Background: Pathological aggregation of islet amyloid polypeptide (IAPP) contributes to β-cell dysfunction in type 2 diabetes. Our previous studies demonstrated that caveolin-1 (Cav-1) deficiency protects β-cells from palmitate-induced apoptosis. Microarray profiling further indicated that Cav-1 silencing alters IAPP expression. This study aimed to investigate the effects of Cav-1 depletion on IAPP secretion and expression and to explore the potential involvement of thioredoxin-interacting protein (TXNIP). Methods: We performed lentiviral-mediated Cav-1 knockdown in NIT-1 cells and isolated murine islets, and simultaneously generated an inducible β-cell-specific Cav-1 knockout (iβ-Cav1 KO) mouse model. IAPP secretion and expression were assessed by ELISA, Western blot, qPCR and immunofluorescence. The expression of IAPP-processing enzymes (PAM, PC1, and PC2) and degradation factors (IDE and BACE2) was examined. Co-immunoprecipitation (Co-IP) and immunofluorescence were performed to investigate the interaction between Cav-1 and TXNIP. Results: Cav-1 depletion significantly reduced both IAPP secretion and expression in vitro and in vivo. High-fat-diet-fed iβ-Cav1 KO mice exhibited the lowest serum IAPP levels. Mechanistically, Cav-1 depletion was associated with downregulation of PAM, PC1, and PC2 and upregulation of IDE and BACE2. Additionally, Cav-1 depletion decreased TXNIP expression. Immunofluorescence revealed co-localization of Cav-1 and TXNIP, and co-immunoprecipitation further demonstrated their direct physical interaction. Conclusions: Cav-1 is essential for IAPP secretion and expression in β-cells. The direct physical interaction between Cav-1 and TXNIP suggests that TXNIP may mediate the regulatory effects of Cav-1 on IAPP processing or secretion. These findings identify the Cav-1-TXNIP axis as a potential target for mitigating IAPP-related β-cell dysfunction.

Lipotoxicity caused β-cell mass decrease and impaired β-cell function in type 2 diabetes mellitus. We previously reported that caveolin-1 (Cav-1) deficiency protected pancreatic β cells against palmitate (PA)-induced apoptosis and dysfunction in both NIT-1 cells and isolated islets. In this study, we firstly established inducible β-cell-specific Cav-1 KO mice model. Next, we investigated whether Cav-1 depletion in vitro or in vivo affected β-cell function and survival through the regulation of autophagy under lipotoxicity. Our results showed that Cav-1 depletion exhibited increased islets size, improved insulin resistance and glucose tolerance in lipid stressing conditions. In addition, KO of Cav-1 also increased β-cell viability through suppression of gene expression of pro-apoptotic molecules (BIM, BID, SMAC, Apaf, Caspase9, Caspase3, AIF, and EndoG). Mechanism studies revealed that Cav-1 depletion protected β cells from PA-induced apoptosis through p38 MAPK signaling pathway. Meanwhile, we also found that Cav-1 depletion enhanced autophagy through elevated expression of Beclin-1 and Lc3b, and increased degradation of p62 protein. Further investigation indicated that mechanistic target of rapamycin (mTOR) signaling pathway was participated in the regulation of PA-induced autophagy. Taking together, our data suggested that Cav-1 depletion ameliorated PA-induced glucose metabolism abnormality, increased cell viability, and improved β-cell apoptosis through the enhancement of autophagy. Our findings would provide valuable clues for developing novel treatments based on Cav-1 inhibition against diabetes under lipotoxicity.

Long noncoding RNA (lncRNA) maternally expressed gene 3 (MEG3) plays an important role in protection of ischemia–reperfusion (I/R) injury in brain and liver. However, role of MEG3 in myocardial I/R injury remains unclear. Here, the role of MEG3 in protection of myocardial I/R injury and its association with microRNA-7-5p (miR-7-5p) was investigated using rat cardiac I/R model and myocardial I/R cell model. Our results showed that MEG3 was significantly up-regulated and miR-7-5p was significantly down-regulated after I/R. Following I/R, the levels of intact PARP and intact caspase-3 were reduced, while the cleaved fragments of PARP and caspase-3 were increased. TUNEL assay showed an increase in cardiomyocyte apoptosis after I/R. The levels of I/R-induced creatine kinase (CK) and lactate dehydrogenase (LDH) were inhibited by knockdown of MEG3 (siMEG3). SiMEG3 increased cell proliferation and inhibited cell apoptosis after I/R. In contrast, overexpression of MEG3 increased the I/R-induced CK and LDH activities and cell apoptosis and decreased cell proliferation. The dual-luciferase reporter system showed a direct binding of MEG3 to miR-7-5p. The level of miR-7-5p was negatively associated with the change in levels of MEG3 in H9c2 cells. The levels of intact RARP1 and caspase-3 were significantly increased by knockdown of MEG3. Co-transfection of miR-7-5p inhibitor with siMEG3 activates CK and LDH, significantly decreased cell proliferation, increased cell apoptosis, and decreased intact poly(ADP-ribose) polymerase 1 (PARP1) and caspase-3. In summary, down-regulation of MEG3 protects myocardial cells against I/R-induced apoptosis through miR-7-5p/PARP1 pathway, which might provide a new therapeutic target for treatment of myocardial I/R injury.

