Indocyanine-Green-Loaded Liposomes for Photodynamic and Photothermal Therapies: Inducing Apoptosis and Ferroptosis in Cancer Cells with Implications beyond Oral Cancer.

Oral cancer represents a global health burden, necessitating novel therapeutic strategies. Photodynamic and photothermal therapies using indocyanine green (ICG) have shown promise due to their distinctive near-infrared (NIR) light absorption characteristics and FDA-approved safety profiles. This study develops ICG-loaded liposomes (Lipo-ICGs) to further explore their potential in oral cancer treatments. We synthesized and characterized the Lipo-ICGs, conducted in vitro cell culture experiments to assess cellular uptake and photodynamic/photothermal effects, and performed in vivo animal studies to evaluate their therapeutic efficacy. Quantitative cell apoptosis and gene expression variation were further characterized using flow cytometry and RNA sequencing, respectively. Lipo-ICGs demonstrated a uniform molecular weight distribution among particles. The in vitro studies showed a successful internalization of Lipo-ICGs into the cells and a significant photodynamic treatment effect. The in vivo studies confirmed the efficient delivery of Lipo-ICGs to tumor sites and successful tumor growth inhibition following photodynamic therapy. Moreover, light exposure induced a time-sensitive photothermal effect, facilitating the further release of ICG, and enhancing the treatment efficacy. RNA sequencing data showed significant changes in gene expression patterns upon Lipo-ICG treatment, suggesting the activation of apoptosis and ferroptosis pathways. The findings demonstrate the potential of Lipo-ICGs as a therapeutic tool for oral cancer management, potentially extending to other cancer types.

Rapid time-lapse 3D oxygen tension measurements within hydrogels using widefield frequency-domain fluorescence lifetime imaging microscopy (FD-FLIM) and image segmentation.

Understanding the influence of oxygen tension on cellular functions and behaviors is crucial for investigating various physiological and pathological conditions. In vitro cell culture models, particularly those based on hydrogel extracellular matrices, have been developed to study cellular responses in specific oxygen microenvironments. However, accurately characterizing oxygen tension variations with great spatiotemporal resolutions, especially in three dimensions, remains challenging. This paper presents an approach for rapid time-lapse 3D oxygen tension measurements in hydrogels using a widely available inverted fluorescence microscope. Oxygen-sensitive fluorescent microbeads and widefield frequency-domain fluorescence lifetime imaging microscopy (FD-FLIM) are utilized for oxygen tension estimation. To incorporate the third dimension, a motorized sample stage is implanted that enables automated image acquisition in the vertical direction. A machine learning algorithm based on K-means clustering is employed for microbead position identification. Using an upside-down microfluidic device, 3D oxygen gradients are generated within a hydrogel sample, and z-stack images are acquired using the FD-FLIM system. Analyses of the acquired images, involving microbead position identification, lifetime calculation, and oxygen tension conversion, are then performed offline. The results demonstrate the functionality of the developed approach for rapid time-lapse 3D oxygen tension measurements in hydrogels. Furthermore, the 3D oxygen tension adjacent to a tumor spheroid within a hydrogel during media exchange is characterized. The results further confirm that the 3D spatiotemporal oxygen tension profiles can be successfully measured quantitatively using the established setup and analysis process and that the approach may have great potential for investigating cellular activities within oxygen microenvironments.

Study Hypoxic Response under Cyclic Oxygen Gradients Generated in Microfluidic Devices Using Real-Time Fluorescence Imaging.

Oxygen plays important roles in regulating various biological activities under physiological and pathological conditions. However, the response of cells facing temporal variation in oxygen microenvironments has seldom been studied due to technical limitations. In this paper, an integrated approach to studying hypoxic response under cyclic oxygen gradients is developed. In the experiments, a cell culture system based on a microfluidic device is constructed to generate cyclic oxygen gradients with desired periods by alternately introducing gases with specific compositions into the microfluidic channels next to the cell culture channel separated by thin channel walls. Observation of the hypoxic responses is performed using real-time fluorescence imaging of dyes sensitive to extra- and intracellular oxygen tensions as well as intracellular calcium concentrations. Cellular hypoxic responses of human aortic smooth muscle cells (AoSMCs) and lung carcinoma epithelium (A549) cells, including intracellular oxygen and calcium levels, are measured. The results show that the two types of cells have different hypoxic responses to the applied cyclic oxygen gradients. With the capability of real-time cellular response monitoring under cyclic oxygen gradients, the developed approach provides a useful scheme to investigate hypoxic responses in vitro under microenvironments mimicking various in vivo physiological and pathological conditions.

