Metformin regulates the LIN28B­mediated JNK/STAT3 signaling pathway through miR­140­3p in subretinal fibrosis.

Subretinal fibrosis (SF) is an important cause of submacular neovascularization that leads to permanent vision loss, but has no effective clinical treatment. The present study examined the influence of metformin on SF, and investigated whether the mechanism involves the microRNA (miR)-140-3p/LIN28B/JNK/STAT3-mediated regulation of oxidative stress, angiogenesis and fibrosis-associated indicators. A mouse model of laser-induced SF was established. In addition, an ARPE-19 fibrotic cell model was established using TGF-ß1. A Cell Counting Kit-8 assay was used to examine cell viability. Flow cytometry was used to measure reactive oxygen species levels, and western blotting was used to detect the levels of proteins associated with epithelial-mesenchymal transition (EMT), signaling and fibrosis. The levels of superoxide dismutase, malondialdehyde, glutathione-peroxidase and catalase were measured using kits. Scratch assays and Transwell assays were used to assess cell migration and invasion, respectively, and reverse transcription-quantitative PCR was used to determine the levels of miR-140-3p and LIN28B. Dual-luciferase assays were used to verify the targeting relationship between miR-140-3p and LIN28B, and coimmunoprecipitation was used to confirm the interaction between LIN28B and JNK. Masson staining and hematoxylin and eosin staining were used to examine collagenous fibers and the histopathology of eye tissue. In ARPE-19 cells induced by TGF-ß1, metformin promoted miR-140-3p expression and inhibited LIN28B expression and JNK/STAT3 pathway activation, thereby inhibiting oxidative stress, EMT and fibrosis in ARPE-19 cells. The overexpression of LIN28B or treatment with the JNK/STAT3 agonist anisomycin partially reversed the inhibitory effect of metformin on oxidative stress and fibrosis in ARPE-19 cells. The dual-luciferase reporter assay and coimmunoprecipitation assay showed that miR-140-3p targeted the 3' untranslated region of LIN28B mRNA and inhibited LIN28B expression. LIN28B targeted and bound to JNK and regulated the JNK/STAT3 pathway. Therefore, it may be concluded that metformin can promote miR-140-3p expression, inhibit LIN28B and then inhibit the JNK/STAT3 pathway to alleviate SF.

Handyfuge Microfluidic for On-Site Antibiotic Susceptibility Testing.

Low-cost, rapid, and accurate acquisition of minimum inhibitory concentrations (MICs) is key to limiting the development of antimicrobial resistance (AMR). Until now, conventional antibiotic susceptibility testing (AST) methods are typically time-consuming, high-cost, and labor-intensive, making them difficult to accomplish this task. Herein, an electricity-free, portable, and robust handyfuge microfluidic chip was developed for on-site AST, termed handyfuge-AST. With simply handheld centrifugation, the bacterial-antibiotic mixtures with accurate antibiotic concentration gradients could be generated in less than 5 min. The accurate MIC values of single antibiotics (including ampicillin, kanamycin, and chloramphenicol) or their combinations against Escherichia coli could be obtained within 5 h. To further meet the growing demands of point-of-care testing, we upgraded our handyfuge-AST with a pH-based colorimetric strategy, enabling naked eye recognition or intelligent recognition with a homemade mobile app. Through a comparative study of 60 clinical data (10 clinical samples corresponding to six commonly used antibiotics), the accurate MICs by handyfuge-AST with 100% categorical agreements were achieved compared to clinical standard methods (area under curves, AUCs = 1.00). The handyfuge-AST could be used as a low-cost, portable, and robust point-of-care device to rapidly obtain accurate MIC values, which significantly limit the progress of AMR.

Near-infrared light-responsive multifunctional hydrogel releasing peptide-functionalized gold nanorods sequentially for diabetic wound healing.

