Effect of high-fructose consumption in pregnancy on the bone growth of offspring rats.

Growing evidence suggests that bone health is programmed in early life. Maternal diet may influence the skeletal development of offspring. We aimed to determine the possible effects of high-fructose intake during pregnancy on different aspects of long bone morphology in the offspring of rats and to initially explore the possible mechanisms. Pregnant Sprague-Dawley rats were randomly divided into four groups and intragastrically administered the same dose of distilled water (CON, n = 12), 20 g/kg/day glucose (GLU, n = 12), 10 g/kg/day fructose (LFRU, n = 12), or 20 g/kg/day fructose (HFRU, n = 12) for 21 days during gestation. Computed tomography was used to analyze the cortical and cancellous bones of the distal femur of the offspring rats, and circulating bone metabolic biomarkers were measured using enzyme immunoassay. The results showed that high-fructose intake during pregnancy could decrease body weight, impair glucose metabolism, and increase serum leptin and uric acid in offspring. The offspring in the HFRU group had higher levels of the N-terminal propeptide of type I procollagen (PINP) and the C-telopeptide of type I collagen (CTX). The bone mean density (BMD), the total cross-sectional area inside the periosteal envelope (Tt.Ar), cortical bone area (Ct.Ar), medullary (or marrow) area (Ma.Ar), and trabecular mean density of the offspring in the HFRU group were lower than those in the CON group. Tartrate-resistant acid phosphatase (Trap) staining showed that high-fructose intake during pregnancy could increase the number of osteoclasts and increase the absorption area. Our results suggested that excessive fructose intake during pregnancy could inhibit skeletal development in offspring. Thus, attention to fructose intake during pregnancy is important for bone development in offspring.

Analysis of Factors Influencing Depression in Elderly People Living with HIV/AIDS Based on Structural Equation Model: A Cross-Sectional Study in Guangxi, China.

Purpose: The purpose of our study is to understand the current status of depression and medical social support in elderly HIV/AIDS, as well as the role of social support on depression, so as to provide a certain reference for reducing the occurrence of depression in the population. Methods: A total of 115 participants with PLWHA (people living with HIV/AIDS) aged 50 years or older were collected in Guilin from December 2021 to July 2022. Depression and medical social support were assessed using the Center for Streaming Depression Scale (CES-D) and the Medical Social Support Scale (MOS-SSS). The structural equation model was used to examine the relationship between medical social support and depression. Results: Sixty-one of 115 participants developed depressive symptoms with a prevalence of 53.0%. The results of univariate analysis showed that ethnicity, health status, mean monthly income, antiviral treatment status, and medical social support influenced PLWHA depression (P<0.05). Simple linear regression showed that health status (95% CI: -9.901~-2.635), and antiviral treatment status (95% CI: -12.969~-3.394) influent depression (P<0.05). There were associations between total medical social support, practical support dimension, message and emotional support dimension, social interactive cooperation dimension, emotional support dimension and depression (unadjusted and adjusted for contextual factors) (P < 0.05). Using multiple linear regression analyses, we found that medical-social support was negatively associated with depression with a standardized effect value of -0.223. PLWHA with higher medical social support had lower prevalence of depression. Conclusion: The results indicate that the prevalence of depression among HIV/AIDS patients in Guilin is high. So the joint efforts of individuals, families, and society are needed to improve the physical and mental health of the PLWHA.

Let7b-5p inhibits insulin secretion and decreases pancreatic ß-cell mass in mice.

MicroRNAs are crucial regulators for the development, mass and function of pancreatic ß-cells. MiRNA dysregulation is associated with ß-cell dysfunction and development of diabetes. The members of let7 family are important players in regulating cellular growth and metabolism. In this study we investigated the functional role of let7b-5p in the mouse pancreatic ß-cells. We generated pancreatic ß-cell-specific let7b-5p transgenic mouse model and analyzed the glucose metabolic phenotype, ß-cells mass and insulin secretion in vivo. Luciferase reporter assay, immunofluorescence staining and western blot were carried out to study the target genes of let7b-5p in ß-cells. Let7b-5p overexpression impaired the insulin production and secretion of ß-cells and resulted impaired glucose tolerance in mice. The overexpressed let7b-5p inhibited pancreatic ß-cell proliferation and decreased the expression of cyclin D1 and cyclin D2. Our findings demonstrated that let7b-5p was critical in regulating the proliferation and insulin secretion of pancreatic ß-cells.

