The transcript ENST00000444125 of lncRNA LINC01503 promotes cancer stem cell properties of glioblastoma cells via reducing FBXW1 mediated GLI2 degradation.

LINC010503 is a novel oncogenic lncRNA in multiple cancers. In this study, we further explored the expression of LINC010503 transcripts and their regulations on the glioblastoma (GBM) stem cell (GSC) properties. LINC01503 transcription patterns in GBM and normal brain tissues were compared using RNA-seq data from Genotype-Tissue Expression (GTEx) and The Cancer Genome Atlas (TCGA)-GBM. GBM cell lines (U251 and U87) were used as in vitro cell models for cellular and molecular studies. The results showed that ENST00000444125 was the dominant transcript of LINC01503 in both normal and tumor tissues. Its expression was significantly elevated in the tumor group and associated with poor survival outcomes. LINC01503 had both cytoplasmic and nuclear distribution. It positively modulated the expression of multiple GSC markers, including CD133, SOX2, NESTIN, ALDH1A1, and MSI1, and tumorsphere formation in U251 and U87 cells. RNA pull-down and RIP-qPCR assay confirmed an interaction between ENST00000444125 and GLI2. ENST00000444125 positively regulated the half-life of the GLI2 protein in GBM cells. ENST00000444125 overexpression reduced GLI2 ubiquitination and partially attenuated FBXW1 overexpression induced GLI2 ubiquitination. ENST00000444125 overexpression could activate Wnt/ß-catenin signaling in GBM cells. However, these activating effects were remarkedly hampered when GLI2 was knocked down. In conclusion, this study revealed that LINC01503 might have isoform-specific dysregulation in GBM. Among the two major transcripts expressed in GBM cells, ENST00000444125 might be the major functional transcript. Its upregulation might enhance the GSC properties of GBM cells via reducing FBXW1-mediated proteasomal degradation of GLI2.

Cu/Ag Complex Modified Keggin-Type Coordination Polymers for Improved Electrochemical Capacitance, Dual-Function Electrocatalysis, and Sensing Performance.

Different metal-organic units were introduced into the {PMo12} polyoxometalate (POM) system to yield three porous coordination polymers with distinct characteristics, {Cu(pra)2}[{Cu(pra)2}3{PMo11VIMoVO40}] (1), [{Ag5(pz)6(H2O)0.5Cl}{PMo11VIMoVO40}] (2), and [{Cu3(bpz)5(H2O)}{PMo12O40}] (3) (pra = pyrazole; pz = pyrazine; bpz = benzopyrazine), via an in situ hydrothermal method. In comparison with the maternal Keggin cluster and most reported POM electrode materials, compounds 1-3 exhibit larger specific capacitances (672.2, 782.1, and 765.2 F g-1 at a current density of 2.4 A g-1, respectively), superior cyclic stability (91.5%, 89.3%, and 87.8% of cycle efficiency after 5000 cycles, respectively), and boosted conductivity, which may be attributed to the introduction of metal-organic units. The result indicates that metal-organic units can effectively enhance the capacitance performance of POMs. This may be due to the fact that they provide additional redox centers, induce the formation of stable porous structures, and improve ion/electron transfer efficiency. Compounds 1-3 present excellent electrocatalytic activity in reducing peroxide (H2O2) and oxidizing ascorbic acid (AA). In addition, compound 2 shows an outstanding sensing performance detection of AA and H2O2.

Cell-recruited microspheres for OA treatment by dual-modulating inflammatory and chondrocyte metabolism.

