Stress Induces Dynamic, Cytotoxicity-Antagonizing TDP-43 Nuclear Bodies via Paraspeckle LncRNA NEAT1-Mediated Liquid-Liquid Phase Separation.

Despite the prominent role of TDP-43 in neurodegeneration, its physiological and pathological functions are not fully understood. Here, we report an unexpected role of TDP-43 in the formation of dynamic, reversible, liquid droplet-like nuclear bodies (NBs) in response to stress. Formation of NBs alleviates TDP-43-mediated cytotoxicity in mammalian cells and fly neurons. Super-resolution microscopy reveals distinct functions of the two RRMs in TDP-43 NB formation. TDP-43 NBs are partially colocalized with nuclear paraspeckles, whose scaffolding lncRNA NEAT1 is dramatically upregulated in stressed neurons. Moreover, increase of NEAT1 promotes TDP-43 liquid-liquid phase separation (LLPS) in vitro. Finally, we discover that the ALS-associated mutation D169G impairs the NEAT1-mediated TDP-43 LLPS and NB assembly, causing excessive cytoplasmic translocation of TDP-43 to form stress granules, which become phosphorylated TDP-43 cytoplasmic foci upon prolonged stress. Together, our findings suggest a stress-mitigating role and mechanism of TDP-43 NBs, whose dysfunction may be involved in ALS pathogenesis.

Controlling the Emulsion Type Using Adjustable Polyelectrolyte-Surfactant Complexes.

The combination of polyelectrolytes and ionic surfactants in precise proportions presents the possibility of producing a new class of emulsifiers with tunable emulsification properties. We use chitosan along with dioctyl sulfosuccinate sodium, also known as aerosol-OT (AOT), to demonstrate that emulsion types can be varied, and phase inversion emulsification (PIE) can be induced via changes in the water-phase pH and the molar ratio of the surfactant to the repeat unit of the polyelectrolyte. Confocal microscopy of the emulsions shows that the morphology can be changed from O/W to O/W/O to W/O by varying the surfactant to polyelectrolyte molar ratio at a fixed aqueous-phase pH while maintaining droplet sizes in the range of micrometers to tens of micrometers. Measurements of the oil (toluene)-water partition coefficient suggest that controlling the emulsion type relies on the ability of the surfactants to partition from the bulk oil to the bulk water phase and to induce polyelectrolyte-surfactant aggregation. We confirm this hypothesis using different combinations of polyelectrolytes and surfactants. Changes in the water-phase pH in situ induce phase inversion only in a particular direction, which suggests that the complexes at the interface are in a kinetically trapped state. Changes in the molar ratio in situ by addition of an oppositely charged surfactant also can induce phase inversion.

microRNA-199a counteracts glucocorticoid inhibition of bone marrow mesenchymal stem cell osteogenic differentiation through regulation of Klotho expression in vitro.

Osteogenic differentiation (OD) of bone marrow mesenchymal stem cells (BMSCs) is critically important for mitigation of osteoporosis. Glucocorticoids (GCs) are extensively used for treating chronic inflammation, although long-term exposure to GCs is capable of triggering osteoporosis. microRNAs (miRNAs) have been reported to play a critical role in bone diseases. In the present study, we treated BMSCs with dexamethasone (DEX) during OD to stimulate GC-mediated osteoporosis. Microarray and quantitative polymerase chain reaction (Q-PCR) assays demonstrated that miR-199a was upregulated during OD of BMSCs, while DEX treatment caused a significant reduction in miR-199a. Alkaline phosphatase (ALP) activity, Alizarin red (AR) staining, and Q-PCR were applied to assess the role of miRNA-199a overexpression in DEX-triggered OD inhibition. miR-199a was able to rescue OD and ALP activity, which were inhibited by DEX. Additionally, we observed that ALP, BMP2, COL1A1, and Runx2 were increased after transfection of miRNA-199a mimics. Furthermore, we confirmed that miRNA-199a facilitates OD of BMSCs through direct inhibition of Klotho protein and messenger RNA expression affecting the downstream fibroblast growth factor receptor 1/extracellular-signal-regulated kinase and Janus kinase 1/signal transducer and activator of transcription 1 pathways. This study indicates that miR-199a plays a critical role in preventing GC-mediated osteoblast differentiation and may function as a promising miRNA biomarker for osteoporosis.