Elevated serum cytokeratin-18 concentration in patients with type 2 diabetes mellitus and non-alcoholic fatty liver disease.

BACKGROUND: Serum cytokeratin-18 is believed to be a marker of hepatic cell damage. However, few studies have discussed about the serum cytokeratin-18 concentration in type 2 diabetes mellitus patients and investigated its association with non-alcoholic fatty liver disease as well as metabolic biomarkers. METHODS: Healthy participants and type 2 diabetes mellitus patients were enrolled. Physical and metabolic factors were recorded, and non-alcoholic fatty liver disease was screened by abdominal ultrasound and the fatty liver index. The cytokeratin-18 concentration was detected using two commercially available immunoassay kits (M30 and M65 ELISA kit, Previa AB, Sweden). RESULTS: Overall, 22.8% (29/127) and 35.9% (42/117) of the participants were diagnosed with non-alcoholic fatty liver disease in the non-diabetes mellitus group and type 2 diabetes mellitus group, respectively. In the non-diabetes mellitus group and type 2 diabetes mellitus group, our result showed that participants with non-alcoholic fatty liver disease had a higher serum cytokeratin-18 M30 and cytokeratin-18 M65 concentration as compared with participants without non-alcoholic fatty liver disease. Interestingly, as compared with healthy participants without non-alcoholic fatty liver disease, our result also demonstrated that type 2 diabetes mellitus patients without non-alcoholic fatty liver disease had a higher serum cytokeratin-18 M30 (108.4 ± 66.2 vs. 87.1 ± 34.6 U/L; P = 0.038) and cytokeratin-18 M65 concentration (285.4 ± 115.3 vs. 248.5 ± 111.3 U/L; P = 0.031). The independent relationship between type 2 diabetes mellitus and cytokeratin-18 was further strengthened by the significant positive association between fasting plasma glucose and serum cytokeratin-18 concentration via multivariate regression analyses (cytokeratin-18 M30: ß = 0.034, P = 0.029; cytokeratin-18 M65: ß = 0.044, P = 0.002). CONCLUSIONS: Independent of non-alcoholic fatty liver disease, our results suggested that the cytokeratin-18 concentration is closely associated with the hyperglycaemic milieu. The association between serum cytokeratin-18 and type 2 diabetes mellitus may be worthy of further investigation.

Escherichia coli Fiber Sensors Using Concentrated Dielectrophoretic Force with Optical Defocusing Method.

A sensitive tapered optical fiber tip combined with dielectrophoretic (DEP) trapping was used for rapid and label-free detection of bacteria in water. The angular spectrum of the optical field at the fiber tip was changed with the surrounding refractive index (RI). By measuring far-field intensity change at the defocus plane, the intensity sensitivity was up to 95 200%/RIU (RI unit), and the detection limit was 5.2 × 10-6 RIU at 0.5% intensity stability. By applying an AC voltage to a Ti/Al coated fiber tip and an indium-tin-oxide glass, the DEP force effectively trapped the Escherichia coli ( E. coli) near the fiber tip. Those bacteria can be directly measured from optical intensity change due to the increase of surrounding RI. By immobilizing the antibody on the Ti/Al fiber tip, the tests for specific K12 bacteria and nonspecific BL21 bacteria verified the specificity. The antibody-immobilized Ti/Al coated fiber tip with DEP trapping can detect bacteria at a concentration about 100 CFU/mL.

Renal function preservation with pioglitazone or with basal insulin as an add-on therapy for patients with type 2 diabetes mellitus.