Components characterisation of Berchemia lineata (L.) DC. by UPLC-QTOF-MS/MS and its metabolism with human liver microsomes.

Investigations were performed on the determination of the main components in Berchemia lineata (L.) DC. (BL) and its metabolism with human liver microsomes (HLM). A total of 35 compounds were detected in BL extracts and 25 of them including 6 naphthopyrones, 10 flavonoids, 2 phenolic acids, 2 phenols, 4 fatty acids and 1 quinone were unambiguously or tentatively identified by UPLC-QTOF-MS/MS. Among them, naphthopyrones were first identified in BL extracts and labelled in chromatography. In addition, the weak inhibitory effects of BL extracts (IC50＝149.25 µg/mL) and rubrofusarin-6-O-α-L-rhamnosyl-(1-6)-O-ß-D-glu-copyranside (the main component of BL extracts, M0; IC50＝82.14 µM) on CYP3A4 were also proved using testosterone as specific probe drug. The main metabolic pathway of M0 by HLM was hydroxylation in its aglycone, the metabolite was tentatively identified as 10-hydroxy-rubrofusarin-6-O-α-L-rhamnosyl-(1-6)-O-ß-D-glucopyranside. Components characterisation and the metabolism with HLM could help the further development and application of BL.

Silencing of lncRNA DLEU1 inhibits tumorigenesis of ovarian cancer via regulating miR-429/TFAP2A axis.

Long non-coding RNAs (lncRNAs) are known as crucial regulators in the development of OC. In the current study, we aim to explore the function and molecular mechanism of lncRNA DLEU1 in OC. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to determine the expression of DLEU1, miR-429, and TFAP2A in OC cells and tissues. The relationship among DLEU1, miR-429, and TFAP2A was tested by dual-luciferase reporter (DLR) assay. Besides, the proliferative, migratory and invasive abilities of OC cells were analyzed by MTT, wound healing, and transwell assays, respectively. Western blot was performed to determine the protein expression of TFAP2A. The expression of lncRNA DLEU1 and TFAP2A were upregulated, and miR-429 was downregulated in OC tissues. Silencing of DLEU1 inhibited the proliferation, migration, and invasion of OC cells. Bioinformation and DLR assay showed that DLEU1 acted as the sponge for miR-429. Moreover, miR-429 could directly target TFAP2A and inhibit the proliferation, migration, and invasion of OC cells. Moreover, we observed a negative correlation between miR-429 and DLEU1, and between miR-429 and TFAP2A in OC tissues. The transfection of miR-429 inhibitor or pcDNA-TFAP2A reversed the inhibitory effects of si-DLEU1 on the proliferation, migration, and invasion of OC cells. Silencing of DLEU1 inhibited the proliferation, migration, and invasion of OC cells by regulating miR-429/TFAP2A axis, indicating a potential therapeutic target for OC.

Altered Inflammatory Pathway but Unaffected Liver Fibrosis in Mouse Models of Nonalcoholic Steatohepatitis Involving Interleukin-1 Receptor-Associated Kinase 1 Knockout.

