A maternal low-protein diet impaired glucose metabolism and altered the lncRNA profiles of islets in adult offspring.

A maternal low-protein diet during pregnancy can increase children's susceptibility to diabetes mellitus in adulthood. However, whether long noncoding RNAs (lncRNAs) in islets participate in the development of diabetes in adult offspring following maternal protein restriction is not fully understood. Female mice were fed a low-protein (LP) diet or control diet throughout gestation and lactation. The male offspring were then randomly divided into two groups according to maternal diet: offspring from control diet group dams (Ctrl group) and offspring from LP group dams (LP group). We observed the glucose metabolism of adult offspring. A lncRNA microarray was constructed for the islets from the LP group and Ctrl group to explore the differently expressed lncRNAs. Gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes analyses were subsequently used to predict the functions of the differently expressed lncRNAs. The body weight from birth to 12 weeks of age was significantly lower in the LP offspring. Adult LP offspring exhibited impaired glucose tolerance and decreased insulin secretion, consistent with the reduction in ß-cell proliferation. According to the lncRNA microarray, four lncRNAs, three upregulated lncRNAs, and one downregulated lncRNA were differently expressed in LP offspring islets compared with Ctrl offspring. Gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that these differentially expressed lncRNAs were mostly associated with the hypoxia-inducible factor-1α signaling pathway. Additionally, we validated the expression of these four differentially expressed lncRNAs via quantitative real-time polymerase chain reaction. Our findings demonstrated the expression patterns of lncRNAs in islets from adult offspring of mothers who consumed a maternal low-protein diet.

Efficacy and safety of Android artificial pancreas system use at home among adults with type 1 diabetes mellitus in China: protocol of a 26-week, free-living, randomised, open-label, two-arm, two-phase, crossover trial.

INTRODUCTION: Do-it-yourself artificial pancreas system (DIY APS) is built using commercially available insulin pump, continuous glucose monitoring (CGM) and an open-source algorithm. Compared with commercial products, DIY systems are affordable, allow personalised settings and provide updated algorithms, making them a more promising therapy for most patients with type 1 diabetes mellitus (T1DM). Many small and self-reported observational studies have found that their real-world use was associated with potential metabolic and psychological benefits. However, rigorous-designed studies are urgently needed to confirm its efficacy and safety. METHODS AND ANALYSIS: In this 26-week randomised, open-label, two-arm, two-phase, crossover trial, participants aged 18-75 years, with T1DM and glycated haemoglobin (HbA1c) 7-11%, will use AndroidAPS during one 12-week period and sensor-augmented pump during another 12-week period. This study will recruit at least 24 randomised participants. AndroidAPS consists of three components: (1) real-time CGM; (2) insulin pump; (3) AndroidAPS algorithm implemented in Android smartphone. The primary endpoint is time in range (3.9-10.0 mmol/L) derived from CGM. The main secondary endpoints include percentage of sensor glucose values below, within and above target range; mean sensor glucose value; measures of glycaemic variability and centralised HbA1c. Safety endpoints mainly include the frequency of hypoglycaemia events, diabetic ketoacidosis and other serious adverse events. ETHICS AND DISSEMINATION: This study has been approved by the Ethics Committee of the Third Affiliated Hospital of Sun Yat-sen University. There will be verbal and written information regarding the trial given to each participant. The study will be disseminated through peer-reviewed publications and conference presentations. OVERALL STATUS: Recruiting. STUDY START: 11 February 2023. PRIMARY COMPLETION: 31 July 2024. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov Registry (NCT05726461).

Epidemiology and survival of patients with malignant carotid body tumors in the SEER database.

OBJECTIVE: The objective of this study was to investigate population-based epidemiology, survival outcomes, and prognostic factors of malignant carotid body tumors (CBTs). METHODS: Patients with malignant CBTs who were diagnosed between 1975 and 2018 were screened from nine registries of the Surveillance, Epidemiology, and End Results (SEER) database. Cases that were coded as "carotid body tumor, malignant" or malignant tumors with the primary site recorded as "carotid body" were screened for inclusion in the study. The incidence of malignant CBT was calculated with SEER∗Stat software. Survival outcomes were analyzed using the Kaplan-Meier method and log-rank tests. RESULTS: A total of 72 patients with malignant CBT were screened for inclusion in the study, including 41 females (56.9%) and 31 males (43.1%). Based on the SEER program data, the incidence of malignant CBT was found to fluctuate between 0 to 0.02 cases per 100,000 people per year, with a slow but noticeable uptick after 1990. The most commonly affected populations included women and patients between the ages of 35 and 44, which accounted for 59.9% and 27.8% of patients in the study, respectively. During a median follow-up of 82 months, four patients were lost to follow-up, and 28 deaths were identified. Of those, 20 were considered disease-specific deaths. Further analysis found that the 5-year and 10-year overall survival rates were 78.9% and 67.8%, respectively, whereas the 5-year and 10-year disease-specific survival rates were 84.5% and 75.2%, respectively. The Kaplan-Meier method and log-rank tests indicated that age <50 years, sex, race, tumor number, and surgical treatment were unrelated to both overall survival and disease-specific survival. CONCLUSIONS: A retrospective review of the SEER database found that the incidence of malignant CBT was extremely rare and prone to fluctuation, but that it slowly trended upward over time. Malignant CBT was found to more likely affect females, and it could be diagnosed at any age. The overall prognosis for malignant CBT appeared to be good, with acceptable 5-year and 10-year survival rates. Due to a number of factors complicating malignant CBT surgery, surgical treatment should be considered with caution.

