary epithelial cells apoptosis through MAPK/ERK/Bcl-3 pathway.

Serotonin (5-HT) is a monoamine and it could regulate cell growth by its receptors working on signaling pathways. 5-HTP is the precursor of 5-HT that help 5-HT synthesis. B cell leukemia/lymphoma 3 (Bcl-3) involved in cell death and proliferation through mitogen activated protein kinase (MAPK) pathway. However, there is little information about the effects of MAPK/Bcl-3 on apoptosis of goat mammary gland epithelial cells (GMECs). The aim of this study is to explore the interaction among 5-HTP, MAPK and Bcl-3 in GMEC apoptosis. In this study, 5-HTP treatment decreased cell apoptosis and promoted phosphorylation of ERK1/2 in GMEC. We also found that the activation and inhibition of ERK1/2 could affect GMEC apoptosis. The Annexin V-FITC/PI staining and western blotting results suggested that 5-HTP decreased GMEC apoptosis through ERK1/2 signaling pathway. And the results of RT-qPCR and western blotting demonstrated that both 5-HTP and ERK1/2 positively regulated Bcl-3 expression. Sum up all the results, we could draw the conclusion that 5-HTP decreased GMEC apoptosis through MAPK/ERK/Bcl-3 pathway.

Selenium Exerts Protective Effects Against Fluoride-Induced Apoptosis and Oxidative Stress and Altered the Expression of Bcl-2/Caspase Family.

Fluoride is widely distributed in nature, and at high concentrations, it targets the kidney and especially proximal tubule epithelial cells. Selenium is a typical trace element beneficial to humans, and the role of selenium in the prevention and treatment of fluoride-induced organ damage is an important research topic. The purpose of this study was to investigate the possible protective effects of selenium against fluoride-induced oxidative stress and apoptosis in rat renal tubular epithelial cells. We showed that the activity of antioxidant enzymes (superoxide dismutase and glutathione peroxidase) and total antioxidant capacity were significantly reduced in NaF-treated normal rat kidney cells (NRK-52E), whereas the levels of nitrogen monoxide (NO) and malondialdehyde (MDA) were significantly increased. Moreover, the number of apoptotic cells, mRNA expression of Bax, Bad, caspase-3, caspase-8, and caspase-9, and protein expression of Bax were elevated, while mitochondrial membrane potential and the protein expression of Bcl-2 were reduced. Compared with the NaF group, pretreatment with selenium enhanced the activity of antioxidant enzymes, mitochondrial membrane potential, and protein expression of Bcl-2, while the levels of NO and MDA, number of apoptotic cells, mRNA expression of Bax, Bad, caspase-3, caspase-8, and caspase-9, and protein expression of Bax were decreased. In conclusion, selenium exerted remarkable protective effect against NaF-induced oxidative stress and apoptosis and altered the expression of Bcl-2/caspase family.

Chaiqi decoction ameliorates vascular endothelial injury in metabolic syndrome by upregulating autophagy.

OBJECTIVE: The present study aimed to investigate the protective effect of the Chaiqi decoction on vascular endothelial injury in metabolic syndrome and to determine whether the underlying mechanism was associated with autophagy. METHODS: Chaiqi formula granules were administered to a rat model of metabolic syndrome established by feeding with a high-salt-sugar-fat diet (HSSFD). The drug-containing serum was used in a hyperglycemia cell model established using HUVECs cultured with palmitic acid PA. The influence of the Chaiqi decoction on metabolic syndrome-related vascular endothelial injury and autophagy was investigated. Autophagy flux was assessed in vitro by transfecting cells with GFP-mRFP-LC3 adenoviruses or incubating with DALGreen and DAPRed. RESULTS: The metabolic syndrome model rats displayed adiposity, hyperglycemia, dyslipidemia, hypertension, thickened intima, deposition of various forms of collagen and lipid droplets, downregulated levels of phosphorylated endothelial nitric oxide synthase and nitric oxide, upregulated expression of endothelin 1, and dysfunctional autophagy. All these abnormalities were ameliorated by administration of the Chaiqi decoction to the metabolic syndrome rats. Furthermore, the Chaiqi-containing serum could upregulate autophagy similarly to rapamycin, in a time-dependent manner. CONCLUSION: The Chaiqi decoction could ameliorate vascular endothelial injury by improving autophagy in metabolic syndrome.

5-Hydroxy-l-tryptophan Promotes the Milk Calcium Level via the miR-99a-3p/ATP2B1 Axis in Goat Mammary Epithelial Cells.

5-Hydroxy-l-tryptophan (5-HTP) is the primary product that converts l-tryptophan into 5-hydroxytryptamine by a rate-limiting enzyme. Our previous study found that 5-HTP could promote the intracellular calcium level in goat mammary epithelial cells (GMECs). Herein, first, dairy goats were injected with 5-HTP or saline daily from 7 days before delivery, and the calcium level in colostrum of 5-HTP-injected goats was significantly higher than that of saline-injected goats. Moreover, miR-99a-3p expression was significantly increased after 5-HTP treatment from transcriptome sequencing analysis and quantitative real-time polymerase chain reaction. In addition, it was found that ATP2B1 is one of the target genes of miR-99a-3p predicted by bioinformatic methods, which plays a crucial role in the maintenance of intracellular calcium homeostasis of mammary epithelial cells. Next, we confirmed that miR-99a-3p could increase the intracellular calcium level via decreasing ATP2B1 in GMECs. Taken together, we draw the conclusion that 5-HTP promotes the calcium level in colostrum possibly by increasing intracellular calcium of mammary epithelial cells induced by the miR-99a-3p/ATP2B1 axis.

