[The efficacy and safety of ticagrelor versus clopidogrel in elderly patients with acute coronary syndrome undergoing percutaneous coronary intervention].

Objective: To compare the efficacy and safety of ticagrelor and clopidogrel in elderly Chinese patients with acute coronary syndrome (ACS) underwent percutaneous coronary intervention (PCI) in the real world. Methods: This study is a post-hoc analysis of a single center, retrospective cohort study. Between March 2016 and March 2018, elderly (age≥65) ACS patients who underwent PCI in the General Hospital of Northern Theater Command were included in the study. The patients were grouped according to P2Y12 receptor inhibitor. The primary endpoints of this study were ischemic events during the 2-year follow-up, which were defined as the composite of cardiac death, myocardial or stroke. The secondary efficiency endpoints included all-cause death and BARC 2, 3, 5 bleeding events. Results: A total of 4 022 elderly (mean age: (71.5±5.3) years) ACS patients were included in this study. Based on the choice of P2Y12 receptor inhibitor, patients were divided into clopidogrel (n=3 201) and ticagrelor (n=821) groups. Incidences of ischemic events (3.2% (26/821) vs. 5.6% (179/3 201), P=0.005) at 2 years were significantly lower in ticagrelor group compared to clopidogrel group. BARC 2, 3, 5 bleeding events (1.7% (14/821) vs. 1.6% (52/3 201), P=0.818) were comparable between the two groups. The incidence of all-cause death (1.5% (12/821) vs. 4.1% (132/3 201), P=0.005) were also lower in the ticagrelor group compared to the clopidogrel group. Clinical outcomes were consistent after adjusting for confounding factors, the incidence of ischemic events (HR= 0.637, 95%CI 0.409-0.991, P=0.046) and all-cause mortality (HR=0.402, 95%CI 0.213-0.758, P=0.005) was significantly lower in the ticagrelor group compared with the clopidogrel group. Risk of BARC 2, 3, 5 bleeding events were similar between the two groups (HR=0.957, 95%CI 0.496-1.848, P=0.897). Conclusion: In real-world clinical practice, for elderly patients with ACS undergoing PCI, ticagrelor use might reduce the incidence of long-term ischemic events and all-cause death without increasing the risk of bleeding.

[Five-year clinical outcomes of the second generation biodegradable polymer sirolimus-eluting stent (EXCROSSAL) in treating patients with de novo coronary lesions].

Objective: To evaluate the five-year safety and efficacy of the second generation biodegradable polymer sirolimus-eluting stent (EXCROSSAL) in treating patients with de novo coronary artery diseases. Methods: Patients with coronary artery disease (CAD)who were implanted with EXTROSSAL stents in CREDIT Ⅱ and CREDIT Ⅲ study were included. CREDIT Ⅱ was a randomized trial, and CREDIT Ⅲ was a single-arm study. From November 2013 to December 2014, 833 CAD patients with de novo coronary lesions implanted with EXTROSSAL stents were selected from 33 centers in China. The primary outcome was 5-year target lesion failure (TLF), a composite of cardiac death, target vessel myocardial infarction and clinically indicated target lesion revascularization. Secondary endpoints was patient-oriented composite endpoint (PoCE), including all-cause death, all myocardial infarction, or any revascularization within 5 years post stenting and stent thrombosis according to Academic Research Consortium's (ARC) definition. Kaplan Meier method was used to calculate the incidence of TLF and PoCE within 5 years after operation. Univariate Cox regression analysis was used to analyze the impacts of diabetes, small vessel disease (vessel diameter ≤ 2.74 mm), lesion length ≥ 16.7 mm and multivessel disease on the incidence of TLF within 5 years after operation. Results: A total of 833 patients were included in this study including 579 males (69.5%), the age was (59.3±9.1) years. And 832 (99.9%) patients completed 5-year clinical follow-up. The incidence of TLF and PoCE in the 5-year follow-up were 10.6%(86/811) and 15.5%(126/811), respectively. Stent thrombosis occurred in 1.0%(8/811) of patients. Univariate Cox regression analysis showed that vessel diameter ≤ 2.74 mm (HR=3.20,95%CI 1.90-5.39,P<0.001), lesion length ≥ 16.7 mm (HR=1.88,95%CI 1.18-2.99,P=0.007) and multivessel disease (HR=2.44,95%CI 1.60-3.72,P<0.001) were related factors of TLF within 5 years after operation. Conclusion: EXCROSSAL stent is effective and safe in treating CAD patients with de novo coronary lesions, with low incidence of TLF and PoCE within 5 years after operation.

