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Abstract: Fenvalerate is a potential endocrine disruptor. Several studies have demonstrated that fenvalerate disrupts testosterone (T) synthesis in testes. T and estradiol (E2) are de novo synthesized in the developing brain. Thus, the aim of the present study was to investigate the effects of pubertal fenvalerate exposure on the synthesis of T and E2 and the expression of androgen receptor (AR) and estrogen receptors (ERs) in cerebral cortex. CD-1 mice were orally administered daily with either vehicle or fenvalerate (7.5 or 30mg/kg) from postnatal day (PND) 28 to PND56. The level of T and E2 in cerebral cortex was significantly decreased in males exposed to fenvalerate. In agreement with the decrease in T and E2 syntheses, the expression of 17β-HSD, a key enzyme for T synthesis, was significantly reduced in cerebral cortex of fenvalerate-exposed males. Conversely, in females, the expression of 17β-HSD in cerebral cortex was mildly up-regulated by fenvalerate and the level of T and E2 was mildly increased. Pubertal fenvalerate exposure had no effect on the expression of StAR, P45017α and P450scc, the key enzymes for T synthesis, and P450 aromatase, the key enzyme for E2 synthesis, in cerebral cortex of males and females. Interestingly, the expression of AR in cerebral cortex was up-regulated in male and female mice exposed to fenvalerate, whereas pubertal fenvalerate exposure did not affect the level of ERα and ERβ in cerebral cortex. Taken together, these results suggest that pubertal fenvalerate exposure disrupts T and E2 synthesis and the expression of AR in cerebral cortex. These changes of steroid status in the developing brain might be deleterious for neurobehavioral development. [Copyright &y& Elsevier]

Abstract: para-Nitrophenol (PNP) degradation cluster was cloned from a newly isolated Arthrobacter sp. strain NyZ415 by genome walking extending from a conserved region of gene encoding the oxygenase component of PNP monooxygenase. Sequence analysis indicated it contained 12 open reading frames including genes npdA1 and npdA2 which encoded the reductase and oxygenase components of PNP monooxygenase, respectively. Both NpdA1 and NpdA2 were expressed and purified to homogeneity. Hydroquinone and hydroxyquinol were captured when PNP was transformed by purified NpdA1A2 and para-benzoquinone was also captured for the first time as one of the products of PNP transformation by monooxygenase involved in the hydroxyquinol pathway. Sequence analysis showed four conserved residues (Arg100, Gln158, Arg161 and Thr193) related to the binding between the oxygenase component and substrate. Substitutions of any of above four residues to Ala have resulted in the complete loss of its activity on PNP transformation, indicating these four residues play important roles in the catalytic activity of PNP monooxygenase. [Copyright &y& Elsevier]

Summary: Human interferon alpha2b (IFN-α2b) is a pleiotropic cytokine used for the treatment of various cancers. Antibacterial peptide CM4 is a small peptide that can strongly inhibit many kinds of bacteria, fungi, and tumor cells, but it does no harm to normal cells. Here, we describe a protein expression system for the production of IFN-α2b/CM4 fusion protein in insoluble form in Escherichia coli, coupled to an efficient dialysis refolding and histidine-tag purification protocol. The expressed IFN-α2b/CM4 fusion protein not only displays significantly improved antitumor activity, but also retains the antibacterial–antifungal activity of CM4. [Copyright &y& Elsevier]

The aim of this study was to investigate the possible relationship between changes in neuronal and endothelial nitric oxide synthase (nNOS and eNOS) and arginase expression in the vestibular nucleus complex and the hippocampus (CA1, CA2/3 and the dentate gyrus (DG) at 10 h or 2 weeks following a unilateral vestibular deafferentation (UVD) in rats. There were no significant differences in nNOS or arginase II expression in the ipsilateral or contralateral VNC at either 10 h or 2 weeks post-UVD. For eNOS, there was only a significant decrease in expression in the ipsilateral VNC at 2 weeks post-UVD (P<0.01). In the hippocampus, the only significant difference in nNOS expression was a decrease in the ipsilateral DG at 2 weeks post-UVD (P<0.05). There was a significant decrease in eNOS expression in the contralateral CA2/3 region at 10 h post-UVD (P<0.01). The only other significant change in eNOS was an increase in the contralateral DG at 10 h post-UVD (P<0.01). Although arginase II was expressed in all regions of the hippocampus, there were no significant differences in arginase II expression at any time point following UVD. These results suggest that the changes in NOS expression that occur in the VNC and hippocampus following UVD are not correlated with one another or with changes in arginase II. [Copyright &y& Elsevier]