Prevalence and risk factors of depression in patients with lung cancer: protocol for a systematic review and meta-analysis.

INTRODUCTION: Patients with lung cancer often experience heavy psychological distress, especially depression, which results in poorer quality of life, shorter survival time and greater mortality. Our aim is to summarise data on the prevalence and risk factors of depression in patients with lung cancer. METHODS AND ANALYSIS: We will search PubMed, EMBASE, MEDLINE, Cochrane Library, Web of Science, China National Knowledge Infrastructure, Wanfang and Chinese Biomedicine Literature Database (SinoMed) for studies on the prevalence and risk factors of depression in patients with lung cancer, which should be published from 1 January 1975 to 25 November 2018 in English/Chinese. Two reviewers will independently screen studies, extract data and assess the risk of bias. We will use RevMan V.5.0 and STATA V.12.0 software for statistical analysis. The I² test will be used to identify the extent of heterogeneity. Publication bias will be assessed by generating a funnel plot and performing the Begg and Egger test. The quality of the systematic review will be evaluated using the AMSTAR ('A Measurement Tool to Assess Systematic Reviews') criteria and 'The Grading of Recommendations Assessment, Development and Evaluation'. ETHICS AND DISSEMINATION: Since this is a review involving analysis of publicly available data, ethical approval is not required. The final results of this study will be published in a peer-reviewed journal. PROSPERO REGISTRATION NUMBER: CRD42018118167.

Prevalence and risk factors of anxiety and depression in Chinese patients with lung cancer：a cross-sectional study.

Background: Lung cancer is very common in China. The low cure rate, limited overall survival, and continuous therapies lead the patients to experience considerable psychological distress. Traditional Chinese medicine therapy is one unique treatment method in China. Nevertheless, most patients in the existing studies on anxiety and depression were treated in western medical hospitals. Therefore, it is necessary to identify the prevalence and risk factors of these emotional disorders in lung cancer patients treated in traditional Chinese medical hospitals. These findings may assist in clinical intervention. Patients and methods: A total of 315 patients with lung cancer were enrolled. Individuals completed the Hospital Anxiety and Depression Scale to assess their levels of anxiety and depression. Demographic and clinical data were also collected. Binary logistic regression analysis was used to identify factors that significantly predicted anxiety and depression. Results: The anxiety and depression prevalence rates of lung cancer patients were 43.5% and 57.1%, respectively. In the univariate analysis, patients without surgery, who were young, or who received radiotherapy were more likely to experience anxiety. Patients without surgery, who were young, or who had late-stage cancer, were more likely to experience depression. Binary logistic regression analysis showed that the risk factors of both anxiety and depression were lack of surgery and young and middle age (＜65, especially 45-65 years). Conclusion: Anxiety and depression were very common in lung cancer patients. Lack of surgery, young, and middle age, were independent risk factors for anxiety and depression. Therefore, medical workers should pay close attention to the emotional changes of young or middle-aged patients, or patients without the chance to undergo surgery.

Ursodeoxycholic acid after common bile duct stones removal for prevention of recurrence: A systematic review and meta-analysis of randomized controlled trials.

INTRODUCTION: The recurrence rate of common bile duct stones (CBDS) after removal has been reported to exceed 10% and no established pharmacologic treatment exists for the prevention of recurrent CBDS. Many studies indicated ursodeoxycholic acid (UDCA) has the potential to prevent the recurrence of CBDS. The aim of this systematic review is to evaluate the effects of UDCA for prevention of recurrence after common bile duct stones removal. METHODS AND ANALYSIS: We will systematically screen all randomized controlled trials (RCTs) published through electronically and hand searching. The following search engines including Ovid Medline, EMBASE, Cochrane CENTRAL, Proquest, Scopus, Web of Science, Pubmed, the Chinese Biomedical Literature Database, the China National Knowledge Infrastructure, VIP Information, Wanfang Data. Supplementary sources will be searched including gray literature, conference proceedings, and potential identified publications in OpenGrey.eu and Google Scholar databases. Two reviewers will independently conduct the trial inclusion, data extraction and assess the quality of studies. The recurrence rate of CBDS will be assessed as the primary outcomes. The adverse event that required discontinuation of UDCA intervention and the drop-outs (lost to follow-up) before the end of the study will be measured as secondary outcomes. Methodological quality will be evaluated according to the Cochrane risk of bias. All analyses will be applied by RevMan (version 5.3). RESULTS: This systemic review and meta-analysis will evaluate the effects of UDCA for prevention of recurrence after CBDS removal in RCTs. CONCLUSION: Our study will provide evidence to judge whether UDCA is an effective intervention to prevent the recurrence after CBDS removal.

MiR-330-5p negatively regulates ovine preadipocyte differentiation by targeting branched-chain aminotransferase 2.

The differentiation of preadipocytes into adipose tissues is tightly regulated by various factors including microRNAs and cytokines. This article aims to study the effect of miR-330-5p on expression of BCAT2 in ovine preadipocytes. Ovine preadipocytes were isolated, and we found that the miR-330-5p expression decreased gradually during the early differentiation of ovine preadipocytes, while BCAT2 expression increased. BCAT2 was identified as a direct target of miR-330-5p, ectopic expression of miR-330-5p could change the expression of both BCAT2 mRNA and protein. Silencing BCAT2 had the same inhibition effects as overexpressing miR-330-5p on the preadipocyte differentiation, but overexpressing BCAT2 had the converse effects. Taken together, we demonstrated that miR-330-5p is a negative regulator of differentiation by targeting BCAT2, and clarified the role of BCAT2 and miR-330-5p during preadipocyte differentiation.