Treatment for chronic diabetic wounds remains a clinical challenge. Wound healing process occurs in three phases: inflammation, proliferation and remodeling. Several factors including bacterial infection, decreased local angiogenesis and diminished blood supply delay wound healing. There is an urgent need to develop wound dressings with multiple biological effects for different stages of diabetic wound healing. Here, we develop a multifunctional hydrogel with two-stage sequential release upon near-infrared (NIR) stimulation, antibacterial activity and pro-angiogenic efficacy. This hydrogel consists of covalently crosslinked bilayer structure, with the lower thermoresponsive poly(N-isopropylacrylamide)/gelatin methacrylate (NG) layer and the upper highly stretchable alginate/polyacrylamide (AP) layer embedding different peptide-functionalized gold nanorods (AuNRs) in each layer. Antimicrobial peptide-functionalized AuNRs released from NG layer exert antibacterial effects. After NIR irradiation, the photothermal transition efficacy of AuNRs synergistically enhances bactericidal efficacy. The contraction of thermoresponsive layer also promotes the release of embedded cargos during early stage. The pro-angiogenic peptide-functionalized AuNRs released from AP layer promote angiogenesis and collagen deposition by accelerating fibroblast and endothelial cell proliferation, migration and tube formation during the subsequent healing phases. Therefore, the multifunctional hydrogel with effective antibacterial activity, pro-angiogenic efficacy and sequential release behaviors is a potential biomaterial for diabetic chronic wound healing.

HRCT quantitative analysis of airway remodeling and airway trapping in the small airway asthma phenotype and its correlation with pulmonary function.

OBJECTIVES: We aimed to explore whether large airway remodeling and small airway structural changes exist in subjects with small airway asthma phenotype and to evaluate the relationships between quantitative high-resolution computed tomography (qHRCT) parameters and lung function. METHODS: We enrolled 15 subjects with small airway asthma phenotype and 18 healthy controls. The two groups were matched by age, sex and body square area (BSA) with propensity score matching (PSM). Pulmonary function and qHRCT parameters [wall thickness (WT), wall area (WA), lumen area (LA), wall area percentage (WA%) of the 4th-6th generations in the right upper lobe apical segmental bronchus (RB1), adjusted by BSA, WT/BSA, WA/BSA, and LA/BSA, relative volume change -860 HU to -950 HU (RVC-860 to -950) and the expiration to inspiration ratio of mean lung density (MLDE/I)) were compared between the groups. Correlation analysis was employed to assess the relationship between qHRCT parameters and pulmonary function. RESULTS: The small airway asthma phenotype had significantly higher WA%, RVC-860 to -950 and MLDE/I and lower LA/BSA than the healthy control. Additionally, we found moderate to strong correlations between impulse oscillation (IOS) indices and WA6% and WT6/BSA. No significant correlation was found between bronchial parameters and air trapping parameters (p > 0.05). CONCLUSIONS: Combining physiological tests with imaging approaches can lead to better evaluation of small airway disfunction (SAD) in asthmatic patients. Additionally, despite nonexistent airflow obstruction in patients with small airway asthma phenotype, large airway remodeling and small airway structural changes may appear simultaneously in the early stage of disease.

Preparation, physicochemical characterization, and in vitro and in vivo osteogenic evaluation of a bioresorbable, moldable, hydroxyapatite/poly(caprolactone-co-lactide) bone substitute.

