YB-1 transferred by gastric cancer exosomes promotes angiogenesis via enhancing the expression of angiogenic factors in vascular endothelial cells.

BACKGROUND: Angiogenesis is important for the progression of gastric cancer (GC). Y-box binding protein 1 (YB-1) predicts advanced disease and indicates neovasculature formation in GC tissues, while the related mechanisms remain elusive. Exosomes mediate intercellular communications via transferring various molecules including proteins, lipids, mRNAs, and microRNAs, while the cargos of GC exosomes and the related mechanisms in GC angiogenesis were rarely reported except for several microRNAs. METHODS: In this study, human umbilical vein endothelial cells (HUVECs) were, respectively, treated by the exosomes isolated from the YB-1 transfected and the control SGC-7901 cells (SGC-7901-OE-Exo and SGC-7901-NC-Exo), and their apoptosis, proliferation, migration, invasion, and angiogenesis were, sequentially, compared. The levels of angiogenic factors including VEGF, Ang-1, MMP-9 and IL-8 in the exosome-treated HUVECs and the GC-derived exosomes were, separately, detected using PCR and Western blotting as well as RNA sequencing assays. RESULTS: We observed the consistent level of YB-1 in the exosomes and their originated GC cells, and the internalization of exosomes into HUVECs. Comparing with SGC-7901-NC-Exo, SGC-7901-OE-Exo significantly inhibited the apoptosis but promoted the proliferation, migration, invasion, and angiogenesis of HUVECs, within which the increased mRNA and protein levels of VEGF, Ang-1, MMP-9 and IL-8 were demonstrated. Meanwhile, mRNA levels of VEGF, Ang-1, MMP-9 and IL-8 showed no significant difference between SGC-7901-NC-Exo and SGC-7901-OE-Exo, although statistically higher mRNA of YB-1 was detected in the SGC-7901-OE-Exo. CONCLUSIONS: Our findings illustrate YB-1 as the key component of exosome to promote GC angiogenesis by upregulating specific angiogenic factors in the exosome-treated endothelial cells but not in the exosomes themselves.

Primary adult primitive neuroectodermal tumor of the bladder: A case report and literature review.

RATIONALE: A primary primitive neuroectodermal tumor (PNET) is a rare and highly malignant tumor that often occurs in the central nervous system of children and young adults. This tumor is rarely observed in the bladder. PATIENT CONCERNS: In this paper, we describe the case of a 64-year-old man with a PNET of the bladder. He experienced dull pain in the lower left abdomen for 5 months (without any obvious inducement), which gradually became aggravated and intolerable. DIAGNOSES: Partial cystectomy was performed, and a PNET of the bladder, which is extremely rare, was confirmed. INTERVENTIONS: Following cystectomy, the patient's general postoperative state was poor and he could not tolerate chemotherapy. Thus, he was subjected to pelvic radiotherapy for 2 weeks. OUTCOMES: His physical condition did not improve significantly after radiotherapy; however, we still plan to continue it. If the patient's physical condition improves, chemotherapy will be considered. LESSONS: Most cases of PNETs are intravesical or at least mainly endophytic. However, in this case, the mucosal layer was barely involved, and the tumor mainly grew out of the bladder, which is very rare. The present case provides reference for the diagnosis of PNET.

Repair and regeneration of small intestine: A review of current engineering approaches.

The small intestine (SI) is difficult to regenerate or reconstruct due to its complex structure and functions. Recent developments in stem cell research, advanced engineering technologies, and regenerative medicine strategies bring new hope of solving clinical problems of the SI. This review will first summarize the structure, function, development, cell types, and matrix components of the SI. Then, the major cell sources for SI regeneration are introduced, and state-of-the-art biofabrication technologies for generating engineered SI tissues or models are overviewed. Furthermore, in vitro models and in vivo transplantation, based on intestinal organoids and tissue engineering, are highlighted. Finally, current challenges and future perspectives are discussed to help direct future applications for SI repair and regeneration.

Calcifying fibrous tumor of the mediastinum: A case report.

BACKGROUND: Calcifying fibrous tumor (CFT) is a rare benign mesenchymal tumor that often occurs in deep soft tissue of children and young adults. CFT rarely occurs in the mediastinum. CASE SUMMARY: In this paper, we describe a 31-year-old male patient with CFT in the mediastinum. The patient did not have any symptoms, and the posterior mediastinal lesion was unintentionally found during routine re-examination of thyroid cancer. The tumor had no adhesion to the surrounding tissue and was successfully and completely removed. Pathology showed a large amount of collagen-rich fibrous connective tissue. There was scattered dystrophic calcification and gravel in the fibrous tissue and a small amount of lymphocyte and plasma cell infiltration and lymphoid follicle formation in the interstitial fluid. In addition, findings showed 20 IgG4+ plasma cells per high-powered field of the diseased tissue, an IgG4+/IgG ratio of about 20%, and normal serum IgG4 levels. The final diagnosis was CFT of the mediastinum (CFTM). No evidence of tumor recurrence was observed by computed tomography at 3 mo after surgery. CONCLUSION: IgG4+ plasma cell enlargement may occur in CFTM, but clinical manifestations and serological tests suggest that it is not IgG4-related disease. We speculate that it may be an independent tumor subtype.