[Medication compliance for secondary prevention and long-term outcome among patients with acute coronary syndrome after percutaneous coronary intervention in different regions].

Objective: To explore the medication compliance for secondary prevention drugs and long-term prognosis of patients with acute coronary syndrome (ACS) undergoing percutaneous coronary intervention (PCI) between hospitals in different regions of China. Methods: The Optimal Antiplatelet Therapy for Chinese Patients with Coronary Artery Disease (OPT-CAD) study was a prospective, multi-center and registered study. Patients diagnosed as ACS and underwent PCI in OPT-CAD study were selected. Taking the Yangtze River as the dividing line between the south and the north of China, these patients were divided into two groups according to the hospitals where the patients visited, namely the southerns region group (n=1 958) and the northerns region group (n=5 091). In order to reduce selection bias and potential confounding factors, the patients in the two groups were matched by the tendency score, and the patients in the two groups were matched by the 1: 1 nearest match method according to the tendency score. The main endpoint of this study was the major adverse cardiovascular and cerebrovascular events (MACCE) occurring within 5 years after discharge, namely the composite endpoint of cardiac death, myocardial infarction, and/or ischemic stroke. Secondary endpoints were all-cause death, cardiac death, myocardial infarction, ischemic stroke, and type 2, 3, and 5 bleeding events defined by the Academic Research Consortium on Hemorrhage (BARC) within 5 years. The secondary preventive drugs was recorded, including antiplatelet drugs, statins, beta blockers, angiotensin converting enzyme inhibitors/angiotensinⅡreceptor blockers (ACEI/ARB), etc. Before and after the matching, the secondary preventive medication and the incidence of clinical events of the two groups were compared. Results: A total of 7 049 ACS patients, including 1 958 patients in the southern region group and 5 091 patients in the northern region group were enrolled in this study. There were 5 319 males (37.9%), and the aged was (60.7±6.7) years. After propensity score matching, there were 1 324 cases in each group. Before matching, in the northern region group, the proportion of smoking, hypertension and diabetes, previous history (myocardial infarction, PCI and stroke) and family history of coronary heart disease were higher (all P<0.05). The proportion of complex lesions, diffuse lesions, small vessel lesions and thrombotic lesions in the northern region group was higher than that in the southern region group (all P<0.05). Sixty months after discharge, the antiplatelet patterns were quite different between patients in the northern and southern region group (P<0.001). The proportion of clopidogrel monotherapy in the southern region group was higher than that in the northern region group (9.8% (130/1324) vs. 1.1% (14/1324)), while the proportion of aspirin monotherapy in the northern region group was higher than that in the southern region group (67.4% (893/1324) vs. 46.5% (616/1324)). As for the use of other secondary prophylactic drugs, the proportion of patients in southern region group receiving beta blockers (24.5% (325/1324) vs. 16.8% (222/1324), P<0.001) and ACEI/ARB (19.4% (257/1324) vs. 10.0% (133/1324), P<0.001) was higher than that in northern region group. After matching, the incidence of MACCE (8.4%(111/1 324) vs.6.2% (82/1 324), P=0.030) and BARC 2, 3 and 5 bleeding (6.0% (80/1 324) vs. 4.0% (53/1 324), P=0.020) was higher in patients in northern region group. Conclusions: ACS patients who undergo PCI in northern area hospital is at higher prevalence of comorbidities and complicated coronary artery lesions compared to patients in the southern area hospital, and the drug compliance is worse than that in southern area, and the prognosis is also relatively poor.

