Currently, the characteristics of solid-state phase transformation corresponding to octacalcium phosphate (OCP) sintering is still not completely understood. Although it was known that Ca-deficient ...(Ca-def) hydroxyapatite (HA) is usually unstable at high temperature, the thermal stability and sintering behavior of OCP have been rarely reported. The objective of this study is to systematically investigate the thermal stability and sintering behavior of OCP powders by comprehensive characterization techniques including in-situ high temperature X-ray diffraction (XRD), transmission electron microscopy (TEM), differential thermal analysis (DTA) and scanning electron microscopy (SEM). In-situ XRD results showed that the “collapsed OCP”, Ca-def HA and β-tricalcium phosphate (TCP) crystal structures formed by heating OCP at 200–250 °C, 300–600 °C and 800–900 °C, respectively. Furthermore, DTA analysis revealed that the peak temperatures of the phase transitions of OCP to Ca-def HA and Ca-def HA to β-TCP were ∼170 °C and ∼720 °C, respectively. SEM examination indicated that a large number of pores were introduced during sintering OCP powders due to the release of structurally contained water at 200–250 °C. Therefore, a porous β-TCP ceramic can be directly prepared by sintering OCP powders at 900 °C. This phase transition induced foaming method may be extended for preparing porous bioceramics by sintering calcium phosphate phases as long as they contain crystalline water, which could provide guidelines for better utilizing the phase transformations of calcium phosphate to produce special bioceramics for biomedical applications.
•“Collapsed octacalcium phosphate (OCP)” formed by heating OCP at 200–250 °C.•Ca-deficient (Ca-def) hydroxyapatite (HA) formed by heating OCP at 300–600 °C.•β-tricalcium phosphate (TCP) formed by heating OCP at 800–900 °C.•Peak temperatures of OCP to Ca-def HA and Ca-def HA to β-TCP were ∼170 °C and ∼720 °C.•Many pores were introduced during sintering OCP due to crystalline water release.
Timely diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is a prerequisite for treatment and prevention. The serology characteristics and complement diagnosis value ...of the antibody test to RNA test need to be demonstrated.
Serial sera of 80 patients with PCR-confirmed coronavirus disease 2019 (COVID-19) were collected at the First Affiliated Hospital of Zhejiang University, Hangzhou, China. Total antibody (Ab), IgM and IgG antibodies against SARS-CoV-2 were detected, and the antibody dynamics during the infection were described.
The seroconversion rates for Ab, IgM and IgG were 98.8%, 93.8% and 93.8%, respectively. The first detectible serology marker was Ab, followed by IgM and IgG, with a median seroconversion time of 15, 18 and 20 days post exposure (d.p.e.) or 9, 10 and 12 days post onset (d.p.o.), respectively. The antibody levels increased rapidly beginning at 6 d.p.o. and were accompanied by a decline in viral load. For patients in the early stage of illness (0-7 d.p.o), Ab showed the highest sensitivity (64.1%) compared with IgM and IgG (33.3% for both; p<0.001). The sensitivities of Ab, IgM and IgG increased to 100%, 96.7% and 93.3%, respectively, 2 weeks later. When the same antibody type was detected, no significant difference was observed between enzyme-linked immunosorbent assays and other forms of immunoassays.
A typical acute antibody response is induced during SARS-CoV-2 infection. Serology testing provides an important complement to RNA testing in the later stages of illness for pathogenic-specific diagnosis and helpful information to evaluate the adapted immunity status of patients.
The electrical microenvironment plays an important role in bone repair. However, the underlying mechanism by which electrical stimulation (ES) promotes bone regeneration remains unclear, limiting the ...design of bone microenvironment–specific electroactive materials. Herein, by simple co-incubation in aqueous suspensions at physiological temperatures, biocompatible regenerated silk fibroin (RSF) is found to assemble into nanofibrils with a β-sheet structure on MXene nanosheets, which has been reported to inhibit the restacking and oxidation of MXene. An electroactive hydrogel based on RSF and bioencapsulated MXene is thus prepared to promote efficient bone regeneration. This MXene/RSF hydrogel also acts as a piezoresistive pressure transducer, which can potentially be utilized to monitor the electrophysiological microenvironment. RNA sequencing is performed to explore the underlying mechanisms, which can activate Ca2+/CALM signaling in favor of the direct osteogenesis process. ES is found to facilitate indirect osteogenesis by promoting the polarization of M2 macrophages, as well as stimulating the neogenesis and migration of endotheliocytes. Consistent improvements in bone regeneration and angiogenesis are observed with MXene/RSF hydrogels under ES in vivo. Collectively, the MXene/RSF hydrogel provides a distinctive and promising strategy for promoting direct osteogenesis, regulating immune microenvironment and neovascularization under ES, leading to re-establish electrical microenvironment for bone regeneration.