Excessive free fatty acids (FFAs) accumulation is a leading risk factor for the pathogenesis of insulin resistance (IR) in metabolic tissues, including the liver. Ethanolamine-phosphate phospho-lyase (ETNPPL), a newly identified metabolic enzyme, catalyzes phosphoethanolamine (PEA) to ammonia, inorganic phosphate, and acetaldehyde and is highly expressed in hepatic tissue. Whether it plays a role in regulating FFA-induced IR in hepatocytes has yet to be understood. In this study, we established an in vitro palmitic acid (PA)-induced IR model in human HepG2 cells and mouse AML12 cells with chronic treatment of PA. Next, we overexpressed ETNPPL by using lentivirus-mediated ectopic to investigate the effects of ETNPPL per se on IR without PA stimulation. We show that ETNPPL expression is significantly elevated in PA-induced IR and that silencing ETNPPL ameliorates this IR in hepatocytes. Inversely, overexpressing ETNPPL under normal conditions without PA promotes IR, reactive oxygen species generation, and ARG2 activation in both HepG2 and AML12 cells. Moreover, ETNPPL depletion markedly down-regulates ARG2 expression in hepatocytes. Besides, silencing ARG2 prevents ETNPPL-induced ROS accumulation and inhibition of autophagic flux and IR in hepatocytes. Finally, we found that phytopharmaceutical disruption of ETNPPL by quercetin ameliorates PA-induced IR in hepatocytes. Our study discloses that ETNPPL inhibiting autophagic flux mediates insulin resistance triggered by PA in hepatocytes via ARG2/ROS signaling cascade. Our findings provide novel insights into elucidating the pathogenesis of obesity-associated hepatic IR, suggesting that targeting ETNPPL might represent a potential approach for T2DM therapy. [Display omitted] • ETNPPL is aberrantly upregulated in PA-induced insulin resistance in hepatocytes. • ETNPPL impairing hepatic autophagy via the ARG2-ROS signaling axis contributes to PA-induced hepatic insulin resistance. • Quercetin disrupting ETNPPL ameliorates PA-induced IR in hepatocytes. [ABSTRACT FROM AUTHOR]

Abstract: Withangulatin A (WA), a bioactive withanolide isolated from Physalis angulata L., has a strong immunosuppressive activity. We have demonstrated that WA significantly suppressed the over-proliferation of ConA-induced T lymphocytes in a dose- and time-dependent manner, which modulated Th1/Th2-type balance dramatically. Moreover, WA-induced heme oxygenase 1 (HO-1) expression in mice T lymphocytes through the ERK/MAPK pathway and inhibited ConA-induced nuclear factor κB (NF-κB p65) nuclear translocation, directly or indirectly mediated by HO-1. Notably, WA affected the expression of COX-2 and PPAR-γ in ConA-induced T lymphocytes. The synergistic manner in which WA enhanced HO-1 expression through the ERK/MAPK pathway and NF-κB nuclear translocation and other regulatory proteins, thereby restricting T lymphocytes from over-expression and modulating Th1/Th2-type balance, could be the mechanism for the immunosuppressive properties of WA, the immunosuppressive effect of which may protect against autoimmune disorder diseases. [Copyright &y& Elsevier]

Abstract: Upregulation of the inducible nitric oxide synthase (iNOS) gene is associated with many pathological conditions such as endoplasmic reticulum (ER) stress, and X-box binding protein 1 (XBP1) is critical in mediating ER-stress responsive genes, including iNOS. Nonetheless, the mechanism by which XBP1 regulates iNOS during ER stress remains unexplored. Here we show that the active/spliced form of XBP1 protein, XBP1S, directly binds to the AABS (A-activator-binding site) in the iNOS promoter in vitro and in living cells. XBP1S exhibits dose-dependent activation of iNOS-specific reporter gene activity and endogenous iNOS expression. XBP1S is elevated whereas the unspliced form of XBP1, XBP1U, reduced in ER stress in HepG2 cells. In addition, XBP1U binds to XBP1S and this complex is associated with the iNOS promoter in response to ER stress. Furthermore, XBP1U acts as a negative mediator and suppresses XBP1S-mediated induction of iNOS. Collectively, we present the first evidence demonstrating the regulation of iNOS gene induction by the interaction between the spliced and unspliced forms of XBP1 in response to ER stress. [Copyright &y& Elsevier]