Objective: This study aimed to explore the insulin requirement during continuous subcutaneous insulin infusion (CSII) treatment in patients with type 2 diabetic kidney disease (T2DKD). Methods: This study retrospectively analyzed the clinical data of 150 T2DKD patients in the Department of Endocrinology and Metabolism of the Third Affiliated Hospital of Sun Yat-sen University from January 2018 to December 2021. All patients received short-term CSII treatment and achieved the blood glucose target by adjusting insulin infusion. The patients' daily insulin requirements during the treatment were recorded and analyzed. Result: There were 91 males and 59 females with an average age of 60.6 ± 10.6 years. Total daily insulin dose (TDD), total daily insulin dose per kilogram (TDD kg-1), total basal insulin dose per kilogram (TBa kg-1), and total bolus insulin dose per kilogram (TBo kg-1) fell with the decline of eGFR when achieving the blood glucose target except for the ratio of total basal insulin dose (TBD) to TDD (%TBa). Insulin requirement was less in patients with eGFR < 60 mL/min/1.73 m2 compared to those with eGFR ≥ 60 mL/min/1.73 m2 (p < 0.05). When achieving the blood glucose target, eGFR ≥ 60 mL/min/1.73 m2 group and eGFR < 60 mL/min/1.73 m2 group were 0.71 ± 0.19 U/kg and 0.60 ± 0.24 U/kg, respectively. Multiple linear regression analyses showed that glycated hemoglobin and eGFR were independent factors associated with TDD kg-1. Conclusion: In patients with T2DKD who received short-term CSII therapy, the insulin requirement decline with the decrease of eGFR, while the %TBa did not change significantly. The dosage of insulin should be adjusted according to the level of eGFR in patients with T2DKD treated with CSII.

Background: To investigate the effect of glucokinase activation on glucose and lipid metabolism in diet-induced obese mice. Methods: Mice fed with High-fat-diet (HFD) or control diet for 16 weeks were subjected to glucokinase activator (GKA) or vehicle treatment by gavage for 4 weeks. Glucose tolerance tests were performed to evaluate the glucose-lowering effect of GKA. Hepatic lipid accumulation was assessed by H&E, Oil-red’O staining, and transmission electronic microscopy. The underlying mechanism and subsequent effects of glucokinase activation-induced lipid accumulation was analyzed in liver. Protein levels of related genes were analyzed in mouse livers or AML12 cells. Results: Glucokinase activation improved glucose tolerance and insulin sensitivity in obese mice. Moreover, increased liver weight and TG Content were found in GKA-treated obese mice. Significant hepatic lipid accumulation was observed by multiple assays. RNA sequencing analysis indicated the increased expression of lipogenesis genes and activated PERK-UPR pathway in the liver. Inhibition of the lipogenesis-related gene Acc reduced the lipid levels caused by GKA treatment and alleviated PERK-UPR pathway in AML12 cell line. Conclusions: Glucokinase activation improved glucose tolerance while induced hepatic lipid accumulation through increased lipogenesis, which subsequently triggered PERK-UPR signaling pathway. Disclosure N.Cai: None. L.Zeng: None. X.Chen: None. J.Liu: None. Z.Wen: None. S.Wen: None. W.Zeng: None. S.Lin: None. Y.Chen: None. G.Shi: None. Funding National Natural Science Foundation of China (81770826, 82070811); National Key R&D Program of China (2017YFA0105803); Key Area R&D Program of Guangdong Province (2019B020227003); Sci-Tech Research Development Program of Guangzhou City (202201020497); Natural Science Foundation of Guangdong Province (2018B030311012)

