Protective effect of the mixture of Lactiplantibacillus plantarum KC3 and Leonurus Japonicas Houtt extract on respiratory disorders.

Air pollutants, such as particulate matter (PM) and diesel exhaust particles (DEP), are associated with respiratory diseases. Therefore, preventive and therapeutic strategies against PM-and DEP (PM10D)-induced respiratory diseases are needed. Herein, we evaluate the protective effects of a mixture of Lactiplantibacillus plantarum KC3 and Leonurus Japonicas Houtt (LJH) extract against airway inflammation associated with exposure to PM10D. To determine the anti-inflammatory effects of the LJH extract, reactive oxygen species (ROS) production and the expression of inflammatory pathways were determined in PM10-induced MH-S cells. For the respiratory protective effects, BALB/c mice were exposed to PM10D via intranasal injection, and a mixture of L. plantarum KC3 and LJH extract was administered orally for 12 days. LJH extract inhibited ROS production and the phosphorylation of downstream factors of NF-κB in PM10-stimulated MH-S cells. The mixture of L. plantarum KC3 and LJH repressed the infiltration of neutrophils, reduced the immune cells number, and suppressed the proinflammatory mediators and cyclooxygenase (COX)-2 expressions in PM10D-induced airway inflammation with reduced phosphorylation of downstream factors of NF-κB. In addition, these effects were not observed in an alveolar macrophage depleted PM10D-induced mouse model using clodronate liposomes. The extract mixture also regulated gut microbiota in feces and upregulated the mRNA expression of Foxp3, transforming growth factor (TGF)-ß1, and interleukin (IL)-10 in the colon. The L. plantarum KC3 and LJH extract mixture may inhibit alveolar macrophage- and neutrophil-mediated inflammatory responses and regulate gut microbiota and immune response in PM10D-induced airway inflammation, suggesting it is a potential remedy to prevent and cure airway inflammation and respiratory disorders.

Lateral Size Effect of Graphene Oxide on Its Surface-Enhanced Raman Scattering Property.

The lateral size effect of graphene oxide (GO) on surfaced-enhanced Raman scattering (SERS) property is systematically investigated by using size-fractionalized GO. For the size fractionalization without changes of chemical structure, large-sized GO (LGO) and small-sized GO (SGO) are separated from the as-synthesized GO (AGO) by centrifugation and membrane filtration, respectively. The size-fractionalized GO sheets are immobilized on a solid substrate for the parallel comparison of their SERS property. As a result, we find that LGO shows considerably higher SERS property than SGO for typical Raman probes such as rhodamine 6G and crystal violet. Furthermore, the lateral size effect of GO derivatives is consistently observed when they are hybridized with plasmonic silver nanoparticles. These results indicate that LGO is superior to AGO and SGO as a SERS platform, and it is also quantitatively confirmed by calculating their enhancement factor.

Cyclase-associated protein 1 is a binding partner of proprotein convertase subtilisin/kexin type-9 and is required for the degradation of low-density lipoprotein receptors by proprotein convertase subtilisin/kexin type-9.

AIMS: Proprotein convertase subtilisin/kexin type-9 (PCSK9), a molecular determinant of low-density lipoprotein (LDL) receptor (LDLR) fate, has emerged as a promising therapeutic target for atherosclerotic cardiovascular diseases. However, the precise mechanism by which PCSK9 regulates the internalization and lysosomal degradation of LDLR is unknown. Recently, we identified adenylyl cyclase-associated protein 1 (CAP1) as a receptor for human resistin whose globular C-terminus is structurally similar to the C-terminal cysteine-rich domain (CRD) of PCSK9. Herein, we investigated the role of CAP1 in PCSK9-mediated lysosomal degradation of LDLR and plasma LDL cholesterol (LDL-C) levels. METHODS AND RESULTS: The direct binding between PCSK9 and CAP1 was confirmed by immunoprecipitation assay, far-western blot, biomolecular fluorescence complementation, and surface plasmon resonance assay. Fine mapping revealed that the CRD of PCSK9 binds with the Src homology 3 binding domain (SH3BD) of CAP1. Two loss-of-function polymorphisms found in human PCSK9 (S668R and G670E in CRD) were attributed to a defective interaction with CAP1. siRNA against CAP1 reduced the PCSK9-mediated degradation of LDLR in vitro. We generated CAP1 knock-out mice and found that the viable heterozygous CAP1 knock-out mice had higher protein levels of LDLR and lower LDL-C levels in the liver and plasma, respectively, than the control mice. Mechanistic analysis revealed that PCSK9-induced endocytosis and lysosomal degradation of LDLR were mediated by caveolin but not by clathrin, and they were dependent on binding between CAP1 and caveolin-1. CONCLUSION: We identified CAP1 as a new binding partner of PCSK9 and a key mediator of caveolae-dependent endocytosis and lysosomal degradation of LDLR.