Downregulation of Kcnq1ot1 attenuates ß-cell proliferation and insulin secretion via the miR-15b-5p/Ccnd1 and Ccnd2 axis.

AIM: To examine the effect of lncRNA Kcnq1ot1 on pancreatic ß cells in the development of diabetes. METHODS: The expression levels of Kcnq1ot1 were detected in the islets of diabetes mouse models and the serum of patients with type 2 diabetes by qRT-PCR. CCK8, Ki67 staining, immunohistochemical analyses, glucose-stimulated insulin secretion and intraperitoneal glucose tolerance test were performed to detect the effect of Kcnq1ot1 on ß-cell proliferation and insulin secretion in vitro and in vivo. The relationship between Kcnq1ot1 and miR-15b-5p was predicted by bioinformatics prediction, which was confirmed by luciferase reporter assay. RESULTS: Kcnq1ot1 was more abundant in the pancreas. The expression of Kcnq1ot1 was decreased in the islets of db/db mice and diet-induced obese mice and in the serum of patients with type 2 diabetes. Silencing Kcnq1ot1 inhibited the ß-cell proliferation concomitant with a reduction in the levels of Ccnd1 and Ccnd2. Insulin synthesis and secretion were impaired, along with the decreased expression of Ins1, Ins2, and insulin-related transcription factors. Moreover, Kcnq1ot1 knockdown in vivo reduced glucose tolerance and decreased insulin secretion, consistent with the reduction in the relative islet area and Ki67-positive ß-cells detected by immunochemistry and immunofluorescence staining, respectively. Mechanistically, Kcnq1ot1 directly targeted miR-15b-5p which regulated ß-cell proliferation and insulin secretion through Ccnd1 and Ccnd2. Notably, the suppression of miR-15b-5p attenuated the inhibition of Min6 proliferation and insulin production induced by Kcnq1ot1 knockdown. CONCLUSION: Kcnq1ot1 regulated ß-cell proliferation and insulin secretion via the miR-15b-5p/Ccnd1 and Ccnd2 axis, which is worthy of further investigation considering its potential in diabetes treatment.

Identifying Needs and Barriers to Diabetes Dietary Education in Chinese People with Type 2 Diabetes and Their Family Members in Guangzhou: A Qualitative Study.

PURPOSE: The purpose of this exploratory qualitative study was to identify the needs and barriers to diabetes dietary education issues in a Chinese context among people with type 2 diabetes and also their family members. PATIENTS AND METHODS: Nineteen patients with type 2 diabetes and 15 family members were randomly selected from a larger study in Guangzhou. Descriptive phenomenological qualitative inquiry guided this study. RESULTS: Both patients and their family members claimed a variety of needs regarding diabetes dietary information and that it should be patient-centered. In addition, both groups identified the effectiveness and helpfulness of diabetes diet-related health education, but family members reported a lack of professional education. Patients reported that the barriers to diabetes diet-related health education were: 1) patients' different faculties of memory and acceptance, 2) educators' methods of explaining the information, 3) lack of advertising and intensity of publicity both inside and outside of the hospital. Family members identified that diabetes diet-related health education assisted them with taking care of patients with diabetes and it was also beneficial for themselves to gain more dietary knowledge and develop healthy dietary habits. CONCLUSION: It is necessary to make some adjustment to traditional diabetes dietary education. It may be a good strategy to investigate the needs and faculties of memory and acceptance of people with diabetes before starting a diabetes education program. Hospitals should intensify their publicity of diabetes education and make it more attractive to patients with diabetes and their family members. Educators should add information specifically for family members while conducting diabetes education. Tertiary hospitals could provide standardized training regarding diabetes-related health education to community family physicians; in this way, diabetes education could gradually be transferred to them and both groups of clinicians could share in the education.

Short-term GLP-1 receptor agonist exenatide ameliorates intramyocellular lipid deposition without weight loss in ob/ob mice.