Use of bioresorbable artificial bone substitutes is anticipated for bone augmentation in dental implant surgery because they are relatively economical and uniform in quality compared to heterogeneous bone. In this study, a new shapable, rubbery, bioresorbable bone substitute was developed. The material was prepared by ultrasonically dispersing hydroxyapatite (HA) particles throughout a poly (caprolactone-co-lactide) (PCLLA) rubbery matrix. Physiochemical properties of the bone substitute including its composition, deformability, anti-collapse ability, degradation behavior, and in vitro and in vivo osteogenic ability were evaluated. Results revealed that HA/PCLLA, which consists of homogeneously dispersed HA particles and a rubbery matrix composed of PCLLA, possesses a deformable capacity. The result of the mass retention rate of the material indicated an excellent durability in an aqueous environment. Further, the effects of HA/PCLLA on cell functions and bone-regenerated performance were evaluated in vitro and in vivo. The results showed that HA/PCLLA had enhanced proliferative capacity, and ability to undergo osteogenic differentiation and mineralization in vitro. It was also found that HA/PCLLA had an appropriate degradation rate to induce consecutive new bone formation without collapse at the early stage in vivo, as well as the ability to maintain the contour of the bone-grafting area, which is comparable to the deproteinized bovine bone mineral. These results indicated that HA/PCLLA is a promising bioresorbable bone substitute with properties that meet clinical requirements, including deformability, resistance to collapse in an aqueous environment, appropriate early-stage degradation rate, biocompatibility, osteogenic bioactivity and the capacity to regenerate bone tissue with favorable contour.

Micro/Nanostructured Topography on Titanium Orchestrates Dendritic Cell Adhesion and Activation via ß2 Integrin-FAK Signals.

Background and Purpose: In clinical application of dental implants, the functional state of dendritic cells (DCs) has been suggested to have a close relationship with the implant survival rate or speed of osseointegration. Although microscale surfaces have a stable osteogenesis property, they also incline to trigger unfavorable DCs activation and threaten the osseointegration process. Nanoscale structures have an advantage in regulating cell immune response through orchestrating cell adhesion, indicating the potential of hierarchical micro/nanostructured surface in regulation of DCs' activation without sacrificing the advantage of microscale topography. Materials and Methods: Two micro/nanostructures were fabricated based on microscale rough surfaces through anodization or alkali treatment, the sand-blasted and acid-etched (SA) surface served as control. The surface characteristics, in vitro and in vivo DC immune reactions and ß2 integrin-FAK signal expression were systematically investigated. The DC responses to different surface topographies after FAK inhibition were also tested. Results: Both micro/nano-modified surfaces exhibited unique composite structures, with higher hydrophilicity and lower roughness compared to the SA surface. The DCs showed relatively immature functional states with round morphologies and significantly downregulated ß2 integrin-FAK levels on micro/nanostructures. Implant surfaces with micro/nano-topographies also triggered lower levels of DC inflammatory responses than SA surfaces in vivo. The inhibited FAK activation effectively reduced the differences in topography-caused DC activation and narrowed the differences in DC activation among the three groups. Conclusion: Compared to the SA surface with solely micro-scale topography, titanium surfaces with hybrid micro/nano-topographies reduced DC inflammatory response by influencing their adhesion states. This regulatory effect was accompanied by the modulation of ß2 integrin-FAK signal expression. The ß2 integrin-FAK-mediated adhesion plays a critical role in topography-induced DC activation, which represents a potential target for material-cell interaction regulation.

Hand-powered vacuum-driven microfluidic gradient generator for high-throughput antimicrobial susceptibility testing.

The growth of bacterial resistance to antimicrobials is a serious problem attracting much attention nowadays. To prevent the misuse and abuse of antimicrobials, it is important to carry out antibiotic susceptibility testing (AST) before clinical use. However, conventional AST methods are relatively laborious and time-consuming (18-24 h). Here, we present a hand-powered vacuum-driven microfluidic (HVM) device, in which a syringe is used as the only vacuum source for rapid generating concentration gradient of antibiotics in different chambers. The HVM device can be preassembled with various amounts of antibiotics, lyophilized, and stored for ready-to-use. Bacterial samples can be loaded into the HVM device through a simple suction step. With the assistance of Alamar Blue, the AST assay and the minimum inhibitory concentration (MIC) of different antibiotics can be investigated by comparing the growth results of bacteria in different culture chambers. In addition, a parallel HVM device was proposed, in which eight AST assays can be performed simultaneously. The results of MIC of three commonly used antibiotics against E. coli K-12 in our HVM device were consistent with those obtained by traditional method while the detection time was shortened to less than 8 h. We believe that our platform is high-throughput, cost-efficient, easy to use, and suitable for POCT applications.