Antiphospholipid syndrome (APS) is an autoimmune disorder while adrenal hemorrhage could be its rare complication. Herein, we report the case of a 32-year-old unmarried woman with a history of systemic lupus erythematosus (SLE) who was hospitalized after complaints of upper abdominal pain, limb weakness, and loss of appetite for 2 weeks. Laboratory examination revealed hyponatremia, low plasma cortisol levels, increased adrenocorticotropic hormone levels, and a positive anticardiolipin antibody status. Furthermore, computed tomography (CT) revealed the presence of bilateral adrenal masses. Ultimately, based on dynamic changes in CT images, these masses were diagnosed as adrenal hemorrhage owing to APS. A computer-assisted literature search was conducted to identify cases of primary adrenal insufficiency associated with APS and/or SLE. The clinical features, laboratory examination, treatments, and outcomes of these cases were summarized. Our findings emphasize the importance of screening for adrenal insufficiency in patients with SLE or APS who present with abdominal complaints, asthenia, and hyponatremia. It is also recommended to test for APS all patients with adrenal hemorrhage.

Objective:To explore the potential target of unfolded protein response-protein kinase R-like endoplasmic reticulum kinase (UPR-PERK) pathway in the process of ER stress in liver, we also tested whether the UPR-PERK pathway could regulate the oxidative stress-induced growth inhibitor 1 ( Osgin1) in the liver endoplasmic reticulum (ER) stress models. Methods:Using GEO database as the source of analysis data, the differentially expressed genes were selected by GEO2R software, using Venn diagram, we identified Osgin1 as the gene with significant variation in the three physiological factors of fasting-refeeding, exercise, and age. The direct interaction between the UPR-PERK pathway and Osgin1 was verified using the GENEMANIA database. Subsequently, we used three different liver endoplasmic reticulum stress models (acute endoplasmic reticulum stress model, non-alcoholic fatty liver disease model and liver cancer model) in GEO Database and TCGA database to further verify the regulatory relationship of UPR-PERK pathway to Osgin1. GraphPadPrism 9.0.0 software was used for statistical analysis. Pearson correlation analysis was used to analyze the correlation between the transcriptional levels of UPR-PERK pathway genes and Osgin1. Results:Compared with the control group, fasting-refeeding, increased exercise and aging significantly changed the transcription level of Osgin1 ( Osgin1 mRNA was up-regulated by 700%, down-regulated by 156% and 229%, respectively, P<0.05). The GENEMANIA database verified the direct interaction between the UPR-PERK pathway and Osgin1. In the model of endoplasmic reticulum (ER) stress, compared with control group, inhibition of UPR-PERK pathway related gene ( Eif2ak3) could significantly reverse the up-regulation of Osgin1 mRNA induced by Tunicamycin ( Osgin1 mRNA was downregulated by 87%, P<0.001). In the non-alcohol fatty liver model, compared with control group, when the UPR-PERK pathway was alleviated by the administration of drugs, the Osgin1 transcription level was also down-regulated by 48%, 43.3% and 56.6% with BI4556906, EX10970 and IXA4 interventions ( P<0.05). In hepatic cancer cell tissues, compared with control group, the transcription levels of UPR-PERK pathway-related genes and Osgin1 were significantly up-regulated 109% ( P<0.001). Conclusions:The activation of UPR-PERK pathway upregulates Osgin1 transcription in physiological conditions and liver endoplasmic reticulum stress models.

Lipotoxicity-induced pancreatic β cell damage is a strong predictor of type 2 diabetes mellitus (T2DM). Our previous work showed that Caveolin-1 (Cav-1) depletion decreased β-cell apoptosis and improved β-cell viability. Further microarray analysis indicated significant changes in the expression of genes related to fatty acid metabolism and inflammation. The objective of this study was to explore the role of Cav-1 in intracellular lipid accumulation and inflammation in β cells under lipotoxic conditions. Here, we established a β-cell-specific Cav-1 knockout (β-Cav-1 KO) mouse model and a CAV-1 depleted β cell line (NIT-1). We found that Cav-1 silencing significantly reduced palmitate (PA)-induced intracellular triglyceride (TG) accumulation and decreased proinflammatory factor expression in both the mouse and cell models. Further mechanistic investigation revealed that amelioration of lipid metabolism was achieved through the downregulation of lipogenic markers (SREBP-1c, FAS and ACC) and upregulation of a fatty acid oxidation marker (CPT-1). Meanwhile, decrease of inflammatory cytokines (IL-6, TNF-α, and IL-1β) secretion was found with the involvement of the IKKβ/NF-κB signaling pathways. Our findings suggest that Cav-1 is of considerable importance in regulating lipotoxicity-induced β-cell intracellular lipid accumulation and inflammation.

