A two-step neural network-based guiding system for obtaining reliable radiographs for critical shoulder angle measurement.

Background: The critical shoulder angle (CSA) has been reported to be highly associated with rotator cuff tears (RCTs) and an increased risk of RCT re-tears. However, the measurement of the CSA is greatly affected by the malpositioning of the shoulder. To address this issue, a two-step neural network-based guiding system was developed to obtain reliable CSA radiographs, and its feasibility and accuracy was evaluated. Methods: A total of 1,754 shoulder anteroposterior (AP) radiographs were retrospectively acquired to train and validate a two-step neural network-based guiding system to obtain reliable CSA radiographs. The study included patients aged 18 years or older who underwent X-rays and/or computed tomography (CT) scans of the shoulder. Patients who had undergone shoulder surgery, had a confirmed fracture, or were diagnosed with a musculoskeletal tumor or glenoid defect were excluded from the study. The system consisted of a two-step neural network that in the first step, localized the region of interest of the shoulder, and in the second step, classified the radiography according to type [i.e., 'forward' when the non-overlapping coracoid process is above the glenoid rim, 'backward' when the non-overlapping coracoid process is below or aligned with the glenoid rim, a ratio of the transverse to longitudinal diameter of the glenoid projection (RTL) ≤0.25, or a RTL >0.25]. The performance of the model was assessed in an offline, prospective manner, focusing on the sensitivity and specificity for the forward, backward, RTL ≤0.25, or RTL >0.25 types (denoted as SensF, B, -, + and SpecF, B, -, +, respectively), and Cohen's kappa was also reported. Results: Of 273 cases in the offline prospective test, the SensF, SensB, Sens-, and Sens+ were 88.88% [95% confidence interval (CI): 50.67-99.41%], 94.11% (95% CI: 82.77-98.47%), 96.96% (95% CI: 91.94-99.02%), and 95.06% (95% CI: 87.15-98.40%), respectively. The SpecF, SpecB, Spec-, and Spec+ were 98.48% (95% CI: 95.90-99.51%), 99.55% (95% CI: 97.12-99.97%), 95.04% (95% CI: 89.65-97.81%), and 97.39% (93.69-99.03%), respectively. A high classification rate (93.41%; 95% CI: 89.14-96.24%) and almost perfect agreement (Cohen's kappa: 0.903, 95% CI: 0.86-0.95) were achieved. Conclusions: The guiding system can rapidly and accurately classify the types of AP shoulder radiography, thereby guiding the adjustment of patient positioning. This will facilitate the rapid obtainment of reliable CSA radiography to measure the CSA on proper AP radiographs.

The Longitude-Latitude-Loop Used for Complex Bankart Lesion Repair: An All-Arthroscopic Technique.

The most frequent operation for anterior shoulder instability is the arthroscopic Bankart repair, which has a positive outcome and a low rate of complications. A variety of restoration procedures have been reported to reconstruct labral height and reproduce a dynamic concavity-compression reaction. The longitude-latitude loop is a knotless high-strength suture method that simultaneously tightens the joint capsule in the warp and weft direction and resists tearing. The suture method is safe and reproducible. This study aimed to present a longitude-latitude loop suture for joint capsule labral complex repair during Bankart arthroscopy surgery.

Arthroscopic Autologous Coracoacromial Ligament Augment Technique for Particle Thickness of Rotator Cuff with at Least Two Years Follow-Up.

OBJECTIVE: Patch technology has been the new technique in the treatment of partial thickness of the rotator cuff tear (PTRCTs) to address the limitation of traditional techniques. Compared with allogeneic patches and artificial materials, coracoacromial ligament is obviously closer to their own biology. The purpose of the study was to evaluate the functional and radiographic outcomes following arthroscopic autologous coracoacromial ligament augment technique for treatment of PTRCTs. METHOD: This study included three female patients with PTRCTs who underwent arthroscopy operation in 2017 with an average age of 51 years (range from 50 to 52 years). The coracoacromial ligament implant was attached to the bursal side surface of the tendon. The clinical results were evaluated by American Shoulder and Elbow Surgeons (ASES) score, Simple Shoulder Test (SST), acromiohumeral distance (AHD) and muscle strength before and 12 months after operation. Magnetic resonance imaging (MRI) was performed 24 months after operation to assess the integrity of the anatomical structure of the original tear site. RESULT: The average ASES score improved significantly from 57.3 preoperatively to 95.0 at 1-year follow-up. The strength improved significantly from grade 3 preoperatively to grade 5 at 1 year. Two of three patients underwent the MRI at 2-year follow-up. Radiographic released the complete healing of rotator cuff tear. No implant-related serious adverse events were reported. CONCLUSION: The new technique of using autogenous coracoacromial ligament patch augment provides good clinical results on patients with PTRCTs.

