GSDMA at the crossroads between pyroptosis and tumor immune evasion in glioma.

Pyroptosis, an inflammatory and programmed cell death process, has been controversial in its role in tumor immunity. However, as the first molecule in the gasdermin family, the mechanism of GSDMA in glioma growth is not well understood. We identified the differentially expressed gene GSDMA from Treg cells-related genes using the TCGA database. The biological functions of GSDMA and the relationship between GSDMA expression and tumor immune cell infiltration and cancer patient survival were investigated using open-source databases and platforms. Additionally, flow cytometry analysis was used to examine the effect of GSDMA on tumor immune cell infiltration. Our study showed that GSDMA expression played an important role in immune evasion in glioma. Patients with high GSDMA expression had a worse prognosis. In vivo studies demonstrated that GSDMA knockdown could enhance the infiltration level of CD8+ T cells. High GSDMA expression was also positively correlated with poor anti-PD-L1 treatment outcomes in GBM patients, suggesting that GSDMA may be a potential biomarker that should be considered in combination with anti-PD-L1 therapy for glioma patients. In conclusion, our study demonstrates that high GSDMA expression in gliomas is associated with immune-infiltrating cells CD8+ T cells and Treg cells, and indicates a worse prognosis in glioma. Therefore, GSDMA may serve as a therapeutic target for glioma progression and should be applied in immunotherapy for glioma patients.

Targeting the COP9 signalosome for cancer therapy.

The COP9 signalosome (CSN) is a highly conserved protein complex composed of 8 subunits (CSN1 to CSN8). The individual subunits of the CSN play essential roles in cell proliferation, tumorigenesis, cell cycle regulation, DNA damage repair, angiogenesis, and microenvironmental homeostasis. The CSN complex has an intrinsic metalloprotease that removes the ubiquitin-like activator NEDD8 from cullin-RING ligases (CRLs). Binding of neddylated CRLs to CSN is sensed by CSN4 and communicated to CSN5 with the assistance of CSN6, thus leading to the activation of deneddylase. Therefore, CSN is a crucial regulator at the intersection between neddylation and ubiquitination in cancer progression. Here, we summarize current understanding of the roles of individual CSN subunits in cancer progression. Furthermore, we explain how the CSN affects tumorigenesis through regulating transcription factors and the cell cycle. Finally, we discuss individual CSN subunits as potential therapeutic targets to provide new directions and strategies for cancer therapy.

Smartphone Positioning and Accuracy Analysis Based on Real-Time Regional Ionospheric Correction Model.

As one of the main errors that affects Global Navigation Satellite System (GNSS) positioning accuracy, ionospheric delay also affects the improvement of smartphone positioning accuracy. The current ionospheric error correction model used in smartphones has a certain time delay and low accuracy, which is difficult to meet the needs of real-time positioning of smartphones. This article proposes a method to use the real-time regional ionospheric model retrieved from the regional Continuously Operating Reference Stations (CORS) observation data to correct the GNSS positioning error of the smartphone. To verify the accuracy of the model, using the posterior grid as the standard, the electron content error of the regional ionospheric model is less than 5 Total Electron Content Unit (TECU), which is about 50% higher than the Klobuchar model, and to further evaluate the impact of the regional ionosphere model on the real-time positioning accuracy of smartphones, carrier-smoothing pseudorange and single-frequency Precise Point Positioning (PPP) tests were carried out. The results show that the real-time regional ionospheric model can significantly improve the positioning accuracy of smartphones, especially in the elevation direction. Compared with the Klobuchar model, the improvement effect is more than 34%, and the real-time regional ionospheric model also shortens the convergence time of the elevation direction to 1 min. (The convergence condition is that the range of continuous 20 s is less than 0.5 m).

Disulfiram inhibits inflammation and fibrosis in a rat unilateral ureteral obstruction model by inhibiting gasdermin D cleavage and pyroptosis.