Usefulness of the Hematopoietic Stem Cell Donor Pool as a Source of HLA-Homozygous Induced Pluripotent Stem Cells for Haplobanking: Combined Analysis of the Cord Blood Inventory and Bone Marrow Donor Registry.

Induced pluripotent stem cells (iPSCs) have opened up unprecedented opportunities for novel therapeutic options for precision medicine. Hematopoietic stem cell (HSC) donor pools with previously determined HLA types may be ideal sources for iPSC production. Based on the HLA distribution of cryopreserved cord blood units (CBUs) and registered bone marrow (BM) donors, we estimated how much of the Korean population could be covered by HLA-homozygous iPSCs. We analyzed a total of 143,866 Korean HSC donors (27,904 CBUs and 115,962 BM donors). Each donor sample was typed for the HLA-A, -B, and -DRB1 alleles at low to intermediate resolution by DNA-based molecular techniques: PCR sequence-specific oligonucleotide (PCR-SSOP), PCR with sequence-specific primers (PCR-SSP) and PCR with sequence-based typing (PCR-SBT). We also identified individuals possessing homozygous HLA haplotypes by direct counting. The matching probabilities for zero-mismatch transplantation were calculated for 143,866 Koreans and 50 million potential Korean patients. Among the HSC donor pool, 17 HLA-A alleles, 41 HLA-B alleles, and 13 HLA-DRB1 alleles, as well as 128 homozygous HLA-A-B-DRB1 haplotypes, were identified at serologic equivalents, and those haplotypes cumulatively matched 93.20% of the 143,866 Korean donors as zero HLA-mismatch iPSC sources. Among the combinations of 2,056 haplotypes with frequencies ≥ 0.001% in a population of 50 million, those 128 homozygous haplotypes can provide 93.65% coverage for potential Korean recipients. Haplobanking of a reasonable number of HLA-A, -B, and -DRB1 homozygous iPSC lines derived from CBUs and cells of registered BM donors may be an efficient option for allogenic iPSC therapy.

Inside-Out Repair of the Meniscus in Concomitant Anterior Cruciate Ligament Reconstruction: Absorbable Versus Nonabsorbable Sutures.

PURPOSE: To compare the clinical and magnetic resonance imaging (MRI) outcomes of meniscal repair using absorbable versus nonabsorbable sutures in patients undergoing concomitant anterior cruciate ligament reconstruction. METHODS: Data of 142 patients who underwent meniscal repair with concomitant anterior cruciate ligament reconstruction using either absorbable or nonabsorbable sutures for longitudinal meniscal tear were retrospectively reviewed. Inside-out suture technique was used for all meniscal repairs. Weight bearing and flexion (>90°) were allowed after 6 weeks postoperatively. Clinical evaluations were assessed by the International Knee Documentation Committee subjective score, Lysholm score, and Tegner activity score preoperatively and at 2-year follow-up. MRI outcomes at 1-year follow-up were compared to identify the successful healing (complete or partial healing) rate and incidence of additional meniscal tears. Subgroup analysis was performed to evaluate the results of medial or lateral meniscus. RESULTS: Eighty patients underwent meniscal repair using absorbable sutures (mean age, 26.3 ± 11.9 years) and 62 patients with nonabsorbable sutures (mean age, 27.2 ± 10.0 years). There were no differences in zone and length of meniscal tears and stability tests between the groups. At a 2-year follow-up, all clinical scores had improved in both groups but did not differ significantly between the groups. Successful healing rate based on 1-year postoperative MRI was not significantly different between the absorbable and nonabsorbable sutures (93.7% vs 96.8%, P = .469). However, the absorbable sutures showed a lower additional tear incidence than the nonabsorbable sutures (2.5% vs 9.6%, P = .031). Subgroup analysis showed that the successful healing rate was not significantly different between the suture materials in both the medial and lateral menisci. CONCLUSIONS: The use of absorbable sutures leads to comparable healing rates to and lower incidence of additional tears than nonabsorbable sutures in patients undergoing meniscal repair with anterior cruciate ligament reconstruction. LEVEL OF EVIDENCE: Level III, retrospective comparative therapeutic trial.