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•MXene nanosheets could direct the selective growth of silk nanofibrils.•Prepared MXene/RSF hydrogel exhibited good conductivity and sensing ability.•The electroactive hydrogel could promote osteogenic differentiation of BMSCs by activating the Ca2+/CALM signaling pathway.•The conductive system created an osteoblast–macrophage–endotheliocyte virtuous circle for bone microenvironment.
Organic-rich black shale of the Upper Yangtze Basin from the Late Ordovician and Early Silurian is considered an excellent source rock in South China. The formation and preservation conditions of ...this resource are revealed by its geochemical characteristics in this study. Geochemical indices, including redox indices (V/(V + Ni), V/Cr, V/Sc, and Ni/Co) and primary productivity indices (P/Ti and Ba/Al), and paleoclimate, clastic flux and sedimentary rate analyses are presented to investigate the accumulation mechanism of organic matter. Redox indices suggest that a stagnant, anoxic environment predominated in the Upper Yangtze Basin during accumulation of Wufeng and Longmaxi formations. In contrast, ventilated and oxygenated marine conditions pervaded the Upper Yangtze Basin during deposition of Linxiang and Guanyinqiao formations. The concentrations of V and U demonstrate that accumulation of organic matter was mainly controlled by redox conditions. Besides, such factors as clastic fluxes, fresh water inflows or a mixed deposition with a rapid sedimentary rate cannot be ignored due to their influences on organic matter enrichment and preservation. However, weak co-variance relationship of TOC content and productivity proxies, including P/Ti and Ba/Al, demonstrates that the accumulation of organic matter was not controlled by primary productivity. Results of the present study suggest a depositional model that stresses the importance of tectonic movements and glacial events on the accumulation and preservation of organic matter. The model shows that the Upper Yangtze Basin was a semi-restricted basin system influenced by the isolation of Xuefeng, but also it implies that oxygen-depleted bottom water of the basin favored the accumulation and preservation of sedimentary organic matter, resulting in the formation of organic-rich black shale.
•A stratified water column and dysoxic to anoxic bottom water conditions played a dominant role in the preservation of organic matter.•A model of the black shale's formation is established.•Tectonic movement and glacial dominate stagnation and stratification of seawater.
Microfluidic electric impedance flow cytometry (IFC) devices have been applied in single cell analysis, such as cell counting, volume discrimination, cell viability, etc. A cell’s shape provides ...specific information about cellular physiological and pathological conditions, especially in microorganisms such as yeast. In this study, the particle orientation focusing was theoretically analyzed and realized by hydrodynamics. The pulse width (passing time for the particles) of the conductance signal was used to discriminate particle shapes. Spherical and rod-shaped particles with similar volumes/lengths were differentiated by the IFC device, using the impedance pulse parameters of the events. Then, typical late-budding, early budding, and unbudded yeast cells were distinguished by the width, amplitude, and ratio of width to amplitude (R) of the impedance pulse. The pulse amplitude and the R combination gate for identifying the late-budding yeast was estimated through the statistic results. Using the gate, the late-budding rates under different conditions were calculated. Late-budding rates obtained using our method showed a high correlation (R 2 = 0.83) with the manual cell counting result and represented the budding status of yeast cells under different conditions proficiently. Thus, the late-budding rate calculated using the above method can be used as a qualitative parameter to assess the reproductive performance of yeast and whether a yeast culturing environment is optimal. This IFC device and cell shape discrimination method is very simple and could be applied in the fermentation industry and other microorganisms’ discrimination as a rapid analysis technique in the future.
Mitochondria, the only semiautonomous organelles in mammalian cells, possess a circular, double-stranded genome termed mitochondrial DNA (mtDNA). While nuclear genomic DNA compaction, chromatin ...compartmentalization and transcription are known to be regulated by phase separation, how the mitochondrial nucleoid, a highly compacted spherical suborganelle, is assembled and functions is unknown. Here we assembled mitochondrial nucleoids in vitro and show that mitochondrial transcription factor A (TFAM) undergoes phase separation with mtDNA to drive nucleoid self-assembly. Moreover, nucleoid droplet formation promotes recruitment of the transcription machinery via a special, co-phase separation that concentrates transcription initiation, elongation and termination factors, and retains substrates to facilitate mtDNA transcription. We propose a model of mitochondrial nucleoid self-assembly driven by phase separation, and a pattern of co-phase separation involved in mitochondrial transcriptional regulation, which orchestrates the roles of TFAM in both mitochondrial nucleoid organization and transcription.