Abstract: A cytosolic manganese superoxide dismutase (cMnSOD) cDNA was cloned from the haemocytes of swimming crab Portunus trituberculatus using reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) method. Analysis of the nucleotide sequence revealed that the cMnSOD full-length cDNA consisted of 1106bp with an open reading frame of 861bp encoding a protein of 286 amino acids. Four conserved amino acids responsible for binding manganese (H110, H158, D243 and H247) and MnSOD signatures from 243 to 250 (DVWHHAYY) were observed. Sequence comparison showed that the cMnSOD deduced amino acid sequence of P. trituberculatus had similarity of 92%, 78%, 77% and 75% to that of blue crab Callinectes sapidus, white shrimp Litopenaeus vannamei, giant tiger prawn Penaeus monodon and freshwater prawn Macrobrachium rosenbergii, respectively. Quantitative real-time RT-PCR analysis revealed that P. trituberculatus cMnSOD transcript was strongly detected in haemocytes, hepatopancreas, heart, stomach, intestine, gill, ovary and muscle. RT-PCR analysis indicated that cMnSOD transcripts both in haemocytes and hepatopancreas increased at 3h and 6h after infection. These facts indicate that cMnSOD is potentially involved in the acute response against invading bacteria in P. trituberculatus. [ABSTRACT FROM AUTHOR]

Cholestatic Liver Fibrosis (CLF) is a hepatobiliary disease that typically arises as a late-stage complication of cholestasis, which can have multiple underlying causes. There are no satisfactory chemical or biological drugs for CLF. Total Astragalus saponins (TAS) are considered to be the main active constituents of the traditional Chinese herb Astragali Radix (AR), which has the obvious improvement effects for treating CLF. However, the mechanism of anti-CLF effects of TAS is still unclear. The present study was undertaken to investigate the therapeutic effects of TAS against bile duct ligation (BDL) and 3, 5-diethoxycarbonyl-1,4-dihydroxychollidine (DDC) -induced CLF models and to reveal the potential mechanism to support its clinic use with scientific evidence. In this study, BDL-induced CLF rats were treated with TAS (20 mg/kg, 40 mg/kg) and DDC-induced CLF mice were treated with 56 mg/kg TAS. The therapeutic effects of TAS on extrahepatic and intrahepatic CLF models were evaluated by serum biochemical analysis, liver histopathology and hydroxyproline (Hyp). Thirty-nine individual bile acids (BAs) in serum and liver were quantified by using UHPLC-Q-Exactive Orbitrap HRMS. qRT-PCR, Western blot and immunohistochemistry analysis were used to measure the expression of liver fibrosis and ductular reaction markers, inflammatory factors and BAs related metabolic transporters, along with nuclear receptor farnesoid X receptor (FXR). The serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bilirubin (TBiL), direct bilirubin (DBiL) and contents of liver Hyp were dose-dependently improved after treatment for TAS in BDL and DDC- induced CLF models. And the increased levels of ALT and AST were significantly improved by total extract from Astragali radix (ASE) in BDL model. The liver fibrosis and ductular reaction markers, α-smooth muscle actin (α-SMA) and cytokeratin 19 (CK19), were significantly ameliorated in TAS group. And the liver expression of inflammatory factors: interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α) and interleukin 1β (IL-1β) were significantly decreased after TAS treatment. In addition, TAS significantly ameliorated taurine-conjugated BAs (tau-BAs) levels, particularly α-TMCA, β-TMCA and TCA contents in serum and liver, which correlated with induced expressions of hepatic FXR and BAs secretion transporters. Furthermore, TAS significantly improved short heterodimer partner (SHP), cholesterol 7α-hydroxylase (Cyp7a1), Na+ taurocholate cotransport peptide (NTCP) and bile-salt export pump (BSEP) mRNA and protein expression. TAS exerted a hepatoprotective effect against CLF by ameliorating liver injury, inflammation and restoring the altered tau-BAs metabolism to produce a positive regulatory effect on FXR-related receptors and transporters. [Display omitted] • TAS improve cholestatic liver fibrosis in BDL and DDC models. • TAS regulate tau-BAs contents, especially α-TMCA, β-TMCA and TCA. • FXR signaling pathways are critical for the treatment of TAS in CLF. [ABSTRACT FROM AUTHOR]