Co-Mabiq Flies Solo: Light-Driven Markovnikov-Selective C- and N-Alkylation of Indoles and Indazoles without a Cocatalyst.

Indoles and indazoles are common moieties in pharmaceuticals and naturally occurring bioactive compounds. The development of light-driven methods using earth-abundant transition-metal catalysts offers an attractive route for functionalization of such compounds. Herein, we report a visible-light-induced method for the C3- and N-alkylation of indoles and indazoles with styrenes, catalyzed by Co complexes based on the macrocyclic Mabiq ligand (Mabiq = 2-4:6-8-bis(3,3,4,4-tetramethyldihydropyrrolo)-10-15-(2,2'-biquinazolino)-[15]-1,3,5,8,10,14-hexaene-1,3,7,9,11,14-N6). The photochemical behavior of two CoIII catalysts was examined: Co(Mabiq)Cl2 and the newly synthesized Co(MabiqBr)Cl2, which contains the Br-modified ligand. Both complexes undergo visible-light-induced homolysis that is significant to their activity but exhibit differences in reactivity. The alkylation reactions are regioselective, furnishing the alkylated indole and indazole products in a Markovnikov fashion with excellent yields of up to 96% across a broad range of substrates. Notably, in contrast to dual-transition-metal and photoredox-catalyzed cross-coupling reactions, our studies reveal that the Co complex plays a dual role─as a photosensitizer and catalytically active metal center with the Mabiq ligand offering regiocontrol.

The Value of Impulse Oscillometric Parameters and Quantitative HRCT Parameters in Differentiating Asthma-COPD Overlap from COPD.

PURPOSE: To evaluate the value of impulse oscillometry (IOS) and quantitative HRCT parameters for differentiating asthma-COPD overlap (ACO) in COPD patients. PATIENTS AND METHODS: We enrolled 44 controls and 66 COPD patients, divided into the pure COPD group (n=40) and the ACO group (n=26). Spearman correlation analysis was utilized to assess the relationship between the quantitative HRCT and IOS parameters. A binary logistic regression analysis was performed to analyze the associations between the different variables and the risk of ACO. Receiver operating characteristic (ROC) curves were employed to identify the optimal cutoff and assess the diagnostic value of relative volume change -856 HU to -950 HU (RVC-856 to -950), decrease in the resistance from 5 Hz to 20 Hz (R5-R20) and their combination in predicting ACO. Bootstrapping validation was used to evaluate the internal validation. The concordance index (C-index) and calibration plot were calculated to assess the discrimination and calibration of the prediction model. RESULTS: Binary logistic regression analysis indicated that RVC-856 to -950 and the IOS parameters (R5-R20, R5, X5) were independently correlated with a higher risk of developing ACO after adjusting for age, sex, body mass index (BMI), history of smoking, exacerbation and atopy or allergic rhinitis. A correlation analysis showed a good correlation between the pulmonary function parameters and RVC-856 to -950, with a weaker correlation with the % area of low attenuation (LAA%) in ACO patients. Combining RVC-856 to -950 and R5-R20 to predict ACO, the AUC was 0.909, and the optimal cutoff value was >-0.62 for RVC-856 to -950 and >0.09 for R5-R20. Good calibration and favorable discrimination were displayed with a higher C-index. CONCLUSION: More serious small airway impairment exists in ACO patients. The combination of RVC-856 to -950 and R5-R20 could be applied to differentiate ACO from COPD patients.

Advanced technologies in periodontal tissue regeneration based on stem cells: Current status and future perspectives.

Periodontitis is a progressive inflammation disease, the clinical management of which remains a challenge. The traditional management may control periodontal inflammation, but failed to regenerate functional periodontium. This review summarizes the most advancing regenerative techniques regarding stem cell culture and scaffold fabrication, such as cell sheeting, spheroid culture, electrospinning and 3D printing. The applications of different techniques manifest tremendous potential of regenerating the complete and functional periodontium. Albeit promising, new technologies have met with their own drawbacks such as insufficient vascularization and precision, which necessitate deeper modification. Thus, this review also points out the potential perspectives and methods aiming at their disadvantages, illuminating the directions of future researches to successful clinical scenarios.