AIMS: Clinical outcome may differ owing to the distinct pharmacological characteristics of insulin sensitizers and insulin. This study was performed to compare the metabolic and renal function changes with add-on pioglitazone treatment versus basal insulin in patients with type 2 diabetes mellitus (DM) in whom sulfonylurea and metformin regimens failed. METHODS: Patients who were consecutively managed in the diabetes comprehensive program with add-on pioglitazone or detemir/glargine treatment for at least 2 years following sulfonylurea and metformin treatment failure were included. RESULTS: A total of 1002 patients were enrolled (pioglitazone: 559, detemir: 264, glargine: 179). After propensity score matching, there were 105 patients with matchable baseline characteristics in each group. After a mean of 3.5 years of follow-up, the pioglitazone group showed a greater HbA1c reduction than the detemir group and the glargine group. Despite patients in all three groups exhibiting significant body weight gain, those in the pioglitazone group and the glargine group showed greater body weight increases than the patients in the detemir group (2.1, 1.6 and 0.8 kg, respectively, p < 0.05). Interestingly, Cox regression analysis indicated that patients under detemir or glargine treatment had a higher probability of CKD progression as compared with the pioglitazone group, with hazard ratios of 2.63 (95% CI 1.79-3.88) and 3.13 (95% CI 2.01-4.87), respectively. CONCLUSIONS: Our study first showed that treatment with both pioglitazone and basal insulin improved glycemic control, while only pioglitazone treatment was observed to be advantageous in terms of preserving renal function when used as an add-on therapy for patients with type 2 DM in whom sulfonylurea and metformin regimens failed.

Sensitive Detection of Small Particles in Fluids Using Optical Fiber Tip with Dielectrophoresis.

This work presents using a tapered fiber tip coated with thin metallic film to detect small particles in water with high sensitivity. When an AC voltage applied to the Ti/Al coated fiber tip and indium tin oxide (ITO) substrate, a gradient electric field at the fiber tip induced attractive/repulsive force to suspended small particles due to the frequency-dependent dielectrophoresis (DEP) effect. Such DEP force greatly enhanced the concentration of the small particles near the tip. The increase of the local concentration also increased the scattering of surface plasmon wave near the fiber tip. Combined both DEP effect and scattering optical near-field, we show the detection limit of the concentration for 1.36 µm polystyrene beads can be down to 1 particle/mL. The detection limit of the Escherichia coli (E. coli) bacteria was 20 CFU/mL. The fiber tip sensor takes advantages of ultrasmall volume, label-free and simple detection system.

Serum uric acid level as an indicator for CKD regression and progression in patients with type 2 diabetes mellitus-a 4.6-year cohort study.

BACKGROUND: To investigate the association of serum uric acid level with renal function change in patients with type 2 diabetes mellitus (T2DM). METHODS: T2DM patients who had been followed-up for at least 3 years were included. Participants were categorized into stable, progression, or regression groups according to their change in chronic kidney disease (CKD) stage. During the follow-up period, all numeric values of metabolic factors, including the uric acid level and the medication possession rate, were calculated in order to investigate their associations with CKD development. Multivariate Cox regression analyses were used to identify independent factors associated with change in CKD. RESULTS: A total of 2367 T2DM patients were enrolled in this study and followed-up for a mean of 4.6 years. The numbers of patients in the stable, progression and regression groups were 1133 (47.9%), 487 (20.6%), and 747 (31.5%), respectively. The progression group had the highest serum uric acid level (6.9 ± 1.8 mg/dL), and the regression group had the lowest uric acid level (5.4 ± 1.5 mg/dL). In addition, we found that the serum uric acid level was an independent factor associated with CKD progression when the value exceeded 6.3 mg/dL. A lower uric acid level could be beneficial for CKD improvement in T2DM patients with stage 3-5 CKD. CONCLUSIONS: Our data indicated that the serum uric acid level is associated with CKD regression and progression and suggested that a high normal serum uric acid level should be closely monitored in patients with T2DM. Copyright © 2015 John Wiley & Sons, Ltd.

Paradoxical negative HDL cholesterol response to atorvastatin and simvastatin treatment in Chinese type 2 diabetic patients.

