<b>Evaluation of Some Proteins as Potential Markers in Different Durations of Diabetic Patients Type 2 </b>

Diabetes mellitus is a globally prevalent disease with several parameters that may be involved in diagnosis or altered throughout disease progression. In this study, 120 patients with type 2 diabetes and healthy controls were recruited, with ages ranging from 30-65 years old. The research focused on determining levels of Protein Z (PROZ), Laminin Subunit Alpha 2 (LAMA2), Mixed Lineage Leukemia 4 (MLL4), and Plexin Domain Containing 2 (PLXDC2). Participants were divided into four groups: group I (n=30) with long-term metformin treatment, group II (n=30) with short-term metformin treatment, group III (n=30) newly diagnosed without medication, and group IV (n=30) healthy controls. Protein markers were measured by ELISA, while fasting blood glucose, lipid profiles, and HbA1c were determined by spectrophotometry and HPLC. ROC curve analysis revealed PLXDC2 had high sensitivity and specificity as a potential biomarker for diabetes diagnosis. LAMA2, MLL4, and PROZ also distinguished between diabetes patients and controls. Overall, these results identify promising biomarkers for diabetes detection and monitoring.

Objective: This study aimed to assess the role of Fas/Fas ligand (FasL) system in the progression of type 2 diabetes mellitus (T2DM). Materials and Methods: This study included 100 participants – 30 persons as a control group and 70 patients with T2DM (35 males and 35 females) and their ages were ranged from 40 to 70 years. The patients were distributed into two groups according to gender and duration of the disease: newly diagnosed group for short duration ≤5 years and chronic diagnosed group for long duration >5 years. Serum sFas and sFasL levels were measured by enzyme-linked immunosorbent assay technique, and also, lipid and glucose profile were measured by COBAS analyzer. Results: The results revealed a significant (P ≤ 0.05) decrement in the levels of FasL in T2DM than controls while the levels of Fas were increasing significantly (P ≤ 0.05) in T2DM than controls. Hemoglobin A1c (HbA1c) and fasting blood glucose were negatively correlated with FasL, while high-density lipoprotein was positively correlated with it, and whereas HbA1c positively correlated with Fas, the gender and duration of disease did not show any correlation with the disease. Conclusion: Our findings suggest that hyperglycemia causes increase in Fas levels which lead to dysfunction of pancreatic β-cell in T2DM.

Reliable protein markers for pre-diabetes in humans are not clinically available. In order to identify novel and reliable protein markers for pre-diabetes in humans, healthy volunteers and patients diagnosed with pre-diabetes and stroke were recruited for blood collection. Blood samples were collected from healthy and pre-diabetic subjects 12 h after fasting. BMI was calculated from body weight and height. Fasting blood glucose (FBG), glycated hemoglobin (HbA1C), triglyceride (TG), total cholesterol, high-density lipoprotein, low-density lipoprotein (LDL), insulin and albumin were assayed by automated clinical laboratory methods. We used a quantitative proteomics approach to identify 1074 proteins from the sera of pre-diabetic and healthy subjects. Among them, 500 proteins were then selected using Mascot analysis scores. Further, 70 out of 500 proteins were selected via volcano plot analysis according to their statistical significance and average relative protein ratio. Eventually, 7 serum proteins were singled out as candidate markers for pre-diabetes due to their diabetic relevance and statistical significance. Immunoblotting data demonstrated that laminin subunit alpha 2 (LAMA2), mixed-lineage leukemia 4 (MLL4), and plexin domain containing 2 (PLXDC2) were expressed in pre-diabetic patients but not healthy volunteers. Receiver operating characteristic curve analysis indicated that the combination of the three proteins has greater diagnostic efficacy than any individual protein. Thus, LAMA2, MLL4 and PLXDC2 are novel and reliable serum protein markers for pre-diabetic diagnosis in humans.