Pore Structure and Gas Content Characteristics of Lower Jurassic Continental Shale Reservoirs in Northeast Sichuan, China.

The Jurassic shale in the northeastern Sichuan Basin is one of the main target intervals for continental shale gas exploitation. Research on the pore structure and gas-bearing properties of shales is the key issue in target interval optimization. Through core observation, geochemistry, bulk minerals, scanning electron microscopy, nitrogen adsorption, and isothermal adsorption experiments, various lithofacies with different pore structure characteristics were clarified. In addition, the factors that control gas-bearing properties were discussed, and a continental shale gas enrichment model was finally established. The results show that the Jurassic continental shale in the northeastern Sichuan Basin can be classified into six lithofacies. Organic pores, intergranular pores, interlayer pores in clay minerals, intercrystalline pores in pyrite framboids, and dissolution pores can be observed in shale samples. Pore structures varied in different shale lithofacies. The contact angle of shales is commonly less than 45°, leading to complex wettability of pores in the shales. Free gas content is mainly controlled by the organic matter (OM) content and the brittleness in the Jurassic shale. The adsorbed gas content is mainly controlled by the OM content, clay mineral type, and water saturation of the shales. The enrichment mode of the Lower Jurassic continental shale gas in the northeastern Sichuan Basin is established. Paleoenvironments control the formation of organic-rich shales in the center part of lakes. The "baffle" layer helps the confinement and high pressure, and the complex syncline controls the preservation, forming the enrichment pattern of the complex syncline-central baffle layer.

Allogeneic Tendon Transplantation for the Treatment of Pathological Patellar Ligament Defect in Children: Technical Note and 4-Year Follow-Up.

OBJECTIVE: The absence of patellar ligament will bring about a severe negative impact on daily life. Many reconstruction techniques have been described in adults. However, there is a lack of technical introduction regarding the reconstruction of the patellar ligament in children. The purpose of this study was to report a surgical technique for reconstructing the patellar ligament in children. METHOD: A retrospective analysis of the clinical data on a patellar ligament (tendon sheath fibroma) patient with allogeneic tendon reconstruction. An 8-year-old child with postoperative recurrence of left patellar ligament tumor was enrolled in our study. Anterior tibialis tendon allograft was used to reconstruct the patellar ligament after complete resection of the patellar ligament for the tumor. The tunnels were constructed on the deep surface of the tibial tubercle and the root of the quadriceps tendon (to decrease the harmful impact on patella development), respectively. The allogeneic tendon was passed through the tunnels above in the shape of "8," and the two ends of the tendon were attached to the bleeding bone bed at the inferior edge of the patella with suture anchors to achieve better bone-tendon healing. During the follow-up, the knee's range of motion and imaging manifestations were recorded. RESULT: Postoperative pathology suggests chondromesenchymal hamartoma, a rare benign soft tissue tumor different from the previous operation (tendon sheath fibroma). During the 4-year follow-up, the patient's active range of motion of the knee achieved 0° to 120°; and the patient could walk normally without any external help. Physical examinations (the apprehension sign and J sign) showed no ligamentous instability or patellar ligament tenderness. Imaging analysis showed that the ratio length of the patellar ligament to the patella was almost normal. The integrity, continuity, and shape of the allogeneic ligament showed excellent results in MRI. Combined with clinical and imaging findings, allogeneic tendon patellar ligament reconstruction was deemed successful. CONCLUSION: Allogeneic ligament reconstruction technique can provide a treatment option by reconstructing the extensor mechanism, minimizing the impact on patellar development, and augmenting biological healing for children with the absence of the patellar ligament.

Fibrosis in frozen shoulder: Activation of IL-6 through PI3K-Akt signaling pathway in synovial fibroblast.