BACKGROUND Interleukin-1 receptor-associated kinases (IRAKs) are crucial mediators in the signaling pathways of Toll-like receptors (TLRs)/IL1Rs. Targeting the IRAK4/IRAK1/TRAF6 axis and its associated pathway has therapeutic benefits in liver fibrosis. However, the function of IRAK1 itself in the development of liver fibrosis remains unknown. MATERIAL AND METHODS Irak1 global knockout (KO) mice were generated to study the functional role of Irak1 in liver fibrosis. Male Irak1 knockout and control mice were challenged with chronic carbon tetrachloride (CCl4) or fed a methionine- and choline-deficient diet (MCDD) to generate models of nonalcoholic steatohepatitis (NASH). Liver inflammation and collagen deposition were assessed by histological examination, quantitative real-time PCR (qRT-PCR), and western blotting of hepatic tissues. RESULTS The mRNA expression of the downstream inflammatory gene Il1b was significantly lower in Irak1-KO than in control mice. Irak1 ablation had little effect on inflammatory cell infiltration into livers of mice with NASH. Collagen deposition and the expression of genes related to fibrogenesis were similar in the livers of Irak1-KO and control mice exposed to CCl4 and MCDD. The loss of Irak1 did not affect lipid or glucose metabolism in these experimental models of steatohepatitis. CONCLUSIONS Irak1 knockout reduced the expression of inflammatory genes but had no effect on hepatic fibrogenesis. The Irak1-related pathway may regulate liver fibrosis via other pathways or be compensated for by other factors.

YY1 deficiency in ß-cells leads to mitochondrial dysfunction and diabetes in mice.

BACKGROUND: The transcription factor YY1 is an important regulator for metabolic homeostasis. Activating mutations in YY1 lead to tumorigenesis of pancreatic ß-cells, however, the physiological functions of YY1 in ß-cells are still unknown. Here, we investigated the effects of YY1 ablation on insulin secretion and glucose metabolism. METHODS: We established two models of ß-cell-specific YY1 knockout mice. The glucose metabolic phenotypes, ß-cell mass and ß-cell functions were analyzed in the mouse models. Transmission electron microscopy was used to detect the ultrastructure of ß-cells. The flow cytometry analysis, measurement of OCR and ROS were performed to investigate the mitochondrial function. Histological analysis, quantitative PCR and ChIP were performed to analyze the target genes of YY1 in ß-cells. RESULTS: Our results showed that loss of YY1 resulted in reduction of insulin production, ß-cell mass and glucose tolerance in mice. Ablation of YY1 led to defective ATP production and mitochondrial ROS accumulation in pancreatic ß-cells. The inactivation of YY1 impaired the activity of mitochondrial oxidative phosphorylation, induced mitochondrial dysfunction and diabetes in mouse models. CONCLUSION: Our findings demonstrate that the transcriptional activity of YY1 is essential for the maintenance of mitochondrial functions and insulin secretion in ß-cells.

Effects of 7,8-dihydroxyflavone on rat jejunal dynamics subjected to ischaemia-reperfusion injury.

It is known that 7,8-dihydroxyflavone (7,8-DHF), a synthetic agonist specific for TrkB, promotes intestinal cholinergic contraction. However, after intestinal ischaemia-reperfusion (IR) injury, how 7,8-DHF affects intestinal contractile dynamics is unknown. In this study, an IR injury model was prepared with rats subjected to 45 minutes clamping of the superior mesenteric artery. The IR injury decreased postoperative food intake and body weight, delayed defecation time, lowered intestinal propulsive rate and decreased cholinergic contraction of jejunal muscle strips, indicating the occurrence of injured jejunal contraction after IR. Feeding rats with 7,8-DHF improved these intestinal activities injured by IR, which exhibited the in vivo effect of 7,8-DHF. To explore its molecular mechanism, the expression and phosphorylation of TrkB, PLC γ1, Akt, and ERK1/2 in the jejunal strips were examined with western blots. The IR injury significantly decreased the expression and phosphorylation levels of all factors studied here. However, 7,8-DHF feeding specifically enhanced the phosphorylation of TrkB, PLC γ1 and Akt factors in both sham- and IR-operated rats, indicating that 7,8-DHF may have activated TrkB which then activated its downstream PLC γ1 and Akt. Finally, we found that 7,8-DHF augmented cholinergic receptor M3 expression somehow. These results imply a possibility that 7,8-DHF might be capable of alleviating the jejunal contractile damage caused by IR through activation of TrkB and augmentation of M3 expression.