Influence of Posterior Tibial Slope on Clinical Outcomes and Survivorship After Anterior Cruciate Ligament Reconstruction Using Hamstring Autografts: A Minimum of 10-Year Follow-Up.

PURPOSE: To investigate the influence of medial and lateral posterior tibial slope (PTS) on long-term clinical outcomes and survivorship after anterior cruciate ligament (ACL) reconstruction using hamstring autografts. METHODS: A total of 232 patients (mean age, 28.2 ± 8.9 years) who underwent primary ACL reconstruction from October 2002 to July 2007 were retrospectively reviewed. Patients with multiple ligament reconstruction, total meniscectomy, contralateral knee surgery before ACL reconstruction, open growth plate, and less than 10-year follow-up were excluded in the study. The medial and lateral PTS were measured from preoperative magnetic resonance imaging. Based on Li et al.'s previous study, the patients were divided into 2 groups according to their medial PTS (≤5.6° vs >5.6°) and lateral PTS (≤3.8° vs >3.8°), respectively. Clinical outcomes (clinical scores, stability tests and failure rate) were compared between the groups at the last follow-up. Furthermore, survival analysis was performed using the Kaplan-Meier method. RESULTS: All clinical scores (International Knee Documentation Committee subjective, Lysholm, and Tegner activity scores) and stability tests (physical examinations and side-to-side difference in Telos stress radiographs) were insignificantly different between the 2 groups classified based on medial or lateral PTS. However, the failure rate was significantly higher in patients with medial PTS >5.6° (16.1% vs 5.1%, P = .01) or lateral PTS >3.8° (14.5% vs 4.7%; P = .01). The odds ratios of graft failure due to increased medial and lateral PTS were 3.18 (95% confidence interval, 1.22-8.28; P = .02) and 3.43 (95% confidence interval, 1.29-9.09; P = .01), respectively. In addition, the 10-year survivorship was significantly lower in patients with medial PTS >5.6° (83.9% vs 94.9%, P = .01) or lateral PTS >3.8° (85.5% vs 96.0%; P = .01). CONCLUSIONS: Increased medial (>5.6°) and lateral (>3.8°) PTS were associated with higher failure rate and lower survivorship at a minimum of 10-year follow-up after primary ACL reconstruction using hamstring autografts. LEVEL OF EVIDENCE: Level III, retrospective comparative trial.

Re-revision anterior cruciate ligament reconstruction showed more laxity than revision anterior cruciate ligament reconstruction at a minimum 2-year follow-up.

PURPOSE: This study aimed to compare patient demographics, associated lesions (concurrent meniscal and chondral injuries), and clinical outcomes between revision and re-revision anterior cruciate ligament reconstructions. METHODS: Patients who underwent revision or re-revision anterior cruciate ligament reconstruction between 2008 and 2016 with a minimum 2-year follow-up were retrospectively evaluated. Detailed patient demographic data, radiographic preoperative tunnel diameters, posterior tibia slope, and concurrent meniscal and chondral lesion were reviewed. Clinical scores and laxity tests' results were compared between the groups at the last follow-up. RESULTS: Eighty-two patients (mean age, 33.8 ± 9.9 years; revision group, n = 62; re-revision group, n = 20) were included. The re-revision group showed a higher grade for preoperative arthritis (P < 0.001); more severe preoperative bone defects of the femoral (13.8 ± 2.6 vs 11.7 ± 2.7 mm, P = 0.004) and tibial tunnels (14.6 ± 2.4 vs 13.0 ± 2.3 mm, P = 0.010); and a higher prevalence of subtotal medial meniscectomy (P = 0.008) and chondral defects of the medial (P = 0.006) and lateral femoral condyles (P < 0.001), patella (P = 0.040), and trochlea (P = 0.036). At the final follow-up, the clinical scores did not differ significantly between the groups. However, the re-revision group showed more instability in the anterior drawer (P = 0.001), Lachman (P < 0.001), and pivot-shift (P < 0.001) tests, while a side-to-side difference was observed on the Telos stress radiographs (7.1 ± 4.7 vs 4.9 ± 3.7 mm, P = 0.038). CONCLUSION: These findings showed that the patients who underwent re-revision had poor prognostic factors as compared with those who underwent revision anterior cruciate ligament reconstruction. Although the clinical scores did not differ significantly between the groups, the re-revision group showed more laxity at the 2-year follow-up. LEVEL OF EVIDENCE: Cohort study; IV.