Fibrosis is the main cause of limited range of motion (ROM) of shoulder in patients with frozen shoulder (FS). Overexpression of Interleukin 6 (IL-6) has been correlated with pathogenesis of FS. However, the underlying mechanism remains largely unexplored. In the current study, we focused on isolating synovial fibroblasts of FS and determining the influence of IL-6 as well as PI3K-Akt signaling pathway on the fibrotic process of synovial fibroblasts in FS by using RNA Sequencing (RNA-seq) and other molecular biology techniques. Synovial fibroblasts of FS express more extra cellular matrix (ECM) than that of control. RNA-seq results and bioinformatic analysis indicate that PI3K-Akt signaling pathway play an important role in the fibrotic process of FS, and IL-6 is the most related gene among those related to this process. The expression levels of IL-6 / IL-6R in FS synovial fibroblasts and IL-6 in culture supernatant were both significantly increased. siRNA interference with the expression of IL-6 attenuates the fibrosis level of FS as well as phosphorylation level of Akt. The findings suggest that synovial fibroblasts are key effector cells of fibrosis of FS. Activation of PI3K-Akt pathway can promote fibrosis of synovial fibroblasts in FS. IL-6 is up-regulated in synovial fibroblasts of FS and promoted the FS fibrosis through PI3K-Akt signaling pathway.

Oxygen-Carrying and Antibacterial Fluorinated Nano-Hydroxyapatite Incorporated Hydrogels for Enhanced Bone Regeneration.

Insufficient oxygen availability in tissue engineering is one of the major factors for the failure of clinical transplantation. One potential strategy to conquer this limitation is the fabrication of spontaneous and continuous oxygen supplying scaffolds for in situ tissue regeneration. In this work, a versatile fluorine-incorporating hydrogel is designed which can not only timely and continuously supply oxygen for mesenchymal stem cells (MSCs) to overcome deficient oxygen before vascularization in scaffolds, but can present a higher antibacterial capability to avoid bacterial infections. The HAp@PDA-F nanoparticles are first prepared and then incorporated with the quaternized and methacrylated chitosan forming CS/HAp@PDA-F by photo-crosslinking. In vitro results indicate that CS/HAp@PDA-F hydrogel has outstanding mechanical performance, moreover, it also has the oxygen-carrying ability to prolong survival ability, enhance proliferation activity, and preserve osteogenic differentiation potency and promote osteogenic-related genes expression of rat bone mesenchymal stem cells (rBMSCs) under hypoxic environment. Furthermore, the CS/HAp@PDA-F hydrogel can inhibit the growth of Staphylococcus aureus and Escherichia coli, providing a good antibacterial activity. Additionally, in vivo experiments demonstrate higher bone volume and bone mineral density, and more new bone tissue generation in CS/HAp@PDA-F group than in CS/HAp@PDA group. These results indicate that the rational design of fluorinated hydrogel possesses a good clinical application prospect for bone regeneration.

Proanthocyanidin A1 promotes the production of platelets to ameliorate chemotherapy-induced thrombocytopenia through activating JAK2/STAT3 pathway.

BACKGROUND: Chemotherapy-induced thrombocytopenia (CIT) is a severe adverse drug reaction, and the main reason for CIT is the destruction of megakaryocytes (MKs, precursor cells of platelet) in bone marrow by chemotherapy. Peanut skin, the seed coat of Arachis hypogaea L., is a traditional Chinese medicine commonly used to treat thrombocytopenia. However, its active compounds and the mechanisms remain unclear. PURPOSE: This study aims to clarify the active compounds of peanut skin to exhibit thrombogenic effects against CIT and their underlying mechanisms in vitro and in vivo. STUDY DESIGN: The bioassay-guided isolation based on the proliferation of MKs was used to explore the possible platelet-enhancing ingredients in peanut skin. HSCCC technique coupled with preparative HPLC was used to separate the active compounds. Dami cells and carboplatin-treated mice model were used to evaluate the thrombogenic effects of PS-1. Network pharmacology, molecular docking, dynamics simulation studies, kinase activity, surface plasmon resonance (SPR), cellular thermal shift assay (CETSA), isothermal dose-response fingerprint (ITDRFCETSA) and western blot analysis were performed to investigate the mechanisms of PS-1. RESULTS: Proanthocyanidin A1 (PS-1) and its stereoisomers (PS-2-4) were demonstrated to promote the proliferation of MKs (Dami cells), especially PS-1 (EC50 = 8.58 µM). Further studies demonstrated that PS-1 could induce the differentiation of Dami cells in dose/time-dependent manner. Biological target analysis showed that PS-1 directly bound to JAK2 (KD = 2.06 µM) to exert potent activating effect (EC50 = 0.66 µM). Oral administration of PS-1 (25 or 50 mg/kg) significantly improved CIT, but this effect was confirmed to be inhibited by JAK2 inhibitor AG490, indicating that PS-1 exerted its efficacy through JAK2 in vivo. CONCLUSION: Proanthocyanins (PS-1-4) derived from peanut skin were first clarified as platelet-enhancing ingredients to improve CIT. The underlying mechanism of PS-1 was proved to promote the proliferation and differentiation of MKs via JAK2/STAT3 pathway both in vitro and in vivo.