MiR-221/222 Inhibit Insulin Production of Pancreatic ß-Cells in Mice.

Microribonucleic acids (miRNAs) are essential for the regulation of development, proliferation, and functions of pancreatic ß-cells. The conserved miR-221/222 cluster is an important regulator in multiple cellular processes. Here we investigated the functional role of miR-221/222 in the regulation of ß-cell proliferation and functions in transgenic mouse models. We generated 2 pancreatic ß-cell-specific-miR-221/222 transgenic mouse models on a C57BL/6J background. The glucose metabolic phenotypes, ß-cell mass, and ß-cell functions were analyzed in the mouse models. Adenovirus-mediated overexpression of miR-221/222 was performed on ß-cells and mouse insulinoma 6 (MIN6) cells to explore the effect and mechanisms of miR-221/222 on ß-cell proliferation and functions. Luciferase reporter assay, histological analysis, and quantitative polymerase chain reaction (PCR) were carried out to study the direct target genes of miR-221/222 in ß-cells. The expression of miR-221/222 was significantly upregulated in ß-cells from the high-fat diet (HFD)-fed mice and db/db mice. Overexpression of miR-221/222 impaired the insulin production and secretion of ß-cells and resulted in glucose intolerance in vivo. The ß-cell mass and proliferation were increased by miR-221/222 expression via Cdkn1b and Cdkn1c. MiR-221/222 repressed insulin transcription activity through targeting Nfatc3 and lead to reduction of insulin in ß-cells. Our findings demonstrate that miR-221/222 are important regulators of ß-cell proliferation and insulin production. The expression of miR-221/222 in ß-cells could regulate glucose metabolism in physiological and pathological processes.

MicroRNA-935 acts as a prognostic marker and promotes cell proliferation, migration, and invasion in colorectal cancer.

Colorectal cancer (CRC) is a common cause of cancer-related deaths worldwide. MicroRNA-935 (miR-935) has been reported to be involved in several cancers. In the present study, we aimed to investigate the role of miR-935 in the progression of CRC. The expression levels of miR-935 in CRC tissues and cells were detected using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The prognostic significance of miR-935 was identified using Kaplan-Meier survival analysis and Cox regression assay. Cell proliferation capacity was detected by Cell Counting Kit-8 (CCK-8) assay. Cell migration and invasion capacities were determined by Transwell assay in CRC cells. Bioinformatics analysis and luciferase reporter assays were used to confirm the direct target of miR-935. The expression of miR-935 was increased in CRC tissues and cells. Overexpression of miR-935 was significantly associated with lymph node metastasis and TNM stage. Overexpression of miR-935 in patients predicted shorter overall survival compared to low miR-935 expression. The expression of miR-935 was indicated to be an independent prognostic factor for CRC patients. In addition, overexpression of miR-935 in CRC cells promoted cell proliferation, migration, and invasion, whereas inhibition of miR-935 suppressed cell proliferation, migration, and invasion. Bioinformatics and luciferase assays revealed that INPP4A is a direct target of miR-935. Our findings suggest that miR-935 functions as an oncogene and promotes CRC progression. INPP4A is a potential target of miR-935. And miR-935 may represent a prognostic biomarker and potential therapeutic strategy for CRC treatment.

The distribution and accumulation of mercury and methylmercury in surface sediments beneath the East China Sea.