OBJECTIVES: There is extensive but controversial evidence on the diverse effects of statins on the level of high-density lipoprotein cholesterol (HDL-C). Some of these effects may limit the benefits of statins in terms of cardiovascular risk reduction. To identify the conditions for beneficial effects, this study investigated the response to atorvastatin and simvastatin treatment in type 2 diabetic patients with elevated low-density lipoprotein cholesterol (LDL-C). METHODS: 2,872 subjects with type 2 diabetes from a disease management program were investigated. Patients with LDL-C ≥130 mg/dl or total cholesterol ≥200 mg/dl were put onto statin therapy by the National Health Insurance system in Taiwan. RESULTS: 1,080 patients who completed 1 year of statin treatment were analyzed. There were significant reductions in LDL-C in both the atorvastatin (37.1%) and simvastatin (34.3%) group after one year of treatment compared with baseline levels. Unexpectedly, the majority of diabetic patients who received atorvastatin or simvastatin did not show an increase in HDL-C levels. 59.8% of patients had a significant HDL-C reduction (ΔHDL-C ≤ -3%) after atorvastatin treatment. Multivariate logistic regression analysis showed that the following patients were at higher risk of HDL-C reduction after 12 months: (i) patients in whom statin therapy was initiated aged <65 years and who had a BMI ≥24 kg/m(2), (ii) male patients with a baseline HDL-C >40 mg/dl, and (iii) female patients with a baseline HDL-C >50 mg/dl. However, diabetic patients with severe atherogenic dyslipidemia (LDL-C ≥130, TG ≥204, and HDL-C ≤34 mg/dl) obtained more benefits in terms of HDL-C change after statin therapy. CONCLUSIONS: Diabetic patients, except those with severe atherogenic dyslipidemia, are prone to a decrease in serum HDL-C level after statin treatment, particularly after atorvastatin treatment.

Visfatin in overweight/obesity, type 2 diabetes mellitus, insulin resistance, metabolic syndrome and cardiovascular diseases: a meta-analysis and systemic review.

There are controversies regarding the association of visfatin with overweight/obesity, type 2 diabetes mellitus, insulin resistance (IR), metabolic syndrome and cardiovascular disease in published articles. A meta-analysis was performed to identify the significance of visfatin in these diseases. We searched for relevant articles in Pubmed, Scopus and SCIE. A total of 1035 articles were surveyed and 46 articles were identified, with 14 reports reporting more than one of our investigated diseases. A total of 13 (n = 644), 19 (n = 2405), 20 (n = 2249), 5 (n = 527) and 5 (n = 851) articles/(participants) were included in each meta-analysis regarding the association of visfatin and overweight/obesity, type 2 diabetes mellitus, insulin resistance, metabolic syndrome and cardiovascular diseases, respectively. Plasma visfatin concentrations were increased in participants diagnosed with overweight/obesity, type 2 diabetes mellitus, metabolic syndrome and cardiovascular diseases, with pooled log odds ratios of 1.164 [95% confidence interval (CI): 0.348 to 1.981, p = 0.005], 1.981 (95% CI: 1.377 to 2.584, p < 0.001), 1.094 (95% CI: 0.678 to 1.511, p < 0.001), and 2.902 (95% CI: 0.924 to 4.879, p < 0.005), respectively. The circulating visfatin level was positively associated with insulin resistance, with a Fisher's z of 0.089 (95% CI: 0.013 to 0.165, p = 0.022). No single study was found to affect the overall result of each analysis by sensitivity testing. No publication bias was found by the Egger test. Our study suggests that the use of visfatin may be promising for predicting obesity, diabetes status, insulin resistance, metabolic syndrome and cardiovascular disease.

The role of angiotensin-converting enzyme gene insertion/deletion polymorphism for blood pressure regulation in areca nut chewers.

Areca tannin has been suggested as having a blood pressure regulatory effect through its ability to inhibit the pressor response to both angiotensin I and II. As genetic and environmental factors determine the susceptibility and development of diseases and no report has been published concerning the genetic interaction of metabolic effects in areca nut/betel quid (BQ) chewers, it is proposed that the cardiovascular effects of chronic BQ usage can be affected by the polymorphism of the angiotensin converting enzyme (ACE) gene. This is a cross-sectional community-based study. A total of 189 BQ chewing subjects and 256 non-chewing controls were studied. BQ chewing status was determined by using a structured questionnaire, and insertion/deletion (I/D) polymorphisms were determined by a polymerase chain reaction. BQ chewers with the DD genotype had significantly lower blood pressure, pulse pressure, and prevalence of hypertension compared with those of chewers with II or ID genotypes. Multiple stepwise regression analysis confirmed that the ACE I/D genotype was independently associated with systolic blood pressure (SBP). BQ chewers with the DD genotype accounted for a significant age, sex, and waist-to-hip ratio adjusted decrease in SBP (-5.9 mm Hg, P=0.021). This finding suggests that BQ chewing may be related to blood pressure regulation, which supports the hypothesis that concomitant genetic susceptibility and environmental factors determine the level of blood pressure.

Association of peripheral total and differential leukocyte counts with metabolic syndrome and risk of ischemic cardiovascular diseases in patients with type 2 diabetes mellitus.

