Novel HNF1A gene mutation in maturity-onset diabetes of the young: A case report.

BACKGROUND: Maturity-onset diabetes of the young 3 (MODY3), caused by mutations in the HNF1A gene, is the most common subtype of MODY. The diagnosis of MODY3 is critical because a low dose of sulfonylurea agents can achieve glucose control. CASE SUMMARY: We describe a patient with MODY3 involving a novel splicing mutation, in whom low-dose gliclazide was sufficient to control clinically significant hyperglycemia. Sanger sequencing identified a splicing HNF1A mutation in 12q24 NM_000545.5 Intron5 c.1108-1G>A. Glycemic control has been maintained without insulin therapy for 28 mo after the diagnosis of diabetes. CONCLUSION: This case report highlights a novel HNF1A gene mutation in MODY3 that is responsive to sulfonylurea therapy.

High Maternal Serum Estradiol in First Trimester of Multiple Pregnancy Contributes to Small for Gestational Age via DNMT1-Mediated CDKN1C Upregulation.

High maternal serum estradiol (E2) levels in the first trimester of pregnancy are associated with a high incidence of low birth weight (LBW) and small for gestational age (SGA). This study aimed to investigate the effect of first-trimester high maternal serum E2 levels on fetal growth and the underlying mechanisms in multiple pregnancies. Maternal serum E2 levels of women at 8 weeks of gestation were measured. The expression levels of imprinted genes and DNMT1 were determined by RT-qPCR, and KvDMR1 methylation in embryo tissue, placenta, and newborn cord blood samples was examined by bisulfite sequencing PCR. The effect of E2 on CDKN1C expression was investigated in HTR8 cells. The incidence of SGA was significantly higher in multiple pregnancies reduced to singleton than that in primary singleton pregnancies (11.4% vs. 2.9%) (P < 0.01) and multiple pregnancies reduced to twins than primary twins (38.5% vs. 27.3%) (P < 0.01). The maternal serum E2 level at 8 weeks of gestation increased with the number of fetuses and was negatively correlated with offspring birth weight. CDKN1C and DNMT1 expression was significantly upregulated in embryo tissue, placenta, and cord blood from multiple pregnancies. Furthermore, there was a positive correlation between CDKN1C mRNA expression and KvDMR1 methylation levels. In HTR8 cells, DNMT1 mediated the estrogen-induced upregulation of CDKN1C, which might contribute to SGA. To minimize the risks of LBW and SGA, our findings suggest that abnormally high maternal serum E2 levels should be avoided during the first trimester of multiple pregnancies from assisted reproductive technology (ART).

Comprehensive overview of human serum albumin glycation in diabetes mellitus.

The presence of excess glucose in blood is regarded as a sweet hurt for patients with diabetes. Human serum albumin (HSA) is the most abundant protein in human plasma, which undergoes severe non-enzymatic glycation with glucose in patients with diabetes; this modifies the structure and function of HSA. Furthermore, the advanced glycation end products produced by glycated HSA can cause pathological damage to the human body through various signaling pathways, eventually leading to complications of diabetes. Many potential glycation sites on HSA have different degrees of sensitivity to glucose concentration. This review provides a comprehensive assessment of the in vivo glycation sites of HSA; it also discusses the effects of glycation on the structure and function of HSA. Moreover, it addresses the relationship between HSA glycation and diabetes complications. Finally, it focuses on the value of non-enzymatic glycation of HSA in diabetes-related clinical applications.

Relationship between serum Dickkopf-1 and albuminuria in patients with type 2 diabetes.