Metabotropic Glutamate Receptor 8 Is Regulated by miR-33a-5p and Functions as an Oncogene in Breast Cancer.

It has been reported that glutamate metabotropic receptor 8 (GRM8) is closely implicated in the progression of human neuroblastoma, lung cancer, and glioma, but its role in breast cancer remains unknown. Thus, the present study was performed to uncover it. Immunohistochemistry, real-time PCR (RT-PCR), and western blotting experiments were performed to test GRM8 expression levels in tissues and cells. Cell functions were assessed by Cell Count Kit 8 (CCK-8), flow cytometry, wound healing, transwell chambers, and in vivo xenotransplantation experiments. The relationship between miR-33a-5p and GRM8 was evaluated by luciferase gene reporter and western blotting assay. The results showed that GRM8 expression was increased in breast cancer tissues and cells, which was closely associated with lower overall survival rate. Ectopic expression of GRM8 significantly enhanced cell growth, migration, and invasion and tumorigenesis and repressed cell apoptosis. In addition, GRM8 was under the negative regulation of miR-33a-5p, which was downregulated in breast cancer tissues and served as a tumor suppressor. Moreover, overexpression of GRM8 abrogated the inhibitive role of miR-33a-5p played in breast cancer. Collectively, this study reveals that GRM8 functions as an oncogene in breast cancer and is regulated by miR-33a-5p.

Young's Modulus of Bilateral Infraspinatus Tendon Measured in Different Postures by Shear Wave Elastography Before and After Exercise.

OBJECTIVE: To investigate the Young's modulus value of infraspinatus tendons using shear wave elastography (SWE) technique in normal adults, and to analyze the influence of gender, postures, exercise, and dominant side on Young's modulus of infraspinatus tendons. METHODS: This is a prospective cross-sectional study. From January 2019 to July 2020, 14 healthy subjects were identified, including seven males and seven females aged between 24 to 34, with a mean age of 27.67 ± 3.08 years. The Young's modulus of their infraspinatus tendons was measured by two operators using SWE in neutral and maximum external rotation positions of both sides before exercise and the dominant side after exercise. The Young's modulus values in different sexes, different postures, before vs after exercise, and dominant vs non-dominant side were statistically analyzed. RESULTS: All 14 subjects completed the data collection process. The mean Young's modulus values of infraspinatus tendon for dominant sides in neutral position were 33.04 ± 3.01 kPa for males and 28.76 ± 3.09 kPa for females. And for non-dominant sides in the neutral position, the values were 33.02 ± 2.38 kPa for males and 28.86 ± 2.47 kPa for females. In the maximum external rotation position, the values for dominant sides were 50.19 ± 4.86 kPa for males and 42.79 ± 4.44 kPa for females, and for non-dominant sides were 50.95 ± 3.24 kPa for males and 42.42 ± 3.66 kPa for females. After exercise, the mean Young's modulus values of infraspinatus tendon for dominant sides in neutral position were 54.56 ± 3.76 kPa for males and 46.66 ± 5.99 kPa for females. And for the maximum external rotation position, the values were 59.13 ± 3.78 kPa for males and 54.49 ± 5.67 kPa for females. The Young's modulus of infraspinatus tendon in the neutral and maximum external rotation positions showed statistically significant differences in males and females, as well as before and after exercise (P < 0.05). However, the difference in Young's modulus between the dominant and non-dominant sides was not statistically significant (P > 0.05). Intergroup reliability between both operators was excellent (ICC > 0.85). CONCLUSION: There are gender-related differences and post-exercise increase in Young's modulus, yet such a difference cannot be witnessed between the dominant and non-dominant sides.

The Ratio of the Transverse to Longitudinal Diameter of the Glenoid Projection Is of Good Predictive Value for Defining the Reliability of Critical Shoulder Angle in Nonstandard Anteroposterior Radiographs.