China is a massive mercury emitter, responsible for a quarter of the world's mercury emissions, which transit the atmosphere and accumulate throughout its watercourses. The Changjiang (Yangtze) River is the third largest river in the world, integrating mercury emissions over its 1.8 × 106 km2 catchment and channelling them to the East China Sea where they can be buried. Despite its potential global significance, the importance of the East China Sea as a terminal mercury sink remains poorly known. To address this knowledge gap, total mercury and methylmercury concentrations were determined from 51 surface sediment samples revealing their spatial distribution, whilst demonstrating the overall pollution status of the East China Sea. Sedimentary mercury distributions beneath the East China Sea are spatially heterogeneous, with high mercury concentrations (> 25 ng g-1) corresponding to areas of fine-grained sediment accumulation. In contrast, some sites of fine-grained sediment deposition have significantly lower values of methylmercury (< 15 ng g-1), such as the Changjiang estuary and some isolated offshore areas. Fine-grained particles and organic matter availability appear to exert the dominant control over sedimentary mercury distribution in the East China Sea, whereas in situ methylation serves as an additional control governing methylmercury accumulation. Estimated annual sedimentary fluxes of mercury in the East China Sea are 51 × 106 g, which accounts for 9% of China's annual mercury emissions.

Targeting hepatic miR-221/222 for therapeutic intervention of nonalcoholic steatohepatitis in mice.

BACKGROUND: Effective targeting therapies for common chronic liver disease nonalcoholic steatohepatitis (NASH) are in urgent need. MicroRNA-targeted therapeutics would be potentially an effective treatment strategy of hepatic diseases. Here we investigated the functional role of miR-221/222 and the therapeutic effects of antimiRs-221/222 in NASH mouse models. METHODS: We generated the miR-221/222flox/flox mice on a C57BL/6 J background and the hepatic miR-221/222 knockout (miR-221/222-LKO) mice. The mice were challenged with the methionine and choline deficient diet (MCDD) or chronic carbon tetrachloride (CCl4) treatment to generate experimental steatohepatitis models. Adenovirus-mediated re-expression of miR-221/222 was performed on the MCDD-fed miR-221/222-LKO mice. The MCDD and control diet-fed mice were treated with locked nucleic acid (LNA)-based antimiRs of miR-221/222 to evaluate the therapeutic effects. Histological analysis, RNA-seq, quantitative PCR and Western blot of liver tissues were carried out to study the hepatic lipid accumulation, inflammation and collagen deposition in mouse models. FINDINGS: Hepatic deletion of miR-221/222 resulted in significant reduction of liver fibrosis, lipid deposition and inflammatory infiltration in the MCDD-fed and CCl4-treated mouse models. The hepatic steatosis and fibrosis were dramatically aggravated by miR-221/222 re-expression in MCDD-fed miR-221/222-LKO mice. AntimiRs of miR-221/222 could effectively reduce the MCDD-mediated hepatic steatosis and fibrosis. Systematically mechanistic study revealed that hepatic miR-221/222 controlled the expression of target gene Timp3 and promoted the progression of NASH. INTERPRETATION: Our findings demonstrate that miR-221/222 are crucial for the regulation of lipid metabolism, inflammation and fibrosis in the liver. LNA-antimiRs targeted miR-221/222 could reduce steatohepatitis with prominent antifibrotic effect in NASH mice. FUND: This work is supported by the Natural Science Foundation of China (81530020, 81390352 to Dr. Ning and 81522032 to Dr. Cao and 81670793 to Dr. Jiang); National Key Research and Development Program (No. 2016YFC0905001 and 2017YFC0909703 to Dr. Cao); the Shanghai Rising-Star Program (15QA1402900 to Dr. Cao); Shanghai Municipal Education Commission-Gaofeng Clinical Medicine Grant (20171905 to Dr. Jiang).

[Factors associated with anastomotic leakage after anterior resection in rectal cancer].