Beclin1, also known as ATG6, has been shown to be closely related to coronavirus, however, the link between Beclin1 and nephropathogenic infectious bronchitis virus (NIBV) has been poorly investigated and there are no available antibodies specifically targeting the chicken Beclin1 protein. The study aimed to prepare and assay a polyclonal antibody to Beclin1, enabling a deeper understanding of the mechanism of action of Beclin1 in NIBV. In this study, we amplified the chicken Beclin1 target gene and constructed a recombinant plasmid using prokaryotic expression techniques, then obtained the recombinant target protein by induced expression. Finally, the serum is obtained by immunizing rabbits with the purified and concentrated protein. The results show that the antiserum potency of the ELISA assay was >1:204800. By western blotting and immunofluorescence, the antibodies we prepared specifically recognized the chicken Beclin1 protein, which is mainly found in the nucleus of trachea, lung, kidney, spleen and fabricant cells. NIBV infection significantly decreased the expression of Beclin1 in the trachea, but increased in others. We have successfully prepared specific rabbit anti-chicken Beclin1 polyclonal antibodies, and detected changes in tissues of diseased chickens infected with NIBV, laying the foundation for further studies on the role of Beclin1 in avian diseases. [ABSTRACT FROM AUTHOR]

• A comprehensive single cell miRNA activities map in BCG vaccine injected mice modeling tuberculous meningitis (TBM) brain infection; • Cell type enriched miRNAs were identified in TBM brain infection; • The functional miRNA-mRNA networks of macrophages and microglia, neurons, ependymal cells, and vascular smooth muscle cell was established. • Elevated miR-21a-3p activity is found in Microglia of TBM tissue. Our previous study revealed the transcriptome atlas of specific cell types in tuberculous meningitis (TBM) model mice injected with the BCG vaccine via scRNA sequencing. However, the activities of miRNAs in TBM at single-cell resolution remain to be explored. Cell type-specific miRNA activities were investigated by using motif enrichment analyses (miReact) on the transcriptome data of 15 cell types. The target mRNAs of miRNAs were predicted and subjected to enrichment analysis. Furthermore, miRNAs and their target mRNAs with opposite expression trends were chosen to construct functional networks. Besides, qRT-PCR and RNA scope were performed to verify the expression level of representative miRNA. The tSNE dimensionality reduction presented 15 cell types in TBM model mice, in which microglia and endothelial cells accounted for the majority. Target mRNAs of each cell type were predicted for verification or network construction. The immune and inflammation-related miRNA-mRNA networks of macrophages and microglia, oxidative phosphorylation-related miRNA-mRNA networks of neurons, ion and protein transport-related networks of epididymal cells, and angiogenesis-related miRNA-mRNA networks of VSMCs were constructed. The miRNA activity analysis revealed that miR-21a-3p activity was increased in microglia, macrophages, neurons and epididymal cells. The result of qRT-PCR and RNA scope indicate that miR-21a-3p was significantly higher-expressed in TBM brain tissue compared with normal brain tissue. In our study, an in-depth exploration of the mRNA expression and miRNA activity of macrophages, microglia, epididymal cells, neurons and vascular smooth muscle cells during TBM progression was conducted using scRNA-Seq, which provided novel insights into the immune cell engagement in TBM patients. [ABSTRACT FROM AUTHOR]

In the present study, we investigated the changes of microRNA (miRNA) expression upon depression and electro-acupuncture (EA) intervention in chronic unpredictable mild stress (CUMS) rats using microarray analysis. Results showed that EA intervention remarkably improved behavioral indexes in terms of crossing number, rearing number, sucrose preference and body weight of CUMS rats. Microarray analysis revealed that a total of 153 differentially expressed miRNAs were regulated by CUMS, and the expression of 180 differentially expressed miRNAs was changed after EA intervention. Among these miRNAs, two miRNAs were significantly up-regulated and four miRNAs were significantly down-regulated by CUMS. Moreover, four miRNAs were significantly up-regulated and 12 miRNAs were significantly down-regulated after EA intervention. The expressions of miR-383-5p and miR-764-5p were up-regulated after CUMS, while their expressions were down-regulated by EA intervention. Further analysis showed that 1260 possible target genes were predicted for miR-383-5p and miR-764-5p, and 97 pathways and 137 gene ontology (GO) were involved. Among these pathways and GO, about 20 pathways and 21 GO were related to depression. Changes of miR-383-5p and miR-764-5p indicated that EA might exert its therapeutic effect on depression through promoting the neurotrophy and inhibiting the abnormal apoptosis of neurons as well as other correlative signal pathways. In conclusion, our present study enriched the understanding of pathological process of depression and revealed possible mechanisms of EA on depression. [ABSTRACT FROM AUTHOR]