OBJECTIVE: Ectopic lipid deposition is closely associated with type 2 diabetes (T2D). Accumulating evidence shows that GLP-1 receptor agonists (GLP-1 RAs) improve obesity and liver steatosis. However, it remains unknown whether and how they ameliorate lipid deposition in skeletal muscle. This study aimed to investigate the effect of exenatide (a GLP-1 RA) on intramyocellular lipid deposition in the skeletal muscle of T2D models and its dependence on weight loss. METHODS: Ob/ob mice and diet-induced obese (DIO) mice were treated with exenatide (24 nmol/kg), leptin (1 mg/kg), or saline control intraperitoneally once daily for 4 weeks. Phenotypic evaluations were performed during and after the intervention. PA-induced myoblast C2C12 cells were used as an in vitro model. The expression of key enzymes involved in lipid metabolism was assessed in the skeletal muscle of ob/ob mice and DIO mice. RESULTS: In ob/ob mice, 4-week exenatide treatment did not improve the body weight and fat mass, but modestly ameliorated intramyocellular lipid deposition and lipid profiles. In DIO mice, it remarkably alleviated the body weight, lipid profiles, and intramyocellular lipid deposition. In the skeletal muscle of these two models, exenatide treatment activated the AMP-activated protein kinase (AMPK) signaling pathway, stimulated lipid oxidation enzymes, and upregulated the insulin signaling pathway. In vitro, exendin-4 activated the AMPK signaling pathway and stimulated lipid metabolism to improve lipid accumulation in palmitate-induced myoblast C2C12 cells. CONCLUSIONS: Exenatide ameliorated intramyocellular lipid deposition without body weight reduction in ob/ob mice, but alleviated body weight and intramyocellular lipid deposition in DIO mice. The underlying mechanism included the activation of AMPK signaling pathway and improvement in insulin sensitivity, independent of weight loss in ob/ob mice.

GLP-1 receptor agonist promotes brown remodelling in mouse white adipose tissue through SIRT1.

AIMS/HYPOTHESIS: Accumulating evidence has revealed the significant role of glucagon-like peptide-1 (GLP-1) in weight loss. Sirtuin 1 (SIRT1) plays a vital role in the regulation of lipid metabolism. Here, we investigated the contribution of lipolytic and oxidative changes in white adipose tissue (WAT) to the weight-lowering effect induced by the GLP-1 receptor (GLP-1R) agonist exenatide (exendin-4) in mice. We also looked at the role of SIRT1 in this process. METHODS: C57BL/6J mice and Sirt1 (+/-) mice were treated with exenatide (24 nmol/kg) or an NaCl solution (154 mmol/l) control i.p. for 8 weeks while receiving a high-fat diet (HFD) after a 12 week HFD challenge. Systemic phenotypic evaluations were carried out during and after the intervention. A lentivirus-mediated short hairpin (sh)RNA vector of the Sirt1 gene was transfected into differentiated 3T3-L1 adipocytes. An in vitro model system used adipocytes induced from Sirt1-null mouse embryonic fibroblasts (MEFs). RESULTS: Exenatide reduced fat mass and enhanced the lipolytic and oxidative capacity of WAT in diet-induced obese C57BL/6J mice. However, these effects were significantly impaired in Sirt1 (+/-) mice compared with wild-type controls. In vitro, exendin-4 increased lipolysis and fatty acid oxidation by upregulating SIRT1 expression and activity in differentiated 3T3-L1 adipocytes. Conversely, RNA interference (i)-induced knockdown of SIRT1 attenuated the lipolytic and oxidative responses to exendin-4 in differentiated 3T3-L1 adipocytes. Again, these responses were entirely abolished in Sirt1-null MEFs after induction into adipocytes. CONCLUSIONS/INTERPRETATION: These data highlight that a GLP-1R agonist promotes brown remodelling of WAT in a SIRT1-dependent manner; this might be one of the mechanisms underlying its effect on weight loss.

Diet-induced obesity and insulin resistance are associated with brown fat degeneration in SIRT1-deficient mice.

OBJECTIVE: Recent studies have revealed that SIRT1 gain-of-function could promote adipose tissue browning for the adaptive thermogenesis under normal diet. This study investigated the role of SIRT1 loss-of-function in diet-induced obesity and insulin resistance and the mechanism involved in adipose tissue thermogenesis. METHODS: Male SIRT1(+/-) and wild-type (WT) mice were fed with a high-fat diet (HFD) for 16 weeks to induce obesity and insulin resistance, while mice on a chow diet were used as lean controls. The phenotype data were collected, and different adipose tissue depots were used for mechanism research. RESULTS: Compared with WT mice, SIRT1(+/-) mice exhibited increased adiposity and more severe insulin resistance with less thermogenesis under HFD challenge. Strikingly, SIRT1(+/-) mice displayed an exacerbated brown adipose tissue (BAT) degeneration phenotype, which was characterized by lower thermogenic activity, aggravated mitochondrial dysfunction, and more mitochondrial loss. In addition, SIRT1(+/-) mice showed aggravated inflammation and dysfunction in epididymal adipose tissue after HFD intervention, which also contributed to the systemic insulin resistance. CONCLUSIONS: Diet-induced obesity and insulin resistance are associated with BAT degeneration in SIRT1-deficient mice, which further underlined the beneficial role of SIRT1 in obesity-associated metabolic disorders.