Extracellular production of functional single-chain variable fragment against aflatoxin B1 using Escherichia coli.

scFv-BM3 is a single-chain variable fragment (scFv) against aflatoxin B1 (AFB1 ) engineered by affinity maturation and site-directed mutagenesis, and thus has a 31-fold higher affinity than its wild-type. To apply scFv-BM3 to immunological detection of AFB1 , periplasmic expression in Escherichia coli was attempted to produce a functional form of scFv-BM3. scFv-BM3 accumulated as inactive aggregates in the cells. However, it was found that scFv-BM3 secreted into the culture medium had binding activity to AFB1 . Expression conditions for scFv-BM3 were further manipulated to enhance secretion into the culture medium. This extracellular secretion of functional scFv-BM3 was significantly improved by supplementation with Triton X-100 and optimization of expression conditions. The scFv-BM3 purified from the culture medium exhibited a typical antiparallel ß-sheet structure and adopted a proper conformation to bind AFB1 with high affinity and specificity in various biophysical and biochemical analyses. SIGNIFICANCE AND IMPACT OF THE STUDY: Single-chain variable fragments (scFvs) are recombinant antibodies that are difficult to produce as a functional form in Escherichia coli. This study demonstrates the production of functional scFvs against aflatoxin B1 (AFB1 ) (scFv-BM3) using Escherichia coli by extracellular secretion. While periplasmic expression of scFv-BM3 resulted in formation of inactive aggregates in E. coli, the scFv-BM3 secreted into the culture medium adopted a properly folded structure for specific binding to AFB1 . This study promotes the application of functional scFv-BM3 to the immunological detection of AFB1 in biotechnology fields.

Nafamostat Mesilate Attenuates Ischemia-Reperfusion-Induced Renal Injury.

BACKGROUND: It has been reported that nafamostat mesilate (NM) inhibits inflammatory injury via inhibition of complement activation in ischemic heart, liver, and intestine. However, it is unclear if NM also inhibits apoptosis in ischemia-reperfusion (IR)-injured kidney. We therefore investigated whether NM attenuates IR renal injury that involves inhibition of apoptosis. METHODS: HK-2 cells and male C57BL/6 mice were used for this study. C57Bl/6 mice were divided into 4 groups: sham, NM (2 mg/kg) + sham, IR injury (IR injury; reperfusion 27 minutes after clamping of both the renal artery and vein), and NM + IR injury. Kidneys were harvested 24 hours after IR injury, and functional and molecular parameters were evaluated. For in vitro studies, HK-2 cells were incubated for 6 hours with mineral paraffin oil to induce hypoxic injury, and then treated with various doses of NM to evaluate the antiapoptotic effects. RESULTS: Blood urea nitrogen, serum creatinine levels, and renal tissue injury scores in NM + IR-injured mice were significantly lower than those of control IR mice (all P < .01). NM significantly improved cell survival in hypoxic HK-2 cells (P < .01), significantly decreased renal Bax expression (P < .05), and increased renal Bcl-2 protein levels in IR kidneys and hypoxic HK-2 cells compared with those of the sham and control groups. The numbers of terminal deoxynucleotide transferase-mediated dUTP nick-end labeling- and 8-oxo-2'-deoxyguanosine-positive cells were significantly lower in NM + IR-injured kidneys compared with those in control IR-injured mice (P < .05); NM treatment decreased the expression of inducible and endothelial nitric oxide synthase in IR-injured mice (P < .05). CONCLUSIONS: NM ameliorates IR renal injury via inhibition of apoptosis by, at least in part, lowering nitric oxide overproduction, reducing Bax, and increasing Bcl-2.

Spinning: an arising cause of rhabdomyolysis in young females.

BACKGROUND: 'Spinning' is an indoor cycling regimen. The number of case reports of spinning-induced rhabdomyolysis (SIR) has increased since 2004 in South Korea. AIM: The aim of this study is to evaluate the clinical characteristics of SIR and compare it with other causes of rhabdomyolysis. METHODS: Patients who were diagnosed with rhabdomyolysis from 1 September 2011 to 30 April 2015 were included. We analysed the incidence of rhabdomyolysis, biochemical parameters and forced hospitalisation, which was defined as the days from admission to creatinine phosphokinase < 2000 IU/L. RESULTS: Among 70 included patients, 13 (18.6%) patients were diagnosed with SIR. The mean age of the patients with SIR was 25.69 ± 5.0 years, and most were females under 35 years old (12, 92.3%). Interestingly, the mean duration of spinning exercise before admission was only 59.23 min. Moreover, the patients with SIR showed more severe progress than the all-patients-except-SIR (AESIR) group. The serum creatinine phosphokinase, aspartate transaminase and alanine transaminase levels of the patients with SIR were statistically significantly higher than the patients with AESIR. Additionally, the duration of forced hospitalisation was longer than that of the AESIR (P < 0.01). CONCLUSIONS: Spinning could be an important cause of rhabdomyolysis in young, unfit females, which is typically severe. A graded exercise programme is advised at the first session.