BACKGROUND: There is increasing evidence that leukocytes play a central role in obesity, glucose intolerance, type 2 diabetes mellitus (T2DM), and cardiovascular diseases, but the role of differential leukocytes in metabolic syndrome (MetS) and atherosclerosis is largely unknown. The aim of this study was to examine the relationship between the component features of MetS and peripheral leukocyte counts and to explore whether leukocyte counts are associated with clustering of MetS and macrovascular diseases in patients with T2DM. METHODS: 1872 subjects with T2DM who enrolled in a diabetes disease management program were studied. The definition of MetS was modified from that outlined by the criteria of NCEP-ATP III. Areas under the receiver-operating characteristic curve and odds ratios at various intervals of the WBC counts were computed. RESULTS: The peripheral total leukocyte, monocyte, and neutrophil cell counts are increased parallel to the clustering of components of MetS. When white cell counts were analyzed per quartile and as continuous variables after adjustment for age, sex, and other known risk factors with multiple regression analysis, peripheral total leukocyte, monocyte, neutrophils, and lymphocyte counts were independently and significantly associated with specific features of clustering of MetS and prevalence of ischemic cardiovascular diseases. Leukocyte counts, especially neutrophil/lymphocyte ratio, in addition with MetS is associated with the risk of ischemic cardiovascular diseases. CONCLUSION: Our results indicate that differential leukocyte counts are associated with MetS and that peripheral leukocytes may play a role in the pathogenesis of macrovascular complications in patients with T2DM.

Elevated plasma level of visfatin/pre-B cell colony-enhancing factor in patients with type 2 diabetes mellitus.

CONTEXT: Visfatin (also known as pre-B cell colony-enhancing factor or PBEF) is a cytokine that is highly expressed in visceral fat and whose blood levels correlate with obesity. Originally isolated as a secreted factor that promotes the growth of B cell precursors and recently found to act as an insulin analog on the insulin receptor, its pathophysiological role in humans remains largely unknown. OBJECTIVES: In this study we investigated whether plasma visfatin level is altered in patients with type 2 diabetes mellitus (T2DM). DESIGN AND PATIENTS: Plasma visfatin as well as adiponectin and resistin concentrations were measured through ELISA in type 2 diabetic and nondiabetic subjects. RESULTS: A total of 61 patients with T2DM and 59 sex- and age-matched nondiabetic subjects were studied. Plasma visfatin was found to be elevated in patients with T2DM (31.9 +/- 31.7 vs. 15.8 +/- 16.7 ng/ml, P = 0.002). In contrast, adiponectin was decreased (4.3 +/- 2.5 vs. 30.8 +/- 10.3 microg/ml, P < 0.001), whereas plasma resistin level did not differ between the groups. Increasing concentrations of visfatin were independently and significantly associated with T2DM. Multiple logistic regression analysis revealed visfatin as an independent association factor for T2DM, even after full adjustment of known biomarkers. The association between adiponectin and T2DM was no longer significant after adjustments for body mass index or waist to hip ratio. In a multiple linear regression analysis, only waist to hip ratio was independently associated with plasma visfatin level. CONCLUSION: Our results indicate that visfatin may play a role in the pathogenesis of T2DM.

ENPP1 K121Q polymorphism is not related to type 2 diabetes mellitus, features of metabolic syndrome, and diabetic cardiovascular complications in a Chinese population.

BACKGROUND: Ectoenzyme nucleotide pyrophosphate phosphodiesterase 1 (ENPP1) is known to influence insulin sensitivity by inhibiting insulin receptor signaling. A DNA polymorphism in the ENPP1 gene at exon 4 (K121Q) was demonstrated to be associated with insulin resistance, type 2 diabetes mellitus (T2DM), and a risk of early myocardial infarction, albeit with controversy. Our aim was to investigate any association of ENPP1 K121Q alleles with T2DM, features of the metabolic syndrome, and diabetic cardiovascular complications in a Chinese population of Han origin. METHODS: The ENPP1 K121Q polymorphism was determined by a restriction fragment-length polymorphism-polymerase chain reaction in 1,862 patients with T2DM and 844 non-diabetic subjects. RESULTS: The genotype distributions or Q-allele frequency were not statistically different between the diabetic and non-diabetic groups. The anthropometric parameters, systolic and diastolic blood pressures, lipid profiles, and serum creatinine levels of subjects with different ENPP1 K121Q polymorphisms were not statistically different in the two groups or even in the pooled data. When sub-group analyses of diabetic subjects were stratified according to BMI levels (greater or less than 27), gender, age of diabetes onset (older or younger than 60 years), and the presence or absence of a diabetic family history; this polymorphism was still not associated with T2DM. Nor was the ENPP1 K121Q polymorphism associated with the prevalence of coronary artery disease and ischemic cerebrovascular disease in patients with T2DM. CONCLUSION: The ENPP1 K121Q polymorphism is not related to T2DM, features of the metabolic syndrome, or diabetic macrovascular complications in a Chinese population.