Osteoarthritis (OA) is a degenerative disease potentially exacerbated due to inflammation, cartilage degeneration, and increased friction. Both mesenchymal stem cells (MSCs) and pro-inflammatory macrophages play important roles in OA. A promising approach to treating OA is to modify multi-functional hydrogel microspheres to target the OA microenvironment and structure. Arginyl-glycyl-aspartic acid (RGD) is a peptide widely used in bioengineering owing to its cell adhesion properties, which can recruit BMSCs and macrophages. We developed TLC-R, a microsphere loaded with TGF-ß1-containing liposomes. The recruitment effect of TLC-R on macrophages and BMSCs was verified by in vitro experiments, along with its function of promoting chondrogenic differentiation of BMSCs. And we evaluated the effect of TLC-R in balancing OA metabolism in vitro and in vivo. When TLC-R was co-cultured with BMSCs and lipopolysaccharide (LPS)-treated macrophages, it showed the ability to recruit both cells in substantial numbers. As the microspheres degraded, TGF-ß1 and chondroitin sulfate (ChS) were released to promote chondrogenic differentiation of the recruited BMSCs, modulate chondrocyte metabolism and inhibit inflammation induced by the macrophages. Furthermore, in vivo analysis showed that TLC-R restored the narrowed space, reduced osteophyte volume, and improved cartilage metabolic homeostasis in OA rats. Altogether, TLC-R provides a comprehensive and novel solution for OA treatment by dual-modulating inflammatory and chondrocyte metabolism.

Microenvironment-responsive bilayer hydrogel microspheres with gelatin-shell for osteoarthritis treatment.

Osteoarthritis is a long-term degenerative condition of the joints that is characterized by the breakdown of cartilage and inflammation of the synovial membrane. The presence of an inflammatory microenvironment and the degradation of the extracellular matrix produced by chondrocytes leads to the aggravation of cartilage injury, hindering the treatment of osteoarthritis. A promising approach to address this issue is to apply a combined strategy that is sensitive to the specific conditions in osteoarthritic joints and possesses properties that can reduce inflammation and promote cartilage healing. Here, inspired by the structure of chocolate-covered peanuts, we developed an injectable, environment-responsive bilayer hydrogel microsphere using microfluidics technology. The microsphere applied chondroitin sulfate methacryloyl (ChsMA) as its core and was coated with a methacryloyl gelatin (GelMA) shell that was loaded with celecoxib (CLX) liposomes (ChsMA+CLX@Lipo@GelMA). CLX was released from the liposomes when the GelMA shell rapidly degraded in response to the osteoarthritic microenvironment and suppressed the generation of inflammatory agents, demonstrating a beneficial impact of the outer shell in reducing inflammation. While the inner methacryloyl microsphere core degraded, chondroitin sulfate was released to promote chondrocyte anabolism and facilitate cartilage repair. Thus, the synthesized bilayer hydrogel microspheres hold great potential for treating osteoarthritis.

A TGF-loading hydrogel scaffold capable of promoting chondrogenic differentiation for repairing rabbit nasal septum cartilage defect.

Hydrogel-based tissue engineering has been widely used to repair cartilage injury. However, whether this approach can be applied to treat nasal septum cartilage defects remains unclear. In this study, three gelatin methacrylate-based scaffolds loaded with transforming growth factor (TGF)-ß1 (GelMA-T) were prepared, and their effects on repair of nasal septum cartilage defects were examined. In vitro, the GelMA-T scaffolds showed good biocompatibility and promoted the chondrogenic differentiation of bone mesenchymal stem cells. Among three scaffolds, the 10% GelMA-T scaffold promoted chondrogenic differentiation most effectively, which significantly improved the expression of chondrocyte-related genes, including Col II, Sox9, and ACAN. In vivo, 10% GelMA-T scaffolds and 10% GelMA-T scaffolds loaded with bone mesenchymal stem cells (BMSCs; 10% GelMA-T/BMSCs) were transplanted into a nasal septum cartilage defect site in a rabbit model. At 4, 12, and 24 weeks after surgery, the nasal septum cartilage defects exhibited more complete repair in rabbits treated with the 10% GelMA-T/BMSC scaffold as demonstrated by hematoxylin & eosin, safranine-O, and toluidine blue staining. We showed that GelMA-T/BMSCs can be applied in physiological and structural repair of defects in nasal septum cartilage, providing a potential strategy for repairing cartilage defects in the clinic.

A functionalized collagen-I scaffold delivers microRNA 21-loaded exosomes for spinal cord injury repair.