To clarify the expression of N6-methyladenosine (m6 A) modulators involved in the pathogenesis of type 2 diabetes mellitus (T2DM). We further explored the association of serum insulin-like growth factor 2 mRNA-binding proteins 3 (IGF2BP3) levels and odds of T2DM in a high-risk population. The gene expression data set GSE25724 was obtained from the Gene Expression Omnibus, and a cluster heatmap was generated by using the R package ComplexHeatmap. Differential expression analysis for 13 m6 A RNA methylation regulators between nondiabetic controls and T2DM subjects was performed using an unpaired t test. A cross-sectional design, including 393 subjects (131 patients with newly diagnosed T2DM, 131 age- and sex-matched subjects with prediabetes, and 131 healthy controls), was carried out. The associations between serum IGF2BP3 concentrations and T2DM were modeled by restricted cubic spline and logistic regression models. Two upregulated (IGF2BP2 and IGF2BP3) and 5 downregulated (methyltransferase-like 3 [METTL3], alkylation repair homolog protein 1 [ALKBH1], YTH domain family 2 [YTHDF2], YTHDF3, and heterogeneous nuclear ribonucleoprotein [HNRNPC]) m6 A-related genes were found in islet samples of T2DM patients. A U-shaped association existed between serum IGF2BP3 levels and odds of T2DM according to cubic natural spline analysis models, after adjustment for body mass index, waist circumference, diastolic blood pressure, total cholesterol, and triglyeride. Multivariate logistic regression showed that progressively higher odds of T2DM were observed when serum IGF2BP3 levels were below 0.62 ng/mL (odds ratio 3.03 [95% confidence interval 1.23-7.47]) in model 4. Seven significantly altered m6 A RNA methylation genes were identified in T2DM. There was a U-shaped association between serum IGF2BP3 levels and odds of T2DM in the general Chinese adult population. This study provides important evidence for further examination of the role of m6 A RNA methylation, especially serum IGF2BP3 in T2DM risk assessment.

This study aims to explore whether bilirubin can act as a biomarker of oxidative stress in type 1 diabetes mellitus (T1DM) by analyzing the serum bilirubin levels and possible influencing factors in different disease states and durations in children with T1DM. This is a retrospective study. The medical records of 1652 inpatients with T1DM and 101 healthy children in Shanxi Provincial Children's Hospital from 2014 to 2023 were collected and divided into different subgroups. The relevant indices in different disease states and durations in the T1DM group were statistically analyzed, particularly the serum bilirubin levels and possible influencing factors. Compared to children without diabetic ketoacidosis (DKA)/diabetic ketosis (DK), children with DKA/DK exhibited higher random blood glucose (RBG), HbA1C, total bilirubin (TBil), and indirect bilirubin (IBil) (P < .05). Compared to the control group, the levels of TBil and IBil in the newly-diagnosed and established T1DM children were statistically significantly higher (P < .05). Compared to newly-diagnosed T1DM children, serum TBil and IBil levels were statistically significantly lower in the established T1DM group and subgroups with different disease durations (P < .05). TBil and IBil were correlated with the status of blood glucose control and can be reflected by RBG, HbA1C, and DKA/DK (P < .05), but had no correlation was observed with disease duration (P > .05). Serum bilirubin possesses the potential to be a biomarker of oxidative stress in T1DM children.

