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Aim: The aim of this study was to investigate the relationship between CXCL-6 levels in wound exudates and healing of diabetic foot ulcers (DFU). Materials & methods: One hundred patients with neuropathic DFU were recruited. Wound exudate CXCL-6 levels were measured by enzyme-linked immunosorbent assay. Patients were followed for 24 weeks and divided into rapidly healing and nonhealing groups. Results: Compared with the NH group, the mean CXCL-6 levels in the wound exudates of the rapidly healing group were significantly higher. After adjusting for traditional risk factors, wound exudate CXCL-6 levels were still significantly associated with wound healing. Conclusion: CXCL6 is an independent predictor of wound healing in DFU patients and may be a potentially novel therapeutic target for the treatment of DFU.

Critical Limb Ischemia (CLI) is common but uncommonly diagnosed. Improved recognition and early diagnostic markers for CLI are needed. Therefore, the aim of our study was to identify plasma biomarkers of CLI in patients with type 2 diabetes mellitus (T2DM). In this study, antibody-coated glass slide arrays were used to determine the plasma levels of 274 human cytokines in four matched cases of diabetes with and without CLI. Potential biomarkers were confirmed in an independent cohort by ELISA. After adjusting for confounding risk factors, only plasma level of Siglec-5 remained significantly associated with an increased odds ratio (OR) for diabetes with CLI by binary logistic regression analysis. Receiver operating characteristic (ROC) curve analysis revealed the optimal cut-off points for Siglec-5 was 153.1 ng/ml. After entering Siglec-5, the AUC was 0.99, which was higher than that of confounding risk factors only (AUC = 0.97, P

The purpose of this study was to investigate the effect of glycaemic control on subclinical hypothyroidism (SCH) in Chinese type 2 diabetic patients.The study included 476 diabetic patients with SCH admitted for treatment of type 2 diabetes. The controls were selected euthyroid patients with similar characteristics regarding [age, body mass index (BMI), gender, previous hypertension or duration of diabetes, and smoke]. Total cholesterol (TC), LDL cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglyceride (TG), the homeostatic model assessment of insulin resistance ratio (HOMA-IR), fasting plasma glucose, hemoglobin AThe LDL-C level and HOMA-IR value were significantly higher in the SCH group. Glycaemic control reduced HOMA-IR, HbAOur study shows that glycaemic control may bring some benefits to type 2 diabetic patients with SCH.

Objectives. Chronic foot ulceration is a severe complication of diabetes, driving morbidity and mortality. The aim of our study was to identify novel biomarkers of impaired wound healing in diabetic foot ulcers. Methods. 109 patients with neuropathic diabetic foot ulcers and 30 burn victims otherwise healthy participated. Antibody-coated glass slide arrays were used to determine the levels of 80 human cytokines in pooled plasma or pooled wound exudate of diabetic foot ulcers with rapidly healing (RH, n=12) and matched nonhealing (NH, n=12) patients. Potential biomarkers were confirmed in an independent cohort by enzyme-linked immunosorbent assay (ELISA). Results. Protein array profiling identified 27 proteins or 15 proteins significantly altered in protein profiling of pooled plasma or pooled wound exudate of 12 RH patients compared with 12 matched NH patients, respectively. In an independent cohort, quantitative ELISA validation confirmed a decrease in MCP-2 and ENA-78 levels in NH patients versus RH patients or burn victims. After adjusting for the traditional risk factors (sex, age, body mass index, fasting plasma glucose, ulcer area, HbA1C, diabetes duration, hyperlipidemia, and antibiotic therapy), only wound exudate level of ENA-78 remained having a significant association with an increased odds ratio (OR) for wound healing by binary logistic regression analysis (P<0.05). Conclusion. Decreased wound exudate ENA-78 was independently associated with wound healing of patients with diabetic foot. Exudate ENA-78 level is implicated as a novel predictor of wound healing in patients with diabetic foot ulcers.