MicroRNA (miRNA)-based therapies have shown great potential in the repair of spinal cord injury (SCI). MicroRNA 21 (miR21) has been proven to have an essential protective effect on SCI. However, there are some challenges for miRNAs application due to their easy degradation and ineffective cell penetration. As natural vesicles, exosomes were considered ideal carriers for miRNAs delivery for their advantages of low immunogenicity, inherent stability and tissue/cell penetration. However, poor targeting and the low capacity of specific miRNAs impede their practical applications. This study aims to develop a type of genetically engineered miR21-loaded exosomes that can be entrapped in collagen-I (Col-I) scaffold to repair SCI. The collagen-binding domain (CBD)-fused lysosome-associated membrane glycoprotein 2b (Lamp2b) protein (CBD-LP) and miR21 were overexpressed in host HEK293T (293T) cells that were used to produce engineered miR21-loaded exosomes. The CBD peptide fused in Lamp2b on the exosome surface can stably tether exosomes to Col-I scaffold, facilitate the retention of miR21-loaded exosomes in lesion sites, promote the sustained release of miR21 to cells. Finally, a functionalized Col-I scaffold biomaterial enriched with miR21-loaded exosomes was developed and it could benefit the repair of SCI. STATEMENT OF SIGNIFICANCE: MiRNA-based therapeutics have promising potential in spinal cord injury (SCI) repair. However, easy degradation and ineffective cell penetration impede miRNAs application. Exosomes are natural vehicles for miRNAs delivery but face the challenge of diffusion in vivo. Here, the collagen-binding domain (CBD)-fused Lamp2b and miR21 were overexpressed in HEK293T cells to produce miR21-loaded and CBD-modified exosomes (CBD-LP-miR21-EXOs). The CBD modified on the exosome surface can stably tether exosomes to collagen-I scaffold to form functionalized CBD-LP-miR21-EXO-Col scaffold that can facilitate the retention of miR21-loaded exosomes, promote the sustained release of miR21 to cells and finally benefit SCI repair. Furthermore, this type of functionalized collagen-I materials can be widely applied for other tissue injury repairs by enriching the CBD-LP-EXOs loaded with appropriate miRNAs.

Honeycomb-Like Hydrogel Microspheres for 3D Bulk Construction of Tumor Models.

A two-dimensional (2D) cell culture-based model is widely applied to study tumorigenic mechanisms and drug screening. However, it cannot authentically simulate the three-dimensional (3D) microenvironment of solid tumors and provide reliable and predictable data in response to in vivo, thus leading to the research illusions and failure of drug screening. In this study, honeycomb-like gelatin methacryloyl (GelMA) hydrogel microspheres are developed by synchronous photocrosslinking microfluidic technique to construct a 3D model of osteosarcoma. The in vitro study shows that osteosarcoma cells (K7M2) cultured in 3D GelMA microspheres have stronger tumorous stemness, proliferation and migration abilities, more osteoclastogenetic ability, and resistance to chemotherapeutic drugs (DOX) than that of cells in 2D cultures. More importantly, the 3D-cultured K7M2 cells show more tumorigenicity in immunologically sound mice, characterized by shorter tumorigenesis time, larger tumor volume, severe bone destruction, and higher mortality. In conclusion, honeycomb-like porous microsphere scaffolds are constructed with uniform structure by microfluidic technology to massively produce tumor cells with original phenotypes. Those microspheres could recapitulate the physiology microenvironment of tumors, maintain cell-cell and cell-extracellular matrix interactions, and thus provide an effective and convenient strategy for tumor pathogenesis and drug screening research.

STN versus GPi deep brain stimulation for dyskinesia improvement in advanced Parkinson's disease: A meta-analysis of randomized controlled trials.

