Humanized PD-1 Knock-in Mice Reveal Nivolumab's Inhibitory Effects on Glioblastoma Tumor Progression In Vivo.

BACKGROUND/AIM: Immunotherapy has been considered a promising approach for brain tumor treatment since the discovery of the brain lymphatic system. Glioblastoma (GBM), the most aggressive type of brain tumor, is associated with poor prognosis and a lack of effective treatment options. MATERIALS AND METHODS: To test the efficacy of human anti-PD-1, we used a humanized PD-1 knock-in mouse to establish an orthotopic GBM-bearing model. RESULTS: Nivolumab, a human anti-PD-1, effectively inhibited tumor growth, increased the survival rate of mice, enhanced the accumulation and function of cytotoxic T cells, reduced the accumulation and function of immunosuppressive cells and their related factors, and did not induce tissue damage or biochemical changes. The treatment also induced the accumulation and activation of CD8+ cytotoxic T cells, while reducing the accumulation and activation of myeloid-derived suppressor cells, regulatory T cells, and tumor-associated macrophages in the immune microenvironment. CONCLUSION: Nivolumab has the potential to be a treatment for GBM.

STEMI patients receiving percutaneous coronary intervention for a culprit lesion with coronary artery bifurcation-efficacy and safety of the jailed semi-inflated balloon.

Background: We aimed to evaluate the efficacy and safety of the 'jailed semi-inflated balloon technique' (JSIBT) for side branch (SB) protection in STEMI patients with a culprit lesion involving a coronary artery bifurcation while undergoing emergent percutaneous coronary intervention (PCI). Methods: We treated between Jan, 2011 and Jun, 2020, a total of 264 STEMI patients with a culprit lesion that involved a coronary artery bifurcation using primary PCI. In 30 patients, SB was protected by JSIBT (the JSIBT group). In 234 patients, SB was either protected or not protected by a placed wire (the non-JSIBT group). Results: In both groups, after PCI procedure, TIMI flows of main vessel (MV) and SB were increased significantly compared with their measurements before the procedure. TIMI flows of post-procedural MV were similar between the two groups. In the JSIBT group, TIMI flows of SB both peri-procedure and post-procedure measurements were significantly greater than the non-JSIBT group. Despite a higher incidence of SB dissection in the JSIBT group, no inter-group difference was found in their total SB complications (like SB dissection, SB occlusion, wire entrapment or balloon rupture/entrapment). While JSIBT was an independent predictor for the SB TIMI 3 flow measured at the end of primary PCI, it was not an independent predictor for SB complications. Conclusion: The use of JSIBT as a method of SB protection during primary PCI not only provided better SB protection, but it also had a similar rate of SB complications compared with those with or without prior application of SB wire. This technique may be an effective method of protecting SB for STEMI patients involving coronary artery bifurcation and underwent emergent PCI.

Arabidopsis mRNA decay landscape shaped by XRN 5'-3' exoribonucleases.

5'-3' exoribonucleases (XRNs) play crucial roles in the control of RNA processing, quality, and quantity in eukaryotes. Although genome-wide profiling of RNA decay fragments is now feasible, how XRNs shape the plant mRNA degradome remains elusive. Here, we profiled and analyzed the RNA degradomes of Arabidopsis wild-type and mutant plants with defects in XRN activity. Deficiency of nuclear XRN3 or cytoplasmic XRN4 activity but not nuclear XRN2 activity greatly altered Arabidopsis mRNA decay profiles. Short excised linear introns and cleaved pre-mRNA fragments downstream of polyadenylation sites were polyadenylated and stabilized in the xrn3 mutant, demonstrating the unique function of XRN3 in the removal of cleavage remnants from pre-mRNA processing. Further analysis of stabilized XRN3 substrates confirmed that pre-mRNA 3' end cleavage frequently occurs after adenosine. The most abundant decay intermediates in wild-type plants include not only the primary substrates of XRN4 but also the products of XRN4-mediated cytoplasmic decay. An increase in decay intermediates with 5' ends upstream of a consensus motif in the xrn4 mutant suggests that there is an endonucleolytic cleavage mechanism targeting the 3' untranslated regions of many Arabidopsis mRNAs. However, analysis of decay fragments in the xrn4 mutant indicated that, except for microRNA-directed slicing, endonucleolytic cleavage events in the coding sequence rarely result in major decay intermediates. Together, these findings reveal the major substrates and products of nuclear and cytoplasmic XRNs along Arabidopsis transcripts and provide a basis for precise interpretation of RNA degradome data.