In the article “Type 2 diabetes mellitus, brain atrophy, and cognitive decline,” Moran et al. examined longitudinal relationships between type 2 diabetes mellitus (T2DM), cortical thickness, and cognitive function in older people with normal cognition, mild cognitive impairment (MCI), and Alzheimer disease (AD) in the Alzheimer's Disease Neuroimaging Initiative (ADNI) cohort. They concluded that T2DM contributed to cognitive decline via neurodegeneration (lower baseline cortical thickness) and that previous brain and cognitive reserve (reflected by greater education) may protect against this effect. In response, Ba et al. argue that it is unclear whether the authors properly controlled for other risk factors for cognitive decline such as hypertension, hyperlipidemia, white matter lesion burden, and depression and note that incorporating additional T2DM-related factors such as fasting blood glucose or disease duration would be relevant. They also critique the study's reliance on clinical diagnoses for determination of MCI and AD, which can often result in misattribution of symptoms to AD when supportive biomarkers are not included. Replying to these comments, Dr. Moran acknowledges the absence of detailed T2DM-specific information as a limitation of repurposing the ADNI dataset for their work but notes that the study reported data on fasting glucose, diabetes medication use, blood pressure, and measures of obesity. Although ADNI participants were selected to have a low burden of cerebrovascular disease, Dr. Moran acknowledges that there are additional potential confounders of the relationship between T2DM and brain health, such as other vascular risk factors, some of which were explored in other work using samples with a greater burden of cerebrovascular disease. He also agrees on the importance of incorporating biomarkers of underlying pathology into future work in this area. In the article “Type 2 diabetes mellitus, brain atrophy, and cognitive decline,” Moran et al. examined longitudinal relationships between type 2 diabetes mellitus (T2DM), cortical thickness, and cognitive function in older people with normal cognition, mild cognitive impairment (MCI), and Alzheimer disease (AD) in the Alzheimer's Disease Neuroimaging Initiative (ADNI) cohort. They concluded that T2DM contributed to cognitive decline via neurodegeneration (lower baseline cortical thickness) and that previous brain and cognitive reserve (reflected by greater education) may protect against this effect. In response, Ba et al. argue that it is unclear whether the authors properly controlled for other risk factors for cognitive decline such as hypertension, hyperlipidemia, white matter lesion burden, and depression and note that incorporating additional T2DM-related factors such as fasting blood glucose or disease duration would be relevant. They also critique the study's reliance on clinical diagnoses for determination of MCI and AD, which can often result in misattribution of symptoms to AD when supportive biomarkers are not included. Replying to these comments, Dr. Moran acknowledges the absence of detailed T2DM-specific information as a limitation of repurposing the ADNI dataset for their work but notes that the study reported data on fasting glucose, diabetes medication use, blood pressure, and measures of obesity. Although ADNI participants were selected to have a low burden of cerebrovascular disease, Dr. Moran acknowledges that there are additional potential confounders of the relationship between T2DM and brain health, such as other vascular risk factors, some of which were explored in other work using samples with a greater burden of cerebrovascular disease. He also agrees on the importance of incorporating biomarkers of underlying pathology into future work in this area.

Background Vitamin D deficiency can lead to the increased severity and prevalence of metabolic disorders. However, the relationship between levels of 25-hydroxyvitamin D (25(OH)D) and peripheral arterial disease (PAD) is controversial. Therefore, the purpose of our study was to explore the relationship between 25(OH)D levels and PAD in middle-aged and elderly type 2 diabetes mellitus (T2DM) patients in China. Methods In this study, a total of 183 patients with T2DM were enrolled and categorized into groups with or without PAD. Clinical and biochemical parameters were assessed, and a Pearson analysis was used to identify a possible association between levels of 25(OH)D and glycated hemoglobin (HbA1c). Some biochemical parameters were also assessed in the T2DM patients with PAD according to vitamin D status. Interactions were also explored among HbA1c control, 25(OH)D levels, and PAD. The possible risk factors for PAD were measured by multivariable logistic regression analyses. Results Firstly, the parameters including age, HbA1c, and disease duration between T2DM and T2DM+PAD groups showed significantly different. In addition, the frequency of smoking in the group of T2DM patients was significantly less than that in the T2DM patients with the PAD group, while the frequency of well-controlled HbA1c in the patients with T2DM was significantly higher. There is a trend that the levels of 25(OH)D and HbA1c are correlated, but no interactions among vitamin D deficiency, HbA1c control, and PAD were found. However, HbA1c significantly differed between groups with vitamin D deficiency and insufficiency in the T2DM patients with PAD. According to the multivariate logistic regression analyses, the PAD risk factors of T2DM patients were family history of diabetes, smoking, age, disease duration, HbA1c, and LDL. Conclusions The findings demonstrate that the deficiency of vitamin D level is not related to PAD, but HbA1c may be linked to the presence of PAD in middle-aged and elderly patients with T2DM in China.