BACKGROUND: Diabetic kidney disease is a microvascular complication of diabetes with complex pathogenesis. Wingless signaling-mediated renal fibrosis is associated with diabetic kidney disease. Dickkopf-1, a negative regulator of Wingless, has been proven to participate in renal fibrosis, glucose metabolism, and inflammation. However, whether serum Dickkopf-1 levels are associated with diabetic kidney disease remains unclear. AIM: To assess the relationship between serum Dickkopf-1 levels and albuminuria in individuals with type 2 diabetes. METHODS: Seventy-three type 2 diabetes patients and 24 healthy individuals were enrolled in this case-control study. Diabetic individuals were separated into normal albuminuria, microalbuminuria, and macroalbuminuria groups based on their urinary albumin/creatinine ratios (UACRs). Clinical characteristics and metabolic indices were recorded. Serum Dickkopf-1 levels were determined by enzyme-linked immunosorbent assay. RESULTS: No significant difference in serum Dickkopf-1 levels was found between healthy individuals and the normal albuminuria group. However, the levels in the microalbuminuria group were significantly lower than those in the normal albuminuria group (P = 0.017), and those in the macroalbuminuria group were the lowest. Bivariate analysis revealed that serum Dickkopf-1 levels were positively correlated with hemoglobin A1c level (r = 0.368, P < 0.01) and estimated glomerular filtration rate (r = 0.339, P < 0.01), but negatively correlated with diabetes duration (r = -0.231, P = 0.050), systolic blood pressure (r = -0.369, P = 0.001), serum creatinine level (r = -0.325, P < 0.01), and UACR (r = -0.459, P < 0.01). Multiple and logistic regression showed that serum Dickkopf-1 levels were independently associated with UACR (odds ratio = 0.627, P = 0.021). CONCLUSION: Serum Dickkopf-1 levels are negatively associated with UACR. Lower serum Dickkopf-1 levels could be a critical risk factor for albuminuria in diabetes.

Association between serum Sestrin2 level and diabetic peripheral neuropathy in type 2 diabetic patients.

BACKGROUND: Diabetic peripheral neuropathy (DPN) is a chronic and serious microvascular complication of diabetes linked to redox imbalance. Sestrin2, a novel inducible stress protein, participates in glucose metabolic regulation and redox homeostasis. However, the association between serum Sestrin2 and DPN is unknown. AIM: To explore the association between serum Sestrin2 and DPN in patients with type 2 diabetes mellitus (T2DM). METHODS: A total of 96 T2DM patients and 39 healthy volunteers, matched by age and sex, participated in this cross-sectional study. Clinical features and metabolic indices were identified. Serum Sestrin2 was measured by ELISA. The association between Sestrin2 and DPN was studied. Correlation and logistic regression analyses were used to evaluate the associations of different metabolic indices with Sestrin2 and DPN. RESULTS: The 96 patients with T2DM were divided into DPN (n = 47) and patients without DPN (n = 49). Serum Sestrin2 was significantly lower in healthy volunteers than in all T2DM patients combined [9.10 (5.41-13.53) ng/mL vs 12.75 (7.44-23.80) ng/mL, P < 0.01]. T2DM patients without DPN also had significantly higher levels of Sestrin2 than healthy volunteers [14.58 (7.93-26.62) ng/mL vs 9.10 (5.41-13.53) ng/mL, P < 0.01]. However, T2DM patients with DPN had lower circulating Sestrin2 levels compared to T2DM patients without DPN [9.86 (6.72-21.71) ng/mL vs 14.58 (7.93-26.62) ng/mL, respectively, P < 0.01]. Bivariate correlation analysis revealed that serum Sestrin2 was positively correlated with body mass index (r = 0.672, P = 0.000), hemoglobin A1c (HbA1c) (r = 0.292, P = 0.000), serum creatinine (r = 0.206, P = 0.016), triglycerides (r = 0.731, P = 0.000), and fasting glucose (r = 0.202, P = 0.040), and negatively associated with estimated glomerular filtration rate (r = -0.230, P = 0.007). After adjustment for sex, age, HbA1c, and diabetes duration, multiple regression analysis revealed that Sestrin2 was independently correlated with body mass index and triglyceride levels (P = 0.000). Logistic regression analyses indicated that Sestrin2, diabetes duration, and high-density lipoprotein were strongly associated with DPN (odds ratio = 0.855, 1.411, and 0.041, respectively). CONCLUSION: Our results show Sestrin2 is decreased in T2DM patients with DNP. As lower Sestrin2 is independently associated with DPN, Sestrin2 may contribute to progression of DPN in T2DM patients.

Novel insight into perirenal adipose tissue: A neglected adipose depot linking cardiovascular and chronic kidney disease.

Obesity is associated with adverse metabolic diseases including cardiovascular disease (CVD) and chronic kidney disease (CKD). These obesity-related diseases are highly associated with excess fat accumulation in adipose tissue. However, emerging evidence indicates that visceral adiposity associates more with metabolic and cardiovascular risk factors. Perirenal adipose tissue, surrounding the kidney, is originally thought to provides only mechanical support for kidney. However, more studies demonstrated perirenal adipose tissue have a closer association with renal disease than other visceral fat deposits in obesity. Additionally, perirenal adipose tissue is also an independent risk factor for CKD and even associated more with CVD. Thus, perirenal adipose tissue may be a connection of CVD with CKD. Here, we will provide an overview of the perirenal adipose tissue, a neglected visceral adipose tissue, and the roles of perirenal adipose tissue linking with CVD and CKD and highlight the perirenal adipose tissue as a potential strategy for future therapeutics against obesity-related disease.