Incidence and risk factors for ocular GVHD after allogeneic hematopoietic stem cell transplantation.

We analyzed the incidence and risk factors for ocular GVHD in patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) in Korea. In this retrospective, noncomparative, observational study, 635 subjects were included who had at least 2 years of follow-up ophthalmological examinations after allo-HSCT from 2009 to 2012 at Seoul St Mary's Hospital, Seoul, Korea. The mean duration between allo-HSCT and onset of ocular GVHD was 225.5±194.3 days. The adjusted incidence for acute ocular GVHD was 1.33% and that for chronic GVHD was 33.33%. In the multivariate analysis, preexisting diabetes mellitus (odds ratio (OR): 4.22, 95% confidence interval (CI): 1.66-10.72), repeated allo-HSCT (OR: 29.10, 95% CI: 1.02-8.28) and the number of organs that chronically developed GVHD by stage I (OR: 14.63, 95% CI: 9.81-21.84) increased risk of ocular GVHD. Careful monitoring of ocular GVHD is needed in patients with chronic GVHD in multiple organs and preexisting diabetes.

Bone marrow-derived clonal mesenchymal stem cells inhibit ovalbumin-induced atopic dermatitis.

Mesenchymal stem cells (MSCs) possess immunomodulatory activities, including suppression of T- and B-cell activation. However, their effects on atopic dermatitis (AD) have not yet been studied. Using an ovalbumin-induced AD mouse model, we investigated whether MSCs can be used as therapeutics in AD. We isolated both allogeneic and syngeneic clonal MSCs (cMSCs) from mouse bone marrow according to the subfractionation culturing method. Our cMSCs suppressed both T- and B-cell activation. T-cell proliferation and cytokine production, including interferon (IFN)-γ and interleukin (IL)-4, were suppressed by inhibition of transcription factors, such as T-bet, GATA-3, and c-Maf. Those transcription factors were nitric oxide dependent. Immunoglobulin E (IgE) suppression occurred through downregulation of AID and BLIMP-1, important regulators for isotype class switch and B-cell differentiation. The cMSCs were injected intravenously into ovalbumin-induced AD mouse model, and the therapeutic effects were analyzed. Injection of both allogeneic and syngeneic cMSCs in an AD mouse model inhibited cell infiltration in skin lesions and decreased the serum level of IgE. IL-4 expression was also suppressed by cMSCs in both the lymph node and skin. The cMSCs migrated to skin lesions and draining lymph nodes. Taken together, these data demonstrated that cMSCs, which suppressed T- and B-cell functions, can be used for the treatment of AD in mice.

Serum adipokine concentrations in dogs with naturally occurring pituitary-dependent hyperadrenocorticism.