Mechanically-Activated Microcapsules for 'On-Demand' Drug Delivery in Dynamically Loaded Musculoskeletal Tissues.

Delivery of biofactors in a precise and controlled fashion remains a clinical challenge. Stimuli-responsive delivery systems can facilitate 'on-demand' release of therapeutics in response to a variety of physiologic triggering mechanisms (e.g. pH, temperature). However, few systems to date have taken advantage of mechanical inputs from the microenvironment to initiate drug release. Here, we developed mechanically-activated microcapsules (MAMCs) that are designed to deliver therapeutics in an on-demand fashion in response to the mechanically loaded environment of regenerating musculoskeletal tissues, with the ultimate goal of furthering tissue repair. To establish a suite of microcapsules with different thresholds for mechano-activation, we first manipulated MAMC physical dimensions and composition, and evaluated their mechano-response under both direct 2D compression and in 3D matrices mimicking the extracellular matrix properties and dynamic loading environment of regenerating tissue. To demonstrate the feasibility of this delivery system, we used an engineered cartilage model to test the efficacy of mechanically-instigated release of TGF-ß3 on the chondrogenesis of mesenchymal stem cells. These data establish a novel platform by which to tune the release of therapeutics and/or regenerative factors based on the physiologic dynamic mechanical loading environment, and will find widespread application in the repair and regeneration of numerous musculoskeletal tissues.

One-Step Generation of Salt-Responsive Polyelectrolyte Microcapsules via Surfactant-Organized Nanoscale Interfacial Complexation in Emulsions (SO NICE).

Polyelectrolyte microcapsules are versatile compartments for encapsulation, protection, and controlled/triggered release of active agents. Conventional methods of polyelectrolyte microcapsule preparation require multiple steps or do not allow for efficient encapsulation of active agents in the lumen of the microcapsule. In this work, we present the fabrication of hollow polyelectrolyte microcapsules with a salt-responsive property based on surfactant organized nanoscale interfacial complexation in emulsions (SO NICE). In SO NICE, polyelectrolyte microcapsules are templated by water-in-oil-in-water (W/O/W) double emulsions. One polyelectrolyte is dissolved in the inner water droplet of the W/O/W double emulsions, whereas the second polyelectrolyte is dissolved in the organic phase by hydrophobic ion paring with an oppositely charged hydrophobic surfactant. Interfacial complexation of the two polyelectrolytes generates a few hundred-nanometer thick film at the inner water-oil interface of the W/O/W double emulsions. SO NICE microcapsules can be triggered to release their cargo by increasing the ionic strength of the solution, which is a hallmark of polyelectrolyte-based microcapsules. By enabling dissolution and interfacial complexation of polyelectrolytes in organic solvents, SO NICE widens the pallet of polymers that can be used to generate functional polyelectrolyte microcapsules with high encapsulation efficiency for applications in encapsulation and controlled/triggered release.

Seroprevalence survey of avian influenza A (H5) in wild migratory birds in Yunnan Province, Southwestern China.

BACKGROUND: Highly pathogenic avian influenza virus (HPAIV) is a highly contagious disease which is a zoonotic pathogen of significant economic and public health concern. The outbreaks caused by HPAIV H5N1 of Asian origin have caused animal and human disease and mortality in several countries of Southeast Asia, such as Bangladesh, Cambodia, China, India, Indonesia, Laos, Myanmar, Thailand and Viet Nam. For the first time since 1961, this HPAIV has also caused extensive mortality in wild birds and has sparked debate of the role wild birds have played in the spread of this virus. Other than confirmed mortality events, little is known of this virus in wild birds. There is no report on the seroprevalence of avian influenza H5 infection in wild migratory birds in Yunnan Province. In this study we examined live wild birds in Yunnan Province for H5 specific antibody to better understand the occurrence of this disease in free living birds. METHODS: Sera from 440 wild birds were collected from in Kunming and Northern Ailaoshan of Yunnan Province, Southwestern China, and assayed for H5 antibodies using the hemagglutination inhibition (HI) assays. RESULTS: The investigation revealed that the seroprevalence of avian influenza H5 was as following: Ciconiiformes 2.6%, Strigiformes 13.04%, Passeriformes 20%, Cuculiformes 21.74%, Gruiformes 0%, Columbiformes 0%, Charadriiformes 0% and Coraciiformes 0%. Statistical analyses showed that there was a significant difference of prevalence between the orders (P < 0.01). Specific avian influenza H5 antibodies were detected in 23 of 440 (5.23%) sera. Mean HI titer 23 positive sera against H5 were 5.4 log2. CONCLUSIONS: The results of the present survey indicated that the proportion of wild birds had previously infected AIV H5 at other times of the year. To our knowledge, this is the first seroprevalence report of avian influenza H5 infection in wild migratory birds in China' s southwestern Yunnan Province. The results of the present survey have significant public health concerns.