BACKGROUND: Deep brain stimulation (DBS) of the subthalamic nucleus (STN) and the globus pallidus internus (GPi) are currently the most common and effective surgical targets for advanced Parkinson's disease (APD). Herein, we conducted a meta-analysis to evaluate the comprehensive efficacy of STN-DBS and GPi-DBS in patients with APD. METHODS: We conducted a systematic search for relevant articles written in English in the Cochrane Library, PubMed, and EMBASE databases through January 2020. Studies comparing the efficacy and clinical outcomes of GPi-DBS and STN-DBS for APD were included and analyzed. RESULTS: Ten eligible trials with a total of 857 patients were included in this meta-analysis. The results showed no significant difference between the STN-DBS and GPi-DBS groups in Unified Parkinson's Disease Rating Scale (UPDRS) III scores during the on and off-medication phases(SMD, 0.1; 95 % CI, -0.04 to 0.25; p = 0.17, on-med), (SMD,-0.12;95 % CI -0.37 to 0.13, p = 0.33,off-med). Dyskinesia scores and the activities of daily living (ADLs) scores during the on-medication phase showed significant differences in favor of GPi stimulation (SMD, 0.16; 95 % CI, 0.01-0.32; P < 0.05)/(SMD, 0.18; 95 % CI, 0.01-0.34; P < 0.05). The ADLs score during the off-medication phase showed no significant difference between the STN-DBS and GPi-DBS groups (SMD, -0.11; 95 % CI, -0.32-0.11; P = 0.33). The LED showed significant differences in favor of STN stimulation (SMD, -0.57; 95 % CI, -0.74-0.40; P < 0.00001). CONCLUSIONS: Both STN and GPi-DBS were equally effective in improving motor dysfunction. STN-DBS was superior for medication reduction, whereas GPi-DBS perhaps led to less dyskinesia and improved the postoperative ADLs (on-medication) in APD patients. Hence, the goals of DBS can be important in the target selection. More studies comparing the adverse events and quality of life between the two targets are needed.

A functional extracellular matrix biomaterial enriched with VEGFA and bFGF as vehicle of human umbilical cord mesenchymal stem cells in skin wound healing.

The construction of microvascular network is one of the greatest challenges for tissue engineering and cell therapy. Endothelial cells are essential for the construction of network of blood vessels. However, their application meets challenges in clinic due to the limited resource of autologous endothelium. Mesenchymal stem cells can effectively promote the angiogenesis in ischemic tissues for their abilities of endothelial differentiation and paracrine, and abundant sources. Extracellular matrix (ECM) has been widely used as an ideal biomaterial to mimic cellular microenvironment for tissue engineering due to its merits of neutrality, good biocompatibility, degradability, and controllability. In this study, a functional cell derived ECM biomaterial enriched with VEGFA and bFGF by expressing the collagen-binding domain fused factor genes in host cells was prepared. This material could induce endothelial differentiation of human umbilical cord mesenchymal stem cells (hUCMSCs) and promote angiogenesis, which may improve the healing effect of skin injury. Our research not only provides a functional ECM material to inducing angiogenesis by inducing endothelial differentiation of hUCMSCs, but also shed light on the ubiquitous approaches to endow ECM materials different functions by enriching different factors. This study will benefit tissue engineering and regenerative medicine researches.

The Extracellular Matrix Enriched With Exosomes for the Treatment on Pulmonary Fibrosis in Mice.

Pulmonary fibrosis (PF) is a severe respiratory disease caused by lung microenvironment changes. TGF-ß/Smad3 signaling pathway plays a critical role in the fibrotic process. MicroRNA-29 (miR-29) has proved to alleviate the occurrence of PF by downregulating TGF-ß/Smad3 signaling pathway. The miRNA application encounters obstacles due to its low stability in body and no targeting to lesions. Exosomes can be used for therapeutic delivery of miRNA due to their favorable delivery properties. However, low efficiency of separation and production impedes the therapeutic application of exosomes. In this study, we developed a liquid natural extracellular matrix (ECM) enriched with miR-29-loaded exosomes for PF treatment. The collagen-binding domain (CBD)-fused Lamp2b (CBD-Lamp2b) and miR-29 were overexpressed in human foreskin fibroblast (HFF) host cells for the entrapment of miR-29-loaded exosomes in ECM of the cells. The repeated freeze-thaw method was performed to prepare the liquid ECM enriched with exosomes without destroying the exosomal membrane. In summary, this study developed a novel functional ECM biomaterial for therapy of PF, and also provided a promising gene therapy platform for different diseases by treatment with liquid ECM that is, enriched with exosomes loaded with different functional miRNAs.

Immunopolarization-regulated 3D printed-electrospun fibrous scaffolds for bone regeneration.