A two-portal technique using a flexible reamer system is a safe and effective method for transportal anterior cruciate ligament reconstruction.

INTRODUCTION: A flexible reamer system (FRS) for transportal anterior cruciate ligament reconstruction (ACLR) has been developed to overcome the technical challenges of a rigid reamer system. The purpose of this study was to investigate the safety and effectiveness of the two-portal technique using an FRS by evaluating femoral tunnel geometry. METHODS: This study included 30 patients (mean age 30 ± 12.1) who underwent transportal single-bundle ACLR. Operations were performed with the two-portal technique using an FRS. Three-dimensional computed tomography was performed for all patients 2 days after the operation. The femoral tunnel position, femoral graft bending angle, femoral tunnel length, and posterior wall breakage were evaluated. These radiologic outcomes were compared to previous literature-reported outcomes. RESULTS: The mean distances (measured as a percentage) from the posterior wall and the intercondylar notch roof to the femoral tunnel center were 29.6 ± 5.5% and 20.1 ± 6.7%, respectively. The femoral graft bending angle (108.4° ± 6.9°) was similar to that associated with the traditional transportal technique using a rigid reamer system, but it was less acute than that associated with the three-portal technique using an FRS. The femoral tunnel length (32.8 ± 4.5 mm) was also similar to the results of the traditional transportal technique using a rigid reamer system, but it was shorter than that of three-portal technique using an FRS. The prevalence of posterior wall breakage was as low as the reported outcomes of the outside-in technique (2 cases, 6.6%). CONCLUSIONS: The two-portal technique for transportal ACLR using an FRS can achieve comparable femoral graft bending angle and femoral tunnel length compared with the conventional three-portal technique using the rigid reamer system and had a low risk of posterior wall breakage. Therefore, the two-portal technique using the FRS can be considered a safe and effective method for transportal ACLR. LEVEL OF EVIDENCE: Retrospective case series; level of evidence, 4.

Is Lateral Posterior Tibial Slope Correlated With Clinical Outcomes of Lateral Meniscus Allograft Transplantation?

PURPOSE: To investigate (1) the correlation between lateral posterior tibial slope (PTS) and clinical outcomes of lateral meniscus allograft transplantation (MAT) and (2) the difference of lateral PTS between the extrusion and nonextrusion groups or between the failure and nonfailure groups in lateral MAT. METHODS: Between January 2001 and February 2016, we retrospectively evaluated 61 patients (mean age, 29.1 ± 12.2 years) who underwent postoperative magnetic resonance imaging (MRI) and were followed for a minimum of 2 years after primary lateral MAT. The lateral PTS and graft extrusion in the coronal and sagittal planes were assessed by using MRI performed at 1 year postoperatively. Clinical scores and graft failure were evaluated at the last follow-up visit. The correlation between lateral PTS and clinical outcomes (clinical scores, graft extrusion) was analyzed. Lateral PTS was compared between the extrusion and nonextrusion groups and between the failure and nonfailure groups. RESULTS: Mean lateral PTS on MRI was 6.6° ± 3.1° (range, 0.8° to 15.7°). A significant correlation was not identified between lateral PTS and clinical outcomes (clinical scores, graft extrusion in the coronal and sagittal planes). A significant difference in lateral PTS was not identified between the extrusion and nonextrusion groups in the coronal (6.2° ± 2.5° vs 7.0° ± 3.4°, P = .400) and sagittal (anterior horn, 6.5° ± 2.3° vs 6.7° ± 3.7°, P = .988; posterior horn, 6.8° ± 3.5° vs 6.5° ± 2.7°, P = .771) planes. Moreover, a significant difference was not identified between the failure and nonfailure groups (7.5° ± 3.3° vs 6.4° ± 3.0°, P = .388). CONCLUSIONS: A significant correlation between lateral PTS and clinical or radiologic outcomes of lateral MAT was not identified. LEVEL OF EVIDENCE: Level IV, therapeutic case series with subgroup analysis.