Development of Cryogel-Based Guidance Conduit for Peripheral Nerve Regeneration.

Development of a nerve guidance conduit (NGC) is critically required for replacing the gold standard, autograft, in the repair of peripheral nerve (PN) injury with a long transecting gap. In the present study, an engineered cryogel-based NGC composed of methacrylated gelatin (MeGel), methacrylated hyaluronic acid (MeHA), and 4 arm poly(ethylene glycol) acrylate (4Arm-PEG-acrylate) was fabricated by utilizing a free radical cryopolymerization method. In vitro studies showed that the MeGel/MeHA/4Arm-PEG-acrylate hybrid cryogel provided a favorable surface and microenvironment for the adhesion, proliferation, and phenotype maintenance of rabbit Schwann cells. A 10 mm long transection in the rat sciatic nerve was created for in vivo studies. The results demonstrated that after the 16 weeks of repairing, the cryogel-based NGC supported axon regeneration and remyelination with comparable myelinating diameter and density of the regenerated nerve fibers to the autograft group. The rats with implanted cryogel-based NGC also showed sensory function recovery. This study demonstrated that our cryogel-based NGC is effective at assisting the repair of damaged PN because of its excellent structural stability, great handling convenience, and regeneration guidance capacity.

Establishment of a Human iPSC- and Nanofiber-Based Microphysiological Blood-Brain Barrier System.

The blood-brain barrier (BBB) is an active and complex diffusion barrier that separates the circulating blood from the brain and extracellular fluid, regulates nutrient transportation, and provides protection against various toxic compounds and pathogens. Creating an in vitro microphysiological BBB system, particularly with relevant human cell types, will significantly facilitate the research of neuropharmaceutical drug delivery, screening, and transport, as well as improve our understanding of pathologies that are due to BBB damage. Currently, most of the in vitro BBB models are generated by culturing rodent astrocytes and endothelial cells, using commercially available transwell membranes. Those membranes are made of plastic biopolymers that are nonbiodegradable, porous, and stiff. In addition, distinct from rodent astrocytes, human astrocytes possess unique cell complexity and physiology, which are among the few characteristics that differentiate human brains from rodent brains. In this study, we established a novel human BBB microphysiologocal system, consisting of a three-dimensionally printed holder with a electrospun poly(lactic- co-glycolic) acid (PLGA) nanofibrous mesh, a bilayer coculture of human astrocytes, and endothelial cells, derived from human induced pluripotent stem cells (hiPSCs), on the electrospun PLGA mesh. This human BBB model achieved significant barrier integrity with tight junction protein expression, an effective permeability to sodium fluorescein, and higher transendothelial electrical resistance (TEER) comparing to electrospun mesh-based counterparts. Moreover, the coculture of hiPSC-derived astrocytes and endothielial cells promoted the tight junction protein expression and the TEER value. We further verified the barrier functions of our BBB model with antibrain tumor drugs (paclitaxel and bortezomib) and a neurotoxic peptide (amyloid ß 1-42). The human microphysiological system generated in this study will potentially provide a new, powerful tool for research on human BBB physiology and pathology.

Mechanically robust cryogels with injectability and bioprinting supportability for adipose tissue engineering.

Bioengineered adipose tissues have gained increased interest as a promising alternative to autologous tissue flaps and synthetic adipose fillers for soft tissue augmentation and defect reconstruction in clinic. Although many scaffolding materials and biofabrication methods have been investigated for adipose tissue engineering in the last decades, there are still challenges to recapitulate the appropriate adipose tissue microenvironment, maintain volume stability, and induce vascularization to achieve long-term function and integration. In the present research, we fabricated cryogels consisting of methacrylated gelatin, methacrylated hyaluronic acid, and 4arm poly(ethylene glycol) acrylate (PEG-4A) by using cryopolymerization. The cryogels were repeatedly injectable and stretchable, and the addition of PEG-4A improved the robustness and mechanical properties. The cryogels supported human adipose progenitor cell (HWA) and adipose derived mesenchymal stromal cell adhesion, proliferation, and adipogenic differentiation and maturation, regardless of the addition of PEG-4A. The HWA laden cryogels facilitated the co-culture of human umbilical vein endothelial cells (HUVEC) and capillary-like network formation, which in return also promoted adipogenesis. We further combined cryogels with 3D bioprinting to generate handleable adipose constructs with clinically relevant size. 3D bioprinting enabled the deposition of multiple bioinks onto the cryogels. The bioprinted flap-like constructs had an integrated structure without delamination and supported vascularization. STATEMENT OF SIGNIFICANCE: Adipose tissue engineering is promising for reconstruction of soft tissue defects, and also challenging for restoring and maintaining soft tissue volume and shape, and achieving vascularization and integration. In this study, we fabricated cryogels with mechanical robustness, injectability, and stretchability by using cryopolymerization. The cryogels promoted cell adhesion, proliferation, and adipogenic differentiation and maturation of human adipose progenitor cells and adipose derived mesenchymal stromal cells. Moreover, the cryogels also supported 3D bioprinting on top, forming vascularized adipose constructs. This study demonstrates the potential of the implementation of cryogels for generating volume-stable adipose tissue constructs and provides a strategy to fabricate vascularized flap-like constructs for complex soft tissue regeneration.

