Sirtuin-6 knockout causes exacerbated stalled healing of diabetic ulcers in mice.

BACKGROUND: Diabetic ulcers (DUs) are characterized by chronic inflammation and delayed re-epithelialization, with a high incidence and weighty economic burden. The primary therapeutic strategies for refractory wounds include surgery, non-invasive wound therapy, and drugs, while the optimum regimen remains controversial. Sirtuin-6 (SIRT6) is a histone deacetylase and a key epigenetic factor that exerts anti-inflammatory and pro-proliferatory effects in wound healing. However, the exact function of SIRT6 in DUs remains unclear. METHODS: We generated tamoxifen-inducible SIRT6 knockout mice by crossing SIRT6flox/flox homozygous mice with UBC-creERT2+ transgenic mice. Systemic SIRT6 null mice, under either normal or diabetic conditions, were utilized to assess the effects of SIRT6 in DUs treatment. Gene and protein expressions of SIRT6 and inflammatory cytokines were measured by Western blotting and RT-qPCR. Histopathological examination confirmed the altered re-epithelialization (PCNA), inflammation (NF-κB p50 and F4/80), and angiogenesis (CD31) markers during DUs restoration. RESULTS: Knockout of SIRT6 inhibited the healing ability of DUs, presenting attenuated re-epithelialization (PCNA), exacerbated inflammation responses (NF-κB p50, F4/80, Il-1ß, Tnf-α, Il-6, Il-10, and Il-4), and hyperplasia vascular (CD31) compared with control mice. CONCLUSIONS: SIRT6 could boost impaired wound healing through improving epidermal proliferation, inflammation, and angiogenesis. Our study highlighted the therapeutic potential of the SIRT6 agonist for DUs treatment.

Risk factors and incidence of deep vein thrombosis in lower extremities among critically ill patients.

AIMS AND OBJECTIVES: To investigate how many critically ill patients developed deep vein thrombosis (DVT) during their admission to intensive care units (ICU) and to compare the characteristics of patients with and without deep vein thrombosis. BACKGROUND: Critically ill patients are a high-risk group for deep vein thrombosis because they typically have multiple risk factors, such as prolonged immobility, mechanical ventilation and old age. DESIGN: A prospective observational study was employed. METHODS: The subjects were 90 patients who were older than 18 years of age, who were admitted to an intensive care unit for more than five days and were not provided any prophylactic measures. Data were collected at a university hospital for five months. A duplex scan was performed on day 2.4 on average and repeated between days 5-7 to diagnose deep vein thrombosis. The iliac, femoral, popliteal and tibial veins were examined by compression and colour Doppler methods of the duplex scan by one technician. RESULTS: Age, gender and body mass index were significant factors for deep vein thrombosis development (p < 0.05). Ten patients (11.1%) developed deep vein thrombosis during their stay in the intensive care units. CONCLUSIONS: The incidence was lower than in Western studies in which patients were not provided prophylaxis, but may increase with an extended observation period as in previous studies. RELEVANCE TO CLINICAL PRACTICE: The results of this study could allow ICU nurses to recognise the DVT incidence in critically ill patients. This result could lead to more active prevention and monitoring of DVT by ICU nurses, especially for high-risk patients, such as older or obese patients.

Quality of life and psychological impact in patients with noncardiac chest pain.

BACKGROUND: Chest pain is common and data regarding noncardiac chest pain (NCCP) in Asia are lacking. AIM: To determine the differences in clinical presentations, psychologic impact, and quality of life between patients with NCCP and cardiac chest pain (CCP), and to identify any factors that impacted on these patients. METHODS: Consecutive patients undergoing coronary angiography for the evaluation of chest pain were recruited in Hong Kong and Wuhan, China. One hundred and forty patients with abnormal and 141 patients with normal angiography were included in the study. The validated gastroesophageal reflux disease questionnaire, the Hospital Anxiety-Depression Scale, and the 12-item Short Form Health Survey (SF-12) were used for assessment. RESULTS: NCCP patients reported similar days-off work and impairment of their social life compared with those with CCP. No difference was found in the anxiety and depression scores between the 2 groups. NCCP patients with reflux symptoms had higher anxiety score (7.19 vs. 5.74, P=0.044), reported more interruption of their social life (26% vs. 5%, P<0.0001), and had taken more sick leaves (17% vs. 5%, P=0.018) compared with those without gastroesophageal reflux disease. CONCLUSIONS: The quality of life and psychologic impact of patients with NCCP were as significant as those with CCP. NCCP patients with reflux symptoms were more anxious and were impaired in their productivity and social life.