Validation of a non-invasive, real-time, human-in-the-loop model of intracortical brain-computer interfaces.

Objective. Despite the tremendous promise of invasive brain-computer interfaces (iBCIs), the associated study costs, risks, and ethical considerations limit the opportunity to develop and test the algorithms that decode neural activity into a user's intentions. Our goal was to address this challenge by designing an iBCI model capable of testing many human subjects in closed-loop.Approach. We developed an iBCI model that uses artificial neural networks (ANNs) to translate human finger movements into realistic motor cortex firing patterns, which can then be decoded in real time. We call the model the joint angle BCI, or jaBCI. jaBCI allows readily recruited, healthy subjects to perform closed-loop iBCI tasks using any neural decoder, preserving subjects' control-relevant short-latency error correction and learning dynamics.Main results. We validated jaBCI offline through emulated neuron firing statistics, confirming that emulated neural signals have firing rates, low-dimensional PCA geometry, and rotational jPCA dynamics that are quite similar to the actual neurons (recorded in monkey M1) on which we trained the ANN. We also tested jaBCI in closed-loop experiments, our single study examining roughly as many subjects as have been tested world-wide with iBCIs (n= 25). Performance was consistent with that of the paralyzed, human iBCI users with implanted intracortical electrodes. jaBCI allowed us to imitate the experimental protocols (e.g. the same velocity Kalman filter decoder and center-out task) and compute the same seven behavioral measures used in three critical studies.Significance. These encouraging results suggest the jaBCI's real-time firing rate emulation is a useful means to provide statistically robust sample sizes for rapid prototyping and optimization of decoding algorithms, the study of bi-directional learning in iBCIs, and improving iBCI control.

The Procedural and Clinical Outcomes of Rotational Atherectomy in Patients Presenting With Acute Myocardial Infarction.

Background: Rotational atherectomy (RA) is an indispensable tool used for calcified lesion preparation in percutaneous coronary intervention (PCI). However, use of RA in the setting of acute myocardial infarction (AMI) is challenged with limited clinical data. Objectives: This study aims to retrospectively investigate the procedural results, periprocedural complications, and clinical outcomes of RA in patients with AMI. Methods: All possible consecutive patients who received RA in AMI from January 2009 to March 2018 in a single tertiary center were analyzed retrospectively. Patients without AMI during the study period were also enrolled for comparison. Results: A total of 121 patients with AMI (76.0 ± 10.8 years, 63.6% males) and 290 patients without AMI were recruited. Among the AMI group, 81% of patients had non-ST-elevation myocardial infarction (NSTEMI) and 14% presented with cardiogenic shock. RA could be completed in 98.8% of patients in the AMI group and 98.3% in the non-AMI group (p = 1.00). The periprocedural complication rates were comparable between the AMI and non-AMI groups. The risks of in-hospital, 30-day, 90-day, and 1-year cardiovascular major adverse cardiac events (CV MACE) were significantly higher in the AMI group compared with the non-AMI group (in-hospital 13.2 vs. 2.8%, p < 0.001; 30-day 14.2 vs. 4.5%, p < 0.001; 90-day 20.8 vs. 6.9%, p < 0.001; 1-year 30.8 vs. 19.1%, p = 0.01). AMI at initial presentation and cardiogenic shock were predictors for both in-hospital CV MACE and 1-year CV MACE in multivariable binary logistic regression analysis. Other predictors for 1-year CV MACE included serum creatinine level and triple vessel disease. Conclusion: RA in patients with AMI is feasible with a high procedural completion rate and acceptable periprocedural complications. Given unstable hemodynamics and complex coronary anatomy, the in-hospital and 1-year MACE rates remained higher in patients with AMI compared with patients without AMI.

Restoring Osteochondral Defects through the Differentiation Potential of Cartilage Stem/Progenitor Cells Cultivated on Porous Scaffolds.