OBJECTIVE: To investigate the factors associated with the anastomotic leakage after anterior resection in rectal cancer. METHODS: From January 2014 to January 2017 471 patients underwent Dixon procedure for rectal cancer in The Affiliated Hospital of Qingdao University. The data of those patients was collected and reviewed retrospectively. Inclusion criteria included: 1) rectal cancer confirmed by preoperative electron colonoscopy; 2) the standard of total mesorectal excision followed by the surgeon during the surgery; and 3) elective surgery. Exclusion criteria included multi-primary rectal cancer, secondary surgery for tumor recurrence, palliative surgery, Miles procedure, Hartmann procedure, hormone drugs used, presence of rheumatic and immune diseases, and distant metastasis of rectal cancer. The variables, including demograpic characteristics, ASA score, diabetes mellitus, preoperative radiochemotherapy, histopathologic grade, pathological T stage, laparoscopic or open surgery, distance of the tumor from the anal verge ≤5 cm, were analyzed to identify the risk factors for anastomotic leakage. RESULTS: Of 471 patients, 285 and 186 were men and women, respectively, with a mean age of 61 years (range, 31-92) years. Symptomatic clinically anastomotic leakage occurred in 31 patients (6.6%, 31/471) after Dixon procedure for rectal cancer. On univariate analysis, the occurrence of anastomotic leakage was associated with diabetes (χ2=10.972, P=0.001), serum albumin level <35 g/L (χ2=9.784, P=0.002), neoadjuvant chemoradiotherapy (χ2=6.867, P=0.009), distance ≤5 cm between the tumor and anal edge (χ2=5.993, P=0.014), preventive colostomy (χ2=5.630, P=0.018), and the use of double-perfusion cannula for abdominal flushing (χ2=4.232, P=0.040). Multivariate analysis revealed that diabetes (OR=3.632, 95%CI: 1.620-8.145, P=0.002), neoadjuvant chemoradiotherapy (OR=3.177, 95%CI: 1.283-7.867, P=0.012) and distance ≤5 cm between the tumor and anal edge(OR=2.444, 95%CI: 1.172-5.059, P=0.017) were independent risk factors for anastomotic leakage, while preventive colostomy (OR=0.138, 95%CI: 0.056-0.345, P=0.000) and the use of double-perfusion cannula for abdominal flushing (OR=0.223, 95%CI: 0.086-0.575, P=0.002) were independent protective factors for anastomotic leakage. CONCLUSIONS: For patients with rectal cancer with diabetes, undergoing neoadjuvant chemoradiotherapy, or distance ≤5 cm between the tumor and anal edge, anastomotic leakage after anterior resection of rectal cancer must be paid attention. When necessary, preventive colostomy or use of double-perfusion cannula for abdominal flushing should be considered.

RAGE and its emerging role in the pathogenesis of Parkinson's disease.

Receptor for advanced glycation end products (RAGE) is a multiligand receptor belonging to the immunoglobulin superfamily and plays crucial roles in the development of many human diseases such as neurodegenerative diseases, diabetes, cardiovascular diseases, osteoarthritis and cancer. RAGE involves in a number of cell processes such as neuroinflammation, apoptosis, proliferation and autophagy. In CNS, RAGE was primarily expressed in neurons, microglia and vascular endothelial cells. Interacting with ligands, RAGE induces a series of signal transduction cascades and leads to the activation of transcription factor NF-κB as well as increased expression of cytokines like TNF-α, IL-1. Moreover, binding to RAGE can also stimulate the generation of reactive oxygen species (ROS), which is implicated in neuron death. It was reported that RAGE were highly expressed in PD patients when compared to age-matched controls. And RAGE ablation protected nigral dopaminergic neurons against cell death in MPTP treated mice. Here we review this article to elucidate the role of RAGE in PD pathogenesis and highlight the anti-RAGE strategies in the treatment of PD.

Correlation between TSC1 gene polymorphism and epilepsy.