Clinical outcomes and survival rate of autologous chondrocyte implantation with and without concomitant meniscus allograft transplantation: 10- to 15-year follow-up study.

PURPOSE: The purpose of this study was to compare the clinical outcomes and survival rate of autologous chondrocyte implantation (ACI) with or without concomitant meniscus allograft transplantation (MAT). METHODS: Patients who underwent ACI of the medial or lateral femoral condyle with or without concomitant MAT were retrospectively reviewed. There were 14 patients (mean age, 31.2 ± 9.9 years) who underwent isolated ACI and 19 patients who underwent ACI with concomitant MAT (mean age, 34.8 ± 8.4 years). The International Knee Documentation Committee (IKDC) subjective score, Lysholm score, Tegner activity score, and 10- to 15-year survival rate were compared between groups. RESULTS: All clinical scores showed significant improvement postoperatively in both groups. At final follow-up, the IKDC subjective score was superior in isolated ACI (75.8 ± 18.4) compared to ACI with MAT (61.0 ± 16.6, p = 0.024). The Lysholm score was also higher in isolated ACI (77.5 ± 19.1) than ACI with MAT (62.5 ± 18.1, p = 0.029). The Tegner activity score did not differ between treatments (isolated ACI, 5.3 ± 1.1; ACI with MAT, 4.5 ± 1.3; p = 0.072). The 15-year survival rate for isolated ACI was higher than that of ACI with concomitant MAT (69.6% vs 50.2%), but this difference was not statistically significant (p = 0.19). CONCLUSIONS: ACI with concomitant MAT did not restore clinical outcomes as much as isolated ACI. There was a trend for the long-term survival rate to be greater in isolated ACI than ACI with MAT. These results should be considered in planning for the treatment of focal chondral defect with meniscus deficiency. LEVEL OF STUDY: Retrospective comparative trial; level of evidence, 3.

Direct conversion of adult skin fibroblasts to endothelial cells by defined factors.

BACKGROUND: Cell-based therapies to augment endothelial cells (ECs) hold great therapeutic promise. Here, we report a novel approach to generate functional ECs directly from adult fibroblasts. METHODS AND RESULTS: Eleven candidate genes that are key regulators of endothelial development were selected. Green fluorescent protein (GFP)-negative skin fibroblasts were prepared from Tie2-GFP mice and infected with lentiviruses allowing simultaneous overexpression of all 11 factors. Tie2-GFP(+) cells (0.9%), representing Tie2 gene activation, were detected by flow cytometry. Serial stepwise screening revealed 5 key factors (Foxo1, Er71, Klf2, Tal1, and Lmo2) that were required for efficient reprogramming of skin fibroblasts into Tie2-GFP(+) cells (4%). This reprogramming strategy did not involve pluripotency induction because neither Oct4 nor Nanog was expressed after 5 key factor transduction. Tie2-GFP(+) cells were isolated using fluorescence-activated cell sorting and designated as induced ECs (iECs). iECs exhibited endothelium-like cobblestone morphology and expressed EC molecular markers. iECs possessed endothelial functions such as Bandeiraea simplicifolia-1 lectin binding, acetylated low-density lipoprotein uptake, capillary formation on Matrigel, and nitric oxide production. The epigenetic profile of iECs was similar to that of authentic ECs because the promoters of VE-cadherin and Tie2 genes were demethylated. mRNA profiling showed clustering of iECs with authentic ECs and highly enriched endothelial genes in iECs. In a murine model of hind-limb ischemia, iEC implantation increased capillary density and enhanced limb perfusion, demonstrating the in vivo viability and functionality of iECs. CONCLUSIONS: We demonstrated the first direct conversion of adult fibroblasts to functional ECs. These results suggest a novel therapeutic modality for cell therapy in ischemic vascular disease.