BACKGROUND: An excess of intra-abdominal fat is observed frequently in dogs with hyperadrenocorticism (HAC). Adipokine dysregulation is a possible cause of complications related to visceral obesity, but little information is available on adipokine in dogs with naturally occurring HAC. OBJECTIVES: To examine the differences in the circulating adipokines concentrations in overweight dogs with and without pituitary-dependent HAC (PDH). ANIMALS: Thirty healthy dogs and 15 client-owned dogs with PDH. METHODS: Case-controlled observational study, which enrolled 15 overweight dogs diagnosed with PDH and 30 otherwise healthy dogs of similar body condition score. Nine of 15 dogs with PDH were treated with low-dose trilostane twice daily and reassessed after treatment. RESULTS: The serum leptin (P < .0001) and insulin (P < .0001) concentrations were significantly higher in the PDH group (leptin, 22.8 ± 8.8 [mean ± SD]; insulin, 9.1 ± 6.1) than the healthy group (leptin, 4.9 ± 3.7; insulin, 1.9 ± 0.9). However, there were no significant differences in the adiponectin, resistin, tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6, IL-10, and IL-18 levels between the 2 groups. In the PDH group, the serum cortisol concentrations had a linear association with the leptin concentrations, and there were significant decreases in the leptin (P = .0039) and insulin (P = .0039) levels after trilostane treatment. However, the leptin and insulin levels remained higher after trilostane treatment than in healthy control dogs with similar body condition score. CONCLUSIONS AND CLINICAL IMPORTANCE: Hypercortisolemia in dogs with PDH might upregulate the circulating leptin levels. However, a large population-based study will be necessary to determine whether the upregulation of leptin is involved directly with the complications caused by HAC.

A retrospective analysis of 29 isolated sphenoid fungus ball cases from a medical centre in Korea (1999-2012).

BACKGROUND: Isolated sphenoid sinus disease (ISSD) is rare. Fungus ball (FB) is the third most common ISSD. We analysed the characteristics of isolated sphenoid FB based on demographic data, presenting symptoms, preoperative computed tomography (CT), magnetic resonance imaging (MRI), and treatment outcomes. METHODOLOGY: From 1999 to 2012, 29 patients were identified with isolated sphenoid FB. Demographic data; clinical characteristics; endoscopic, CT, and MRI findings and treatment outcomes were retrospectively analysed. RESULTS: The most common symptom was headaches, which were localized in various regions of the brain. Other symptoms were uncommon. The most common CT findings were sclerosis, calcification, enlarged sinus and total opacification. On T2-weighted MRI images, we most commonly observed signal void. Endoscopic transnasal paraseptal sphenoidotomy was performed in all patients, and for most, this was performed under local anaesthesia. No recurrence was observed in any patient. CONCLUSION: Isolated sphenoid FB is predominantly observed in older women, and it is characterised by headaches and sclerosis of the sinus wall observed on CT scans. In cases of isolated sphenoid FB, endoscopic transnasal paraseptal sphenoidotomy can be successfully performed under local anaesthesia, which may facilitate rapid recovery and a low morbidity rate.

Alteration in E-cadherin/ß-catenin expression in canine melanotic tumors.

ß-Catenin, encoded by the ctnnb1 gene, plays a critical role in intercellular adhesion, and its altered expression has been implicated in tumor progression in humans and animals. The aims of this study were to examine the alterations in ß-catenin expression in canine melanoma as well as the causes of these changes (eg, E-cadherin or exon 3 mutations) and to compare identified changes between skin and oral melanomas. Forty-two primary canine skin and oral melanoma tissue samples were used in the study. The expression levels of ctnnb1 and the levels of E-cadherin/ß-catenin complex in the tissues were determined by semiquantitative RT-PCR and immunohistochemistry, respectively. The mutational status of ß-catenin exon 3 was examined by DNA sequencing. RT-PCR revealed higher levels of ctnnb1 expression in oral melanoma tissues compared with normal melanocytes, irrespective of sex or histopathological appearance of the tissue (ie, amelanotic vs melanotic). Immunohistochemistry revealed simultaneous loss of membrane E-cadherin/ß-catenin complex and cytoplasmic accumulation of both proteins in 37 cases (84%). Intranuclear ß-catenin was also detected in all tissues with reduced membrane ß-catenin expression. In mutational analyses, one amelanotic oral melanoma showed 13 single nucleotide polymorphisms (SNPs); however, after protein translation, all the SNPs were silent mutations. The present study demonstrates that dysregulation of E-cadherin/ß-catenin complexes is involved in both types of canine melanotic tumors and that the disruption of E-cadherin/ß-catenin complexes and increased ß-catenin may induce tumor progression and malignancy.

Efficacy of low- and high-dose trilostane treatment in dogs (< 5 kg) with pituitary-dependent hyperadrenocorticism.