The study aimed to determine the relationships between the basal metabolic rate (BMR) and body composition of overweight and obese Chinese adults with type 2 diabetes mellitus (DM). This cross-sectional clinical study enrolled 193 Chinese adults with type 2 DM who were overweight (24 kg/m(2)=BMI≤28 kg/m(2), n=99), or obese (BMI ≥28 kg/m(2), n=94). Ninety-seven adults with normal BMIs, including 50 DM patients and 47 healthy adults, were recruited as a control group. BMR was measured by indirect calorimetry; predicted BMR was calculated according to the Schofield equation; and the relationships between BMR, body composition, and biochemical results were determined by the Pearson correlation. The results showed that obese DM patients had significantly higher BMRs than both overweight patients (P

The aim of this study was to investigate whether osteopontin(OPN)variants are associated with susceptibility to ankylosing spondylitis (AS) in a Chinese population. Polymorphisms at the 9175th position in exon 7 ofOPNand rs17524488 were genotyped using direct sequencing in 186 unrelated AS patients and 188 ethnically matched healthy controls. Serum concentration of OPN was measured by enzyme-linked immunosorbent assay (ELISA) in all participants. AS patients displayed significantly higher OPN serum levels than the controls (P<001). A heterozygous, novel 9175 T>A in exon 7 of theOPNgene was found in this study. In healthy controls, subjects carrying the rs17524488 G/G genotype of theOPNdisplay significantly higher OPN serum levels than the GG/GG genotype (P<0.05). Plasma OPN level is implicated as an early diagnostic marker of AS. The novel 9175th- (exon 7) position polymorphism ofOPNand rs17524488 were related to susceptibility to AS in a Chinese population, the rs17524488 G/G genotype may be involved in the pathogenesis of AS, and the precise molecular mechanism underlying the influence ofOPNpolymorphisms on the development of AS remains to be determined in the further prospective studies.

Critical limb ischemia (CLI) is the most severe manifestation of peripheral artery disease, which is common but rarely diagnosed. Noninvasive biomarkers are urgently required to assist in the diagnosis of CLI. Accumulating evidence indicates that miRNAs play an important role in the development of various diseases. In this study, microarray profiling revealed 11 miRNAs with significantly altered expression in four T2DM patients with CLI compared with that in four sex- and age-matched T2DM patients without CLI. In independent cohorts, qRT-PCR validation confirmed the increased miRNA-4739 level in patients with CLI versus patients without CLI. miRNA-4739 levels increased with FPG and HbA1c (all P < 0.05). After adjusting for the risk factors, miRNA-4739 levels were found to be associated with an increased odds ratio (OR) of T2DM with CLI (OR =12.818, 95% confidence intervals (CI) 1.148 to 143.143, P = 0.038). ROC curve analysis revealed that the area under the curve (AUC) of miR-4739+confounding risk factors was 0.94 (95% CI 0.891 to 0.998, P < 0.001), which was higher than that of confounding risk factors (AUC 0.94 vs. 0.91, 95% CI -0.122 to 0.060, P > 0.05) and of miR-4739 (AUC 0.94 vs. 0.69, 95% CI -0.399 to -0.101, P < 0.001), respectively. We conclude that elevated plasma miRNA-4739 levels are independently associated with CLI in T2DM patients. miRNA-4739 is implicated as a novel diagnostic marker and a potential therapeutic target for CLI in diabetes.