Effects of tunable, 3D-bioprinted hydrogels on human brown adipocyte behavior and metabolic function.

Obesity and its related health complications cause billions of dollars in healthcare costs annually in the United States, and there are yet to be safe and long-lasting anti-obesity approaches. Using brown adipose tissue (BAT) is a promising approach, as it uses fats for energy expenditure. However, the effect of the microenvironment on human thermogenic brown adipogenesis and how to generate clinically relevant sized and functioning BAT are still unknown. In our current study, we evaluated the effects of endothelial growth medium exposure on brown adipogenesis of human brown adipose progenitors (BAP). We found that pre-exposing BAP to angiogenic factors promoted brown adipogenic differentiation and metabolic activity. We further 3D bioprinted brown and white adipose progenitors within hydrogel-based bioink with controllable physicochemical properties and evaluated the cell responses in 3D bioprinted environments. We used soft, stiff, and stiff-porous constructs to encapsulate the cells. All three types had high cell viability and allowed for varying levels of function for both white and brown adipocytes. We found that the soft hydrogel constructs promoted white adipogenesis, while the stiff-porous hydrogel constructs improved both white and brown adipogenesis and were the optimal condition for promoting brown adipogenesis. Consistently, stiff-porous hydrogel constructs showed higher metabolic activities than stiff hydrogel constructs, as assessed by 2-deoxy glucose uptake (2-DOG) and oxygen consumption rate (OCR). These findings show that the physicochemical environments affect the brown adipogenesis and metabolic function, and further tuning will be able to optimize their functions. Our results also demonstrate that 3D bioprinting of brown adipose tissues with clinically relevant size and metabolic activity has the potential to be a viable option in the treatment of obesity and type 2 diabetes. STATEMENT OF SIGNIFICANCE: One promising strategy for the treatment or prevention of obesity-mediated health complications is augmenting brown adipose tissues (BAT), which is a specialized fat that actively dissipate energy in the form of heat and maintain energy balance. In this study, we determined how pre-exposing human brown adipose progenitors (BAP) to angiogenic factors in 2D and how bioprinted microenvironments in 3D affected brown adipogenic differentiation and metabolic activity. We demonstrated that white and brown adipogenesis, and thermogenesis were regulated by tuning the bioprintable matrix stiffness and construct structure. This study not only unveils the interaction between BAP and 3D physiological microenvironments, but also presents a novel tissue engineered strategy to manage obesity and other related metabolic disorders.

3D Bioprinting of Breast Cancer Models for Drug Resistance Study.

Adipose-derived mesenchymal stem/stromal cells (ADMSC) are one of the major stromal cells in the breast cancer microenvironment that promote cancer progression. Previous studies on the effects of ADMSC on breast cancer metastasis and drug resistance, using two-dimensional (2D) cultures, remained inconclusive. In the present study, we compared cocultured ADMSC and human epidermal receptor 2 positive breast primary breast cancer cells (21PT) in 2D and three-dimensional (3D) cultures and then examined their response to doxorubicin (DOX). We examined 3D bioprinted constructs with breast cancer cells in the middle and ADMSC in the edge region, which were made by using dual hydrogel-based bioinks. We found that the percentage of cleaved Caspase-3 positive cells was significantly lower in the bioprinted constructs with ADMSC and 21PT than that in the cancer cell alone constructs, in response to low DOX dose. We further increased the thickness of the ADMSC layers to mimic the status of obesity and then examined the effect of ADMSC thickness on DOX resistance and lysyl oxidase (LOX) secretion. In the moderate and thick-layered ADMSC constructs, significantly more cells were stained negative for cleaved Caspase-3, indicating less apoptosis. Both ADMSC and 21PT intrinsically expressed LOX, regardless of changes in thickness or DOX administration. Notably, treatment with a LOX inhibitor significantly decreased the stiffness in the ADMSC region but did not affect the stiffness in the 21PT region. In addition, LOX inhibitor treatment enhanced DOX sensitivity of 21PT in the bioprinted constructs, as seen by a decrease in LOX secretion and downregulation of adenosine triphosphate-binding cassette transporter gene expression. Taken together, we demonstrate that 3D bioprinted these breast cancer models faithfully reproduce in vivo conditions and should provide better models for examining breast cancer biology and for screening for drug discoveries.