The correlation between tuberous sclerosis complex 1 (TSC1) gene polymorphism and epilepsy was studied. In total, 38 patients with epilepsy treated in People's Hospital of Rizhao from May 2015 to June 2016 were selected as study subjects, as the observation group, 38 healthy people in the same period were selected as the control group. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to study the polymorphism of TSC1 gene in the above study subjects. The mRNA expression of TSC1 gene in the observation group and the control group was measured by fluorescence quantitative PCR, the expression of TSC1 protein in the control and observation group was measured by western blotting and ELISA. The polymorphisms of TSC1 gene in control group and observation group were analyzed by PCR-RFLP. There were three genotypes of TCS1 gene locus 142 in healthy population: CC (79.3%), CA (13.9%) and AA (6.8%), there were also three genotypes at locus 142 in the observation group: CC (21.3%), CA (26.4%) and AA (52.3%), there was significant difference in the genotypes at locus 142 between healthy population and the patients with epilepsy (P<0.05). It was observed by fluorescence quantitative PCR that there was no significant difference in the mRNA expression of TSC1 gene between the control group and the observation group (P>0.05). The expression of TSC1 gene was detected by western blot method. Western blotting showed no significant difference in TSC1 protein expression between the two groups (P>0.05). However, by determining the activity of TSC1 protein in the observation group and the control group by ELISA, it was found that TSC1 activity in healthy human body (8.95±2.41 U/ml) was much lower than that in the patients with epilepsy (29.27±4.06 U/ml), the difference was statistically significant (P<0.05). It was found that locus 142 may be located at the active center of TSC1 enzyme by homology modeling of SWISS-MODEL, the mutation of locus 142 could lead to the change of TSC1 activity. The polymorphism of locus 142 in TSC1 gene is correlated with epilepsy, that is, the increase of CA and AA content in locus 142 leads to the occurrence of epilepsy.

Metformin Suppresses Proliferation and Viability of Rat Pheochromocytoma Cells.

BACKGROUND Previous studies have clearly demonstrated that metformin inhibits cell proliferation and cell growth in many types of human cancers. Increased survival rates in patients with breast and lung cancer receiving metformin have also been observed. However, the effect of metformin on pheochromocytoma cells remains unexplored. MATERIAL AND METHODS Rat pheochromocytoma cells (PC12 cells) were cultured and treated with metformin or vehicle control. Cell proliferation, cell-cycle, apoptosis, genes expression, and the signaling pathways involved were analyzed in PC12 cells. RESULTS The metformin treatment reduced cell viability and proliferation in rat pheochromocytoma PC12 cells in a dose- and time-dependent manner. Furthermore, metformin exposure led to an increased apoptosis rate and cell-cycle arrest accompanied with downregulation of Ccna2 and Ccnb2. At the molecular level, the AMPK signaling pathway was activated, whereas the mTOR and ERK1/2 signaling pathways were inhibited by metformin. CONCLUSIONS Our data suggest an antiproliferative role of metformin in pheochromocytoma development, which may provide a novel option for future cancer therapy.

Chemical reactive features of novel amino acids intercalated layered double hydroxides in As(III) and As(V) adsorption.

Layered double hydroxides (LDHs) intercalated with amino acids such as methionine (Met) were synthesized as new adsorbents to remediate arsenic-polluted water. This Zn2Al-Met-LDHs, identified with the formula of Zn0.7Al0.3(OH)2(Met)0.3·0.32H2O, has good thermal stability. Adsorption experiments with Zn2Al-Met-LDHs showed that the residual arsenic in solution could be reduced below the regulation limit, and this adsorption process fitted Langmuir isotherm and the pseudo-second-order kinetics well. A remarkably high removal efficiency and the maximum adsorption capacity for As(III) were achieved, 96.7% and 94.1 mg/g, respectively, at 298 K. The desorption efficiency of As(III) from the arsenic-saturated Zn2Al-Met-LDHs (<8.7%), far less than that of As(V), promises a specific and reliable uptake of As(III) in sorts of solutions. More importantly, a complete and in-depth spectra analysis through FTIR, XPS and NMR was conducted to explain the excellent performance of Zn2Al-Met-LDHs in arsenic removal. Herein, two special chemical reactions were proposed as the dominant mechanisms, i.e., hydrogen bonding between the carboxyl group of the host Met and the hydroxyl group of As(III) or As(V), and the formation of a chelate ring between the guest As(III) and the S, N bidentate ligands of the intercalated Met in the LDHs.