The relationship between type 1 diabetes mellitus (T1DM) and periodontal disease may exhibit by the alteration of bone metabolism. However, evidence for this relationship is scarce and inconclusive. Thus, the aims of the present study were to investigate salivary receptor activator of nuclear factor kappa-β (RANK), receptor activator of nuclear factor kappa-β ligand (RANKL), osteoprotegerin (OPG) gene expression and the RANKL:OPG ratio in T1DM and non-T1DM. Secondary objective was to determine the relationships of RANK, RANKL and OPG gene expression to clinical parameters of T1DM and periodontal disease. Twenty patients with T1DM and twenty age-matched non-T1DM were recruited. Clinical periodontal parameters were measured. Total RNA was isolated from non-stimulated saliva, and the relative gene expressions of RANK, RANKL, OPG and RANKL:OPG ratio were determined by quantitative real-time polymerase chain reaction. The T1DM group had significantly higher mean periodontal parameters than the non-T1DM group, while the mean plaque scores of both groups were not significantly different. There was a trend of higher relative gene expression of RANK, RANKL, and the RANKL:OPG ratio and lower expression of OPG in T1DM group but no statistic significant different when compared to non-T1DM. In the T1DM group, RANKL:OPG correlated with the percentage of bleeding sites, whereas RANK, RANKL, and HbA1c levels correlated with pocket depth. Bone metabolisms demonstrating by decreased OPG gene expression and upregulated of RANK, RANKL, RANKL:OPG with higher pocket depth and bleeding in T1DM may play an important role in periodontal destruction in T1DM.

Background: Glucagon-like peptide-1 receptor agonists (GLP-1 RAs), such as liraglutide, have demonstrated efficacy in weight reduction and glycemic control. However, comparative data across obese individuals with differing metabolic states non-diabetic (ND), prediabetic (pre-DM), and type 2 diabetes mellitus (T2DM) remain limited, particularly regarding hepatic, lipid, and safety outcomes. This study aimed at evaluating liraglutide’s efficacy in reducing body weight and improving metabolic parameters (HbA1c, lipid profile, liver enzymes) and its safety in obese individuals across these groups. Methods: A prospective study was conducted on 120 obese patients receiving Liraglutide for six months (ND: n=7; pre-DM: n=16; T2DM: n=97). Baseline demographic, anthropometric, and biochemical parameters were recorded. Outcomes assessed at 3 and 6 months included weight, BMI, HbA1c, blood pressure, liver enzymes (ALT, AST), lipid profile, and adverse events. Between-group comparisons were performed using ANCOVA, adjusting for age, disease duration, and baseline metabolic variables. Results: At baseline, patients with T2DM were older and had significantly higher HbA1c, ALT, total cholesterol (TC), and triglycerides (TG) compared with ND and pre-DM groups (p&lt;0.05). Over six months, weight and BMI decreased significantly in all groups, with the greatest mean reduction observed in T2DM (–13.28 ± 8.22 kg), followed by pre-DM (–13.19 ± 9.34 kg), and ND (–6.43 ± 6.53 kg). Higher baseline HbA1c predicted greater weight loss in T2DM and pre-DM (p&lt;0.001). Liraglutide was associated with reductions in ALT and AST across all groups, particularly in those with elevated baseline levels. Lipid improvements were most pronounced in T2DM, with significant reductions in LDL and TG. Conclusion: Liraglutide therapy in obese patients led to significant weight loss and favorable effects on glycemic, hepatic, and lipid parameters across ND, pre-DM, and T2DM groups, supporting the broader role of Liraglutide in obesity and metabolic disease management.

Aims. To evaluate the profile of pro- and anti-inflammatory cytokines in type 1 diabetes mellitus (T1DM) and the way they are connected in co-regulated networks and determine whether disease duration influences their pattern.Methods. Plasma levels of 20 cytokines and soluble CD40 (sCD40) from 44 uncomplicated patients and 22 healthy controls (HCs) were measured using enzyme-linked immunosorbent assay (ELISA) and protein array technology.Results. Patients showed significantly higher levels of sCD40, IL-1a, IL-2, IL-4, IL-5, IL-10, granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage inflammatory protein (MIP)-1a, MIP-1b, regulated on activation normal T cell expressed and secreted (RANTES), matrix metalloproteinase (MMP)-9, and a trend to higher IL-6 than did HCs. RANTES and sCD40 discriminated significantly between diabetics and HCs. In patients with disease duration >6 months, cytokines were organized in two clusters mainly regulated by Th17 and Th1/Th2 cells respectively, while in those with disease duration ≤6 months a set of Th1-cytokines was separated apart from the second cluster. Monocyte chemotactic protein (MCP)-1 was revealed as the most discriminant factor between patients with disease duration of more than and less than 6 months.Conclusions. A parallel elevation of both inflammatory and anti-inflammatory cytokines was observed in patients compared with HCs. In T1DM patients with disease duration ≤6 months, Th1-cytokines were organized on a separate cluster, suggesting a possible role of Th1 cells in the progress of beta-cell destruction during the first period of the disease.