BACKGROUND: As an inhibitor of GSDMD, Disulfiram (DSL) can significantly inhibit cell pyroptosis. Cell pyroptosis plays an important role in renal fibrosis. METHODS: HK-2 cells were induced by Lps and ATP to form a pyroptosis model, and the cells were treated by DSL. CCK-8 detected the cell activity. Immunofluorescence (IF) detected the GSDMD. ELISA detected the expression of inflammatory cytokines. Flow cytometry and Western blot detected cell apoptosis and pyroptosis. Collagen type I kit detected collagen secretion, and western blot detected fibrosis marker protein expression. Then, a rat model of unilateral ureteral obstruction (UUO) was established. HE staining detected the degree of renal tissue injury, and Masson staining detected the degree of fibrosis. What's more, the apoptosis level of tissue cells was detected by TUNEL. And the inflammatory factors in peripheral blood and renal tissue were detected by ELISA. Furthermore, the expression of GSDMD was detected by immunohistochemistry (IHC), and Western blot was used to detect the expression levels of apoptosis and pyroptosis-related proteins in tissues. RESULTS: It was found that DSL can inhibit the cell membrane perforation of GSDMD-N by inhibiting the cleavage of GSDMD, hence, it inhibited the occurrence of inflammation, cell pyroptosis, and the fibrosis of HK-2 cells. But if the cell has already undergone pyroptosis, DSL does not provide significant prevention. In vivo experiment, it further verified that pretreated DSL had inhibited renal fibrosis injury. CONCLUSION: Disulfiram can inhibit inflammation and fibrosis in renal fibrosis rats by inhibiting GSDMD.

Mesenchymal stem cell treatment for peripheral nerve injury: a narrative review.

Peripheral nerve injuries occur as the result of sudden trauma and lead to reduced quality of life. The peripheral nervous system has an inherent capability to regenerate axons. However, peripheral nerve regeneration following injury is generally slow and incomplete that results in poor functional outcomes such as muscle atrophy. Although conventional surgical procedures for peripheral nerve injuries present many benefits, there are still several limitations including scarring, difficult accessibility to donor nerve, neuroma formation and a need to sacrifice the autologous nerve. For many years, other therapeutic approaches for peripheral nerve injuries have been explored, the most notable being the replacement of Schwann cells, the glial cells responsible for clearing out debris from the site of injury. Introducing cultured Schwann cells to the injured sites showed great benefits in promoting axonal regeneration and functional recovery. However, there are limited sources of Schwann cells for extraction and difficulties in culturing Schwann cells in vitro. Therefore, novel therapeutic avenues that offer maximum benefits for the treatment of peripheral nerve injuries should be investigated. This review focused on strategies using mesenchymal stem cells to promote peripheral nerve regeneration including exosomes of mesenchymal stem cells, nerve engineering using the nerve guidance conduits containing mesenchymal stem cells, and genetically engineered mesenchymal stem cells. We present the current progress of mesenchymal stem cell treatment of peripheral nerve injuries.

Fusion of GNSS and Speedometer Based on VMD and Its Application in Bridge Deformation Monitoring.

Real-time dynamic displacement and spectral response on the midspan of Jiangyin Bridge were calculated using Global Navigation Satellite System (GNSS) and a speedometer for the purpose of understanding the dynamic behavior and the temporal evolution of the bridge structure. Considering that the GNSS measurement noise is large and the velocity/acceleration sensors cannot measure the low-frequency displacement, the Variational Mode Decomposition (VMD) algorithm was used to extract the low-frequency displacement of GNSS. Then, the low-frequency displacement extracted from the GNSS time series and the high-frequency vibration calculated by speedometer were combined in this paper in order to obtain the high precision three-dimensional dynamic displacement of the bridge in real time. Simulation experiment and measured data show that the VMD algorithm could effectively resist the modal aliasing caused by noise and discontinuous signals compared with the commonly used Empirical Mode Decomposition (EMD) algorithm, which is guaranteed to get high-precision fusion data. Finally, the fused displacement results can identify high-frequency vibrations and low-frequency displacements of a mm level, which can be used to calculate the spectral characteristics of the bridge and provide reference to evaluate the dynamic and static loads, and the health status of the bridge in the full frequency domain and the full time domain.

Novel preparation of Au nanoparticles loaded Laponite nanoparticles/ECM injectable hydrogel on cardiac differentiation of resident cardiac stem cells to cardiomyocytes.

Cardiac tissue engineering offers a facile biomedical technology to develop a cardiac tissue regeneration or tissue repair by involving combinations of effective biomaterials and nanomaterials with engineering strategies. The cardiac regenerative materials were fabricated by electrically active nanoparticles onto the biocompatible matrix to inspire low-resistance electrical signal in the native heart tissues. In the current report, we investigated on the improvement of cardiac functionally inspired conductive injectable hydrogel fabricated from the electroactive gold (Au) and laponite (Lap) nanoparticles loaded myocardial extracellular matrix (ECM) to enhance the functional and biological properties of cardiomyocytes. The incorporation of nanoformulations into the ECM was maintained the functionality behaviors and maintenance of electrical conductivity into cardiomyocytes. The effects of nanoparticles onto the ECM were provided decreasing porous structure and interconnected pores in the hydrogel structure for the favorable environment of cardiac tissues. The biological analysis of cell survival and immunostaining studies of cardiomyocytes established that Au loaded Lap/ECM hydrogel improve cell compatibility and phenotypes maturation of cardiac specific proteins. The combination of electrically active nanoformulations and biologically active ECM was enhanced the cell expression of cardiac specific markers (SAC, cTnl and Cx43), indicating the potential role of nanoparticles loaded ECM hydrogel as an appropriate cardiac regenerative material for the repair of infarcted myocardium.