PURPOSE: To investigate (1) whether measurement of the critical shoulder angle (CSA) is still reliable in some nonstandard anteroposterior radiographs; and (2) whether the ratio of the transverse to longitudinal diameter of the glenoid projection (RTL) could be used to determine the reliability of the CSA in nonstandard anteroposterior radiographs. METHODS: The authors analyzed radiology records from 2017 to 2019 for patients with computed tomography (CT) scans of the shoulder. According to the Suter-Henninger classification system, each CT scan underwent 3-dimensional (3D) reconstructions to obtain 8 digitally reconstructed radiographs (DRRs), including 1 type A1 film and 7 type D1 films with different rotation angles. CSA and RTL were measured on all films, and 2 blinded reviewers evaluated DRRs. The relationship between RTL and CSA was determined by Pearson correlation test. The threshold value was determined by receiver operating characteristic (ROC) analyses using RTL as predictors and defined reliable CSA as criterion in the logistic regression. Intra- and inter-rater reliability were assessed by the intraclass correlation coefficient (ICC). Discriminative capacity was calculated with ROC analyses and area under the curve (AUC). An optimal measurement cutoff with resultant sensitivity and specificity was calculated. RESULTS: A total of 86 patients were included, mean age 40.70 ± 18.25 years, 19 of whom had rotator cuff tears (RCTs) and 67 without RCTs. Spearman's correlation test revealed that the deviation of CSA was positively correlated with RTL, correlation coefficient 0.544 (P < .001) in all patients and 0.686 (P < .001) only in patients with RCTs. ROC analysis of all patients showed that the AUC of the RTL was 0.90, and the optimal cutoff point was 0.25 (sensitivity 88%, specificity 87%). ROC analysis only of patients with RCTs showed that the AUC of the RTL was 0.95, and the optimal cutoff point was 0.22 (sensitivity 95%, specificity 92%). CONCLUSION: The ratio of the transverse to longitudinal diameter of the glenoid projection (RTL) is of good predictive value in defining the reliability of the CSA in malposition films. Based on the results, the CSA can be considered reliable if its RTL is <0.25. LEVEL OF EVIDENCE: III, retrospective cohort study investigating a diagnostic test.

Polyhedral Oligomeric Silsesquioxane-Incorporated Gelatin Hydrogel Promotes Angiogenesis during Vascularized Bone Regeneration.

Many approaches have been made toward the development of scaffolds with good biocompatibility and appreciable physicochemical properties to facilitate stem cell adhesion, osteogenic differentiation, and vascularization in tissue engineering. Nowadays, vascularization is a main bottleneck in tissue engineering strategies that is needed to be overcome and developed. Herein, we construct a series of polyhedral oligomeric silsesquioxane (POSS)-modified porous gelatin hydrogels with different POSS concentrations from 0 to 5 wt %, defined as X% POSS hydrogels (X = 0, 1, 2, 3, 4, 5) to support vascularized bone repair. The introduction of POSS into gelatin effectively promoted adhesive protein adsorption and integrin α5ß1 expression, subsequently leading to enhanced adhesion of both rat bone marrow mesenchymal stem cells and human umbilical vein endothelial cells (HUVECs). In vitro experiments further demonstrated that POSS-containing hybrid hydrogels more effectively support the angiogenic tube and network formation in HUVECs than the 0% POSS hydrogel. Besides, POSS-containing hybrid hydrogels showed desirable performance as a sustained release system of vascular endothelial growth factor (VEGF) and bone morphogenetic protein-2 (BMP-2), and they further accelerated vascular network establishment and the formation of a new bone in defect regions. When the hydrogels were implanted into critical-sized rat calvarial defects in vivo, the VEGF/BMP-2-coupled 3% POSS group gained a higher blood vessel volume in the bone defect regions (5.49 ± 0.35 mm3) than the 3% POSS group (3.12 ± 0.20 mm3) and the 0% POSS group (1.57 ± 0.25 mm3), suggesting that the 3% POSS hydrogel with VEGF/BMP-2 would expedite vascularization. Based on these evaluations, our results indicated that the POSS-incorporated gelatin hydrogel would provide a promising bone graft scheme in potential clinical application of large bone defect repair.

In situ bone regeneration enabled by a biodegradable hybrid double-network hydrogel.