Peroxisome proliferator-activated receptor-δ activates endothelial progenitor cells to induce angio-myogenesis through matrix metallo-proteinase-9-mediated insulin-like growth factor-1 paracrine networks.

AIMS: The roles of peroxisome proliferator-activated receptor (PPAR)-δ in vascular biology are mainly unknown. We investigated the effects of PPAR-δ activation on the paracrine networks between endothelial progenitor cells (EPCs) and endothelial cells (ECs)/skeletal muscle. METHODS AND RESULTS: Treatment of EPCs with GW501516, a PPAR-δ agonist, induced specifically matrix metallo-proteinase (MMP)-9 by direct transcriptional activation. Subsequently, this increased-MMP-9 broke down insulin-like growth factor-binding protein (IGFBP)-3, resulting in IGF-1 receptor (IGF-1R) activation in surrounding target cells. Treatment of conditioned medium from GW501516-stimulated EPCs enhanced the number and functions of human umbilical vein ECs and C2C12 myoblasts via MMP-9-mediated IGF-1R activation. Systemic administration of GW501516 in mice increased MMP-9 expression in EPCs, and augmented IGFBP-3 degradation in serum. In a mouse hindlimb ischaemia model, systemic treatment of GW501516 or local transplantation of GW501516-treated EPCs induced IGF-1R phosphorylation in ECs and skeletal muscle in the ischaemic limbs, leading to augmented angiogenesis and skeletal muscle regeneration. It also enhanced wound healing with increased angiogenesis in a mouse skin punch wound model. These pro-angiogenic and muscle-regenerating effects were abolished by MMP-9 knock-out. CONCLUSION: Our results suggest that PPAR-δ is a crucial modulator of angio-myogenesis via the paracrine effects of EPCs, and its agonist is a good candidate as a therapeutic drug for patients with peripheral vascular diseases.

Bio-inspired interfacial chemistry for the fabrication of a robust and functional graphene oxide composite film.

A strong and functional artificial nacre film is developed by using polyethyleneimine-functionalized GO (PEI-GO) and pyrogallol (PG) inspired by insect exoskeleton sclerotization. PEI-GO is macroscopically assembled into the laminated films and then reacted with PG under the optimized condition for their efficient cross-linking through Schiff-base reactions. The internal structure and physicochemical properties of PG-treated PEI-GO (PG@PEI-GO) films are systematically explored with various analytical tools. The optimized PG@PEI-GO films exhibit excellent tensile strength, modulus, and toughness of 216.0 ± 12.9 MPa, 17.0 ± 1.1 GPa, and 2192 ± 538.5 kJ m-3 which are 2.7, 2.8, and 2.3-fold higher than those of GO films, respectively. Furthermore, silver nanoparticles (AgNPs) are densely immobilized on the PG@PEI-GO films harnessing their abundant amine groups, and the AgNPs immobilized PG@PEI-GO films exhibit a high catalytic activity in the conversion of 4-nitrophenol (4-NP) to 4-aminophenol (4-AP) with maintaining structural integrity. Based on the results, we demonstrate that the rational design of interfaces, inspired by natural materials, is an efficient approach to achieving strong and functional GO laminated composite films.

Durable silver nanowire transparent electrodes enabled by biorenewable nanocoating using chitin and cellulose nanofibers for flexible electronics.

The protection of silver nanowire (AgNW) networks is crucial for enhancing their durability and applicability to flexible electronics. In this study, we present a sustainable and efficient strategy to protect AgNW-based flexible transparent electrodes (FTEs) using a layer-by-layer (LBL) assembly of biorenewable chitin and cellulose nanofibers (Chi and Cell). These uniform LBL-assembled thin films were successfully fabricated on AgNW FTEs due to their opposite surface charges. The resulting (Chi/Cell)n bilayers, where n is the number of bilayers, did not degrade the optoelectrical properties of AgNW FTEs and significantly enhanced their stability under various harsh conditions. The optimized (Chi/Cell)10@Al-AgNW FTEs exhibited comprehensive stability against UV/O3 treatment for 40 min, thermal treatment at 250 °C for 350 min, Na2S (1%), HCl (10%), and NH3 (30%) treatments for 3, 30, and 105 min, respectively, sonication for 300 min, and 10 000 cycles of bending test. Therefore, the (Chi/Cell)10@Al-AgNW FTEs were successfully applied to transparent heaters (TH) and pressure sensors with remarkably improved applicability, durability, and performance compared to pristine AgNW FTEs, providing a reassuring solution to the stability issues of AgNW-based FTEs.