Identification of KCNJ15 as a Susceptibility Gene in Asian Patients with Type 2 Diabetes Mellitus

Objective: To investigate the clinical characteristics of symptoms and signs of ocular surface lesions in dry eye patients with type 2 diabetes mellitus (T2DM) of different disease durations. Methods: A cross-sectional study was conducted. Consecutive T2DM patients complicated with dry eye who visited the Special Clinic for Diabetic Ocular Surface Diseases at Qingdao Eye Hospital, Shandong First Medical University from January 2020 to December 2024 were enrolled as the T2DM group. Dry eye patients without T2DM from the Dry Eye Clinic and age-and gender-matched healthy volunteers were recruited as the non-T2DM group and control group, respectively. The T2DM group was further divided into four stages according to disease duration: Stage Ⅰ (duration≤5 years), Stage Ⅱ (5 years<duration≤10 years), Stage Ⅲ (10 years<duration≤20 years), and Stage Ⅳ (duration>20 years). Ocular symptoms were evaluated using the Ocular Surface Disease Index (OSDI) questionnaire. Ocular surface function was assessed by corneal sensitivity (CS), corneal fluorescein staining (CFS) score, Schirmer Ⅰ test (SⅠT), tear meniscus height (TMH), non-invasive first tear film break-up time (NIKf-BUT), non-invasive average tear film break-up time (NIKav-BUT), lipid layer thickness (LLT), and meibomian gland loss (MGL) grading score. Anterior segment optical coherence tomography was used to measure central corneal epithelial thickness (CCET) and central corneal thickness (CCT). In vivo confocal microscopy was performed to detect corneal nerve fiber parameters. Spearman's rank correlation analysis was used to evaluate the correlations between variables. Results: A total of 278 T2DM patients (556 eyes) were enrolled in the T2DM group, with a mean age of (60.25±8.71) years, a male-to-female ratio of 117∶161, and a mean disease duration of (11.09±8.02) years; among them, 84 cases were Stage Ⅰ, 81 cases Stage Ⅱ, 90 cases Stage Ⅲ, and 23 cases Stage Ⅳ. The non-T2DM group included 60 patients (120 eyes), with a mean age of (58.52±7.93) years and a male-to-female ratio of 23∶37. The control group comprised 53 healthy volunteers (106 eyes), with a mean age of (58.79±5.73) years and a male-to-female ratio of 21∶32. There were no statistically significant differences in age and gender distribution among the three groups (all P>0.05). Compared with the control group, the T2DM group showed significantly higher OSDI score (31.58%±20.56%), MGL grading score (3.59±0.95) and CFS score (1.84±3.01) (all P<0.05); while significantly lower SⅠT [(5.79±3.10) mm], NIKf-BUT [(5.81±3.14)s], NIKav-BUT [(9.31±4.85)s], LLT [(60.48±16.6) nm], CCET [(51.95±5.56) μm], CS [(56.9±8.30) mm], corneal endothelial cell density [(2 596±465.38)/mm²], and all corneal nerve parameters (including paracentral corneal nerve fiber density and length, etc.) (all P<0.05). Compared with the non-T2DM group, the T2DM group had significantly lower LLT, CCET, CS, OSDI score and all corneal nerve parameters, along with significantly higher MGL grading score and CFS score (all P<0.05); no statistically significant differences were found in SⅠT, NIKf-BUT and NIKav-BUT between the two groups (all P>0.05). In T2DM patients with disease duration≤5 years, compared with the control group, the OSDI score (23.98%±18.21%) and MGL grading score (3.35±0.91) were significantly increased, while SⅠT [(5.65±2.93) mm], LLT, NIKf-BUT and all corneal nerve parameters were significantly decreased (all P<0.05); meanwhile, compared with the non-T2DM group, this subgroup had significantly lower OSDI score and significantly higher MGL grading score (all P<0.05). When the disease duration exceeded 10 years, the OSDI score further increased to be comparable with that of the non-T2DM group, and NIKf-BUT [(5.44±2.92)s], CFS score (2.75±3.25), CCET [(51.36±4.17) μm], CS [(55.21±8.02) mm] showed statistically significant differences compared with both the control group and non-T2DM group (all P<0.05). Spearman's correlation analysis indicated that disease duration was significantly positively correlated with OSDI score, MGL grading score and CFS score, and significantly negatively correlated with NIKf-BUT, CCET, CS and all corneal nerve parameters (all P<0.05). Conclusions: Ocular surface lesion characteristics vary among dry eye patients with T2DM of different disease durations: decreased tear secretion, meibomian gland dysfunction and corneal nerve structural changes occur when the disease duration is ≤5 years, and corneal hypoesthesia and epithelial thinning develop when the duration exceeds 10 years, with lesions gradually worsening as the disease progresses. Compared with non-diabetic dry eye, significant differences exist in meibomian gland function, corneal nerves and epithelial barrier between the two groups.