Cartilage stem/progenitor cells (CSPCs) are cartilage-specific, multipotent progenitor cells residing in articular cartilage. In this study, we investigated the characteristics and potential of human CSPCs combined with poly(lactic-co-glycolic acid) (PLGA) scaffolds to induce osteochondral regeneration in rabbit knees. We isolated CSPCs from human adult articular cartilage undergoing total knee replacement (TKR) surgery. We characterized CSPCs and compared them with infrapatellar fat pad-derived stem cells (IFPs) in a colony formation assay and by multilineage differentiation analysis in vitro. We further evaluated the osteochondral regeneration of the CSPC-loaded PLGA scaffold during osteochondral defect repair in rabbits. The characteristics of CSPCs were similar to those of mesenchymal stem cells (MSCs) and exhibited chondrogenic and osteogenic phenotypes without chemical induction. For in vivo analysis, CSPC-loaded PLGA scaffolds produced a hyaline-like cartilaginous tissue, which showed good integration with the host tissue and subchondral bone. Furthermore, CSPCs migrated in response to injury to promote subchondral bone regeneration. Overall, we demonstrated that CSPCs can promote osteochondral regeneration. A monophasic approach of using diseased CSPCs combined with a PLGA scaffold may be beneficial for repairing complex tissues, such as osteochondral tissue.

Dyslipidemia, Not Inflammatory Markers or Adipokines, Contributes Significantly to a Higher SYNTAX Score in Stable Coronary Artery Disease (from the Taichung CAD Study).

BACKGROUND: The SYNTAX score is an index of coronary severity used to determine the revascularization strategy of a patient. Our previous study confirmed that the SYNTAX score is helpful in predicting major adverse cardiac events in patients with stable coronary artery disease (CAD). However, few studies have comprehensively investigated the predictors for SYNTAX scores in patients with stable CAD, including conventional risk factors, lipid parameters, inflammatory markers and adipokines. METHODS: The coronary severities of 181 revascularization-naïve CAD patients who had received coronary angiograms were coded using SYNTAX scores. Conventional risk factors, inflammatory markers, and adipokines were investigated in order to determine the independent predictors for SYNTAX severity in the patients with stable CAD. RESULTS: The SYNTAX severity score was divided according to the generally accepted criterion (low: ≤ 22, intermediate-high: ≥ 23). In univariate comparisons, the intermediate-high SYNTAX group had a significantly higher low-density lipoprotein cholesterol (LDL-C) level compared to the low SYNTAX score group (p = 0.046). In binary logistic regression, LDL-C, total cholesterol, ratio of total cholesterol/high-density lipoprotein cholesterol (HDL-C) and pre- admission statin use were significant predictors for a higher SYNTAX severity score in the patients with stable CAD. In contrast, circulating adipokines, high-sensitivity C-reactive protein and HDL-C alone were not. CONCLUSIONS: In revascularization-naïve CAD patients, dyslipidemia, including elevated LDL-C, total cholesterol, total cholesterol/HDL-C ratio and pre-index admission statin use, were associated with an intermediate-high SYNTAX severity score.

Clinical outcomes of multivessel coronary artery disease patients revascularized by robot-assisted vs conventional standard coronary artery bypass graft surgeries in real-world practice.

ABSTRACT: The treatment of patients with multivessel coronary artery disease (MVD) by coronary stenting (PCI) and the "gold standard" conventional coronary-artery bypass grafting (C-CABG) has been well explored in the literature. However, the clinical outcomes of robot-assisted CABG (R-CABG) vs C-CABG in MVD patients in real-world practice were unknown. We aimed to study the clinical outcomes of MVD patients who underwent R-CABG (robotic MIDCAB) and C-CABG at our institution between January 2005 and December 2013.A total of 516 MVD patients received CABG were recruited into this study. Among them, 281 patients received R-CABG and 235 patients underwent C-CABG. Patients in the R-CABG group were younger, and had fewer vessels with coronary artery disease (CAD), lower prevalence of chronic renal disease (CKD), higher left ventricular ejection fraction (LVEF), as well as lower Euro scores. The in-hospital and long-term mortalities were lower in the R-CABG group, but the incidences of target lesion revascularization (TLR), target vessel revascularization (TVR), myocardial infarction (MI), and stroke were not significantly different between the two groups. The long-term mortality was related to age, lower LVEF, and CKD, but not residual SYNTAX score, or completeness of revascularization. The revascularization modality (R-CABG vs C-CABG) was a borderline significantly independent predictor of long-term mortality (OR 1.76 [0.99-3.14], P = .055).Our study concluded that R-CABG, in comparison with C-CABG, for MVD carried out in younger patients involved fewer clinical complexities was associated with lower in-hospital and long-term mortalities in real-world practice. However, the long-term rates of TLR, TVR, MI, and stroke were similar. The long-term mortality was correlated with age, lower LVEF, and CKD, where R-CABG remained a borderline significant predictor after correcting for confounding factors. R-CABG could be an effective alternative to C-CABG for MVD patients with fewer clinical complexities in real-world practice.