We recently reported that transplantation of autologous bone marrow mononuclear cells (BM-MNCs) may be an effective and promising therapy to treat refractory diabetic sensorimotor polyneuropathy (DSPN) in patients with type 2 diabetes mellitus (T2DM). This study was designed to investigate the potential mechanisms of BM-MNCs therapy, which recruited 60 patients with DSPN, 30 T2DM patients without complications, and 30 healthy control participants. All clinical parameters, the levels of inflammatory markers, and growth factors in the three groups were compared. Patients in DSPN group had higher level of tumor necrosis factor-α (TNF-α) (DSPN vs control, 412.90 ± 64.58 vs 374.81 ± 63.18 pg/mL, P < 0.01) and lower level of vascular endothelial growth factor (VEGF) (DSPN vs control, 140.93 ± 24.78 vs 157.39 ± 25.11 pg/mL, P < 0.01) than those in control group. DSPN group had the highest level of soluble intercellular adhesion molecule-1 (sICAM-1) among three groups (DSPN and DM vs control, 1477.56 ± 228.00 and 1342.17 ± 237.54 vs 1308.00 ± 200.94 ng/mL, P < 0.05). The level of nerve growth factor in the DSPN group was slightly lower than that in the DM group (DSPN vs DM, 3509.11 ± 438.39 vs 3734.87 ± 647.50 pg/mL, P < 0.05). All patients with DSPN received one intramuscular injection of BM-MNCs and clinical follow-ups after the therapy for 2 days, 1, 4, 12, 24, and 48 weeks. Neuropathic symptoms of foot pain, numbness, and weakness were significantly improved within 4 weeks after BM-MNCs injection. Patients with DSPN were divided into the responder ( n = 35) and nonresponder groups ( n = 19) based on the improvement of nerve conduction velocity at 12 weeks post-transplantation. Compared with nonresponders, responders were younger (57.3 ± 5.2 vs 62.0 ± 4.8, P < 0.01), had a shorter history of diabetes (7.1 ± 2.7 vs 11.2 ± 5.4 years, P < 0.01), and had higher numbers of mobilized CD34+cells (17.61 ± 2.64 vs 14.79 ± 1.62 ×105/L, P < 0.01) and BM-MNCs (12.05 ± 2.16 vs 9.84 ± 1.53 ×108/L, P < 0.01). The levels of TNF-α and sICAM-1 decreased just after BM-MNCs injection in both groups and slowly reverted to baseline levels. The duration of the downtrend of TNF-α and sICAM-1 in the responder group lasted longer than that in the nonresponder group. Serum level of VEGF in the responder group increased immediately after BM-MNC therapy and reached the highest point after the injection for 12 weeks. On the other hand, VEGF levels in the nonresponder group only increased slightly. Binary logistic regression was performed to evaluate the corresponding prognostic factors for BM-MNCs treatment. The number of applied CD34+cells and the duration of diabetes were the independent predictors of responding to BM-MNCs therapy. No adverse event associated with the treatment was observed during follow-up observations. These results indicated that BM-MNCs transplantation is an effective and promising therapeutic strategy to treat refractory DSPN. The immune regulation and paracrine function of BM-MNCs may contribute to the improvement of DSPN.Trial registration:Chinese Clinical Trial Registry, ChiCTR-TRC-12002570. URL: http://www.chictr.org.cn/showproj.aspx?proj=6981

Objective To investigate whether height is associated with peripheral arterial disease (PAD) in patients with type 2 diabetes. Research design and methods This was an observational study performed in 4,528 Chinese patients with type 2 diabetes. Anthropometric measures and the ankle-brachial index (ABI) were performed on each subject. PAD was defined as those patients with a history of revascularization or amputation due to ischemia, or an ABI

The role of hydrogen sulfide (H2S) in myocardial infarction (MI) has not been previously studied. We therefore investigated the effect of H2S in a rat model of MI in vivo. Animals were randomly divided into three groups ( n = 80) and received either vehicle, 14 μmol/kg of sodium hydrosulfide (NaHS), or 50 mg/kg propargylglycine (PAG) everyday for 1 wk before surgery, and the treatment was continued for a further 2 days after MI when the animals were killed. The mortality was 35% in vehicle-treated, 40% in PAG-treated, and 27.5% in NaHS-treated ( P < 0.05 vs. vehicle) groups. Infarct size was 52.9 ± 3.5% in vehicle-treated, 62.9 ± 7.6% in PAG-treated, and 43.4 ± 2.8% in NaHS-treated ( P < 0.05 vs. vehicle) groups. Plasma H2S concentration was significantly increased after MI (59.2 ± 7.16 μM) compared with the baseline concentration (i.e., 38.2 ± 2.07 μM before MI; P < 0.05). Elevated plasma H2S after MI was abolished by treatment of animals with PAG (39.2 ± 5.02 μM). We further showed for the first time cystathionine-gamma-lyase protein localization in the myocardium of the infarct area by using immunohistochemical staining. In the hypoxic vascular smooth muscle cells, we found that cell death was increased under the stimuli of hypoxia but that the increased cell death was attenuated by the pretreatment of NaHS (71 ± 1.2% cell viability in hypoxic vehicle vs. 95 ± 2.3% in nonhypoxic control; P < 0.05). In conclusion, endogenous H2S was cardioprotective in the rat model of MI. PAG reduced endogenous H2S production after MI by inhibiting cystathionine-gamma-lyase. The results suggest that H2S might provide a novel approach to the treatment of MI.