Galectin-3 is a β-galactoside-binding lectin involved in inflammation and fibrosis and has been implicated in the pathogenesis of type 2 diabetes mellitus (T2DM) and its microvascular complications. This study aimed to evaluate the association between serum galectin-3 levels and microvascular complications in patients with T2DM and to determine whether this association persists after adjustment for relevant clinical factors. This comparative cross-sectional study included 64 participants recruited from Thumbay University Hospital, Ajman, UAE. Participants were divided into two groups: 32 patients with T2DM and established microvascular complications and 32 healthy controls. Serum galectin-3 levels were measured using enzyme-linked immunosorbent assay (ELISA). Clinical and biochemical parameters, including body mass index (BMI), HbA1c, fasting blood glucose, lipid profile, and microalbuminuria, were assessed. Multivariable linear regression analysis was performed with microalbuminuria as the dependent variable, adjusting for BMI, HbA1c, and age. Serum galectin-3 levels were significantly higher in patients with T2DM and microvascular complications compared with healthy controls (p<0.001). Galectin-3 showed a strong positive correlation with microalbuminuria (p=0.720, p<0.001) and a moderate positive correlation with HbA1c (p=0.599, p<0.001). However, in multivariable regression analysis, galectin-3 was not independently associated with microalbuminuria after adjustment for BMI, HbA1c, and age (p=0.197), whereas HbA1c remained a significant independent predictor (p<0.001). Although serum galectin-3 levels are elevated in patients with T2DM and microvascular complications and demonstrate strong univariate associations with microalbuminuria, galectin-3 does not remain independently associated after adjustment for BMI and glycemic control. These findings suggest that galectin-3 may reflect underlying metabolic and inflammatory burden rather than acting as an independent predictor of microvascular complications in T2DM.

Nailfold capillaroscopy (NC) is a non-invasive tool that can detect microvascular changes in the early stages of vascular disease. To assess capillary microarchitecture in children with type 1 diabetes mellitus (T1DM) and its relationship with clinical characteristics, laboratory findings, and glycemic control. We included children and adolescents with T1DM, aged 6-18 years, and diagnosed for at least one year and an equal number of age- and sex-matched healthy controls. For all patients with T1DM, data on diabetes duration were collected, and the average annual HbA1c value was calculated for the four measurements made at routine follow-up in the preceeding year. In patients using 24-hour continuous glucose monitoring (CGM) devices, glycemic data from the previous three months were analyzed. The capillaroscopic findings were evaluated by two different researchers with experience in the field of pediatric rheumatology. Capillaroscopic parameters were compared based on glycemic control (HbA1c ≥7.5% vs. <7.5%), disease duration (<5 vs. ≥5 years), time in range (TIR≥70% vs. <70%), and glucose variability (CV≤36% vs. >36%). The median age of the 55 patients with T1DM was 14.5 (11.3-17.2) years, with a median disease duration of 3.8 (2.3-6.7) years. Compared to controls, patients with T1DM had significantly lower capillary density and more frequent dilated, tortuous, cross-linked, and abnormal capillaries (p<0.001, p<0.001, p<0.001, p=0.01, and p=0.03, respectively). Capillary density was significantly lower in patients with poor glycemic control (p<0.001) and those with longer disease duration (p=0.02). A negative correlation was observed between capillary density and disease duration (r=-0.3, p=0.02). After adjusting for age, gender, body mass index, and diabetes duration, capillary density remained negatively correlated with average HbA1c (r=-0.4, p=0.004). Among CGM users (n=22), capillary density showed a positive correlation with TIR (r=0.5, p=0.04), even after adjustment for confounders. Children with T1DM exhibited significantly higher microvascular changes, mostly associated with poor glycemic control, compared to healthy controls. NC may be a useful technique for detecting early alterations in the capillary structures of children with T1DM, even in the absence of overt clinical microvascular complications.