Objectives Circulating miRNAs have been reported to have potential in detecting various diseases. However, few studies explored differentially expressed miRNAs in plasma of patients with pulmonary embolism (PE). Our study is to identify plasma miRNAs which can serve as potential biomarkers of PE. Materials and methods Exiqon miRCURY Ready-to-Use PCR Human panel I + II V1.M was conducted to identify differently expressed miRNAs in pooled plasma samples of PE patients compared with normal controls. Expressions of identified miRNAs were assessed in 37 PE patients as well as matched normal individuals followed by validation on six Beagle dogs by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Results Twelve miRNAs were identified from the screening phase. Moreover, miR-134, previously reported related with PE, and hypoxia-induced miR-210 were also submitted to the validation phase. Only miR-28-3p was found significantly elevated in the plasma of PE patients. Compared with the level of plasma miR-28-3p of the dogs before PE, the elevated miR-28-3p did not alter significantly at 1, 2, 4 and 6 h after PE. The area under the receiver operating characteristic (ROC) curve of plasma miR-28-3p was 0.792 (95% confidence interval: 0.689–0.896). KEGG pathway analysis showed that miR-28-3p might involve in PE related pathways such as inositol phosphate metabolism and phosphatidylinositol signaling system. Conclusion Our study indicated that elevated plasma miR-28-3p could be used as a non-invasive and stable biomarker in the detection of PE. Further researches on the miRNA are warranted. [ABSTRACT FROM AUTHOR]

Portunus trituberculatus is an important farmed crustacean in China. Growth traits are important economic traits that are controlled by many genes. The aim of this study was to screen growth-related genes using Illumina paired-end sequencing technology, and to identify growth-related SNPs in P. trituberculatus by association analysis. A total of 36,599,712 and 29,247,160 qualified Illumina read pairs for small size (SG) and large size crabs (LG) were obtained, giving rise to total clean bases of 7.14 and 5.72 G, respectively. A gene expression comparison showed that a total of 117 unigenes were differentially expressed between SG and LG, including 49 up-regulated and 68 down-regulated unigenes in SG and LG, respectively. GO functional enrichment analysis results showed up-regulated genes in LG were involved in three biological processes. These were cell communication, ovarian follicle cell development and cell division. Down-regulated genes in LG were involved in five biological processes, which were chitin metabolism, carbohydrate metabolism, ATP hydrolysis coupled proton transport, the steroid hormone mediated signaling pathway and fatty acid biosynthesis. Association analysis identified one growth-associated SNP, which was located in hemocyanin (comp58070-R31). This research has provided information on key genes, SNPs and biological processes related to growth, which will increase the efficiency of selective breeding programs for this important species. [ABSTRACT FROM AUTHOR]

Calcification of end plate chondrocytes is a major cause of intervertebral disc (IVD) degeneration. However, the underlying molecular mechanism of end plate chondrocyte calcification is still unclear. The aim of this study was to clarify whether autophagy in end plate chondrocytes could protect the calcification of end plate chondrocytes. Previous studies showed that intermittent cyclic mechanical tension (ICMT) contributes to the calcification of end plate chondrocytes in vitro. While autophagy serves as a cell survival mechanism, the relationship of autophagy and induced end plate chondrocyte calcification by mechanical tension in vitro is unknown. Thus, we investigated autophagy, the expression of the autophagy genes, Beclin-1 and LC3, and rat end plate chondrocyte calcification by ICMT. The viability of end plate chondrocytes was examined using the LIVE/DEAD viability/cytotoxicity kit. The reverse transcription-polymerase chain reaction and western blotting were used to detect the expression of Beclin-1; LC3; type I, II and X collagen; aggrecan; and Sox-9 genes. Immunofluorescent and fluorescent microscopy showed decreased autophagy in the 10- and 20-day groups loaded with ICMT. Additionally, Alizarin red and alkaline phosphatase staining detected the palpable calcification of end plate chondrocytes after ICMT treatment. We found that increased autophagy induced by short-term ICMT treatment was accompanied by an insignificant calcification of end plate chondrocytes. To the contrary, the suppressive autophagy inhibited by long-term ICMT was accompanied by a more significant calcification. The process of calcification induced by ICMT was partially resisted by increased autophagy activity induced by rapamycin, implicating that autophagy may prevent end plate chondrocyte calcification. [ABSTRACT FROM AUTHOR]