A tale of too many tasks: task fragmentation in motor learning and a call for model task paradigms.

Motor learning encompasses a broad set of phenomena that requires a diverse set of experimental paradigms. However, excessive variation in tasks across studies creates fragmentation that can adversely affect the collective advancement of knowledge. Here, we show that motor learning studies tend toward extreme fragmentation in the choice of tasks, with almost no overlap between task paradigms across studies. We argue that this extreme level of task fragmentation poses serious theoretical and methodological barriers to advancing the field. To address these barriers, we propose the need for developing common 'model' task paradigms which could be widely used across labs. Combined with the open sharing of methods and data, we suggest that these model task paradigms could be an important step in increasing the robustness of the motor learning literature and facilitate the cumulative process of science.

Plantar thermography predicts freedom from major amputation after endovascular therapy in critical limb ischemic patients.

Although plantar thermography can evaluate the immediate perfusion result after an endovascular therapy (EVT) has been performed, a relevant wound outcome study is still lacking.This study was to investigate whether angiosome-based plantar thermography could predict wound healing and freedom from major amputation after EVT in patients with critical limb ischemia (CLI).All 124 patients with CLI (Rutherford category 5 and 6) who underwent EVT from January 2017 to February 2019 were prospectively enrolled. All patients received thermography both before and after EVT. Both wound healing and freedom from major amputation at the 6-month follow-up period were recorded. There were 61 patients in the healing group and 63 patients in the non-healing group, whereas the major amputation total was 14 patients. The mean pre- and post-EVT temperature of the foot was significantly higher in the healing group than in the non-healing group (30.78 °C vs 29.42 °C, P = .015; and 32.34 °C vs 30.96 °C, P = .004, respectively). DIFF2 was significantly lower in the non-healing group (-1.38 vs -0.90, P = .009). DIFF1 and DIFF2 were significantly lower in the amputation group (-1.85 °C vs -1.11 °C, P = .026; and -1.82 °C vs -1.08 °C, P = .004). Multivariate analysis showed that DIFF2 stood out as an independent predictor for freedom from major amputation (hazard ratio 0.51, P = .045). Receiver operating characteristic curve analysis showed a DIFF2 cut-off value of -1.30 °C, which best predicts freedom from major amputation.Plantar thermography is associated with wound healing and helps predict freedom from major amputation in CLI patients undergoing EVT.

The Characteristics and Clinical Outcomes of Rotational Atherectomy under Intra-Aortic Balloon Counterpulsation Assistance for Complex and Very High-Risk Coronary Interventions in Contemporary Practice: An Eight-Year Experience from a Tertiary Center.

BACKGROUND: The use of Complex and High-risk Coronary Interventions (CHIPs) has increased in recent years. Both rotational atherectomy (RA) and hemodynamic support are important parts of CHIPs. OBJECTIVES: This study aimed to retrospectively investigate the procedure results and clinical outcomes of intra-aortic balloon pump (IABP)-assisted RA in the contemporary drug-eluting stent era. METHODS: All consecutive patients who received RA under in-procedure IABP assistance from April 2010 to March 2018 were analyzed retrospectively. RESULTS: A total of 63 patients (77.7 ± 10.1 years, 69.8% male) were recruited, of whom 51 underwent RA with primary IABP assistance and 12 underwent bailout IABP. RA could be completed in 61 (96.8%) of the patients. Overall, vessel perforation, profound in-procedure shock, and ventricular arrhythmia occurred in 1.6%, 4.8% and 3.2% of the patients, respectively. The in-hospital, 30-day and 90-day major adverse cardiac event (MACE) rates were 22.2%, 27.4% and 36.1%, respectively, mostly driven by mortality. The MACE rates were significantly higher in the bail-out group in the hospital (50.0% vs. 15.7%, p = 0.018) at 30 days (58.3% vs. 20.0%, p = 0.013) and 90 days (66.7% vs. 28.6%, p = 0.020). CONCLUSIONS: Bail-out IABP was associated with increased MACEs, implying that the use of IABP should be implemented at the beginning of RA if a complex procedure is anticipated.

Catheter directed diagnosis of ST-segment elevation myocardial infarction induced by type A aortic dissection: A case report.