Stem cell therapy based on advanced biomaterials provides a promising strategy in bone tissue engineering. Nevertheless, guided bone regeneration which fulfills the criteria in terms of biomechanics, biodegradability and bioactivity is highly appealing but challenging. Inspired by the superior double-network (DN) structure, herein, a biodegradable hybrid DN hydrogel is proposed to promote in situ bone regeneration. The DN hydrogel is constructed by interspersing a methacrylated gelatin (GelMA) network into a well-defined nanocomposite (NC) hydrogel consisting of methacrylated chitosan (CSMA) and polyhedral oligomeric silsesquioxane (POSS) via a two-step photo-crosslinking process. The hybrid DN hydrogel has the following advantageous characteristics: (i) it exhibits enhanced stiffness and toughness benefiting from the inorganic POSS units and unique energy dissipation; (ii) naturally occurring biomacromolecules (chitosan and gelatin) as the hydrogel framework result in an appropriate biodegradation behavior, which can be replaced by newly formed tissues; (iii) it preferentially guides mesenchymal stem cells (MSCs) toward osteogenic differentiation in vitro by detecting the elevated levels of enzyme activity and calcium deposition along with the up-regulated osteogenesis-related genes and proteins; and (iv) accelerated in situ bone regeneration is observed after implanting MSC-loaded hydrogels into rat calvarial defects. Therefore, we provide a new insight to develop functional hydrogels for triggering specific cellular responses toward stem cell therapy and bone-related tissue engineering.

Influence of Crohn's disease related polymorphisms in innate immune function on ileal microbiome.

We have previously identified NOD2 genotype and inflammatory bowel diseases (IBD) phenotype, as associated with shifts in the ileal microbiome ("dysbiosis") in a patient cohort. Here we report an integrative analysis of an expanded number of Crohn's disease (CD) related genetic defects in innate immune function (NOD2, ATG16L1, IRGM, CARD9, XBP1, ORMDL3) and composition of the ileal microbiome by combining the initial patient cohort (Batch 1, 2005-2010, n = 165) with a second consecutive patient cohort (Batch 2, 2010-2012, n = 118). These combined patient cohorts were composed of three non-overlapping phenotypes: 1.) 106 ileal CD subjects undergoing initial ileocolic resection for diseased ileum, 2.) 88 IBD colitis subjects without ileal disease (predominantly ulcerative colitis but also Crohn's colitis and indeterminate colitis, and 3.) 89 non-IBD subjects. Significant differences (FDR < 0.05) in microbiota were observed between macroscopically disease unaffected and affected regions of resected ileum in ileal CD patients. Accordingly, analysis of the effects of genetic and clinical factors were restricted to disease unaffected regions of the ileum. Beta-diversity differed across the three disease categories by PERMANOVA (p < 0.001), whereas no significant differences in alpha diversity were noted. Using negative binomial models, we confirmed significant effects of IBD phenotype, C. difficile infection, and NOD2 genotype on ileal dysbiosis in the expanded analysis. The relative abundance of the Proteobacteria phylum was positively associated with ileal CD and colitis phenotypes, but negatively associated with NOD2R genotype. Additional associations with ORMDL3 and XBP1 were detected at the phylum/subphylum level. IBD medications, such as immunomodulators and anti-TNFα agents, may have a beneficial effect on reversing dysbiosis associated with the IBD phenotype. Exploratory analysis comparing microbial composition of the disease unaffected region of the resected ileum between 27 ileal CD patients who subsequently developed endoscopic recurrence within 6-12 months versus 34 patients who did not, suggested that microbial biomarkers in the resected specimen helped stratify patients with respect to risk of post-surgical recurrence.

Enhanced proliferation and differentiation of retinal progenitor cells through a self-healing injectable hydrogel.

Retinal progenitor cell (RPC)-based transportation therapy is a promising strategy for repairing visual loss caused by retinal degeneration (RD) in people; however, its application is still significantly limited by the low effective delivery, proliferation and differentiation of RPCs. Herein, a self-healing injectable hydrogel (CS-Odex) based on chitosan hydrochloride (CS) and oxidized dextran (Odex) was developed via a dynamic Schiff-base linkage as a bioactive vehicle for the delivery of RPCs. Moreover, its biological effects on the RPC behaviors, including survival, proliferation and differentiation, were systematically evaluated. The CS-Odex hydrogel exhibits good biocompatibility and suitable mechanical stiffness for the growth of RPCs, and the cells can retain a high survival ratio (about 90%) with the protection of the self-healing CS-Odex hydrogels post-injection. In addition, the proliferation of RPCs in the CS-Odex hydrogels was significantly enhanced by activating the Akt and Erk pathways, especially in the hydrogel with higher CS content. Moreover, the differentiation of RPCs was improved by the CS-Odex hydrogel. Particularly, the differentiation of RPCs towards photoreceptors, the most important cell-type for RD, was elevated. Therefore, the self-healing injectable CS-Odex hydrogel would provide a promising platform for the delivery of RPCs and promote the proliferation and differentiation of RPCs towards RPC-based transplantation therapy in the future.