The purpose of the current study is to evaluate the cardioprotective effects of purified Salvia miltiorrhiza extract (PSME) on myocardial ischemia/reperfusion injury in isolated rat hearts. Hearts were excised and perfused at constant flow (7 – 9 ml min−1) via the aorta. Non-recirculating perfusion with Krebs-Henseleit (KH) solution was maintained at 37°C and continuously gassed with 95% O2 and 5% CO2. KH solution with or without PSME (100 mg per liter solution) was used after 30-min zero-flow ischemia for the PSME and control group, respectively. Left ventricular (LV) developed pressure; its derivatives, diastolic pressure, and so on were continuously recorded via a pressure transducer attached to a polyvinylchloride balloon that was placed in the left ventricle through an incision in the left atrium. PSME treated hearts showed significant postischemic contractile function recovery (developed pressure recovered to 44.2 ± 4.9% versus 17.1 ± 5.7%, P

In this study, the cardioprotective effects of nitric oxide (NO)-aspirin, the nitroderivative of aspirin, were compared with those of aspirin in an anesthetized rat model of myocardial ischemia-reperfusion. Rats were given aspirin or NO-aspirin orally for 7 consecutive days preceding 25 min of myocardial ischemia followed by 48 h of reperfusion (MI/R). Treatment groups included vehicle (Tween 80), aspirin (30 mg·kg−1·day−1), and NO-aspirin (56 mg·kg−1·day−1). NO-aspirin, compared with aspirin, displayed remarkable cardioprotection in rats subjected to MI/R as determined by the mortality rate and infarct size. Mortality rates for vehicle ( n = 23), aspirin ( n = 22), and NO-aspirin groups ( n = 22) were 34.8, 27.3, and 18.2%, respectively. Infarct size of the vehicle group was 44.5 ± 2.7% of the left ventricle (LV). In contrast, infarct size of the LV decreased in the aspirin- and NO-aspirin-pretreated groups, 36.7 ± 1.8 and 22.9 ± 4.3%, respectively (both P < 0.05 compared with vehicle group; P < 0.05, NO-aspirin vs. aspirin ). Moreover, NO-aspirin also improved ischemiareperfusion-induced myocardial contractile dysfunction on postischemic LV developed pressure. In addition, NO-aspirin downregulated inducible NO synthase (iNOS; 0.37-fold, P < 0.01) and cyclooxygenase-2 (COX-2; 0.61-fold, P < 0.05) gene expression compared with the vehicle group after 48 h of reperfusion. Treatment with NG-nitro-l-arginine methyl ester (l-NAME; 20 mg/kg), a nonselective NOS inhibitor, aggravated myocardial damage in terms of mortality and infarct size but attenuated effects when coadministered with NO-aspirin. l-NAME administration did not alter the increase in iNOS and COX-2 expression but did reverse the NO-aspirin-induced inhibition of expression of the two genes. The beneficial effects of NO-aspirin appeared to be derived largely from the NO moiety, which attenuated myocardial injury to limit infarct size and better recovery of LV function following ischemia and reperfusion.

We investigated the apoptotic development and apoptotic-related gene expression after myocardial infarction (MI) at different time points in the current study. Bax gene expression was increased at 12 h after MI and peaked at 24 h. Fas gene started to over-express at 12 h after MI as well but it reached maximum at 72 h. In the MI groups, strongest staining of apoptosis was detected in rats 3 days post operation. Our results demonstrate that apoptotic development after MI is time dependent in the ischemic area and there could be some linkage with the over expression of angiotensin II receptors post MI.