Background and aims: Type 2 diabetes mellitus remains a substantial medical problem with increasing global prevalence. Pharmacological research is becoming increasingly focused on personalized treatment strategies. Drug development based on glucokinase (GK) activation is an important strategy for lowering blood glucose. This study aimed to investigate the effect of GK activation on glucose and lipid metabolism in diet-induced obese mice. Materials and methods: Mice were fed with a high-fat diet (HFD) for 16 weeks to induce obesity, followed by a GK activator (GKA, AZD1656) or vehicle treatment by gavage for 4 weeks. The effect of GKA treatment on glucose metabolism was evaluated using glucose and insulin tolerance tests. Hepatic lipid accumulation was assessed by hematoxylin and eosin staining, Oil Red O staining, and transmission electron microscopy. The underlying mechanism of GK activation in glucose and lipid metabolism in the liver was studied using transcriptomic analysis, with a mechanistic study in mouse livers in vivo and AML12 cells in vitro. Results: GK activation by GKA treatment improved glucose tolerance in HFD-fed mice while increasing hepatic lipid accumulation. Transcriptomic analysis of liver tissues indicated the lipogenesis and protein kinase RNA-like endoplasmic reticulum kinase (PERK)-unfolded protein response (UPR) pathway activations in GKA-treated HFD-fed mice. Inhibition of the ACC activity, which is an important protein in lipogenesis, attenuated GKA treatment-induced lipid accumulation and PERK-UPR activation in vitro. Conclusions: GK activation improved glucose tolerance and insulin sensitivity while inducing hepatic lipid accumulation by increasing the lipogenic gene expression, which subsequently activated the hepatic PERK-UPR signaling pathway.

Background: Maternal subclinical hypothyroidism (SCH) has been associated with adverse pregnancy outcomes. This study aimed to explore whether SCH in the first trimester contributed to the development of gestational diabetes mellitus (GDM). Materials and Methods: A total of 8,777 pregnant women who first visited before 13 weeks and 6 days of gestation and accepted routine prenatal service at the Third Affiliated Hospital of Sun Yat-Sen University from January 2015 to September 2018 were recruited in this study. Thyroid functions (thyroid stimulating hormone [TSH], free T4, and thyroid peroxidase antibody [TPOAb]) were measured before 13 weeks and 6 days of gestation and data of 7,536 subjects with TSH ≥0.1 mIU/L were analyzed. A 2-hour 75-g oral glucose tolerance test was performed between 24 and 28 gestational weeks. Chi-square test and multivariate logistic regression analysis were applied to evaluate the relationship between SCH and GDM. Results: The prevalence of SCH in this population was 7.53%. After stratifying the relationship between SCH and GDM according to TSH concentrations (slightly elevated TSH: ≥2.5, <4.0 mIU/L; moderately elevated TSH: ≥4.0, <10.0 mIU/L) and TPOAb status, a moderately elevated TSH combined with positive TPOAb (23.9% vs. normal 13.0%, chi-square = 6.317, p = 0.012) was found to increase the incidence of GDM. Furthermore, after adjusting for confounders (maternal age, educational levels, parity, and pregestational body mass index [preBMI]), the SCH group still exhibited a higher risk of GDM (relative risk [RR] 1.867, 95% confidence interval [CI] 1.018-3.424). Conclusion: Our findings indicated that SCH during early pregnancy, in the presence of moderately elevated TSH levels and positive TPOAb, might lead to an increased risk of GDM.