Mussel-inspired injectable hydrogel and its counterpart for actuating proliferation and neuronal differentiation of retinal progenitor cells.

Biomaterials-mediated retinal progenitor cell (RPC)-based transplantation therapy has shown substantial potential for retinal degeneration (RD), but it is limited by the poor RPC survival, proliferation and differentiation. Herein, the gelatin-hyaluronic acid (Gel-HA)-based hydrogels formed via moderate Michael-type addition reaction with or without the introduction of mussel-inspired polydopamine (PDA), i.e. Gel-HA-PDA and its counterpart Gel-HA hydrogels are developed, and their effects on the biological behaviour of RPCs, including adhesion, survival, proliferation, differentiation, delivery and migration are investigated. The hybrid hydrogels can adopt the intricate structure of the retina with suitable mechanical strength, degradation rate and biological activity to support cellular adhesion, survival and delivery. Meanwhile, Gel-HA hydrogel can remarkably promote RPC proliferation with much larger cell clusters, while Gel-HA-PDA hydrogel significantly enhances RPC adhesion and migration, and directs RPCs to preferentially differentiate toward retinal neurons such as photoreceptors (the most crucial cell-type for RD treatment), which is mainly induced by the activation of integrin α5ß1-phosphatidylinositol-3-kinase (PI3K) pathway. This study demonstrates that Gel-HA hydrogel possesses great potential for RPC proliferation, while mussel-inspired PDA-modified Gel-HA hydrogel with superior biocompatibility can significantly promote RPC neuronal differentiation, providing new insights for developing biomedical materials applied for RPC-based transplantation therapy.

Impaired granulocyte-macrophage colony-stimulating factor bioactivity accelerates surgical recurrence in ileal Crohn's disease.

AIM: To examine the relationship between elevated granulocyte-macrophage colony-stimulating factor (GM-CSF) auto-antibodies (Ab) level and time to surgical recurrence after initial surgery for Crohn's disease (CD). METHODS: We reviewed 412 charts from a clinical database at tertiary academic hospital. Patients included in the study had ileal or ileocolonic CD and surgical resection of small bowel or ileocecal region for management of disease. Serum samples were analyzed for serological assays including GM-CSF cytokine, GM-CSF Ab, ASCA IgG and IgA, and genetic markers including SNPs rs2066843, rs2066844, rs2066845, rs2076756 and rs2066847 in NOD2, rs2241880 in ATG16L1, and rs13361189 in IRGM. Cox proportional-hazards models were used to assess the predictors of surgical recurrence. RESULTS: Ninety six percent of patients underwent initial ileocecal resection (ICR) or ileal resection (IR) and subsequently 40% of patients required a second ICR/IR for CD. GM-CSF Ab level was elevated at a median of 3.81 mcg/mL. Factors predicting faster time to a second surgery included elevated GM-CSF Ab [hazard ratio (HR) 3.52, 95%CI: 1.45-8.53, P = 0.005] and elevated GM-CSF cytokine (HR = 2.48, 95%CI: 1.31-4.70, P = 0.005). Factors predicting longer duration between first and second surgery included use of Immunomodulators (HR = 0.49, 95%CI: 0.31-0.77, P = 0.002), the interaction effect of low GM-CSF Ab levels and smoking (HR = 0.60, 95%CI: 0.45-0.81, P = 0.001) and the interaction effect of low GM-CSF cytokine levels and ATG16L1 (HR = 0.65, 95%CI: 0.49-0.88, P = 0.006). CONCLUSION: GM-CSF bioavailability plays a critical role in maintaining intestinal homeostasis. Decreased bioavailability coupled with the genetic risk markers and/or smoking results in aggressive CD behavior.

Biomimetic Macroporous PCL Scaffolds for Ex Vivo Expansion of Cord Blood-Derived CD34+ Cells with Feeder Cells Support.

Ex vivo expansion of hematopoietic stem cells (HSCs) with most current methods can hardly satisfy clinical application requirement. While in vivo, HSCs efficiently self-renew in niche where they interact with 3D extracellular matrix and stromal cells. Therefore, co-cultures of CD34+ cells and mesenchyme stem cells derived from human amniotic membrane (hAMSCs) on the basis of biomimetic macroporous three-dimensional (3D) poly(ε-caprolactone) (PCL) scaffolds are developed, where scaffolds and hAMSCs are applied to mimic structural and cellular microenvironment of HSCs. The influence of scaffolds, feeder cells, and contact manners on expansion and stemness maintenance of CD34+ cells is investigated in this protocol. Biomimetic scaffolds-dependent co-cultures of CD34+ cells and hAMSCs can effectively promote the expansion of CD34+ cells; meanwhile, indirect contact is superior to direct contact. The combination of biomimetic scaffolds and hAMSCs represents a new strategy for achieving clinical-scale ex vivo expansion of CD34+ cells.