BACKGROUND: Trilostane is commonly used to treat pituitary-dependent hyperadrenocorticism (PDH) in dogs. There are differing opinions regarding the dose and frequency of trilostane administration in dogs with PDH. OBJECTIVES: To compare the efficacy of 2 trilostane protocols in the treatment of dogs with PDH. ANIMALS: Sixteen client-owned dogs with PDH and a body weight <5 kg. METHODS: Prospective observational study. Group A (n=9; low-dose treatment group) received 0.78 ± 0.26 mg of trilostane/kg PO every 12 h and group B (n = 7; high-dose treatment group) 30 mg of trilostane/dog PO every 24 h. All of the dogs were reassessed at 2, 4, 8, 12, 16, and 24 weeks after the initiation of treatment. RESULTS: An improvement in both ACTH-stimulated serum cortisol concentrations and clinical signs occurred more slowly in group A than in group B; however, after 20 weeks of treatment, 2/7 dog in group B had clinical signs and abnormal laboratory findings consistent with hypoadrenocorticism. At 24 weeks, an improvement in the clinical findings of all of the dogs in both groups was detected. CONCLUSIONS AND CLINICAL IMPORTANCE: In dogs with PDH, twice-daily administration of low-dose trilostane is an effective approach to the management of PDH. In addition, our results suggest fewer potential adverse effects if trilostane is administered twice daily in the lower dose.

Effect of estradiol on the expression of renal sodium transporters in rats.

BACKGROUND: Although estradiol has been thought to perform an important role in blood pressure regulation, the effects of estradiol on the expression of renal sodium transporters are not fully understood. METHODS: Female Sprague-Dawley rats were treated with 17ß-estradiol or vehicle for 10 days after ovariectomy, and after both ovariectomy and adrenalectomy to eliminate the effect of aldosterone. RESULTS: In the ovariectomized (OVX) rats, estradiol decreased the abundance of the Na-K-2Cl cotransporter (NKCC2) (31.5% of control (OVX), p < 0.01), Na-Cl cotransporter (NCC) proteins (40.5% of control (OVX), p < 0.01) and α- and γ-subunits of the epithelial sodium channel (ENaC) (44.7% and 11.0% of control (OVX), p < 0.01). Estradiol also reduced plasma aldosterone levels (OVX + 17ß-estradiol vs. OVX, 116.3 ± 44.4 vs. 184.2 ± 33.4 pmol/l, p < 0.05) and systolic blood pressure (OVX + 17ß-estradiol vs. OVX, 115 ± 4 vs. 132 ± 2 mmHg, p < 0.05). In rats having undergone adrenalectomy and ovariectomy, estradiol did not reduce systolic blood pressure, or the expression of sodium transporters. CONCLUSION: Estradiol decreased systolic blood pressure, plasma aldosterone levels, and the expression of renal sodium transporters. After aldosterone was eliminated, estradiol did not affect blood pressure or the expression of sodium transporters, which indicates that the effect of estradiol on the renal sodium transporters is at least partly influenced by aldosterone.

Porcine feasibility and safety study of a new paclitaxel-eluting biliary stent with a Pluronic-containing membrane.

BACKGROUND AND STUDY AIM: Metal stents for malignant biliary obstruction are susceptible to occlusion by tumor ingrowth or overgrowth. Therefore, we previously reported our use of a metal stent covered with a paclitaxel-incorporated membrane giving an antitumor effect to prevent occlusion from tumor ingrowth. We have also developed a new generation of paclitaxel-eluting biliary stent using a membrane containing Pluronic F-127 for effective drug delivery. The aim of this study was to investigate the safety and efficacy of drug delivery for this newly developed stent in the biliary tract. METHODS: Metal stents were coated with paclitaxel and various concentrations of Pluronic F-127 in phosphate-buffered saline solution. Stents containing varying concentrations were placed in the bile ducts of eight pigs divided as follows: group I, 0% Pluronic + 0% paclitaxel; group II, 0% Pluronic + 10% paclitaxel; group III, 10% Pluronic + 10% paclitaxel; group IV, 20% Pluronic + 10% paclitaxel. The histology of the porcine bile duct and the amount of paclitaxel in the porcine serum were examined. The amount of paclitaxel released was also measured in vitro. RESULTS: Histologic changes in the porcine biliary epithelium were acceptable in terms of safety, based on inflammatory cell infiltration and fibrotic reaction. No significant differences in histology were observed between the groups. In the porcine serum analysis, released paclitaxel was detected for 28 days with the 10% Pluronic concentration (group III). However, released paclitaxel was observed for only 7 days in groups II and IV. In the in vitro experiments, long-lasting release of paclitaxel was also noted from the stent with 10% Pluronic. CONCLUSIONS: The new paclitaxel-eluting stent with 10% Pluronic F-127 is safe and provides enhanced local drug delivery.