Complex effects of apoplexy secondary to pituitary adenoma.

Pituitary adenoma apoplexy is a well-known clinical syndrome induced by insulin infusion, cardiac surgery, trauma, and hypothalamic releasing factors. Pituitary apoplexy can cause secondary cerebral infarct and internal carotid artery occlusion. With blockade of tumor perfusion, apoplexy triggers a sudden onset of headache, visual impairment, cranial nerve palsy, disturbances of consciousness, eyelid ptosis, and hemiparesis. However, pituitary adenoma cells with high metabolic demand cannot survive with deficient blood supply and glucose concentrations. Moreover, a number of case reports have shown that spontaneous remission of syndromes, such as acromegaly, may be caused by pituitary adenoma after apoplexy. Therefore, understanding mechanism that underlies the balance between pituitary adenoma apoplexy and subsequent spontaneous remission of syndromes may suggest new approaches for treatment of pituitary adenoma apoplexy.

Complement-C1q TNF-Related Protein 3 Alleviates Mesangial Cell Activation and Inflammatory Response Stimulated by Secretory IgA.

BACKGROUND: Elevated circulating IgA complex deposits in the glomerular mesangium and causes resultant mesangial cell activation, eventually leading to the progression of IgA nephropathy (IgAN). Complement-C1q TNF-related protein 3 (CTRP3) is documented as a potential anti-inflammatory and anti-fibrotic mediator; however, its role in IgAN is unknown. METHODS: Serum polymeric IgA (pIgA) was isolated from patients with IgAN and healthy volunteers. Human mesangial cells (HMCs) were incubated with the isolated pIgA. The expression of CTRP3 and its effects on inflammatory cytokines and extracellular matrix were evaluated. RESULTS: Median circulating CTRP3 decreased in patients with IgAN compared to the controls, accompanied by the elevated serum and urine TGF-ß and IL-6. While urine CTRP3 in IgAN patients were comparable to that in healthy subjects. pIgA dose-dependently inhibited the expression of CTRP3 and HMC tolerated pIgA at 1.0 mg/ml, for which galactose-deficient IgA might be responsible. pIgA markedly increased IL-6 and TGF-ß in HMCs, which were suppressed after Ad-CTRP3 transfection. Additionally, CTRP3 significantly inhibited pIgA-stimulated HMC proliferation, cell cycle progression and synthesis of CTGF and fibronectin. Furthermore, we found that pIgA-treated HMC exhibited higher NF-954;B activity as reflected by the increased levels of nuclear p65. Moreover, Src activation and the subsequent Smad3 phosphorylation were significantly elevated in response to the stimulation with pIgA. However, NF-954;B activity and Smad3 signaling were inhibited after Ad-CTRP3 transfection in HMC. CONCLUSION: CTRP3 potentially targeted to NF-954;B and TGF-ß-Src-Smad3 signaling and exerted anti-inflammatory and anti-fibrotic roles to attenuate the progression of IgAN, which may represent a new strategy for the treatment of IgAN.

[Yeast-based production, purification and bioactivity assay of rainbow trout LECT2].

Leukocyte cell-derived chemotaxin 2 (LECT2) is a secretory cytokine that functions in many physiological and pathological processes. We used a Pichia pastoris expression system for the recombinant expression of rainbow trout LECT2. The recombinant LECT2 was purified by UNOsphere S Cation exchange and size-exclusion chromatography columns. The obtained target protein was highly pure (>96% homogeneity) and the yield was >120 mg/L of yeast cultures. An in vitro chamber assay revealed that recombinant LECT2 could induce chemotactic responses in rainbow trout head kidney-derived macrophages. Recombinant LECT2 not only enhanced macrophage respiratory burst activity and bactericidal activity, but also changed macrophage gene expression. In summary, we established a rapid and efficient method to prepare active recombinant rainbow trout LECT2 using a yeast expression system and column chromatography. Bioactive recombinant LECT2 is essential for studies on protein functions.

LECT2 protects mice against bacterial sepsis by activating macrophages via the CD209a receptor.