[A simulation of microwave brain imaging of hemorrhagic stroke detection].

This paper describes a simulation of microwave brain imaging for the detection of hemorrhagic stroke. Firstly, in the research process, the formula of DebyeⅡwas used to study tissues of brain and blood clot so that microwave frequency band was confirmed for imaging. Then a model with electromagnetic characteristics of brain was built on this basis. In addition, an ultra-wideband (UWB) Vivaldi antenna is designed to use for transmitting and receiving microwave signals of widths 1.7 GHz to 4 GHz. Microwave signals were transmitted and received when the antenna revolved around the brain model. Symmetric position de-noising method was used to eliminate the strong background noise signals, and finally confocal imaging method was applied to get brain imaging. Blood clot was distinguished clearly from result of imaging and position error was less than 1 cm.

Fuzzy logic and its application in football team ranking.

Fuzzy set theory and fuzzy logic are a highly suitable and applicable basis for developing knowledge-based systems in physical education for tasks such as the selection for athletes, the evaluation for different training approaches, the team ranking, and the real-time monitoring of sports data. In this paper, we use fuzzy set theory and apply fuzzy clustering analysis in football team ranking. Based on some certain rules, we propose four parameters to calculate fuzzy similar matrix, obtain fuzzy equivalence matrix and the ranking result for our numerical example, T 7, T 3, T 1, T 9, T 10, T 8, T 11, T 12, T 2, T 6, T 5, T 4, and investigate four parameters sensitivity analysis. The study shows that our fuzzy logic method is reliable and stable when the parameters change in certain range.

[Protective effect of Galla chinensis apozema on adriamycin-induced nephropathy in rats].

OBJECTIVE: To study the protective effect of Galla chinensis apozema on adriamycin-induced nephropathy in rats and its mechanism. METHODS: 40 Wistar rats were randomly divided into four groups: normal control group, model group, Galla chinensis apozema high dose group and Galla chinensis apozema low dose group. Adriamycin-induced nephropathy model was built by injecting adriamycin through the tail vein. The high and low dose groups of Galla chinensis apozema were given Galla chinensis apozema for 6 weeks, the other two groups were given normal saline in the same way. Then we observed the 24 h urine protein at each weekend, and determined blood biochemical indexes, SOD and MDA at the last weekend. RESULTS: Galla chinensis decreased urine protein excretion, but increased total cholesterol and triglyceride in plasma, total protein and albumin in serum of model rats. Galla chinensis also decreased the serum levels of MDA but increased the activity of SOD. CONCLUSION: Galla chinensis can protect against adriamycin-induced nephrototoxicity. Its mechanism possibly associated with the reduction of oxidative stress.

Impact of hospital volume on long-term survival after esophageal cancer surgery.

HYPOTHESIS: The improved survival after esophageal cancer surgery in Sweden during recent years may be attributable to the increased centralization of such surgery. DESIGN: Population-based study. SETTING: All Swedish residents undergoing esophageal cancer surgery from January 1, 1987, through December 31, 2000, were identified from the inpatient and cancer registers and were followed up until October 18, 2004, through nationwide registers. Hospital, tumor, and patient characteristics and preoperative oncological treatment were assessed through the registers and histopathological records. PATIENTS: Among 4904 patients with esophageal cancer, 1199 patients (24.4%) who underwent resection constituted the study cohort. Main Outcome Measure Survival rates and hazard ratios (HRs) relative to hospital volume. Low-volume hospitals (LVHs) conducted fewer than 10 esophagectomies annually, while high-volume hospitals (HVHs) conducted 10 or more. Hazard ratios were adjusted for several potential confounders. RESULTS: Thirty-day survival was 96% at HVHs and 91% at LVHs (P = .09). Survival rates 1, 3, and 5 years after surgery at HVHs were nonsignificantly higher (58%, 35%, and 27%, respectively) compared with those at LVHs (55%, 30%, and 24%, respectively). The adjusted HR was nonsignificantly 10% decreased at HVHs (HR, 0.90; 95% confidence interval, 0.79-1.04). In an analysis restricted to 764 patients (64%) without preoperative oncological treatment (in which the tumor stage was also adjusted for), survival was similar at HVHs and at LVHs (HR, 0.99; 95% confidence interval, 0.84-1.18). CONCLUSIONS: This study revealed no effect of hospital volume on long-term survival after esophageal cancer surgery. Tumor biology apparently has a greater effect on the chances of long-term survival than hospital volume.