Peripheral total and differential leukocyte count in diabetic nephropathy: the relationship of plasma leptin to leukocytosis.

OBJECTIVE: Because of increasing evidence that white blood cells (WBCs) play a role in the development and progression of diabetes complications, this study aimed to investigate the relation of circulating total and differential leukocyte counts to nephropathy in patients with type 2 diabetes. Plasma leptin levels were also measured to investigate their role in peripheral leukocytosis. RESEARCH DESIGN AND METHODS: For this study, 1,480 subjects with type 2 diabetes who were enrolled in a disease management program were stratified according to urinary microalbumin and serum creatinine measurements. The total and differential leukocyte profiles of peripheral blood were measured and plasma leptin was examined by enzyme-linked immunosorbent assay. Demographic and potential metabolic confounding factors were analyzed with linear and logistic regression to calculate the effects of leukocyte count on diabetic nephropathy. RESULTS: The peripheral total WBC, monocyte, and neutrophil counts increased in parallel with the advancement of diabetic nephropathy. In contrast, the lymphocyte count decreased. When WBC counts were analyzed per quartile and as continuous variables after adjusting for age, sex, and other known risk factors with multiple regression analysis, peripheral total WBC, monocyte, neutrophil, and lymphocyte counts were independently and significantly associated with diabetic nephropathy. Plasma leptin levels increased in patients with nephropathy and correlated significantly with total WBC count (r = 0.194, P = 0.014). CONCLUSIONS: Because leukocytes are activated and secrete cytokines in the diabetic state and leptin stimulates leukocyte proliferation and differentiation, our results suggest that circulating leukocytes contribute to the development and progression of nephropathy, partially through the effects of leptin, in patients with type 2 diabetes.

The association of silent coronary artery disease and metabolic syndrome in Chinese with type 2 diabetes mellitus.

OBJECTIVES: Cardiovascular diseases account for approximately 75% of the deaths that occur in patients with diabetes. Because the clinical signs of coronary artery disease (CAD) in diabetic patients are hard to detect and routine screening is costly, it would be of great benefit to try to either prevent CAD from occurring or to detect it early and provide optimal care. Therefore, we analyzed the risk factors that might predict CAD in type 2 diabetes mellitus (T2DM) patients with no classical cardiac ischemic symptoms. METHODS: Using a resting 12-lead ECG, exercise treadmill test, or thallium myocardial scintigraphy with exercise testing and dipyridamole injection, we screened diabetic patients already enrolled in a disease management program for possible CAD. We used diagnostic coronary angiography to confirm its presence. The definition and criteria of metabolic syndrome we used were modified from those outlined by the WHO classification and criteria of NCEP-ATP III. RESULTS: A total of 850 T2DM patients without clinical and electrocardiographic evidence of CAD were studied. Three hundred and sixty-eight asymptomatic patients with normal resting ECG were examined by exercise ECG test or thallium scintigraphy examination. Sixty patients considered to have a strong positive test or significant thallium myocardial ischemia received a diagnostic coronary angiography. Fifty-one were found to have significant coronary artery stenosis; 9 showed no significant ischemic lesion. While gender, patients' age, known diabetes duration, serum uric acid level, smoking status, and the presence of WHO-metabolic syndrome defined hypertension and nephropathy were associated with silent CAD, logistic regression analysis found that the only predictor of silent CAD was the presence of nephropathy. The components of NCEP-ATP III-metabolic syndrome were not found to be associated with silent CAD. CONCLUSIONS: A considerable proportion of T2DM patients have silent CAD. A diabetic patient with incipient or overt nephropathy should be examined for the presence of CAD. The definition of metabolic syndrome may be modified for early detection of CAD in patients with T2DM.

A case of acquired generalized lipodystrophy with cerebellar degeneration and type 2 diabetes mellitus.