Resistin Regulates Inflammation and Insulin Resistance in Humans via the Endocannabinoid System.

Resistin plays an important role in the pathophysiology of obesity-mediated insulin resistance in mice. However, the biology of resistin in humans is quite different from that in rodents. Therefore, the association between resistin and insulin resistance remains unclear in humans. Here, we tested whether and how the endocannabinoid system (ECS) control circulating peripheral blood mononuclear cells (PBMCs) that produce resistin and infiltrate into the adipose tissue, heart, skeletal muscle, and liver, resulting in inflammation and insulin resistance. Using human PBMCs, we investigate whether the ECS is connected to human resistin. To test whether the ECS regulates inflammation and insulin resistance in vivo, we used 2 animal models such as "humanized" nonobese diabetic/Shi-severe combined immunodeficient interleukin-2Rγ (null) (NOG) mice and "humanized" resistin mouse models, which mimic human body. In human atheromatous plaques, cannabinoid 1 receptor (CB1R)-positive macrophage was colocalized with the resistin expression. In addition, resistin was exclusively expressed in the sorted CB1R-positive cells from human PBMCs. In CB1R-positive cells, endocannabinoid ligands induced resistin expression via the p38-Sp1 pathway. In both mouse models, a high-fat diet increased the accumulation of endocannabinoid ligands in adipose tissue, which recruited the CB1R-positive cells that secrete resistin, leading to adipose tissue inflammation and insulin resistance. This phenomenon was suppressed by CB1R blockade or in resistin knockout mice. Interestingly, this process was accompanied by mitochondrial change that was induced by resistin treatment. These results provide important insights into the ECS-resistin axis, leading to the development of metabolic diseases. Therefore, the regulation of resistin via the CB1R could be a potential therapeutic strategy for cardiometabolic diseases.

Effect of a Combination of Lactiplantibacillus plantarum KC3 and Leonurus japonicus Extracts in Respiratory Discomfort: A Randomized, Double-Blind, Placebo-Controlled Trial.

The increased global prevalence of chronic respiratory diseases in recent years has caused a substantial public health burden. Lactiplantibacillus plantarum KC3 and Leonurus japonicus Houtt. (LJH) extracts can alleviate respiratory symptoms and improve lung function in vitro and in vivo. However, the clinical efficacy and safety profile of this combination in patients with respiratory diseases remain unclear. Therefore, this multicenter, randomized, double-blind, placebo-controlled clinical trial aimed to evaluate the efficacy and safety of L. plantarum KC3 and LJH extracts in adults with respiratory discomfort. This mixture was termed 'CKDB-315'. Participants, randomly assigned to the CKDB-315 or placebo groups, were treated for 12 weeks. Assessments included the St. George's Respiratory Questionnaire (SGRQ) and the Chronic Obstructive Pulmonary Disease Assessment Test (CAT). The CKDB-315 group showed considerably improved SGRQ and CAT scores compared with the placebo group. Secondary outcomes, including dyspnea, pulmonary function, total antioxidant status, and inflammatory cytokine levels, were consistent with the primary outcomes. Exploratory analyses of the gut microbiota and short-chain fatty acid contents revealed the potential mechanisms underlying the effects of CKDB-315. Finally, safety analysis indicated that CKDB-315 was well tolerated and caused few adverse events. Our findings indicate that CKDB-315 is a promising therapeutic option for respiratory discomfort in adults.

Mechanism of edge restenosis after drug-eluting stent implantation. Angulation at the edge and mechanical properties of the stent.