Aberrant expression and DNA methylation of lipid metabolism genes in PCOS: a new insight into its pathogenesis.

Background: Polycystic ovary syndrome (PCOS), whose etiology remains uncertain, is a highly heterogenous and genetically complex endocrine disorder. The aim of this study was to identify differentially expressed genes (DEGs) in granulosa cells (GCs) from PCOS patients and make epigenetic insights into the pathogenesis of PCOS. Results: Included in this study were 110 women with PCOS and 119 women with normal ovulatory cycles undergoing in vitro fertilization acting as the control group. RNA-seq identified 92 DEGs unique to PCOS GCs in comparison with the control group. Bioinformatic analysis indicated that synthesis of lipids and steroids was activated in PCOS GCs. 5-Methylcytosine analysis demonstrated that there was an approximate 25% reduction in global DNA methylation of GCs in PCOS women (4.44 ± 0.65%) compared with the controls (6.07 ± 0.72%; P < 0.05). Using MassArray EpiTYPER quantitative DNA methylation analysis, we also found hypomethylation of several gene promoters related to lipid and steroid synthesis, which might result in the aberrant expression of these genes. Conclusions: Our results suggest that hypomethylated genes related to the synthesis of lipid and steroid may dysregulate expression of these genes and promote synthesis of steroid hormones including androgen, which could partially explain mechanisms of hyperandrogenism in PCOS.

Preliminary proteomic analysis on the alterations in follicular fluid proteins from women undergoing natural cycles or controlled ovarian hyperstimulation.

PURPOSE: To study the differences in protein expression profiles of follicular fluid (FF) between controlled ovarian hyperstimulation (COH) and natural ovulatory cycles. METHODS: Twelve infertile women undergoing in vitro fertilization and embryo transfer (IVF-ET), with matched clinical information, were retrospectively recruited in the IVF center of our university hospital, including six undergoing COH and another six with natural cycles. FF was sampled from dominant follicles with mature oocytes. Protein expression profiles in each FF sample were analyzed respectively using two-dimensional gel electrophoresis. Differentially expressed proteins were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and validated by western blotting. Differentially expressed proteins were further analyzed using Ingenuity Pathway Analysis (IPA) software. RESULTS: Two proteins were downregulated and 11 proteins were upregulated (change ≥1.5-fold, P < 0.05) in the COH group. We identified one down-egulated and seven upregulated proteins using MALDI-TOF MS. Four differentially expressed proteins, including transferrin, complement component C3 (C3), haptoglobin and alpha-1-antitrypsin (AAT), were further validated by rate nephelometry and western blotting analyses. The IPA analysis revealed a significant network involved in the humoral immune and inflammatory responses. CONCLUSIONS: The eight differentially expressed proteins were related to immune and inflammatory responses in the ovary. Our results provide new insights into the influence of COH on follicular (spp) development and IVF outcomes.

[Effects of inward rectifier potassium channel blockers on EPCs function].

OBJECTIVE: To investigate the effects of inward rectifier potassium channel blockers (BaCl2, CsCl) on the functions of endothelial progenitor cells (EPCs). METHODS: Density gradient centrifugation-isolated rat hone marrow mononuclear cells were cultured in vitro. EPCs were harvested and seeded on six culture dish when cells grew to 3-5 passages. Before testing the EPCs were synchronized with M199, which contain 2% fetal calf serum. In the end, EPCs were treated with different intervention. The experiment mainly included two parts: (1) BaCl2 (100 micromol/L) and free BaC2 of Tyrodes solution; (2) CsCl (1 mmol/L) and control. Cell pretreated with blockers above mentioned for 12 h, then the gene expression of stromal cell-derived factor-1 (SDF-1), epoprotenol (PGI2) were assessed, beyond that the ability of adhesion, migration were assayed with different tests. In addition, the medium was collected when EPCs were treated for 3 days. The levels of SDF-1 were measured by sandwich enzyme-linked immunosorbent assay (ELISA). Going even further, EPCs were treated with the signal pathway blockers in advance, after repeat the above steps, in order to analyze the change of SDF-1 and then discuss its mechanism. RESULTS: Compared with control group, BaCl2, CsCl could increase EPC adhesion and migration to same extent. Moreover, the gene expression of SDF-1, PGI2 was significantly up-regulated and the production of SDF-1 increased evidently. Furthermore, the mechanism of SDF-1 secretion increasing mainly was associated with eNOS signaling pathways. CONCLUSION: Ba2+ and Cs+ play important roles in increasing EPCs functions, such as adhesion, migration and secretion.