Introduction: Angiotensin II (Ang II) exerts its biological effects by binding to Ang II receptors. Two major subtypes of Ang II receptors, AT1R and AT2R, have been recognized by ligand binding studies. AT1R mediates most of the Ang II effects in the cardiovascular system. Usually, the AT2R only occurs in the growing animal but disappears in adults. The function of the AT2R so far has not been elucidated clearly. Recent studies have suggested that the AT2R is coupled to an antigrowth process that counteracts the growth-promoting program initiated by AT1R activation. We previously reported that gene expression of AT1R and AT2R were regulated differently in a time-dependent manner after myocardial infarction (MI). However, it is still not clear how the transcriptional levels of AT1R and AT2R are regulated as well as their localization after MI. Therefore, the present study is to investigate Ang II receptor expression in rat heart after using specific Ang II receptor antibodies. Materials and Methods: MI was surgically induced in Wistar rats weighing around 200g by permanent ligation of the left anterior descending coronary artery. After 3, 7, 14 and 21 days of surgery, these rats were sacrificed and rat heart was collected. H&E staining was used to evaluate the morphological changes and immunohistochemistry was used to investigate the receptor expression and localization of AT1R and AT2R. Results: H&E Staining: Degraded myocardium and fibrosis were seen in the infarcted area of left ventricle. Immunohistochemistry: Degraded myocytes of infarcted area in the left ventricle demonstrated the strongest AT1R staining. Positive immunostaining for AT1R was also detected in the endocardium, myocardium, and epicardium. Occasionally, positivity was also observed in the coronary vessels, mainly coronary vascular endothelium, but not smooth muscle cells. In the left ventricular wall, AT1R expression increased mainly on day 3 but decreased thereafter as a result of degrading myocytes. On the other hand, in the non-infarcted area, AT1R expression was detected but weaker than the infarcted area. On the contrary, AT2R positive immunostaining was less intense than AT1R. It was mainly expressed in the degraded myocardium of infarcted area. Some positive staining was also in the endocardium and epicardium though very weakly. Occasionally, the endothelium of large coronary arteries was positively stained but not in the small vascular endothelium or vascular smooth muscle. The expression of AT2R was increased over time. The strongest expression of AT2R was found on day 21, post-surgery. There was no AT2 expression detected in the non-infarct area. Conclusion: (1) Ang II expression was changed in the heart after MI. (2) Both AT1R and AT2R expressions are increased after remodeling. (3) Up and down-regulation of AT1R and AT2R respectively in the different time points suggested that there is some interaction between each other but its mechanism needs to be further elucidated.

Myocardial infarction (MI) remains the leading cause of death from cardiovascular diseases over the past decades. Apoptosis is ‘programmed’ cell death which leads to clearance of ‘unwanted’ cells without disruption of tissue structure or function. Recently, we reported that angiotensin receptor subtype AT1 and AT2 mRNA levels were time dependently regulated after MI. AT1 and AT2 receptor mRNA levels markedly increased at 30 min and peaked 24 h post-MI. The time-dependent increase of AT1 and AT2 receptor mRNA is associated with the early remodeling process of the non-infarcted myocardium post MI. There is a further question raised whether the up-regulation of AT2 receptor is linked to apoptosis. Therefore, we investigated the apoptotic development and apoptotic related gene expression after MI at different time points in the current study. MI was induced in Wistar rats by ligation of the left anterior descending coronary artery. Bax gene expression was found to be increased at 12 h after MI and peaked at 24 h (4.3-fold). It declined at 72 h after MI. Fas gene started to over-express at 12 h after MI as well but it reached maximum at 72 h (4.7-fold). Protein levels of Bax and Fas expression were detected in the necrotic area and area at risk (surrounding area) at different time points after MI. Apoptosis was detected in the infarcted areas. No apoptosis was detected in the sham operated rats. In the MI groups, strongest staining of apoptosis was detected in rats 3 days post operation. Weak staining was found 1 day, 7 day post MI. Very fewer apoptotic cells were detected in the rats 2 weeks after MI. Our results demonstrate that apoptotic development after MI is time dependent in the ischemic area and there could be some linkage with the over expression of AT2 receptor post MI.