Background The association between isolated maternal hypothyroxinemia (IMH) and adverse pregnancy outcomes is still controversial. This study aimed to evaluate the association between IMH during the first trimester and adverse pregnancy outcomes in southern Chinese women. Methods This was a hospital-based, retrospective cohort study. The records of 7051 women, including 1337 IMH women and 5714 euthyroid women who had a singleton pregnancy and accepted routine prenatal service at the Third Affiliated Hospital of Sun Yat-Sen University from January 2015 to September 2018, were extracted from the electronic medical records system in this study. Thyroid functions [thyroid-stimulating hormone (TSH), free thyroxine (fT4) and anti-thyroperoxidase autoantibody (TPO-Ab)] had to be measured before 13 weeks and 6 days of gestation. The chi-square test and multivariate logistic regression analysis were applied to evaluate the association between IMH during the first trimester and adverse pregnancy outcomes. Results Prepregnancy obesity [prepregnancy body mass index (preBMI) ≥ 25 kg/m 2 ] was found to be more common in the IMH group (11.2% vs. 6.1%) ( P < 0.05). The prevalence of gestational diabetes mellitus (GDM), postpartum haemorrhage (PPH), macrosomia and large for gestational age (LGA) was higher in the IMH group. However, after using multivariate logistic regression analysis to adjust for confounders (maternal age, educational levels and preBMI), only LGA was shown to be associated with an increased risk in IMH women [adjusted OR: 1.27 (95% CI 1.044–1.566)]. The prevalence of preterm delivery (either < 37 or < 34 weeks), gestational hypertension, preeclampsia, placenta previa, placental abruption, premature rupture of membrane (PROM), intrauterine growth restriction (IUGR), polyhydramnios, stillbirth, small for gestational age (SGA) and low Apgar score did not increase. Conclusion IMH during the first trimester did not increase any risk of adverse pregnancy outcomes in southern Chinese women except LGA.

Purpose: To investigate the dietary knowledge, attitude and practice (KAP) among the family members (FMs) of Chinese type 2 diabetes mellitus (T2DM) patients and its influence on the KAP of T2DM patients. Patients and Methods: Two hundred thirty-six pairs of hospitalized T2DM patients and their FMs (472 in total) in our hospital were enrolled. A pair of self-designed questionnaires on dietary KAP (Cronbach's alpha >= 0.763, I-CVI >= 0.857, S-CVI = 0.964, 0.958) were used to collect data and assess the KAP towards diabetes diets. Results: The mean score for dietary KAP of T2DM patients was 2.33 +/- 0.60, 3.03 +/- 0.44 and 2.77 +/- 0.38, whereas that of their FMs was 2.37 +/- 0.55, 3.08 +/- 0.48 and 2.82 +/- 0.61, respectively. Pearson's correlation analysis showed that the glycosylated hemoglobin (HbAlc) of T2DM patients was negatively correlated to their dietary practice (r = -0.218, P < 0.01). There was a positive correlation between T2DM patients and their FMs for dietary KAP (r = 0.306, P < 0.05). The dietary practice of T2DM patients was positively correlated with the dietary KAP of their FMs (r = 0.305, 0.252 and 0.136, respectively, P < 0.01). Logistic regression analysis revealed that the score for dietary knowledge and attitude, occupation, residence, family history, complications of the T2DM patient, and the sex and dietary knowledge score of the FM were significantly associated with dietary practice for T2DM patients. Conclusion: The dietary attitude of FMs was moderate but dietary knowledge and practice were poor. Dietary KAP was positively correlated with T2DM patients and their FMs.

Objective:To explore whether thyroid stimulating hormone (TSH) treat-to-target during early pregnancy had an impact on the incidence of gestational diabetes mellitus (GDM) in women with hypothyroidism.Methods:A retrospective analysis was performed on naturally conceived singleton pregnant women who had accepted routinely prenatal services at the Third Affiliated Hospital of Sun Yat-sen University from January 2015 to December 2018. The gestational weeks of the first obstetric examination were before 13 weeks and 6 days of gestation. Data of 6 978 subjects were analyzed, including 186 cases in the hypothyroidism group and 6 792 cases in the normal group. In the hypothyroidism group, according to whether TSH levels were<2.5 mU/L in the first prenatal examination, 124 cases were in the under control group and 62 cases in the non-control group. Basic information was collected at first visit, and laboratory tests were performed with fasting samples, including thyroid function [TSH, free thyroxine (FT4), thyroid peroxidase antibody (TPOAb)] and fasting plasma glucose (FPG). The 75-gram oral glucose tolerance test (OGTT) was performed at 24 to 28 weeks. The incidence of GDM and the impact of TSH levels on it between the hypothyroidism group and the normal group were explored by using χ2 test. The risk factors of GDM were analyzed by logistic regression. Results:The incidence of GDM in the hypothyroidism group was higher than that in the under-control group [21.5% (40/186) vs. 13.7% (929/6 792), P=0.002]. The incidences of GDM in the under-control group [20.2% (25/124)] and the non-control group [24.2% (15/62)] were higher than that of the control group (all P<0.05). Multivariate logistic regression analysis for risk factors of incidence of GDM was performed, and finally age, pregestational body mass index, history of hypothyroidism, FPG, positive TPOAb, and triglyceride were considered to be independent risk factors for incidence of GDM ( P<0.05). Whether TSH met the standard was ruled out from the equation. Conclusion:Compared with women with normal thyroid function, women with hypothyroidism still are at higher risk of developing GDM, whether TSH levels are under control or not.