Three-dimension (3D)-printed bioscaffolds are precise and personalized for bone regeneration. However, customized 3D scaffolds may activate the immune response in vivo and consequently impede bone formation. In this study, with layer-by-layer deposition and electrospinning technology to control the physical structure, 3D-printed PCL scaffolds with PLLA electrospun microfibrous (3D-M-EF) and nanofibrous (3D-N-EF) composites were constructed, and their immunomodulatory effect and the subsequent osteogenic effects were explored. Compared to 3D-N-EF scaffolds, 3D-M-EF scaffolds polarized more RAW264.7 cells toward alternatively activated macrophages (M2), as demonstrated by increased M2 and deceased classically activated macrophage (M1) phenotypic marker expression in the cells. In addition, the 3D-M-EF scaffolds shifted RAW264.7 cells to the M2 phenotype through PI3K/AKT signaling and enhanced VEGF and BMP-2 expression. Conditional medium from the RAW264.7 cells seeded in 3D-M-EF scaffolds promoted osteogenesis of MC3T3-E1 cells. Furthermore, in vivo study of repairing rat calvarial defects, the 3D-M-EF scaffolds increased the polarization of M2 macrophages, enhanced angiogenesis, and accelerated new bone formation. Collectively, our data suggested that well-designed 3D-M-EF scaffolds are favorable for osteogenesis through regulation of M2 polarization. Therefore, it is potential to utilize the physical structure of 3D-printed scaffolds to manipulate the osteoimmune environment to promote bone regeneration.

Substrate stiffness modulates bone marrow-derived macrophage polarization through NF-κB signaling pathway.

The stiffness of the extracellular matrix (ECM) plays an important role in regulating the cellular programming. However, the mechanical characteristics of ECM affecting cell differentiation are still under investigated. Herein, we aimed to study the effect of ECM substrate stiffness on macrophage polarization. We prepared polyacrylamide hydrogels with different substrate stiffness, respectively. After the hydrogels were confirmed to have a good biocompatibility, the bone marrow-derived macrophages (BMMs) from mice were incubated on the hydrogels. With simulated by the low substrate stiffness, BMMs displayed an enhanced expression of CD86 on the cell surface and production of reactive oxygen species (ROS) in cells, and secreted more IL-1ß and TNF-α in the supernatant. On the contrary, stressed by the medium stiffness, BMMs expressed more CD206, produced less ROS, and secreted more IL-4 and TGF-ß. In vivo study by delivered the hydrogels subcutaneously in mice, more CD68+CD86+ cells around the hydrogels with the low substrate stiffness were observed while more CD68+CD206+ cells near by the middle stiffness hydrogels. In addition, the expressions of NIK, phosphorylated p65 (pi-p65) and phosphorylated IκB (pi-IκB) were significantly increased after stimulation with low stiffness in BMMs. Taken together, these findings demonstrated that substrate stiffness could affect macrophages polarization. Low substrate stiffness promoted BMMs to shift to classically activated macrophages (M1) and the middle one to alternatively activated macrophages (M2), through modulating ROS-initiated NF-κB pathway. Therefore, we anticipated ECM-based substrate stiffness with immune modulation would be under consideration in the clinical applications if necessary.

Gelatin Templated Polypeptide Co-Cross-Linked Hydrogel for Bone Regeneration.

Polypeptides with short chains of amino acid monomers have been widely applied in the clinic because of their various biological functions. However, the easily-inactivated characteristics and burst releasing of the peptides limit their application in vivo. Here, a novel osteogenic polypeptide hydrogel (GelMA-c-OGP) is created by co-cross-linking template photo-cross-linked gelatin (GelMA) with photo-cross-linkable osteogenic growth peptides (OGP) using ultraviolet radiation. GelMA enables the formation of hydrogel with photo-cross-linkable OGP with good mechanical properties and also promotes bone regeneration. GelMA-c-OGP hydrogel accelerates the bone formation procedure of osteogenic precursor cells by significantly enhancing the expression of osteogenic-related genes BMP-2, OCN, and OPN, and increasing the precipitation of calcium salts in osteoblasts. Similarly, GelMA-c-OGP hydrogel promotes bone regeneration in vivo. Furthermore, it is observed that more collagen fibers connect cortical bones in the GelMA-c-OGP implanted group than the control group by hematoxylin-eosin and immunohistochemical staining of Collagen I and TGF-ß. The co-cross-linked OGP polypeptide converts from liquid to solid hydrogel with transient UV light in situ, which also can strengthen the mechanical property of the defect bone and avoid burst osteogenic peptide, releasing during the bone defect healing period. Overall, this hydrogel delivering system has a significant impact on bone defect healing compared with traditional methods.