Designing metallic glass matrix composites with high toughness and tensile ductility.

The selection and design of modern high-performance structural engineering materials is driven by optimizing combinations of mechanical properties such as strength, ductility, toughness, elasticity and requirements for predictable and graceful (non-catastrophic) failure in service. Highly processable bulk metallic glasses (BMGs) are a new class of engineering materials and have attracted significant technological interest. Although many BMGs exhibit high strength and show substantial fracture toughness, they lack ductility and fail in an apparently brittle manner in unconstrained loading geometries. For instance, some BMGs exhibit significant plastic deformation in compression or bending tests, but all exhibit negligible plasticity (<0.5% strain) in uniaxial tension. To overcome brittle failure in tension, BMG-matrix composites have been introduced. The inhomogeneous microstructure with isolated dendrites in a BMG matrix stabilizes the glass against the catastrophic failure associated with unlimited extension of a shear band and results in enhanced global plasticity and more graceful failure. Tensile strengths of approximately 1 GPa, tensile ductility of approximately 2-3 per cent, and an enhanced mode I fracture toughness of K(1C) approximately 40 MPa m(1/2) were reported. Building on this approach, we have developed 'designed composites' by matching fundamental mechanical and microstructural length scales. Here, we report titanium-zirconium-based BMG composites with room-temperature tensile ductility exceeding 10 per cent, yield strengths of 1.2-1.5 GPa, K(1C) up to approximately 170 MPa m(1/2), and fracture energies for crack propagation as high as G(1C) approximately 340 kJ m(-2). The K(1C) and G(1C) values equal or surpass those achievable in the toughest titanium or steel alloys, placing BMG composites among the toughest known materials.

Smurf1-targeting microRNA-136-5p-modified bone marrow mesenchymal stem cells combined with 3D-printed ß-tricalcium phosphate scaffolds strengthen osteogenic activity and alleviate bone defects.

Suitable biomaterials with seed cells have promising potential to repair bone defects. However, bone marrow mesenchymal stem cells (BMSCs), one of the most common seed cells used in tissue engineering, cannot differentiate efficiently and accurately into functional osteoblasts. In view of this, a new tissue engineering technique combined with BMSCs and scaffolds is a major task for bone defect repair. Lentiviruses interfering with miR-136-5p or Smurf1 expression were transfected into BMSCs. The effects of miR-136-5p or Smurf1 on the osteogenic differentiation (OD) of BMSCs were evaluated by measuring alkaline phosphatase activity and calcium deposition. Then, the targeting relationship between miR-136-5p and Smurf1 was verified by bioinformatics website analysis and dual luciferase reporter assay. Then, a rabbit femoral condyle bone defect model was established. miR-136-5p/BMSCs/ß-TCP scaffold was implanted into the defect, and the repair of the bone defect was detected by Micro-CT and HE staining. Elevating miR-136-5p-3p or suppressing Smurf1 could stimulate OD of BMSCs. miR-136-5p negatively regulated Smurf1 expression. Overexpressing Smurf1 reduced the promoting effect of miR-136-5p on the OD of BMSCs. miR-136-5p/BMSCs/ß-TCP could strengthen bone density in the defected area and accelerate bone repair. SmurF1-targeting miR-136-5p-modified BMSCs combined with 3D-printed ß-TCP scaffolds can strengthen osteogenic activity and alleviate bone defects.