Leukocyte cell-derived chemotaxin 2 (LECT2) is a multifunctional cytokine and reduced plasma levels were found in patients with sepsis. However, precise functions and mechanisms of LECT2 remain unclear. The aim of the present study was to determine the role of LECT2 in modulating immune responses using mouse sepsis models. We found that LECT2 treatment improved outcome in mice with bacterial sepsis. Macrophages (MΦ), but not polymorphonuclear neutrophils, mediated the beneficial effect of LECT2 on bacterial sepsis. LECT2 treatment could alter gene expression and enhance phagocytosis and bacterial killing of MΦ in vitro. CD209a was identified to specifically interact with LECT2 and mediate LECT2-induced MΦ activation. CD209a-expressing MΦ was further confirmed to mediate the effect of LECT2 on sepsis in vivo. Our data demonstrate that LECT2 improves protective immunity in bacterial sepsis, possibly as a result of enhanced MΦ functions via the CD209a receptor. The modulation of MΦ functions by LECT2 may serve as a novel potential treatment for sepsis.

[Expression and significance of ß adrenergic receptors in a model of left ventricular mechanical unloading].

OBJECTIVE: To study the expression and significance of ß adrenergic receptors mRNA in a model of left ventricular mechanical unloading, and explore the change in cardiomyocyte in molecular level after left ventricular unloading. METHODS: Heart failure was reproduced in Lewis rats by ligating left anterior descending (LAD) artery. After 4 weeks, the failing hearts and right lungs were heterotopically transplanted into the abdomen of the recipients by anastomosing their ascending aorta to the recipients' descending aorta in the heart transplantation group. Two weeks after transplantation, heart weight, left ventricular weight, myocyte diameter and myocardial fibrosis were determined , and ß adrenergics receptors mRNA expression was essayed by real-time polymerase chain reaction (PCR). Seven normal Lewis rats served as control. RESULTS: The weight of the enlarged heart, left ventricular weight and myocyte diameter of the failing hearts were decreased to normal after transplantation. The levels of ß1- and ß2- adrenergic receptors mRNA expression were significantly lowered in heart failure group compared with that of normal group(0.09 ± 0.03 vs. 0.18 ± 0.04, 0.07 ± 0.06 vs. 0.12 ± 0.02, both P <0.05). The level of ß2- adrenergic receptor mRNA expression in heart transplantation group (0.11 ± 0.05) rose to normal (P>0.05), but ß1- adrenergic receptor mRNA expression (0.08 ± 0.06) was lower in heart transplantation group than that in normal group (P<0.05). CONCLUSION: Myocardium reverse remodeling after left ventricular unloading is accompanied by the change in cardiomyocyte in molecular level , such as the change in ß adrenergic receptors , which may involve in the improvement of heart function after being supported by left ventricular assist device.

Influence of transplantation of allogenic bone marrow mononuclear cells on the left ventricular remodeling of rat after acute myocardial infarction.

To probe into the influence of transplantation of allogenic bone marrow mononuclear cells (BM-MNCs) on the left ventricular remodeling of rat after acute myocardial infarction (AMI), 60 male Wistar rats were evenly divided into three groups at random: control group 1, control group 2 and transplantation group. In control group 1, chest was opened without ligation of coronary artery; in control group 2 and transplantation group, the left anterior descending branch of coronary artery was ligated to establish AMI model. Prepared culture medium and allogenic BM-MNCs suspension were respectively implanted the surrounding area of infarcted cardiac muscle via epicardium of control group 2 and transplantation group. Four weeks after the operation, the osteopontin gene (OPN mRNA, P<0.01), type I collagen (P<0.01) and angiotensin II (AngII, P<0.01) content in the left ventricular non-infarcted myocardium, and the Ang II density in blood plasma (P<0.05) of transplantation group and control group 2 were all significantly higher than that of control group 1. In the transplantation group, the myocardial OPN mRNA, type I collagen and Ang II content of non-infarcted zone in left ventricle, and the Ang II concentration in blood plasma were all significantly lower than those of control group 2 (P<0.05 for all). It is concluded that allogenic BM-MNCs transplantation may ease left ventricular remodeling after AMI by inhibiting the synthesis of type I collagen in the cardiac muscle and down-regulating the expression of Ang II and OPN gene.

Biological characteristics of human bone marrow mesenchymal stem cell cultured in vitro.

Some biological characteristics of human bone marrow mesenchymal stem cells (MSCs) cultured in vitro were observed. hMSCs were isolated from bone marrow and purified by density gradient centrifugation method, and then cultured in vitro. The proliferation and growth characteristics of hMSCs were observed in primary and passage culture. MSCs of passage 3 were examined for the purify by positive rate of CD29 and CD44 through flow cytometry. Human bone marrow MSCs showed active proliferation capacity in vitro. The purify of MSCs separated by our method was higher than 90%. It was concluded that hMSCs have been successfully cultured and expanded effectively. It provided a foundation for further investigation and application of MSCs.