Acquired generalized lipodystrophy (AGL) is a rare disorder of adipose tissue characterized by loss of fat from large regions of the body, occurring after birth. Its etiology remains unknown. Most AGL patients have had fasting and/or postprandial hyperinsulinemia, diabetes mellitus, hypertriglyceridemia, and fatty liver. We describe the case of a 30-year-old woman with a progressively unsteady gait and a generalized loss of body fat beginning at the age of 7. Cerebellar degeneration was revealed by imaging study, and the patient was eventually bedridden at the age of 15, due to progressive ataxia. She developed diabetes at the age of 25 without the presence of any evidence of ketoacidosis. The glutamic acid decarboxylase antibody was negative, C-peptide level 3.6 ng/ml, HbA1c 13%, triglyceride 412 mg/dl, total cholesterol 196 mg/dl, high-density lipoprotein-cholesterol 28 mg/dl, adiponectin 0.76 microg/ml, and resistin was 22.8 ng/ml at the initial state of diabetes. AGL accompanied by type 2 diabetes and cerebellar degeneration was diagnosed on the basis of the clinical features and metabolic derangements.

Association of a 27-bp repeat polymorphism in intron 4 of endothelial constitutive nitric oxide synthase gene with serum uric acid levels in Chinese subjects with type 2 diabetes.

Nitric oxide (NO) was found to modulate uric acid production through its influence on xanthine oxidase activity, and a close circadian relationship of serum uric acid (SUA) and NO was reported. Studies also revealed that serum NO activity could be determined by endothelial constitutive nitric oxide synthase gene (ecNOS) polymorphism. This study was designed to investigate whether SUA could be influenced by a 27-bp repeat polymorphism in intron 4 of ecNOS gene. A total of 398 nondiabetic subjects and 800 patients with type 2 diabetes were studied. The ecNOS gene intron 4 polymorphism was determined by polymerase chain reaction (PCR). The mean SUA level of patients having type 2 diabetes was significantly lower than that of control subjects (6.1 +/- 1.8 mg/dL v 6.6 +/- 1.8 mg/dL, P<.001); and the mean SUA level of diabetic patients with ecNOS ab/aa genotypes was lower than that of patients with bb genotype (5.7 +/- 1.6 mg/dL v 6.2 +/- 1.8 mg/dL, P=.008). When subgrouped by gender, the SUA of female diabetic subjects was found to be significantly associated with ecNOS genotype. Using Pearson's correlation analysis and multiple linear regression analysis, ecNOS genotype was noticed to be an independent factor in contributing to SUA variability in female diabetic patients. Our results suggest that SUA levels may be associated with NO activity and can be genetically predetermined.

Association of resistin gene 3'-untranslated region +62G-->A polymorphism with type 2 diabetes and hypertension in a Chinese population.

Resistin, a recently discovered polypeptide, antagonizes insulin action and may play a part in the pathogenesis of insulin resistance. This study investigates whether resistin gene polymorphism can be associated with type 2 diabetes. We studied 1102 Chinese type 2 diabetes patients and 743 subjects without diabetes. The resistin 3'-untranslated region (UTR) +62G-->A polymorphism was determined by PCR. Type 2 diabetes subjects had a lower frequency of resistin gene 3'UTR +62A allele (GG:GA/AA, 83.5%:16.5%) than the controls (GG:GA/AA, 75.1%:24.9%; odds ratio, 1.524; 95% confidence interval, 1.268-1.831; P < 0.001). Unexpectedly, diabetic patients with the GG genotype had a higher prevalence of hypertension (GG:GA/AA, 49.8%:36.2%; odds ratio, 1.375; 95% confidence interval, 1.116-1.693; P = 0.001). Logistic regression analysis confirmed that the resistin gene 3'UTR +62G-->A polymorphism acts as an independent contributing factor to type 2 diabetes and hypertension. The mean systolic and diastolic blood pressure levels in diabetic subjects with the GG genotype (144 +/- 21/87 +/- 13 mm Hg) were significantly higher than those in subjects with GA/AA variants (139 +/- 21/84 +/- 14 mm Hg; P = 0.004 and P = 0.002, respectively). Multiple linear regression analysis showed resistin gene polymorphism to be an independent factor associated with systolic and diastolic blood pressures in type 2 diabetes patients. These findings suggest that resistin may play a role in the pathogenesis of type 2 diabetes and insulin resistance-related hypertension.