Insulin treatment was confirmed to reduce insulin resistance, but the underlying mechanism remains unknown. Caveolin-1 (Cav-1) is a functional protein of the membrane lipid rafts, known as caveolae, and is widely expressed in mammalian adipose tissue. There is increasing evidence that show the involvement of Cav-1 in the AKT activation, which is responsible for insulin sensitivity. Our aim was to investigate the effect of Cav-1 depletion on insulin sensitivity and AKT activation in glargine-treated type 2 diabetic mice. Mice were exposed to a high-fat diet and subject to intraperitoneal injection of streptozotocin to induce diabetes. Next, glargine was administered to treat T2DM mice for 3 weeks (insulin group). The expression of Cav-1 was then silenced by injecting lentiviral-vectored short hairpin RNA (shRNA) through the tail vein of glargine-treated T2DM mice (CAV1-shRNA group), while scramble virus injection was used as a negative control (Ctrl-shRNA group). The results showed that glargine was able to upregulate the expression of PI3K and activate serine phosphorylation of AKT through the upregulation of Cav-1 expression in paraepididymal adipose tissue of the insulin group. However, glargine treatment could not activate AKT pathway in Cav-1 silenced diabetic mice. These results suggest that Cav-1 is essential for the activation of AKT and improving insulin sensitivity in type 2 diabetic mice during glargine treatment.

Abstract Background Preeclampsia (PE) is a frequently occurring pregnancy disorder in the placenta, which results in various maternal and fetal complications. The current study aims to evaluate the role of extracellular vesicles (EVs)-encapsulated microRNA (miR)-101 in biological processes of trophoblasts in PE and its underlying mechanism. Methods Human umbilical cord mesenchymal stem cell (HUCMSC) and HUCMSC-derived EVs were isolated and cultured, after which EV characterization was carried out using PKH67 staining. In silico analyses were adopted to predict the downstream target genes of miR-101, and dual luciferase reporter gene assay was applied to validate the binding affinity. Furthermore, loss- and gain-of-function approaches were adopted to determine the role of miR-101 and bromodomain-containing protein 4 (BRD4) in trophoblast proliferation and invasion using EDU staining and transwell assay. In addition, a rat model of PE was established to verify the function of EV-encapsulated miR-101 in vivo. Results Placental tissues obtained from PE patients presented with downregulated miR-101 expression and upregulated BRD4 and CXCL11 expression. EV-encapsulated miR-101 from HUCMSCs could be delivered into the trophoblast HTR-8/SVneo cells, thus enhancing proliferation and migration of trophoblasts. Mechanically, miR-101 targeted and negatively regulated BRD4 expression. BRD4 knockdown promoted the proliferation and migration of trophoblasts by suppressing NF-κB/CXCL11 axis. EV-encapsulated miR-101 from HUCMSCs also reduced blood pressure and 24 h urine protein in vivo, thereby ameliorating PE. Conclusion In summary, EV-encapsulated miR-101 promoted proliferation and migration of placental trophoblasts through the inhibition of BRD4 expression via NF-κB/CXCL11 inactivation.

Intravenous (i.v.) glucocorticoid is recommended for active moderate-to-severe thyroid-associated ophthalmopathy (TAO). However, the details of the treatment schedule are still debatable. The present prospective randomized trial was performed to compare clinical outcomes and serum cytokines between the two regimens. A cohort of 90 patients with active moderate-to-severe TAO was randomized to receive i.v. methyl prednisolone on a weekly protocol or daily scheme. The response rate was evaluated at the 12-week follow-up visit. Serum interleukin (IL)-2, IL-6 and IL-17 levels were measured in 160 patients with TAO, 60 patients with isolated Graves' disease (GD) and 60 normal control (NC) at baseline, as well as patients with active moderate-to-severe TAO at the 12(th) week after treatment. The daily scheme had a higher response rate than the weekly protocol without a significant difference (77.8 vs. 63.6%, P>0.05). No major adverse events were recorded under either regimen. Overall, minor events were more common on the daily scheme (11.36 vs. 4.35%, P<0.05)than on the weekly protocol, whereas the deterioration of eye symptoms (two patients) was only reported on the weekly protocol. At baseline, the IL-17 level in the TAO group was higher than that in the isolated GD and NC groups (P<0.05). In addition, the IL-17 level in the active TAO group was higher than that in the inactive TAO group (P<0.05). Furthermore, the IL-17 level had significantly decreased under the two regimens at the 12-week visit (P<0.05). In conclusion, for patients with active moderate-to-severe TAO, daily i.v. glucocorticoid therapy has a relative higher response rate than the weekly protocol with a few more minor adverse events. These two regimens have their own merits with regard to adverse effects. IL-17 has the potential to be a biomarker for evaluating TAO activity and treatment effects.