BACKGROUND: Edge restenosis is not an unusual finding after implantation of drug-eluting stents (DES). We hypothesized that mechanical stress imposed on the stent edge would cause vessel wall injury and inflammation, which may consequently lead to edge restenosis. METHODS AND RESULTS: In total, 1,496 patients were implanted with a sirolimus-eluting stent (SES), paclitaxel-eluting stent (PES) or zotarolimus-eluting stent (ZES) in Seoul National University Hospital between 2007 and 2009. Binary restenosis occurred in 161 lesions in 119 patients. We retrospectively compared the 3 DES with regard to the percentage of edge stent restenosis among all cases of restenosis. We also evaluated the maximal, minimal, and Δ (maximal angle-minimal angle) angles. The percentage of edge restenosis was higher for SES than for ZES (37.5% vs. 16.7%, P=0.017). Maximal angle at the proximal edge was 64.82°±33.46° for 26 stents with proximal edge restenosis compared with 31.84°±31.51° for 89 stents without proximal edge restenosis (P=0.001). The Δ angle was also significantly different between the 2 groups (14.81°±15.98° vs. 7.60°±8.86°, P=0.035). Similar findings were observed for distal edge restenosis. Both the maximal angle (39.09°±21.04° vs. 22.71°±22.83°, P=0.010) and Δ angle (20.23°±15.39° vs. 9.18°±9.66°, P=0.016) at the distal edge were significantly different between the 2 groups. CONCLUSIONS: Physical stress determined by angulation at the stent edge segment and biomechanical properties of the DES can be considered as one of the plausible mechanisms for edge stent restenosis.

Secondary sphere formation enhances the functionality of cardiac progenitor cells.

Loss of cardiomyocytes impairs cardiac function after myocardial infarction (MI). Recent studies suggest that cardiac stem/progenitor cells could repair the damaged heart. However, cardiac progenitor cells are difficult to maintain in terms of purity and multipotency when propagated in two-dimensional culture systems. Here, we investigated a new strategy that enhances potency and enriches progenitor cells. We applied the repeated sphere formation strategy (cardiac explant → primary cardiosphere (CS) formation → sphere-derived cells (SDCs) in adherent culture condition → secondary CS formation by three-dimensional culture). Cells in secondary CS showed higher differentiation potentials than SDCs. When transplanted into the infarcted myocardium, secondary CSs engrafted robustly, improved left ventricular (LV) dysfunction, and reduced infarct sizes more than SDCs did. In addition to the cardiovascular differentiation of transplanted secondary CSs, robust vascular endothelial growth factor (VEGF) synthesis and secretion enhanced neovascularization in the infarcted myocardium. Microarray pathway analysis and blocking experiments using E-selectin knock-out hearts, specific chemicals, and small interfering RNAs (siRNAs) for each pathway revealed that E-selectin was indispensable to sphere initiation and ERK/Sp1/VEGF autoparacrine loop was responsible for sphere maturation. These results provide a simple strategy for enhancing cellular potency for cardiac repair. Furthermore, this strategy may be implemented to other types of stem/progenitor cell-based therapy.

BORIS/CTCFL-mediated transcriptional regulation of the hTERT telomerase gene in testicular and ovarian tumor cells.

Telomerase activity, not detectable in somatic cells but frequently activated during carcinogenesis, confers immortality to tumors. Mechanisms governing expression of the catalytic subunit hTERT, the limiting factor for telomerase activity, still remain unclear. We previously proposed a model in which the binding of the transcription factor CTCF to the two first exons of hTERT results in transcriptional inhibition in normal cells. This inhibition is abrogated, however, by methylation of CTCF binding sites in 85% of tumors. Here, we showed that hTERT was unmethylated in testicular and ovarian tumors and in derivative cell lines. We demonstrated that CTCF and its paralogue, BORIS/CTCFL, were both present in the nucleus of the same cancer cells and bound to the first exon of hTERT in vivo. Moreover, exogenous BORIS expression in normal BORIS-negative cells was sufficient to activate hTERT transcription with an increasing number of cell passages. Thus, expression of BORIS was sufficient to allow hTERT transcription in normal cells and to counteract the inhibitory effect of CTCF in testicular and ovarian tumor cells. These results define an important contribution of BORIS to immortalization during tumorigenesis.