Wetting behavior and nanotribological properties of silicon nanopatterns combined with diamond-like carbon and perfluoropolyether films.

A large number of silicon (Si) patterns consisting of nanopillars of varying diameter and pitch have been fabricated and further coated with diamond-like carbon (DLC) and perfluoropolyether (Z-DOL) films. The wetting behavior and nano-adhesion/friction of the patterns are investigated experimentally in relation to the nanostructures and the hydrophobicity of the materials. Measurements of water contact angle illustrate that the patterning-enhanced wettability of the Si flat surface, along with two distinct wettings which are in good agreement with the Wenzel and hemi-wicking states, depended on the value of the pitch-over-diameter ratio. In the case of the coated patterns, three wetting states are observed: the Cassie-Baxter, the Wenzel, and a transition from the Cassie-Baxter into the Wenzel, which varies with regard to the hydrophobic properties of the DLC and Z-DOL. In terms of tribological properties, it is demonstrated that a combination of the nanopatterns and the films is effective in reducing adhesive and frictional forces. In addition, the pitch and diameter of the patterns are found to significantly influence their adhesion/friction behaviors.

Dysregulation of the Wnt/beta-catenin signaling pathway in canine cutaneous melanotic tumor.

The Wnt/beta-catenin signal transduction pathway is important in many developmental processes and during tumorigenesis. beta-Catenin acts as a signal transducer. To investigate whether the Wnt/beta-catenin signal transduction pathway is involved in canine cutaneous melanomagenesis, 18 formalin-fixed paraffin-embedded canine cutaneous melanotic tumor tissues were examined. For cloning and sequencing of the full-length canine ctnnb1 gene encoding beta-catenin, conserved sequences of the human and mouse ctnnb1 gene were used to design the primers. For analysis of expression and translocation of beta-catenin in canine cutaneous melanotic tumors, semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were performed. The canine ctnnb1 sequence showed a high degree of similarity to those of human and mouse. Semiquantitative RT-PCR showed a substantial increase in expression of ctnnb1 mRNA in canine cutaneous melanotic tumors compared to normal canine melanocytes, regardless of whether the tumor was benign or malignant. Immunohistochemistry revealed cytoplasmic accumulation of beta-catenin in melanotic tumors. In melanoma tissues, nuclear translocation of beta-catenin was also observed. The present study demonstrated that abnormal intracellular accumulation and substantially increased expression of beta-catenin are involved in canine cutaneous melanotic tumor.

Effective removal of leptin via hemodiafiltration with on-line endogenous reinfusion therapy.

AIMS: Leptin is a middle-molecular weight uremic toxin. Hemodiafiltration with on-line endogenous reinfusion (HFR) is a novel dialytic method combining the processes of diffusion, convection and adsorption. We performed a prospective crossover study of patients with end-stage renal disease to investigate the effect of HFR therapy on the level of leptin as compared to conventional low flux hemodialysis (LHD). METHODS: Eleven stable hemodialysis patients were treated with LHD for 12 weeks and then treated with HFR (SG30 Plus; Sorin Group Italia S.r.1, Mirandola, Italy) for 12 weeks. RESULTS: After 12 weeks of HFR treatment, serum leptin levels significantly decreased (17.1 (2.66 - 39.5) at Week 12 vs. 12.3 (1.80 - 24.3) ng/ml at Week 24, p = 0.014). Although serum adiponectin levels also decreased (1.66 (1.44 - 1.86) at Week 12 vs. 1.12 (0.79 - 1.34) g/ml at Week 24, p = 0.001), the ratio of leptin to adiponectin did not increase after HFR treatment. Serum beta2-microglobulin (beta2M) levels significantly decreased (37.7 (29.8 - 42.6) at Week 12 vs. 28.3 (26.5 - 32.2) mg/dl at Week 24, p = 0.002). Dry weight, Kt/V(urea), normalized protein equivalent of nitrogen appearance, subjective global assessment, and serum albumin levels of the patients were not changed after HFR treatment. There was no difference in the serum levels of C-reactive protein or interleukin-6 between Week 12 and Week 24. CONCLUSIONS: The results of our study indicate that HFR may be a better therapy than LHD for removal of middle-molecular-weight uremic toxins such as leptin and b2M.