Aberrant DNA Methylation: Implications in Racial Health Disparity.

BACKGROUND: Incidence and mortality rates of colorectal carcinoma (CRC) are higher in African Americans (AAs) than in Caucasian Americans (CAs). Deficient micronutrient intake due to dietary restrictions in racial/ethnic populations can alter genetic and molecular profiles leading to dysregulated methylation patterns and the inheritance of somatic to germline mutations. MATERIALS AND METHODS: Total DNA and RNA samples of paired tumor and adjacent normal colon tissues were prepared from AA and CA CRC specimens. Reduced Representation Bisulfite Sequencing (RRBS) and RNA sequencing were employed to evaluate total genome methylation of 5'-regulatory regions and dysregulation of gene expression, respectively. Robust analysis was conducted using a trimming-and-retrieving scheme for RRBS library mapping in conjunction with the BStool toolkit. RESULTS: DNA from the tumor of AA CRC patients, compared to adjacent normal tissues, contained 1,588 hypermethylated and 100 hypomethylated differentially methylated regions (DMRs). Whereas, 109 hypermethylated and 4 hypomethylated DMRs were observed in DNA from the tumor of CA CRC patients; representing a 14.6-fold and 25-fold change, respectively. Specifically; CHL1, 4 anti-inflammatory genes (i.e., NELL1, GDF1, ARHGEF4, and ITGA4), and 7 miRNAs (of which miR-9-3p and miR-124-3p have been implicated in CRC) were hypermethylated in DNA samples from AA patients with CRC. From the same sample set, RNAseq analysis revealed 108 downregulated genes (including 14 ribosomal proteins) and 34 upregulated genes (including POLR2B and CYP1B1 [targets of miR-124-3p]) in AA patients with CRC versus CA patients. CONCLUSION: DNA methylation profile and/or products of its downstream targets could serve as biomarker(s) addressing racial health disparity.

Identification of Candidate Adherent-Invasive E. coli Signature Transcripts by Genomic/Transcriptomic Analysis.

Adherent-invasive Escherichia coli (AIEC) strains are detected more frequently within mucosal lesions of patients with Crohn's disease (CD). The AIEC phenotype consists of adherence and invasion of intestinal epithelial cells and survival within macrophages of these bacteria in vitro. Our aim was to identify candidate transcripts that distinguish AIEC from non-invasive E. coli (NIEC) strains and might be useful for rapid and accurate identification of AIEC by culture-independent technology. We performed comparative RNA-Sequence (RNASeq) analysis using AIEC strain LF82 and NIEC strain HS during exponential and stationary growth. Differential expression analysis of coding sequences (CDS) homologous to both strains demonstrated 224 and 241 genes with increased and decreased expression, respectively, in LF82 relative to HS. Transition metal transport and siderophore metabolism related pathway genes were up-regulated, while glycogen metabolic and oxidation-reduction related pathway genes were down-regulated, in LF82. Chemotaxis related transcripts were up-regulated in LF82 during the exponential phase, but flagellum-dependent motility pathway genes were down-regulated in LF82 during the stationary phase. CDS that mapped only to the LF82 genome accounted for 747 genes. We applied an in silico subtractive genomics approach to identify CDS specific to AIEC by incorporating the genomes of 10 other previously phenotyped NIEC. From this analysis, 166 CDS mapped to the LF82 genome and lacked homology to any of the 11 human NIEC strains. We compared these CDS across 13 AIEC, but none were homologous in each. Four LF82 gene loci belonging to clustered regularly interspaced short palindromic repeats region (CRISPR)--CRISPR-associated (Cas) genes were identified in 4 to 6 AIEC and absent from all non-pathogenic bacteria. As previously reported, AIEC strains were enriched for pdu operon genes. One CDS, encoding an excisionase, was shared by 9 AIEC strains. Reverse transcription quantitative polymerase chain reaction assays for 6 genes were conducted on fecal and ileal RNA samples from 22 inflammatory bowel disease (IBD), and 32 patients without IBD (non-IBD). The expression of Cas loci was detected in a higher proportion of CD than non-IBD fecal and ileal RNA samples (p <0.05). These results support a comparative genomic/transcriptomic approach towards identifying candidate AIEC signature transcripts.