Acute Wnt pathway activation positively regulates leptin gene expression in mature adipocytes.

Genome-wide association studies (GWAS) have revealed the implication of several Wnt signaling pathway components, including its effector transcription factor 7-like 2 (TCF7L2) in diabetes and other metabolic disorders. As TCF7L2 is expressed in adipocytes, we investigated its expression and function in rodent fat tissue and mature adipocytes. We found that TCF7L2 mRNA expression in C57BL/6 mouse epididymal fat tissue was up-regulated by feeding but down-regulated by intraperitoneal insulin injection. In high-fat diet (HFD) fed mice, db/db mice and Zucker (fa/fa) rats, epididymal fat TCF7L2 mRNA levels were lower than the corresponding controls. Treating rat adipocytes with 100nM insulin repressed TCF7L2 mRNA and protein levels, associated with the repression of leptin mRNA level. The treatment with 1nM insulin, however, stimulated TCF7L2 and leptin mRNA levels. This stimulation could be attenuated by iCRT14, an inhibitor of ß-catenin/TCF-responsive transcription. Wnt3a stimulated leptin mRNA level, which was also blocked by iCRT14 co-treatment. Utilizing the leptin-expressing cell line HTR8 as a tool, we defined an evolutionarily conserved CREB binding motif that mediated Wnt3a activation. Although Wnt activation is known to repress the differentiation of 3T3-L1 cells towards mature adipocytes, short-term Wnt3a treatment of differentiated 3T3-L1 cells stimulated leptin mRNA levels. Thus, wnt pathway plays a dual function in adipocytes, including the well-known repressive effect on adipogenesis and the stimulation of leptin production in mature adipocytes in response to nutritional status.

[Effects of insulin on pigment epithelium-derived factor expression in adipocytes of type 2 diabetic mellitus in rats].

OBJECTIVE: To explore the effects of insulin therapy on the expression of pigment epithelium-derived factor (PEDF) in adipocytes of type 2 diabetic mellitus (T2DM) in rats. METHODS: A total of 22 newly diagnosed type 2 diabetics received a 2-week intensive insulin therapy. The levels of fasting plasma glucose (FPG), serum triglyceride and PEDF were measured before and after therapy. T2DM was induced by a high-fat diet and a low-dose streptozotocin (STZ). The Spraque-Dawley rats were divided randomly into diabetic, insulin treatment and gliclazide treatment groups. Another group with a chow diet was designated as normal controls. Differentiated 3T3-L1 adipocytes were then incubated with tumor necrosis factor-alpha (TNF-α) and (or) insulin for 24 h. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression of PEDF in adipose tissue or adipocytes. The PEDF levels in both sera and cell supernatant were measured by enzyme-linked immunosorbent assay (ELISA). Glucose uptake was detected after treatment of PEDF or anti-PEDF antibody simultaneously together with insulin in mature 3T3-L1 adipocytes. RESULTS: Insulin therapy decreased the serum levels of FPG and triglyceride of T2DM patients ((12.9 ± 2.8) vs (5.9 ± 1.4) mmol/L, (3.1 ± 1.8) vs (1.7 ± 0.8) mmol/L, P < 0.05) while the serum level of PEDF decreased significantly after therapy ((22.85 ± 5.73) vs (18.38 ± 5.28) µg/L, P < 0.05). Consistently the serum level of PEDF of diabetic rats was remarkably higher than that of normal controls and insulin-treated group ((28.6 ± 0.5) vs (25.4 ± 0.6) and (25.3 ± 0.6) µg/L, P < 0.05). And the elevated levels of PEDF, TNF-α mRNA and protein in adipose tissue (P < 0.05) could be reduced by insulin treatment (P < 0.05). However, no obvious change was detected in gliclazide treatment group. Further evidences suggested that TNF-α could induce more secretion and expression of PEDF in 3T3-L1 adipocyte while this effect became ameliorated by insulin treatment. Furthermore, decreased capacity of glucose uptake by PEDF might be reversed by anti-PEDF antibody in 3T3-L1 adipocytes (P < 0.05). CONCLUSIONS: Insulin can down-regulate the expression of PEDF in adipocytes of T2DM and improve the glucose uptake of adipocytes. It may be one of the mechanisms through which insulin therapy improves peripheral insulin resistance.