The first diagnosis of Severe Fever with Thrombocytopenia Syndrome caused by tick-borne Severe Fever with Thrombocytopenia Syndrome virus in Chongqing, China: A case report and literature review.

BACKGROUND: Severe Fever with Thrombocytopenia Syndrome (SFTS) is a tick-borne disease caused by the SFTS virus (SFTSV) which has the potential to become a pandemic and is currently a major public health concern. CASE PRESENTATION: We present the case of a 74-year-old female from an urban area of Chongqing, with leukocytopenia, thrombocytopenia, organ function, inflammatory, blood coagulation, and immune abnormalities. SFTSV infection was confirmed through molecular detection and metagenomic next-generation sequencing (mNGS) analysis, indicating a diagnosis of SFTS due to the patient's history of tick bites. The patient received symptomatic and supportive therapy, including antibiotics, antiviral treatment, and antifungal therapy, and finally discharged from the hospital on day 18. CONCLUSIONS: This study highlights the need for increased awareness, early diagnosis, and prompt treatment for tick-borne SFTS. It also provides a comprehensive understanding of the disease's characteristics, pathogenesis, detection methods, and available treatments.

Accumulation and transfer of polystyrene microplastics in Solanum nigrum seedlings.

Microplastic (MP) pollution is lately receiving increasing attention owing to its harmful impact on terrestrial ecosystems. In this microcosm study, we assessed the uptake and transfer of MPs in Solanum nigrum seedlings exposed to 50 mg L-1 of 0.2-µm polystyrene (PS) beads for 30 d. Confocal laser scanning micrographs helped detect highly intense red fluorescence signals from PS-MP beads in S. nigrum root compared with the controls. Confocal images revealed that the PS beads were primarily distributed in the epidermis and xylem of roots and vascular systems of stems and leaves. Scanning electron microscopy showed that PS beads were scattered on the cell walls of the root xylem and leaf vascular system. Few PS beads were transferred from roots to stems and leaves via the vascular system following the transpiration stream. In conclusion, our findings showed that PS beads accumulated in S. nigrum roots and were transferred from the roots to the aerial parts.

Compound Chinese medicine (F1) improves spleen deficiency diarrhea by protecting the intestinal mucosa and regulating the intestinal flora.

Compound Chinese medicine (F1) is a traditional prescription in Chinese medicine that is commonly used to treat spleen deficiency diarrhea (SDD). It has demonstrated remarkable effectiveness in clinical practice. However, the precise mechanism by which it exerts its antidiarrheal effect is still unclear. This study aimed at investigating the antidiarrheal efficacy and mechanism of F1 on senna-induced secretory diarrhea (SDD). Senna was utilized to induce the development of a mouse model of senna-induced secretory diarrhea (SDD) in order to observe the rate of diarrhea, diarrhea index, blood biochemistry, and histopathological changes in the small intestine. Additionally, the levels of sodium and hydrogen exchange protein 3 (NHE3) and short-chain fatty acids (SCFAs) were determined using enzyme-linked immunosorbent assay (ELISA). The impact of F1 on the senna-induced SDD mouse models was evaluated by monitoring changes in the gut microbiota through 16S rRNA (V3-V4) sequencing. The results demonstrated that F1, a traditional Chinese medicine, effectively increased the body weight of SDD mice and reduced the incidence of diarrhea and diarrhea index. Additionally, F1 restored liver and kidney function, reduced the infiltration of inflammatory cells in intestinal tissue, and promoted the growth of intestinal villi. Furthermore, F1 was found to enhance the expression of NHE3 and SCFAs. It also increased the abundance of Firmicutes and Lactobacillus species, while decreasing the abundance of Proteobacteria and Shigella.

Seroprevalence of Toxoplasma gondii antibodies from slaughter pigs in Chongqing, China.