[Relationship between gastrointestinal dyskinesis and gastrointestinal neurons in diabetic mellitus: experiment with rats].

OBJECTIVE: To investigate the relationship between gastrointestinal dyskinesis and histologic changes of gastrointestinal myenteric plexus cholinergic and nitrergic neurons in STZ-induced diabetic rats. METHODS: 45 SD rats were randomly divided into control group, diabetic group and insulin group. 16 weeks after diabetic model established, gastrointestinal motility of rats was measured and histologic changes of myenteric plexus cholinergic neuron and nitrergic neuron was observed. RESULTS: Compared with control group, gastrointestinal motility of diabetic group was markedly slow (P < 0.01), the myenteric plexus cholinergic neuron counting of gastric antrum and small intestine were significantly decreased (6.6 +/- 2.9 vs 15.7 +/- 3.8 15.6 +/- 10.3 vs 22.6 +/- 7.4, P < 0.01), yet the number of nitrergic neuron only markedly reduced in gastric antrum (5.3 +/- 1.2 vs 11.8 +/- 2.2, P < 0.01). The gastrointestinal mobility, gastric antrum nitrergic neuron and small intestine cholinergic neuron counting of insulin group were markedly higher than that of diabetic group (P < 0.05), yet lower than that of control group (P < 0.01). CONCLUSION: The gastrointestinal dyskinesis of STZ-induced diabetic rats might be associated with lesions of gastrointestinal myenteric plexus cholinergic neuron and nitrergic neuron. Insulin intensive therapy can partly ameliorate diabetic gastrointesternal dyskinesis.

[The expression of estrogen receptor alpha and beta in the intervention of different estrogens in rat bone metabolism].

In this present study, female rats were ovariectomized (OVX) as the models of osteoporosis. The aim is to determine the different mechanisms of estrogen receptor(ER) alpha and beta pathway in mediating estrogen to participate in trabecular bone metabolism, and to further explore the distinction of modulation on ER alpha or ER beta between estrogens with different components. Mature female Sprague-Dawley rats (n=40) were randomly divided into four groups: group Control (sham operated), group OVX (only ovariectomized), group CEE (OVX rats treated with conjugated equine estrogens) and group EV (OVX rats treated with estradiol valerate). Sham operation and OVX were performed 48 days (12 estrums) before different liquid diet. The rats in group Control and group OVX were orally administrated with physiological saline solution and the rats in group CEE or group EV were orally administrated with CEE or EV for 12 days (3 estrums) before sacrifice. Relative quantitative reverse transcription- polymerase chain reaction (RT-PCR) and western blot techniques were utilized to compare the levels of ER alpha and ER beta mRNA and proteins in trabecular bone among groups. The results showed that in rat trabecular bone of group Control, the expression of ER alpha protein (1.433 +/- 0.250) was significantly higher than that of ER beta(0.687 +/- 0.120), whereas the ER alpha mRNA (0.285 +/- 0.033) was much lower than ERbeta mRNA(0.590 +/- 0.044). Following OVX, the levels of ER alpha protein (0.685 +/- 0.103) declined significantly, whereas mRNA levels (0.405 +/- 0.036) markedly increased. Both the protein (1.091 +/- 0.078) and mRNA (0.729 +/- 0.030) levels of ER beta significantly increased after OVX. After treatment with CEE, the expression of ER beta protein (0.583 +/- 0.129) and mRNA (0.618 +/- 0.043) were markedly down-regulated compared with group OVX. After treatment with EV, the ER alpha protein expression (1.272 +/- 0.247) was markedly up-regulated, while ERa mRNA (0.277 +/- 0.040) and ER beta protein (0.620 +/- 0.174) were markedly down-regulated compared with group OVX. On the other hand, the loss of bone mass in rat trabecular bone by OVX (0.192 +/- 0.010) was markedly gained after CEE (0.228 +/- 0.012) and EV (0.259 +/- 0.007) treatment. These results demonstrate that ERalpha may be the predominant subtype expressed in female rat trebecular bone and play a key role in the bone metabolism. This study also shows that estrogens with different components protect bone through variant ERs pathway.