RATIONALE: Type A aortic dissection (TAAD) is a life-threatening disorder yet it is hard to diagnose. The dissection might extend to the coronary artery causing ST-segment elevation myocardial infarction (STEMI). Physicians might not recognize this particularly early in its presentation and patients proceed to receive the primary percutaneous coronary intervention. We present such a case and found that the marked pressure difference between the radial and ascending aortae could be a useful clue for diagnosing the aortic dissection-related myocardial infarction. PATIENT CONCERNS: A 58-year-old male was presented to our emergency department for the complaint of left side chest pain that lasted for an hour with concomitant hypotension. STEMI was diagnosed at that time. DIAGNOSIS: The emergent primary percutaneous intervention was performed. When the diagnostic catheter was advanced to the ascending aorta, the systolic aorta pressure became 20 mm Hg higher than radial systolic pressure. Due to the abnormally large pressure differential between the peripheral radial artery and central ascending aorta, TAAD was suspected. INTERVENTIONS: After angiography and computer tomography confirmed the diagnosis of TAAD, the patient was sent for emergent surgery. OUTCOMES: The patient was died because of extensive dissection and shock. LESSONS: We present such a case and found that the marked pressure difference between the radial and ascending aortae during catheterization could be a useful clue for diagnosing the aortic dissection-related myocardial infarction. This clue had hinted our speedy examination of the occluded coronary artery and dissection flap, and led to an early and accurate diagnosis.

Acute Angiographic and Intermediate-Term Clinical Results of Patients with Non-Left Main Coronary Bifurcation Lesions Treated with BVS by Jailed Semi-Inflated Balloon Technique and Provisional Side-Branch Stenting Strategy.

BACKGROUND: To evaluate the acute angiographic and intermediate-term clinical results of patients with non-left main (LM) coronary artery bifurcation disease (CABD) treated with BVS, as compared with those treated with DES, using the jailed semi-inflated balloon technique (JSIBT) for side branch (SB) protection and provisional stenting. METHODS AND RESULTS: Sixty-eight patients with non-LM CABD who had undergone provisional one-stent implantation with SB protection by JSIBT between January 2015 and December 2017 were retrospectively enrolled. Among them, 20 patients received Absorb BVS implantation and 48 patients received DES implantation. Patients in the BVS group were younger and had higher BMI, total cholesterol, low-density lipoprotein cholesterol, and hemoglobin but had lower serum creatinine and lower prevalence of prior PCI and MI. No SB balloon rupture/entrapment occurred in either group. The incidence of SB dissection/occlusion and SB in need of rewiring or stenting was rare in both groups and showed no significant difference between them. Postinterventional TIMI flow significantly increased in both groups. The intermediate-term clinical outcomes were good in terms of incidence of target lesion failure, target lesion revascularization, target vessel revascularization, myocardial infarction, and all-cause death in both groups. CONCLUSION: The use of JSIBT for treating CABD with modern BVS can provide SB protection as similar as those with DES, even with higher incidence of acute SB dissection/occlusion. The immediate angiographic results and acute and intermediate-term clinical outcomes were also similar in both groups. Our study results demonstrate that JSIBT might be a safe and alternative SB protection tool for BVS in patients with complex CABD.

Clinical outcomes of patients with multivessel coronary artery disease treated with robot-assisted coronary artery bypass graft surgery versus one-stage percutaneous coronary intervention using drug-eluting stents.

A number of studies have reported on treatment outcomes of coronary stenting (PCI) for multivessel coronary artery diseases (MVD), and compared them with the conventional coronary artery bypass grafting (CABG). However, the clinical outcomes of robot-assisted CABG (R-CABG) in comparison with PCI in MVD patients have not been investigated.We recruited retrospectively MVD patients receiving R-CABG and PCI with drug-eluting stents for all vessels in one stage between January 2005 and December 2013 at our institution with at least 3 years of outcomes were retrospectively recruited and analyzed.A total of 638 MVD patients were studied. Among them, 281 received R-CABG, and 357 received PCI. Similar complete revascularizations were achieved in both groups (R-CABG: 40.2%, PCI: 41.5%, P = .751). The residual stenosis was 4.1 ±â€Š4.4 in the R-CABG group, and comparably 3.5 ±â€Š3.7 in the PCI group (P = .077). Patients in the R-CABG group were younger, with more severe coronary artery disease (CAD) and had more background risk factors. The in-hospital and long-term mortalities as well as the incidence of TLR, myocardial infarction (MI), stroke were all similar between groups. But the incidence of TVR and any revascularization were lower in the R-CABG group. The long-term mortality was predicted by age, left ventricular ejection fraction, and chronic kidney disease, but not by the revascularization modality, completeness of revascularization, nor residual SYNTAX scores. The last 3 factors were not predictors of long-term TLR, TVR, MI, and stroke.The in-hospital and long-term survival rates of MVD were similar for both the R-CABG and PCI groups. But the R-CABG group had rates of TVR and any revascularization lower than PCI. Revascularization modality, completeness of revascularization, and residual SYNTAX scores were not predictors of in-hospital and long-term mortalities, MI, and stroke in real-world practice. R-CABG was associated with lower rates of TLR and TVR, and is likely a safe and effective treatment and an alternative choice of PCI for MVD patients who have low surgical risks.