Toxoplasmosis is a disease caused by the protozoan Toxoplasma gondii which infects most genera of warm-blooded animals, including humans. The objective of this investigation is to evaluate the seroprevalence of toxoplasmosis in pigs in Chongqing Municipality, southwest China. Slaughterhouse pigs' serum samples collected from six different regions in Chongqing were assayed for T. gondii antibodies by an indirect hemagglutination test. The average seroprevalence of T. gondii were found in 30.6% (278/908) in slaughter pigs, ranging from 21.6% to 40.9% among different sampling sites. The results indicated that toxoplasmosis in swine of Chongqing Municipality was relatively serious, and the pork may be an important source for human infection with T. gondii. Comprehensive measures are needed to strengthen further prevention and control of the disease in Chongqing.

Fat mass and obesity associated (FTO)-mediated N6-methyladenosine modification of Krüppel-like factor 3 (KLF3) promotes osteosarcoma progression.

ARSTRACTN6-methyladenosine (m6A) methylation is the most common and abundant methylation modification of eukaryotic mRNAs, which is involved in tumor initiation and progression. The study aims to explore the potential role and the regulatory mechanism of fat mass and obesity associated (FTO) in osteosarcoma (OS) progression. In this study, we detected the expressions of Krüppel-like factor 3 (KLF3) in OS cells and tissues and found that the mRNA and protein levels of KLF3 were increased in OS cells and tissues and significantly related to tumor size, metastasis, and TNM stage and poor prognosis of OS patients. FTO promoted the proliferation and invasion and suppressed apoptosis of OS cells through cell experiments in vitro. Further mechanism dissection revealed that FTO and YTHDF2 enforced the decay of KLF3 mRNA and decreased its expression. FTO-mediated mRNA demethylation inhibited KLF3 expression in the YTHDF2-dependent manner. Moreover, KLF3 overexpression abrogated FTO-induced oncogenic effects on the proliferation and invasion of OS cells. Overall, our findings showed that FTO-mediated m6A modification of KLF3 promoted OS progression, which may provide a therapeutic target for OS.

Circular RNA_ANKIB1 accelerates chemo-resistance of osteosarcoma via binding microRNA-26b-5p and modulating enhancer of zeste homolog 2.

Osteosarcoma is a common bone malignancy in children and adolescents. Chemotherapeutic drug resistance is the major factor impacting the surgical outcome and prognosis of patients with osteosarcoma. This investigation assessed the role and mechanism of circular RNA_ANKIB1 in the development of osteosarcoma. The circular RNA (circ) _ANKIB1, microRNA (miR)-26b-5p, enhancer of zeste homolog 2 (EZH2) expression in OS samples was investigated through RT-qPCR. The EZH2, multidrug resistance protein 1 (MRP1), P-gp, and lipoprotein receptor-related protein (LRP) protein expressions were analyzed through western blot. The association between circ_ANKIB1 and the occurrence of clinic-pathological features in OS patients was assessed; the circular features of circ_ANKIB1 were analyzed. The hFOB1.19, KHOS, U2-OS OS cells were used to study the semi-inhibitory concentration IC50 of Doxorubicin (DXR)-resistant cells, clone formation, invasion, and apoptosis. The luciferase assay was used to study the binding of circ-ANKIB1 with miR-26b-5p and the targeting of miR-26b-5p with EZH2. In vivo experiments were performed via subcutaneous tumorigenic experiments. MiR-26b-5p in OS tissues and cells and DXR-resistant OS tissues and cells was silenced while circ_ANKIB1 and EZH2 were elevated. Circ_ANKIB1 silencing elevated miR-26b-5p, repressed EZH2, MRP1, P-gp, LRP, IC50, and elevated OS advancement. Circ_ANKIB1 bind miR-26b-5p. Reduced miR-26b-5p revered the influence of silencing circ_ANKIB1 on DXR resistant OS cells. MiR-26b-5p targeted EZH2, and EZH2 elevation reversed the impact of increasing miR-26b-5p on DXR resistant cells. Circ_ANKIB1 silencing suppressed DXR-resistant OS cells in vivo. In conclusion, Circ_ANKIB1 binds miR-26b-5p and modulates EZH2 to accelerate the chemo-resistance of osteosarcoma.

Long non-coding RNA long intergenic non-coding 00641 mediates cell progression with stimulating cisplatin-resistance in osteosarcoma cells via microRNA-320d/myeloid cell leukemia-1 axis.