Pulmonary spindle cell carcinoma with unusual morphology: A rare case report and review of the literature.

RATIONALE: Pulmonary spindle cell carcinoma (PSCC) is a highly malignant carcinoma that often exhibits the histopathological characteristic of cell pleomorphism. PATIENT CONCERNS: Here, we report a PSCC case in a 59-year-old woman with multiple metastases. The pathological manifestation of this case did not show high-grade pleomorphism or atypia, and was easily mimicked by other borderline or low-grade malignant tumors such as an inflammatory myofibroblastic tumor (IMT). DIAGNOSES: Based on the results of immunohistochemistry and fluorescence in situ hybridization after the operation, IMT was ruled out, and PSCC was confirmed. INTERVENTIONS: Resection of the right middle lobe and dissection of the lymph nodes were performed. The patient was given 2 cycles of chemotherapy with cisplatin and 1 cycle of radiotherapy after the operation. OUTCOMES: Metastatic lesions in the left kidney, the right lung, the first thoracic vertebrae, the retroperitoneal lymph nodes, and the mediastinal lymph nodes were identified by positron emission tomography/computed tomography 4 months after the operation, and the patient died 2 months later. LESSONS: PSCC is a highly malignant carcinoma, but it rarely shows a low-grade malignant histological morphology. So, efforts should be focused on differentiating it from other borderline or low malignant tumors (such as IMT) to avoid misdiagnosis.

Primary adenoid cystic carcinoma of the trachea with thyroid invasion: a case report and literature review.

Primary adenoid cystic carcinoma (ACC) of the trachea with thyroid invasion is very rare. In this report, we present a 46-year-old man with primary ACC of the trachea with thyroid invasion. ACC invasion of the thyroid is very rare and is easily misdiagnosed. The patient sought consultation due to a 6-month history of dysphagia and associated dyspnea. A contrast-enhanced computed tomography scan obtained at the time of admission revealed bilateral thyroid masses and tracheal wall thickening. The thyroid masses were fused to the trachea and the esophagus without discernible borders, intraoperatively. Frozen pathology suggested poorly differentiated cancer, and a bilateral partial thyroidectomy was performed. Postoperative pathology revealed primary tracheal ACC with thyroid invasion. The patient died 1 month after surgery. We have also summarized the literature on the clinical presentation, diagnosis, and treatment of thyroid-invasive ACC.

Anorectal melanoma metastatic to the breast: a case report and review of the literature.

Melanoma develops from melanocytes and typically occurs on the skin and mucosa with a high degree of malignancy. Intensive local invasion and distant metastasis of melanoma result in poor patient prognosis, owing to frequent metastases to the lungs, bones, brains, and other parts of the body. In the present study, we report a case of anorectal melanoma in a 56-year-old woman who was admitted to our hospital because of local recurrence 9 months after local resection. She subsequently underwent radical surgery. Metastasis to the left breast occurred within 4 months after radical surgery. Metastasis of anorectal melanoma to the breast is very rare. In the present case report and literature review, we analyzed the clinical manifestation, diagnosis, and treatment of anorectal melanoma metastatic to the breast.

A population-based study on health-related quality of life among urban community residents in Shenyang, Northeast of China.

BACKGROUND: Due to the rising standard of living environment and advances in public health and medical care in China, it has been a tendency in recent years that health-related quality of life (HRQoL) has been increasingly acknowledged in community health management. However, large-scale population-based study on evaluating HQRoL in northeast of China was not conducted. This article aims to investigate the HRQoL in community residents in Northeast China and explore the associated factors. METHODS: Stratified multiple-stage sampling method was used in the cross-sectional survey to investigate HRQoL of community residents in northeast of China. Univariate analysis and multiple linear regressions were used to analyze the factors associated to HRQoL of the community residents. RESULTS: The results were confirmed that HRQoL in general population was well performed for the first time in northeast of China in a large scale population. Community residents had better mental health than physical health. The factors influencing HRQoL included gender, age, educational level, marital status, ethnic group, chronic disease status, having breakfast frequency weekly and sleep quality. However, drinking and smoking habits did not affect residents' HRQoL. CONCLUSIONS: In this study, the result of the large-scale survey was satisfactory in northeast of China, providing HRQoL status of community residents. Policies on specific health management in community public health would emphasize on lifestyle behaviors especially eating habits in order to improving HRQoL.