Objective:To investigate the role of caveolin-1 (Cav1) in palmitic acid-induced survival of pancreatic islet β-cell.Methods:Cav1-deficient NIT-1 cells and mouse primary islets were constructed through lentiviral vector transfection. Control-shRNA was transfected as a control. After fatty-acid-free bovine serum albumin (BSA) and palmitic acid (0.5 mmol/L) incubation for 24 and 48 hours, cell viability and apoptosis werecalculated by methylcyclopentadienyl manganese tricarbonylassay and Hoechst33342/propidium iodide (PI) double staining.Real time PCR and Western blot were used to detect the expression of apoptosis-associated mRNA and protein. The t test and one-way ANOVA were used for statistical analysis. Results:Compared with the control group, the cell survival rate of the palmitic acid treatment group was significantly decreased (0.89±0.10 vs 0.24±0.04, t=13.49, P<0.01), and mRNA and protein expressions of P18 and P19 were up-regulated (1.00±0.09 vs 1.30±0.04, 1.00±0.04 vs 1.37±0.13, t=5.28, 4.71, both P<0.05; 0.87±0.04, 1.48±0.05, 1.02±0.06 vs 1.41±0.07, t=16.50, 7.33, both P<0.01); mRNA expression levels of cysteine-containing aspartic proteolytic enzymes (Caspase-6, Caspase-9, and Caspase-12) were significantly up-regulated (1.00±0.04 vs 1.57±0.08, 1.01±0.15 vs 1.57±0.20, 1.02±0.19 vs 1.57±0.09, t=11.04,3.88, 4.53, all P<0.05), and protein expression of them were also significantly increased (0.86±0.10 vs 1.28±0.11, 0.69±0.01 vs 0.86±0.06, 0.70±0.02 vs 1.18±0.01, t=4.89,4.84, 37.18, all P<0.01). The cell survival rate of Cav1-shRNA+palmitic acid group was significantly higher than that of Ctrl-shRNA+palmitic acid group (0.53±0.10 vs 0.26±0.08, t=4.71, P<0.01). The mRNA and protein expression of P19 were all down-regulated (1.53±0.18 vs 0.66±0.04, 1.48±0.05 vs 0.70±0.02, t=8.17, 25.09, both P<0.01); mRNA expression levels of Caspase-6, Caspase-7, Caspase-9 and Caspase-12 were down-regulated (1.82±0.11 vs 1.03±0.07, 1.43±0.24 vs 0.42±0.15, 1.41±0.22 vs 0.51±0.18, 1.45±0.18 vs 0.42±0.13, t=5.48-10.49, all P<0.01), and protein expression of them were also down-regulated (0.99±0.14 vs 0.63±0.11, 0.90±0.14 vs 0.62±0.04, 0.90±0.02 vs 0.60±0.04, 1.30±0.01 vs 0.65±0.04, t=3.50-27.31, all P<0.01). Conclusion:Cav1 deficiency inhibites palmitic acid-induced pancreatic β-cell apoptosis via regulating Caspase family, and ultimately protects β cell viability.