As a staple chemotherapy medicine, cisplatin (DDP) is extensively applied in cancer patients, but its drug resistance is limited. Numerous studies have elucidated that long non-coding RNA (lncRNA) performs as a pivotal agent in osteosarcoma (OS). Nevertheless, lncRNA long intergenic non-coding 00641 (LINC00641)'s functions in DDP resistance for OS remain obscure. The purpose of this study was to investigate the effect and mechanism of LINC00641 on drug resistance of OS. The tissues of both clinical cancer patients and the normal control were gathered. Detection of LINC00641, microRNA-320d (miR-320d) and myeloid cell leukemia-1 (MCL1) was conducted. After the selection of OS cell lines, the detection of cell advancement was applied. Series of experiments were conducted to verify the interaction of LINC00641, miR-320d and MCL1. Xenografted tumor model in vivo was utilized to determine the function of LINC00641. The data displayed, LINC00641 was prominently elevated in OS tissues and cells, especially in DDP-resistant tumors and cell lines. Knock-down LINC00641 was able to attenuate progression of DDP-resistant OS cells thus dampening their drug resistance toward DDP. Moreover, knock-downing LINC00641 gene was also able to manifest antagonism toward DDP-resistance in vivo. On the grounds of bioinformatics prediction, a direct binding of LINC00641 with miR-320d existed, whose target was MCL1. Meanwhile, LINC00641 modulated MCL1 via targeting miR-320d. Additionally, repressive LINC00641 blocked MCL1 via emulative interaction with miR-320d, thus expediting DDP-sensitivity of OS cells. All in all, it is found that LINC00641 is available to escalate drug resistance of DDP-resistant OS cells via mediation of miR-320d/MCL1 axis.

Abnormal glucose regulation in Chinese patients with coronary artery disease: a gender analysis.

BACKGROUND: Diabetes and impaired glucose regulation are very common in patients with coronary artery disease (CAD). In this study, we aim to investigate the prevalence of abnormal glucose regulation in men and women in Chinese CAD patients. METHODS: In this retrospective study, 4100 patients (male, n = 2873; female, n = 1227)with CAD were enrolled. The mean age of these patients was 63 years. The demographic data, medical history, echocardiography findings and blood investigations were collected and analyzed. RESULTS: In this population, 953 (24%) patients had definite diagnosis of type 2 diabetes mellitus, including 636 males (23%) and 317 females (27%). There was a higher prevalence of diabetes in females than men (p < 0.05). For the remaining patients, 48% (n = 959) undergone an oral glucose tolerance test (OGTT), which revealed that 83 male patients (12%) and 41 female patients (16%) suffered from the type 2 diabetes (p > 0.05). 283 men (40%) and 105 women (41%) had impaired glucose regulation (IGR) (p > 0.05). Only 338 men (25%) and 109 women (19%) showed the normal glucose regulation, implying a higher prevalence of abnormal glucose regulation in females (p < 0.01). The odd ratio (OR) showed that women were more prone to have diabetes mellitus or IGT than men and the OR was 1.44 and 1.43 respectively. CONCLUSION: Abnormal glucose regulation is highly prevalent in CAD patients. The women are more prone to have diabetes mellitus or IGT than men.

Molecular Link in Flavonoid and Amino Acid Biosynthesis Contributes to the Flavor of Changqing Tea in Different Seasons.

The present study was aimed to elucidate the flavor formation mechanism of Changqing tea. High-performance liquid chromatography (HPLC) analysis showed that the total catechins of Changqing tea was 65-160 mg/g, with 16-34 mg/g non-galloyated catechins and 49-126 mg/g galloylated catechins. Tea polyphenols and free amino acids account for 286-312 mg/g and 35-89 mg/g, respectively. Transcriptome of Changqing tea during different seasons revealed 316, 130 and 12 DEGs in comparisons of spring vs. autumn, spring vs. summer, and summer vs. autumn, respectively. Compared to spring, the genes involved in flavonoid biosynthesis and bitter imparted amino acids were up-regulated in summer and autumn. Metabolome analysis was conducted by using HPLC-MS; the result indicated that umami and kokumi contributing amino acids were decreased in summer and autumn compared with spring. It could be concluded that the coordination of flavonoid biosynthesis and amino acids biosynthesis resulted in the special flavor of Changqing tea.