Whole exome sequencing of thymic neuroendocrine tumor with ectopic ACTH syndrome.

OBJECTIVE: Thymic neuroendocrine tumor is the second-most prevalent cause of ectopic adrenocorticotropic hormone (ACTH) syndrome (EAS), which is a rare disease characterized by ectopic ACTH oversecretion from nonpituitary tumors. However, the genetic abnormalities of thymic neuroendocrine tumors with EAS remain largely unknown. We aim to elucidate the genetic abnormalities and identify the somatic mutations of potential tumor-related genes of thymic neuroendocrine tumors with EAS by whole exome sequencing. DESIGN AND METHODS: Nine patients with thymic neuroendocrine tumors with EAS who were diagnosed at Shanghai Clinical Center for Endocrine and Metabolic Diseases in Ruijin Hospital between 2002 and 2014 were enrolled. We performed whole exome sequencing on the DNA obtained from thymic neuroendocrine tumors and matched peripheral blood using the Hiseq2000 platform. RESULTS: We identified a total of 137 somatic mutations (median of 15.2 per tumor; range, 1-24) with 129 single-nucleotide mutations (SNVs). The predominant substitution in these mutations was C:G > T:A transition. Approximately 80% of detected mutations resulted in amino acid changes. However, we failed to discover any recurrent mutations in these nine patients. By functional predictions, HRAS, PAK1 and MEN1, previously reported in neuroendocrine tumors, were identified as candidate tumor-related genes associated with thymic neuroendocrine tumors. CONCLUSIONS: Using whole exome sequencing, we identified genetic abnormalities in thymic neuroendocrine tumors with EAS. Thereby, this study acts as a further supplement of the genetic features of neuroendocrine tumors. Somatic mutations of three potential tumor-related genes (HRAS, PAK1 and MEN1) might contribute to the tumorigenesis of thymic neuroendocrine tumors with EAS.

Dual roles of AQDS as electron shuttles for microbes and dissolved organic matter involved in arsenic and iron mobilization in the arsenic-rich sediment.

Microbially-mediated arsenic (As) metabolism and iron (Fe) bioreduction from sediments play crucial roles in global As/Fe cycle, and their mobilization is associated with the various effects within the alliance of "mediator-bacteria-DOM (Dissolved Organic Matter)". The gradient levels (0.05, 0.10 and 1.00mM) of sodium anthraquinone-2,6-disulphonate (AQDS) as a mediator were investigated for their impact on reductive dissolution of As(V) and Fe(III) from arsenic-rich sediment. For the overall performance of AQDS-mediated reductive dissolution on As(V) and Fe(III), a more positive effect resulting from 0.05mM AQDS was observed compared to 0.10mM, whereas an inhibitory effect was observed with 1.00mM. Compared to the biotic supplementation with acetate as electron donors, approximately 13- and 6-fold increased levels of As(III) were released with 0.05 and 0.10mM, respectively, compared to 1.00mM AQDS (107.51µg/L), and approximately 4- and 3-fold increased Fe(II) levels (40.72mg/L) were observed during the same conditions. Multiple-dynamic effects of "bacteria-AQDS-DOM", which result from AQDS, shifted the microbial community and synchronously derived terrestrial DOM, which potentially changes the DOM substrate and complex formation of As(III)-Fe(II)-humic DOM. High-throughput sequencing results indicated an increase in the abundance of metal-reducing bacteria (e.g., Bacillus (>16%), Lactococcus (>13%), Pseudomonas (>4%) and Geobacter (>3%)) when supplemented with 0.05 and 0.10mM of AQDS. However, a boost increasing the abundance of metal oxidizing bacteria was observed with Alicyclobacillus (>16%), Burkholderia (>7%), and Bradyrhizobium (>5%) upon supplementation with 1.00mM AQDS. These novel insights have profound environmental implications and significance in terms of engineering, not only for understanding the cycle of As/Fe in sediment biochemical processes but for considering future alternative bioremediation treatments.