Abstract: Myasthenia gravis (MG) is a T cell-dependent and B cell-mediated autoimmune disease of neuromuscular junctions and cytokines may play a crucial role in the pathogenesis and perpetuation of MG. MicroRNAs (miRNAs) have been implicated as fine-tuning regulators controlling diverse biological processes at the level of posttranscriptional repression. Dysregulation of miRNAs has been described in various disease states. In this study, miRNA microarrays identified let-7 family to be decreased in peripheral blood mononuclear cells (PBMCs) from MG patients compared to the healthy controls. We next demonstrated the differential expression of let-7 family in larger samples by quantitative real-time PCR. Using a combination of bioinformatics and molecular approaches, we confirmed IL-10 as a target for let-7c. IL-10 expression also showed a negative correlation with let-7c expression in PBMCs from MG patients. Further experiments revealed that induced levels of IL-10 were inversely related to let-7c levels. We also showed that let-7c could regulate IL-10 expression in Jurkat cells. In summary, our results suggest that abnormal expression/regulation of microRNAs may contribute to or be indicative of the initiation and progression of MG. [Copyright &y& Elsevier]

Abstract: Objective: Th1 activation and regulatory T (Treg) cell suppression have been observed in acute coronary syndrome (ACS), including unstable angina (UA) and acute myocardial infarction (AMI). However, the role of Th17 cell or IL-17A remains controversial in ACS patients, and little is known about the role of recently discovered Th17/Th1 cells, a subset of Th17 cells, in coronary artery disease (CAD). The purpose of this study is to investigate functional changes of Th17/Th1, Th17, Th1, Th2 and Treg cells in ACS patients. Methods: The contents of Th17/Th1, Th17, Th1, Th2 and Treg cells, related gene expression, and plasma cytokines from CAD and control patients with normal coronary arteries (NCA) were measured by flow cytometry, real-time quantitative PCR and ELISA. Results: Th17/Th1 and Th1 cell contents and related gene expression (T-bet, IFN-γ, STAT4, RORγt, STAT3 and IL-17) were significantly increased in ACS patients, whereas plasma IFN-γ only increased in CAD patients. In contrast, Treg cell population, Foxp3 levels, and plasma TGF-β1 were decreased in ACS patients compared with stable angina (SA) and NCA patients. Conclusion: The study showed activation of Th17/Th1 and Th1 cell in ACS patients, which may provide insight into the mechanisms underlying culprit plaque relevant T-cell activation in ACS patients. [Copyright &y& Elsevier]

Objective: To evaluate the effect of growth differentiation factor-9 (GDF-9) and bone morphogenetic protein-15 (BMP-15) on the development of follicles among patients with polycystic ovary syndrome (PCOS). Design: Case-control study. Setting: University Hospital. Patient(s): Twenty-two oocytes were obtained from 15 patients with PCOS and 67 oocytes from 58 controls. Cumulus granulosa cells (GC) were obtained from 16 patients with PCOS and controls treated with intracytoplasmic sperm injection. Intervention(s): Immunofluorescence combined with laser scanning confocal microscopy and immunocytochemistry were used to analyze the expression of GDF-9 and BMP-15 in oocytes and cumulus GCs. Main Outcome Measure(s): To detect the protein expression levels. Result(s): No significant difference was found in the expression of GDF-9 and BMP-15 in the oocytes and BMP-15 in the cumulus GCs of patients with PCOS and controls. However, the expression of GDF-9 in cumulus GCs of patients with PCOS was decreased significantly compared with controls (8.88 ± 1.52 vs. 5.01 ± 0.83). Conclusion(s): The expression of GDF-9 and BMP-15 in the oocytes of patients with PCOS who received ovulation induction treatment was in the normal range, but the GDF-9 expression in cumulus GCs from patients with PCOS was significantly lower than the normal. Reduced GDF-9 expression in cumulus GCs of patients with PCOS appears to be associated with decreased long-term developmental potential of the oocytes of patients with PCOS. [Copyright &y& Elsevier]