Reference map of soluble proteins from Salmonella enterica serovar Enteritidis by two-dimensional electrophoresis.

Protein identification by peptide mass fingerprinting using matrix-assisted laser desorption ionization time of fight (MALDI-TOF) mass spectrometry (MS) can analyze unambiguously identity of the spots from a 2-dimensional electrophoresis (2-DE) gel. This study developed a technique for 2-DE of Salmonella enterica serovar Enteritidis (S. enteritidis) by improving the dissolution conditions by 2-DE using a pH 4 - 7 immobilized pH gradient (IPG) strip. This report examines the protein components from the patterns of the S. enteritidis protein. The most abundant protein displayed a great number of clusters within the pH 4.5 - 7 range with a molecular mass ranging from 35-80 kDa. Some of these spots were identified as metabolic related enzymes. The protein fraction was also analyzed using an immobilized pH gradient strip. Different proteins were identified on the spot according to the elongation factors. In addition, this study showed that the 2-DE analysis of S. enteritidis provides useful information regarding the S. enteritidis proteome, and this approach might provide a strategy for identifying bacterial proteins using a proteome technology.

Angiopoietin-2: a promising indicator for the occurrence of severe preeclampsia.

BACKGROUND: The known connection between placental hypoxia and the development of preeclampsia suggests that angiogenic factors in the placenta would be changed and affect the maternal and/or umbilical cord plasma levels in patients with preeclampsia. OBJECTIVE: The aim of this study was to determine the difference and correlation of placental mRNA expression and maternal/umbilical cord plasma concentrations of vascular endothelial growth factor A (VEGF-A), angiopoietin-1, and angiopoietin-2 between women with severe preeclampsia and normal pregnancies. METHODS: Sixteen patients with severe preeclampsia and 29 normotensive pregnant women were studied. The placental mRNA expression was assessed using real-time quantitative RT-PCR analysis. Maternal/umbilical cord plasma levels were measured using an enzyme-linked immunoassay. Nonparametric methods were applied for statistical analysis. RESULTS: . Placental mRNA expression of angiopoietin-2 was significantly increased in patients with severe preeclampsia (p < 0.001). The maternal plasma angiopoietin-2 protein level was also significantly increased in women with severe preeclampsia (p < 0.05) and showed a positive correlation with the placental mRNA expression of angiopoietin-2 (r = 0.54, p < 0.005). For VEGF-A and angiopoietin-1, there were no significant differences between the two groups. A maternal plasma angiopoietin-2 concentration of 8.4 ng/mL had a sensitivity of 63% and a specificity of 83% for predicting severe preeclampsia. CONCLUSION: Placental angiopoietin-2 mRNA expression was increased and correlated with the maternal plasma angiopoietin-2 protein concentration in women with severe preeclampsia. This suggests that the plasma angiopoietin-2 protein level may be a candidate marker for severe preeclampsia.

Increasing access to evidence-based smoking cessation treatment: effectiveness of a free nicotine patch program among Chinese immigrants.

Pharmacotherapy substantially increases smoking cessation rates. However, programs to reduce barriers to this evidence-based treatment may not improve access among high risk immigrant non English speaking populations. This study estimates the effectiveness of a tailored free nicotine patch (NRT) program among Chinese American smokers living in New York City (NYC). Between July 2004 and May 2005 NRT was distributed to 375 smokers through two community-based organizations that serve the Asian American population in NYC. Participants completed an in person baseline survey and a 4-month follow-up telephone survey. Using an intention to treat analysis the abstinence rate at 4 months was 26.7% (100/375). Predictors of cessation included higher levels of self efficacy at baseline, not smoking while using the patch and concern about personal health risks. Distribution through easy to access, culturally competent local community organizations increased the reach of a free nicotine patch program and assisted smokers in quitting.