Survival after neoadjuvant therapy compared with surgery alone for resectable esophageal cancer in a population-based study.

BACKGROUND: Esophagectomy remains the standard treatment for resectable esophageal cancer, but the cure rate is low. Neoadjuvant therapy has been tried in attempts to prolong survival and reduce tumor recurrence. The aim of this study was to assess the surgical outcomes with and without neoadjuvant treatment for resectable esophageal cancer in a population-based setting. METHODS: All 1,155 patients treated with esophagectomy for esophageal cancer in Sweden in 1987 through 2000 with or without neoadjuvant therapy were identified and followed up in nationwide registers up to 18 October 2004. Tumor characteristics and response to neoadjuvant treatment were obtained from histopathological reports. Hazard ratios (HRs) with 95% confidence intervals (CIs) adjusted for potential confounding factors were calculated by a Cox proportional hazards regression model. RESULTS: Overall survival was similar in the groups with and without neoadjuvant therapy (adjusted HR 0.99, 95% CI 0.86-1.16). The 3-year survival rates were 34.6% and 32.0%, respectively. Survival was better among the 27.6% of the neoadjuvant group with a complete histopathological response (HR 0.71, 95% CI 0.53-0.94) compared with the surgery only group. Patients without complete response to neoadjuvant therapy had seemingly poorer survival (HR 1.10, 95% CI 0.94-1.29). CONCLUSIONS: Surgical outcomes with and without neoadjuvant therapy were equivalent. Only patients with a complete histopathological response after neoadjuvant treatment had better survival.

Survival after surgery for oesophageal cancer: a population-based study.

BACKGROUND: Oesophageal resection alone remains the only potentially curative treatment for oesophageal cancer. However, few population-based studies of survival after surgery have been done. We aimed to assess whether survival after surgery for oesophageal cancer has improved since 1987. METHODS: We identified from the nationwide cancer register 764 patients in Sweden who had undergone resection alone for oesophageal cancer between Jan 1, 1987 and Dec 31, 2000, and followed up these individuals until Oct 18, 2004, through linkage to the nationwide registers of death, emigration, and total population. Tumour stage, location, and histology were recorded by review of histopathological reports. Hazard ratios, with 95% CI, for death adjusted for age; sex; comorbidity; and tumour stage, location, and histology were calculated by Cox proportional-hazards regression model. FINDINGS: Overall, the proportion of deaths up to 30 days after surgery decreased from 10.1% in 1987-91 to 4.9% in 1997-2000. Increased long-term survival was associated with low tumour stage. Survival was 46.5% 1 year after surgery, 24.1% 3 years after surgery, and 19.7% 5 years after surgery in 1987-91; 58.4%, 31.7%, and 24.9%, respectively, in 1992-96; and 61.7%, 39.9%, and 30.7%, respectively, in 1997-2000. The adjusted hazard ratio for death was 0.76 (95% CI 0.62-0.92) in 1992-96 and 0.57 (0.45-0.71) in 1997-2000. INTERPRETATION: Survival after surgery for oesophageal cancer has improved substantially since 1987. Because this improvement cannot be explained by a change in patient characteristics or tumour characteristics, it is probably a result of better surgery.