Clinical outcomes of coronary artery bifurcation disease patients underwent Culotte two-stent technique: a single center experience.

BACKGROUND: Percutaneous coronary intervention for coronary artery bifurcation disease (CABD) remains challenging. In patients of CABD with situations that two-stent strategy is needed, the culotte technique is a widely used method and also as the majority at out institution. We sought to take a look of the clinical outcomes of our culotte stenting patients. METHODS: This retrospective study analyzed 238 consecutive CABD patients who underwent culotte two-stent technique at a tertiary medical center between July 2008 and November 2015. RESULTS: Culotte technique was used in 238 lesions in 238 patients. Of these patients, all DES were implanted for culotte two-stent technique. Most of these patients were elderly, male gender, ACS on admission and multiple vessel disease. The bifurcation lesions were mostly located at left coronary artery (51.3%), categorized as true bifurcation lesion (92%) and calculated less than 70 degree of bifurcation angle (74.4%). During a medium 3.27-year follow up, the angle of bifurcation lesion ≥70° and body mass index were positively independent predictors for target lesion failure (TLF), diabetes mellitus was an independent predictor of target vessel revascularization, and statin therapy for hyperlipidemia, hemoglobin and EF were negatively independent factors associated to total mortality. The rates of in-hospital and total mortalities were 4.2 and 17.6%. CONCLUSION: In this cohort of CABD patients with most left main and left anterior descending artery lesions treated by culotte stenting, the procedural success rate was high and the intermediate clinical outcomes were acceptable. (Reviewer #1, Comment #1) Bifurcation angle (≥ 70°) is an inherently independent predictor of TLF and other two-stent strategy replaced needed to be considered in this situation.

Continuous Passive Motion Promotes and Maintains Chondrogenesis in Autologous Endothelial Progenitor Cell-Loaded Porous PLGA Scaffolds during Osteochondral Defect Repair in a Rabbit Model.

Continuous passive motion (CPM) is widely used after total knee replacement. In this study, we investigated the effect of CPM combined with cell-based construct-transplantation in osteochondral tissue engineering. We created osteochondral defects (3 mm in diameter and 3 mm in depth) in the medial femoral condyle of 36 knees and randomized them into three groups: ED (empty defect), EPC/PLGA (endothelial progenitor cells (EPCs) seeded in the poly lactic-co-glycolic acid (PLGA) scaffold), or EPC/PLGA/CPM (EPC/PLGA scaffold complemented with CPM starting one day after transplantation). We investigated the effects of CPM and the EPC/PLGA constructs on tissue restoration in weight-bearing sites by histological observation and micro-computed tomography (micro-CT) evaluation 4 and 12 weeks after implantation. After CPM, the EPC/PLGA construct exhibited early osteochondral regeneration and prevention of subchondral bone overgrowth and cartilage degeneration. CPM did not alter the microenvironment created by the construct; it up-regulated the expression of the extracellular matrix components (glycosaminoglycan and collagen), down-regulated bone formation, and induced the biosynthesis of lubricin, which appeared in the EPC/PLGA/CPM group after 12 weeks. CPM can provide promoting signals during osteochondral tissue engineering and achieve a synergistic effect when combined with EPC/PLGA transplantation, so it should be considered a non-invasive treatment to be adopted in clinical practices.

Osteochondral Tissue Regeneration Using a Tyramine-Modified Bilayered PLGA Scaffold Combined with Articular Chondrocytes in a Porcine Model.