This study aims to evaluate the relationship between subclinical hypothyroidism (SCH) during the first trimester and gestational diabetes mellitus (GDM) that was happened later on in our cohort. A total of 6530 pregnant women who first visited before 13+6 gestational weeks and accepted routinely prenatal services in the third affiliated hospital of Sun Yat-Sen University from January 2015 to September 2018 were finally met the inclusion criteria and recruited. Thyroid functions were performed at the first visit and a 2h 75-g OGTTwas performed between 24-28 weeks. The glucose levels of OGTT were shown no differences between SCH and euthyroid women [0 hour: 4.17 mmol/L (3.99-4.38) vs. 4.16 mmol/L (3.97-4.39), P = 0.730; 1 hour: 7.61 mmol/L (6.64-8.66) vs. 7.68 mmol/L (664-8.75), P = 0.449; 2 hour: 6.66 mmol/L (5.85-7.68) vs. 6.75 mmol/L (5.93-7.74), P = 0.142]. The incidence of GDM was no change as the TSH level increased in either TPOAb negative or positive SCH women (all P > 0.05). After using the multivariate logistic regression analysis adjusted for confounders (maternal age, educational levels and preBMI), elevated TSH and TPOAb status also had no effect on the incidence of GDM (Table 1). These results suggested maternal SCH in the first trimester was not related to GDM, regardless of the TPOAb status or the TSH levels. Disclosure P. Li: None. S. Lin: None. J. Fan: None. Funding Science and Technology Planning Project of Guangdong Province (2017A020215026); Medical Scientific Research Foundation of Guangdong Province (A2017314)

目的 探讨小鼠胰岛提取方法 的改良及效果.方法根据胰岛提取方法不同,将小鼠随机分为胆总管穿刺组和胆总管穿刺联合胰腺原位注射组(联合注射组),每组各100只.胰岛提取的改良方法采用胆总管穿刺联合胰腺原位注射法灌注胰腺,体视显微镜下挑选并纯化胰岛.鉴定胰岛的形态和纯化情况,统计两组的胰岛产量及胰岛提取成功率.分析胰岛在体外培养1周内的存活情况,并评估体外培养24 h和4 d后胰岛的胰岛素分泌功能.结果 与胆总管穿刺组比较,联合注射组提取胰岛的产量显著增高(P<0.001).两组胰岛提取成功率均为83％,差异无统计学意义(P>0.05).胆总管穿刺联合胰腺原位注射法所提取的胰岛形态完好、纯度高、活性好.新鲜分离的胰岛体外培养24 h后胰岛存活率接近100％;体外培养1～5 d,胰岛细胞存活情况良好;体外培养6 d开始,胰岛出现中心性死亡.体外培养24 h和4 d后,给予高葡萄糖刺激胰岛后,小鼠胰岛功能均正常.结论 胆总管穿刺联合胰腺原位注射法可以提高胰岛的产量,且获取的胰岛细胞活性和功能良好.

Objective: Islet amyloid polypeptide(IAPP) is a major component of islet amyloid deposition in type 2 diabetic patients. Our preliminary study showed that membrane protein Caveolin-1 (Cav1) silencing inhibited β-cell apoptosis and promote insulin secretion. The purpose of this study was to investigate the regulation of Cav1 on islet β-cell IAPP and its mechanism. Materials and Methods: Firstly, IAPP secretion of each group of primary islets was detected by ELISA, then Western blot and real-time quantitative PCR (qPCR) were used to detect the expression levels of IAPP, PAM(Peptidyl-glycine alpha-amidating monooxygenase), PC1(Prohormone Convertase-1), BACE2(Beta-secretase 2), IDE(Insulin-Degrading Enzyme), TXNIP(Thioredoxin interacting protein), and finally, it confirmed that Cav1 interacted with the protein of TXNIP by immunofluorescence confocal and co-immunoprecipitation (Co-IP). Results: The secretion of IAPP was decreased in primary pancreatic islets and Cav1 deficiency significantly down-regulated the expression of IAPP in NIT-1 cells, while over-expression of Cav1 in βTC-6 cells significantly up-regulated the expression of IAPP. This effect correlated with Cav1 silencing leading to down-regulation of PAM expression associated with IAPP synthesis and over-expression of Cav1 leading to up-regulation of PC1 associated with IAPP synthesis. On the other hand,Cav1 deficiency resulted in the up-regulation of the expression of BACE2 and IDE, the enzymes associated with IAPP decomposition. While over-expression of Cav1 leaded to down-regulation of BACE2 and IDE. Further studies indicated that Cav1 co-localized with TXNIP, and it interacted with and regulated the expression of TXNIP, which regulates IAPP gene transcription. Conclusion: Cav-1 can regulate the synthesis and decomposition of IAPP in islet β cell by interacting with TXNIP. This study suggests that Cav-1 may protect diabetic islet beta cells by reducing islet amyloid deposition. Disclosure K. Liu: None. W. Zeng: None. S. Lin: None. C. Lin: None. F. Xu: None. N. Cai: None. L. Zeng: None. Funding National Natural Science Funding of China (51873071); Natural Science Foundation of Guangdong Province (2018B030311012)