Background and purposeGrowth factor receptor-bound protein 2(GRB2), a bridging protein. An animal study showed that downregulation of GRB2 inhibited the activation of PI3K/AKT/NF-kB pathway which improved lipid accumulation and inflammatory infiltration in rats with atherosclerosis (AS), resulting in an anti-AS effect. This was the first study to investigate blood GRB2 levels in type 2 diabetes mellitus(T2DM) patients with carotid atherosclerosis (CAS), exploring its relationship with various metabolic indicators, and further, examining whether GRB2 has an AS effect in patients with T2DM.MethodsA total of 203 participants were recruited in the study, including 69 T2DM patients without CAS (T2DM group), 67 T2DM patients with CAS (CAS group), and 67 in the age-sex-matched healthy subjects (Control group). Serum GRB2 levels were measured using enzyme-linked immunosorbent assay (ELISA) in 203 subjects who had received carotid ultrasonography. In addition, cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), fasting plasma glucose (FPG), glycosylated hemoglobin (HBA1c), fasting insulin (FINS), hypersensitive C-reactive protein (Hs-CRP), and Interleukin 6 (IL-6) were also tested. The correlation between serum GRB2 levels and other indexes was analyzed. Finally, we analyzed the risk factors affecting carotid intima-media thickness (CIMT) in T2DM patients.ResultsSerum GRB2 levels were increased in the T2DM group than in the control group, and further elevated in the CAS group (median 3.05 vs 4.40 vs 7.09 ng/ml, P<0.001). Spearman correlation analysis showed that GRB2 concentrations were negatively correlated with HDL-C, and positively associated with duration of diabetes, waist-to-hip ratio (WHR), TC, HBA1c, FPG, FINS, homeostasis model assessment-insulin resistance index (HOMA-IR), Hs-CRP, IL-6 and CIMT (P<0.01). Furthermore, serum GRB2 levels (P<0.001) remained independently related to CIMT after adjusting for the age, sex, duration of diabetes, and Body Mass Index (BMI) variables. Stepwise multiple linear regression analysis showed that IL-6, HDL-C, HBA1c, and CIMT are independent correlation factors of serum GRB2 (P<0.01). Univariate logistic regression suggested that disease duration, WHR, systolic blood pressure (SBP), TG, HDL-C, HBA1c, FPG, HOMA-IR, IL-6, Hs-CRP, and GRB2 independently associated with T2DM is combined with CAS(P<0.05). And multivariate logistic regression analysis showed that duration of diabetes, IL-6, and serum GRB2 levels were independent risk factors for T2DM combined with CAS (P<0.05), and serum GRB2 levels were a highly sensitive indicator of early AS (OR=1.405, 95% CI: 1.192-1.658 P<0.001). Moreover, the ROC curve AUC area of serum GRB2 expression levels was 0.80 (95%CI: 0.7291-0.8613, P < 0.001), with a sensitivity of 83.58% and specificity of 70.59%. The risk of CAS was substantially higher in patients with T2DM whose serum GRB2 concentration was >4.59 ng/ml.ConclusionsSerum GRB2 concentrations were significantly increased in T2DM combined with CAS, and serum GRB2 levels were linearly correlated with CIMT, suggesting that GRB2 may be involved in the occurrence and development of T2DM with CAS, which can be used as a predictor of whether T2DM is combined with CAS.