Anti-inflammatory flavonolignans from Hydnocarpus anthelminthica seeds.

A new flavonolignan, anthelminthicol A (1), together with four known compounds, was isolated from the EtOAc extracts of the seeds of Hydnocarpus anthelminthica. Their structures were elucidated using extensive spectroscopic techniques. Bioassay showed that compounds 3-5 could inhibit nitric oxide production in LPS-stimulated RAW 264.7 macrophage cell lines, with IC(50) values of 7.81, 9.38, and 10.55 µM, respectively.

Downregulation of MIAT reduces the proliferation and migratory and invasive abilities of retinoblastoma cells by sponging miR-665 and regulating LASP1.

Long non-coding RNAs (lncRNAs) can function as onco-lncRNAs in several types of human cancer, including retinoblastoma (Rb). The present study investigated the potential role and regulatory mechanism of the lncRNA myocardial infarction-associated transcript (MIAT) in Rb. To do so, the expression levels of MIAT, microRNA (miR)-665, and LIM and SH3 protein 1 (LASP1) in Rb tissues from patients or Rb cells were analysed using reverse transcription quantitative PCR. The interactions between miR-665 and MIAT/LASP1 were confirmed by the dual-luciferase reporter assay. MTT, Transwell (to assess migration and invasion) and western blotting assays were used to explore the functions of the MIAT/miR-665/LASP1 axis on Rb progression in vitro. The results of the present study indicated that MIAT targeted miR-665. In Rb tissues and cell lines, high expression of MIAT was observed, whereas miR-665 was downregulated in Rb tissues. Furthermore, the proliferation and migratory and invasive abilities of Rb Y79 and HXO-RB44 cells were decreased following MIAT downregulation or miR-665 overexpression. In addition, LASP1 was identified as a target gene of miR-665. Both the decreased expression of miR-665 and the elevated expression of LASP1 reversed the suppressive effects of MIAT knockdown on the proliferation and migratory and invasive abilities of Y79 cells. Furthermore, MIAT silencing attenuated the development of Rb by regulating the miR-665/LASP1 axis. Taken together, these findings suggested that MIAT may be considered as a possible therapeutic target for Rb.

miR-27a promotes osteogenic differentiation in glucocorticoid-treated human bone marrow mesenchymal stem cells by targeting PI3K.

MicroRNA-27a (miR-27a) modulates osteogenic differentiation (OD); however, the mechanism by which it influences osteoclastic activity in the glucocorticoid (GC)-elicited osteoporotic bone is still unclear. Bone marrow was obtained from the proximal femur of patients (n = 3) with a femoral neck fracture and those (n = 3) with steroid-related osteonecrosis of the femoral head (ONFH). GC was applied to an established ONFH cell model from human bone marrow mesenchymal stem cells (hBMSCs). The miR-27a expression profiles were found to be downregulated in ONFH samples and GC-induced hBMSCs using microarray analysis and real-time quantitative polymerase chain reaction, whereas the OD capacity of hBMSCs was significantly reduced in the GC group compared with the control group. Subsequent transfection of an miR-27a mimic in hBMSCs revealed that the OD capacity of cells was remarkably strengthened in the GC group compared with the miR-control group. Bioinformatics software (TargetScan) predicted that phosphoinositide 3-kinase (PI3K) might be a potential miR-27a target, which was indicated by dual-luciferase reporter assay. Compared with the control group, the GC group exhibited a significantly downregulated protein expression level of PI3K and its downstream protein kinase B (Akt) and mammalian target of rapamycin (mTOR) expression. Furthermore, administration of 10 µM 740 Y-P, a cell-permeable phosphopeptide activator of PI3K, to hBMSCs increased the expression of Akt and mTOR. Treatment with 740 Y-P reversed the effect of miR-27a on OD in hBMSCs. In conclusion, miR-27a is thought to relieve ONFH and the OD repression in GC-induced hBMSCs by targeting the PI3K/Akt/mTOR pathway.