Abstract: The consumption of dietary fibre plays an important role in the prevention of diseases, such as constipation, haemorrhoids. Recently, chemical and physical properties of citrus fibres have been widely studied. In this paper, the polygalacturonase (PG) and cellulase (Cx) gene expression of Cara Cara (Citrus sinensis (L.) Osbeck) navel orange fruit stored on tree (ST) was compared with fruit stored in room (SR). The results showed that the mRNA expression levels of PG increased significantly in the fruits of ST, in contrast, the expression levels of Cx increased slightly only in peel of ST. Total pectin (TP) and protopectin of ST fruits pulp were higher than those of SR at every time point. The contents of insoluble dietary fibre (IDF), hemicellulose (HC), cellulose (CEL) and lignin of ST fruits were less than that in SR. However, in fruits from ST, a significant increase of soluble dietary fibre (SDF) and water soluble pectin (WSP) occurred, compared with fruits of SR. Our studies indicated that fruit stored on tree is quite useful for regulating the gene expression and controlling contents of dietary fibre on Cara Cara navel orange. [Copyright &y& Elsevier]

Cholestasis caused by bile secretion and excretion disorders is a serious manifestation of liver disease. With limited treatment methods, it affects millions of people worldwide. Huangqi decoction (HQD), an effective traditional Chinese medicine, is used to treat chronic cholestatic liver diseases. However, the action mechanisms of it were not fully elucidated. We aim to investigate the therapeutic effect of HQD, and its active component, astragalosides, against α-naphthylisothiocyanate (ANIT)-induced cholestasis in rats based on targeted metabolomics analysis and revel the potential mechanism. The therapeutic effect of HQD and astragalosides on ANIT-induced cholestasis model rats were evaluated by serum biochemical analysis. Liver damage was identified by histopathology. The levels of bile acids (BAs) and free fatty acids (FFAs) in serum and liver tissues were measured by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-TQMS). qRT-PCR and Western blot analysis were used to measure the expression of nuclear hormone receptor, membrane receptor and BA transporter protein in cholestatic rats before and after HQD and astragalosides treatment. The obtained data showed that the administration of ANIT caused obvious cholestasis with significantly increased intrahepatic retention of hydrophobic BAs and altered FFAs, which were consistent with the liver histopathological and serum biochemical findings. HQD and astragalosides treatment were able to attenuate ANIT-induced BAs and FFAs perturbation, ameliorate the impaired liver function, histopathological ductular reaction, and lipid peroxidation damage by ANIT. Elevated mRNA and protein expression of transporters related to BA metabolism and genes related to lipogenesis and lipid oxidation metabolism in cholestasis were attenuated or normalized by HQD and astragalosides treatment. Intervention by ANIT can significantly change the homeostasis of BAs and FFAs. HQD and astragalosides exerted a hepatoprotective effect against cholestatic liver injury by restoring the altered BA and FFA metabolism through the improvement of BA transporter, nucleus hormone receptor, and membrane receptor. Image 1 [ABSTRACT FROM AUTHOR]

Gut microbiome dysbiosis is closely associated with cholestatic liver disease. Huangqi decoction (HQD), a traditional herbal formula, has protection against cholestatic liver injury. However, the effect of HQD on gut microbiome remains unknown. To investigate the effect of HQD on 3, 5-diethoxycarbonyl-1, 4-dihydrocollidine (DDC) induced cholestatic liver injury and its effect on the gut microbiome profiles. Mice with DDC-induced cholestatic liver injury were treated with low and high doses of HQD for 8 weeks. Fecal samples were analyzed by 16 S ribosomal DNA sequencing. Barrier function as well as intestinal and hepatic inflammation was analyzed by real-time PCR and western blotting. HQD treatment ameliorated the DDC-induced liver injury and collagen deposition around hepatic bile ducts. Moreover, decreased diversity, reduced richness, and abnormal composition of intestinal microbiota of cholestatic mice were remarkably attenuated by HQD supplementation. Differences in bacterial abundance, including levels of Prevotellaceae_NK3B31_group , Alistipes , and Gordonibacter , were increased in DDC-induced mice, as compared with control mice, and were decreased after HQD treatment. Moreover, intestinal dysbiosis promoted disruption of the intestinal barrier in cholestatic mice. However, HQD treatment alleviated intestinal barrier dysfunction. Importantly, increased hepatic expression of pro-inflammatory factors and the NLRP3 inflammasome, which have a positive correlation with differential bacteria, were characteristics found in DDC-induced cholestatic mice that were alleviated upon treatment with HQD. HQD treatment alleviated gut microbiota dysbiosis, ameliorated the intestinal barrier dysfunction, inhibited liver inflammation, and protected against DDC-induced cholestatic liver injury. Image 1 [ABSTRACT FROM AUTHOR]