Pretreatment with the tumor nerosis factor-alpha blocker etanercept attenuated ischemia-reperfusion renal injury.

INTRODUCTION: Tumor necrosis factor (TNF)-alpha mediates inflammation and apoptosis in ischemia-reperfusion (IR) injury of the kidneys. Etanercept, a soluble TNF-alpha receptor, has shown anti-inflammatory and anti-apoptotic effects in several animal models of renal injury, including chronic insufficiency and unilateral ureteral obstruction. We evaluated the protective effect of etanercept against experimental renal IR injury. METHODS: Male Sprague-Dawley (SD) rats were divided into 4 groups: saline-treated sham rats, etanercept-treated sham rats, saline-treated IR rats, and etanercept-treated IR rats. Renal messenger RNA (mRNA) levels of TNF-alpha and monocyte chemotactic protein-1 (MCP-1) were measured by real-time polymerase chain reaction (PCR) at 24 hours after IR injury. The protein levels of renal Bcl-2 associated X (Bax), B-cell lymphoma 2 (Bcl), extracellular signal-regulated kinase (ERK), and caspase-3 activation were evaluated using Western blot analysis. The degree of apoptosis of renal tubular cells was determined using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays. RESULTS: At 24 hours after IR injury, the serum levels of blood urea nitrogen (BUN) and creatinine were significantly lower among etanercept-treated than saline-treated IR rats. Renal mRNA levels of TNF-alpha and MCP-1 in saline-treated IR rats were significantly higher than the levels in saline-treated sham rats, and TNF-alpha and MCP-1 mRNA levels in etanercept-treated IR rats were significantly lower than those in saline-treated IR rats. Etanercept pretreatment of IR-injured rats significantly increased EKR phosphorylation and reduced the renal Bcl-2/Bax ratio, the renal caspase-3 activation, and the number of TUNEL-positive apoptotic cells. CONCLUSION: Etanercept improved resistance to renal injury during IR by enhancing the activation of ERK and increasing the Bcl-2/Bax ratio.

HLA-B27 homozygosity has no influence on clinical manifestations and functional disability in ankylosing spondylitis.

OBJECTIVE: There is an established association between the development of ankylosing spondylitis (AS) and the HLA-B27 allele, but whether or not homozygosity for HLA B27 has any additional effects on the clinical manifestations of AS is unclear. The aim of this study was to determine the influence of HLA-B27 homozygosity on the clinical manifestations of AS in Korea. METHODS: A total of 490 patients were enrolled in this study. Genotyping was carried out using polymerase chain reaction-sequence specific primers (PCR-SSP) to assess HLA-B27 homozygosity or heterozygosity. One PCR reaction was undertaken to determine the HLA-B27 carrier status, and 5 group-specific PCR reactions were carried out to determine all of the other HLA-B alleles. Clinical manifestations of AS and BASFI were also evaluated according to homozygosity or heterozygosity for HLA-B27. RESULTS: HLA-B27 positive patients had a significantly younger age at symptom onset, more uveitis, and a higher frequency of hip joint involvement than HLA-B27 negative patients. One hundred and forty-six (29.8%) patients were homozygous for HLA-B27. No significant association between HLA-B27 homozygosity or heterozygosity and a history of peripheral arthritis, acute anterior uveitis, age at onset, or the Bath Ankylosing Spondylitis Functional Index (BASFI) was found. CONCLUSIONS: Homozygosity for HLA-B27 does not affect the clinical manifestations of AS in Korean patients.