Repairing damaged articular cartilage is challenging due to the limited regenerative capacity of hyaline cartilage. In this study, we fabricated a bilayered poly (lactic-co-glycolic acid) (PLGA) scaffold with small (200⁻300 µm) and large (200⁻500 µm) pores by salt leaching to stimulate chondrocyte differentiation, cartilage formation, and endochondral ossification. The scaffold surface was treated with tyramine to promote scaffold integration into native tissue. Porcine chondrocytes retained a round shape during differentiation when grown on the small pore size scaffold, and had a fibroblast-like morphology during transdifferentiation in the large pore size scaffold after five days of culture. Tyramine-treated scaffolds with mixed pore sizes seeded with chondrocytes were pressed into three-mm porcine osteochondral defects; tyramine treatment enhanced the adhesion of the small pore size scaffold to osteochondral tissue and increased glycosaminoglycan and collagen type II (Col II) contents, while reducing collagen type X (Col X) production in the cartilage layer. Col X content was higher for scaffolds with a large pore size, which was accompanied by the enhanced generation of subchondral bone. Thus, chondrocytes seeded in tyramine-treated bilayered scaffolds with small and large pores in the upper and lower parts, respectively, can promote osteochondral regeneration and integration for articular cartilage repair.

Alterations of plant architecture and phase transition by the phytoplasma virulence factor SAP11.

As key mediators linking developmental processes with plant immunity, TCP (TEOSINTE-BRANCHED, CYCLOIDEA, PROLIFERATION FACTOR 1 and 2) transcription factors have been increasingly shown to be targets of pathogenic effectors. We report here that TB/CYC (TEOSINTE-BRANCHED/CYCLOIDEA)-TCPs are destabilized by phytoplasma SAP11 effectors, leading to the proliferation of axillary meristems. Although a high degree of sequence diversity was observed among putative SAP11 effectors identified from evolutionarily distinct clusters of phytoplasmas, these effectors acquired fundamental activity in destabilizing TB/CYC-TCPs. In addition, we demonstrate that miR156/SPLs and miR172/AP2 modules, which represent key regulatory hubs involved in plant phase transition, were modulated by Aster Yellows phytoplasma strain Witches' Broom (AY-WB) protein SAP11. A late-flowering phenotype with significant changes in the expression of flowering-related genes was observed in transgenic Arabidopsis plants expressing SAP11AYWB. These morphological and molecular alterations were correlated with the ability of SAP11 effectors to destabilize CIN (CINCINNATA)-TCPs. Although not all putative SAP11 effectors display broad-spectrum activities in modulating morphological and physiological changes in host plants, they serve as core virulence factors responsible for the witches' broom symptom caused by phytoplasmas.

Enhancement of chondrogenesis of adipose-derived stem cells in HA-PNIPAAm-CL hydrogel for cartilage regeneration in rabbits.

Injectable thermoresponsive hydrogels have the advantages of effective cell delivery and minimal invasion for tissue engineering applications. In this study, we investigated the chondroinductive potential of newly developed hyaluronic acid (HA)-modified thermoresponsive poly(N-isopropylacrylamide) (HA-PNIPAAm-CL) hydrogels on enhancing rabbit ADSC (rADSC) chondrogenesis in vitro and in the synovial cavity of rabbit. The HA-mixed PNIPAAm (HA-PNIPAAm-CP) and HA-cross-linked PNIPAAm (HA-PNIPAAm-CL) were fabricated using physical interaction and chemical cross-linking methods, respectively. The in vitro results showed that, compared to unmodified PNIPAAm, both HA-modified hydrogels significantly increased cell viability, chondrogenic marker gene (aggrecan and type II collagen) expression and sulfide glycosaminoglycan (sGAG) formation in embedded rADSCs. However, HA-PNIPAAm-CL showed the highest rADSC viability and chondrogenesis. The chondrogenic effects of HA-modified hydrogels on rADSCs were confirmed in vivo by the intraarticular injection of hydrogel-embedded rADSC constructs into rabbit synovial cavities for 3 weeks and tracing with CM-DiI labeling. Neocartilage formation in the hydrogels was determined by histomorphological staining of GAG and type II collagen. In vivo injected rADSC/HA-PNIPAAm-CL constructs showed more hyaline cartilage formation than that of rADSC/HA-PNIPAAm-CP and rADSC/PNIPAAm constructs in the synovial cavity of rabbit. These results suggest that the HA-PNIPAAm-CL provides a suitable microenvironment to enhance ADSC chondrogenesis for articular cartilage tissue engineering applications.