• Transcriptomic and metabolic analysis of jujube fruit peel coloring during ripening. • Strong accumulation of malvidin and delphidin correlated with peel reddening. • Differentially expressed genes in the flavonoid synthesis pathway identified. • 3 UFGT genes may play essential functions for the accumulation of anthocyanins. Coloring is an important external quality of jujube fruit (Ziziphus jujuba Mill.), however during long-term storage, its commercial value degrades as the peel color and lustre significantly fade. Here, the gene expression and metabolite concentration were profiled in the fruit peel of jujube harvested at three ripening periods to elucidate the color formation mechanism. A strong accumulation of malvidin 3-O-glucoside and delphinidin 3-O-glucoside correlated with the reddening of jujube peel. At the onset of the fruit ripening, strong activities of the genes in the early steps of the flavonoid biosynthetic pathway were observed. During the last ripening periods, three UDP-glucose flavonoid 3-O-glucosyltransferase (UFGT) involved in the accumulation of anthocyanins were significantly increased and their proper manipulation could delay the peel reddening. This study sheds light on the metabolic pathways and candidate genes underlying the peel coloration in jujube and lays a foundation for the improvement of jujube fruit appeal during long-term storage. [ABSTRACT FROM AUTHOR]

In this study, in order to explore potential depressive biomarkers and potential regulatory targets of KXS on depression, we assessed the effects of Kai-Xin-San (KXS) on lipid metabolism in depressed patients (DPs) and rats exposed to chronic and unpredictable mild stress (CUMS). Serum samples were collected from DPs, DPs with 8 weeks of KXS treatment (KXS) and healthy controls (HCs), and non-targeted lipidomics was used to analyze the effect of KXS on serum lipid metabolites in DPs. Based on UPLC-Q-TOF/MS technology, differential metabolites were validated in a large sample size. The potential regulatory network of KXS was analyzed by bioinformatic analysis, and the expressions of proteins in serum were verified using western boltting analysis. Moreover, effects of KXS on serum lipid and lipid metabolism-related hormone levels in CUMS rats were detected by enzyme-linked immunosorbent assay and enzymatic method. We validated that the levels of six serum lipid metabolites (N-Desmethylcitalopram (HMDB14021), PC(14:1(9Z)/24:0) (HMDB07926), PC(P-18:1(11Z)/20:0) (HMDB11281), PC(O-18:0/20:4(8Z,11Z,14Z,17Z)) (HMDB13420), PC(16:0/P-18:0) (HMDB07995) and PC(16:0/P-18:1(11Z)) (HMDB07996)) between HC/DP groups and between DP/KXS groups were significantly different. Among these six metabolites, HMDB07995, HMDB07996, HMDB13420 and HMDB11281 were highly sensitive and specific for depression and KXS treatment by receiver operating characteristic (ROC) curve analysis. matrix metalloproteinases (MMPs) including MMP2 and MMP9, apolipoproteins (Apo) including APOA1 and APOC1 were up-regulated and apolipoproteins (Apo) including APOB, APOD and APOE, phospholipid transfer protein (PLTP), Paraoxonase 1 (PON1) were down-regulated in DPs, and KXS treatment could reverse these changes. In CUMS rats, KXS could increase the open-field score, sucrose preference and body weight, and reduce immobility time. Furthermore, KXS increased the serum levels of the above-mentioned six metabolites, reduced serum total cholesterol (TCH), triglyceride (TG) and free fatty acid (FFA) levels and increased the serum high-density lipoprotein cholesterol (HDL-C) level in CUMS rats. In addition, leptin and ghrelin were down-regulated by KXS. The results suggested that KXS exerted antidepressant effects by regulating the signaling pathways involved in lipid metabolism disorders. The lipid metabolites might be potential biomarkers of depression and possible targets for KXS-based treatment of depression. Image 1 [ABSTRACT FROM AUTHOR]

Bone morphogenetic protein-15 is expressed in ooctytes and cumulus granulosa cells. It may play an important role in the hCG-induced oocyte final maturation and corpus luteum (CL) formation. [Copyright &y& Elsevier]