An immunological electrospun scaffold for tumor cell killing and healthy tissue regeneration.

Antibody-based cancer immune therapy has attracted lots of research interest in recent years; however, it is greatly limited by the easy distribution and burst release of antibodies. In addition, after the clearance of the tissue, healthy tissue regeneration is another challenge for cancer treatment. Herein, we have developed a specific immunological tissue engineering scaffold using the agonistic mouse anti-human CD40 antibody (CD40mAb) incorporated into poly(l-lactide) (PLLA) electrospun fibers through the dopamine (PDA) motif (PLLA-PDA-CD40mAb). CD40mAb is successfully incorporated onto the surface of the electrospun fibrous scaffold, which is proved by immunofluorescence staining, and the PLLA-PDA-CD40mAb scaffold has an anti-tumor effect by locally releasing CD40mAb. Therefore, this immunological electrospun scaffold has very good potential to be developed as a powerful tool for localized tumor treatment, and this is the first to be reported in this area.

Occurrence of ethyl carbamate in three types of Chinese wines and its possible reasons.

A total of 75 wines including 30 white wines, 31 red wines, and 14 sparkling wines were obtained from several regions in China (Sinkiang, Tonghua, Huailai, Yantai, Changli, Shanxi, Gansu, and Ningxia). Ethyl carbamate (EC) was detected by gas chromatography mass spectrometry. The EC concentration ranged from less than 1.16 to 38.56 µg/L, and the concentrations in 17 wines exceeded the U.S. limit for table wines (15 µg/L). The concentrations of EC increased in the order of white, red, and sparkling wines with the corresponding mean concentrations of 6.12, 9.22, and 14.03 µg/L. The relationship between EC concentration and wine type suggested that EC concentrations in wines might be affected by vinification patterns, most likely due to the difference between EC precursors in different vinification processes. This work provides a novel clue for EC contamination in different wines.

[Analysis about clinical data of intrauterine infection of hepatitis B virus].

OBJECTIVE: To investigate a practical diagnostic method in clinic for fetuses infecting with hepatitis B (HBV) and study the mutual effects between fetal infection and clinical factors. METHODS: Venous blood was drawn from 144 cases of HBV carrier mothers and their neonates. HBV DNA was detected by polymerase chain reaction (PCR) and hybridization, HBV M was detected by enzyme linked immunoadsorbent assay (ELISA), and aspartate aminotransferase/alanine aminotransferase (AST/ALT) was detected by IFCC. Umbilical blood and femoral blood was taken from 40 of 144 neonates for HBV DNA detection. Clinical data, neonatal AST and ALT level were compared between fetal infection group and control group. RESULTS: (1) The fetal infectious rate was 22.9% (33/144). Comparing with peripheral venous blood sample, the sensitivity and positive predictive value of HBV DNA detected in cords was 100.0%, 80.0% respectively. Following up the infants, HBV DNA was found persistently positive in 7 of 28 intrauterine infectious infants 6 approximately 9 months after birth. HBsAg was found changing to be negative 1 month later in the infants with HBsAg positive at birth. (2) The fetal infectious rate in mothers with HBeAg (+) or HBV DNA (+) was 70.5%, 61.1% respectively which was significantly higher than that in mothers with HBeAg (-) or HBV DNA (-). P < 0.01. There was no significantly difference in mothers' age, gestational age, delivery way, birth weight (BW), body length (BL), Apgar score between fetal infectious group and control group. (3) The mean value of AST, ALT in fetal infectious group was (61.2 +/- 31.3) IU, (24.7 +/- 14.9) IU respectively, which was significantly higher than that in control group [(55.2 +/- 37.1) IU, (19.0 +/- 10.1) IU]. P < 0.01. CONCLUSIONS: (1) Detection of HBV DNA in cord blood is a sensitive index for diagnosing fetal infection, however detection of peripheral venous blood is with the significance of making correct diagnosis. (2) HBsAg or HBV DNA positive in mothers is one of the risk factors of intrauterine infection. There is no relationship among fetal infection and mothers' age, gestational age, delivery way, neonates' sex, BW, BL. (3) The liver function of neonates infected with HBV intrauterinely maybe impaired to some extent.