Gastric electrical stimulation using endoscopically placed mucosal electrodes reduces food intake in humans.

BACKGROUND: Implantable gastric stimulation (IGS) has been proposed for treating obesity. The aim of this study was to investigate the effects of temporary mucosal electrical stimulation on water and food intake as well as gastric emptying in healthy humans. METHODS: The study was designed to study the effects of temporary gastric electrical stimulation (GES) on symptoms, gastric accommodation, food intake, and gastric emptying. It was performed in 12 healthy volunteers on 3 consecutive days. GES was performed using mucosal electrodes endoscopically placed in the fundus. RESULTS: The amount of maximum water intake was reduced with GES (894 +/- 326 mL) compared with sham-GES (1,093 +/- 417 mL, p = 0.01). The food intake was also reduced with GES (p = 0.012). In comparison with sham stimulation, GES delayed gastric emptying during the first 45 min after the meal but not during the remaining time. GES with parameters effective in reducing water and food intake and delaying gastric emptying did not induce significant dyspeptic symptoms, compared with sham stimulation. CONCLUSIONS: GES using temporary mucosal electrodes decreases food intake as well as maximum intake of water, and has a tendency of delaying gastric emptying. It may have a potential application for the treatment of obesity.

Keratin 9 gene mutations in five Korean families with epidermolytic palmoplantar keratoderma.

Epidermolytic palmoplantar keratoderma (EPPK) is an autosomal dominant disease characterized clinically by localized palmoplantar thickening and histopathologically by granular degeneration of the epidermis. Recent molecular biological studies have revealed that EPPK is caused by mutations of the keratin 9 gene in sequences mainly encoding the highly conserved 1 A rod domain. Here we demonstrate a novel mutation of N160H (position 8 of the 1 A domain) and two other previously reported mutations, R162W and N160S, in five unrelated Korean families with EPPK. The three-dimensional structure of the 1 A domain of the related vimentin intermediate filament protein chain is now known. Based on its likely similarity to the keratin 9 chain, we predict that inappropriate amino acid substitutions in position 10 of 1 A will likely interfere with coiled-coil dimer stability, and those in position 8 will interfere with tetramer stability. Accordingly, these mutations compromise the structural integrity of the keratin intermediate filaments leading to the pathology of EPPK.

Effect of Three Dimensional Culture Using Alginate-Fibrin Beads on the Chondrogenic Differentiation of Rabbit's Perichondrial Cells / 대한정형외과연구학회지

PURPOSE: To evaluate the usefulness of three dimensional culture using alginate-fibrin beads on the chondrogenic differentiation of rabbit's perichondrial cells. MATERIALS AND METHODS: Rib perichondrial cells from rabbit expanded by monolayer culture were cultured in monolayer, in alginate bead, and in alginate-fibrin beads. Reverse transcription polymerase chain reaction (RT-PCR) for type I, II, X collagen, and aggrecan was performed at 3 weeks after culture. At that time, we removed the alginate component from the alginate-fibrin bead. Then, the cell-fibrin beads were transplanted into the partial physeal defect of proximal tibia, and some beads were cultured on for additional 3 weeks for RT-PCR. Histologic examination was performed at 2, 4, 8, 12, and 16 weeks after operation. RESULTS: At 3 weeks, type II collagen gene expression was maintained regardless of the culture system used. However, at 6 weeks it was maintained only in three-dimensional culture system using alginate beads and fibrin beads. Histologic examination showed that the implanted perichondrial cell-fibrin beads formed small nest composed of chondrocyte-like cells and matrix. CONCLUSION: These results suggest that alginate-fibrin beads may be used as a biodegradable scaffold for cartilage engineering using perichondrial cells.