Background
Chronic primary pain (CPP) is an intractable pain of unknown cause with significant emotional distress and/or dysfunction that is a leading factor of disability globally. The lack of a ...suitable animal model that mimic CPP in humans has frustrated efforts to curb disease progression. 2R, 6R-hydroxynorketamine (2R, 6R-HNK) is the major antidepressant metabolite of ketamine and also exerts antinociceptive action. However, the analgesic mechanism and whether it is effective for CPP are still unknown.
Methods
Based on nociplastic pain is evoked by long-term potentiation (LTP)-inducible high- or low-frequency electrical stimulation (HFS/LFS), we wanted to develop a novel CPP mouse model with mood and cognitive comorbidities by noninvasive low-frequency percutaneous electrical nerve stimulation (LF-PENS). Single/repeated 2R, 6R-HNK or other drug was intraperitoneally (i.p.) or intrathecally (i.t.) injected into naïve or CPP mice to investigate their analgesic effect in CPP model. A variety of behavioral tests were used to detect the changes in pain, mood and memory. Immunofluorescent staining, western blot, reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and calcium imaging of in cultured dorsal root ganglia (DRG) neurons by Fluo-8-AM were used to elucidate the role and mechanisms of 2R, 6R-HNK in vivo or in vitro.
Results
Intrathecal 2R, 6R-HNK, rather than intraperitoneal 2R, 6R-HNK or intrathecal S-Ketamine, successfully mitigated HFS-induced pain. Importantly, intrathecal 2R, 6R-HNK displayed effective relief of bilateral pain hypersensitivity and depressive and cognitive comorbidities in a dose-dependent manner in LF-PENS-induced CPP model. Mechanically, 2R, 6R-HNK markedly attenuated neuronal hyperexcitability and the upregulation of calcitonin gene-related peptide (CGRP), transient receptor potential ankyrin 1 (TRPA1) or vanilloid-1 (TRPV1), and vesicular glutamate transporter-2 (VGLUT2) in peripheral nociceptive pathway. In addition, 2R, 6R-HNK suppressed calcium responses and CGRP overexpression in cultured DRG neurons elicited by the agonists of TRPA1 or/and TRPV1. Strikingly, the inhibitory effects of 2R, 6R-HNK on these pain-related molecules and mechanical allodynia were substantially occluded by TRPA1 antagonist menthol.
Conclusions
In the newly designed CPP model, our findings highlighted the potential utility of intrathecal 2R, 6R-HNK for preventing and therapeutic modality of CPP. TRPA1-mediated uprgulation of CGRP and neuronal hyperexcitability in nociceptive pathways may undertake both unique characteristics and solving process of CPP.
Highlights
1) We first establish the CPP mouse model by LF-PENS to mimic nociplastic pain.
2) Intrathecal 2R, 6R-HNK effectively alleviates LF-PENS-induced CPP in mouse models by inhibiting TRPA1 in DRG neurons.
Background/Aims: Uncaria rhynchophylla, known as “Gou-teng”, is a traditional Chinese medicine (TCM) used to extinguish wind, clear heat, arrest convulsions, and pacify the liver. Although U. ...rhynchophylla has a long history of being often used to treat central nervous system (CNS) diseases, its efficacy and potential mechanism are still uncertain. This study investigated neuroprotective effect and the underlying mechanism of U. rhynchophylla extract (URE) in MPP+-induced SH-SY5Y cells and MPTP-induced mice. Methods: MPP+-induced SH-SY5Y cells and MPTP-induced mice were used to established Parkinson’s disease (PD) models. Quantitative proteomics and bioinformatics were used to uncover proteomics changes of URE. Western blotting was used to validate main differentially expressed proteins and test HSP90 client proteins (apoptosis-related, autophagy-related, MAPKs, PI3K, and AKT proteins). Flow cytometry and JC-1 staining assay were further used to confirm the effect of URE on MPP+-induced apoptosis in SH-SY5Y cells. Gait analysis was used to detect the behavioral changes in MPTP-induced mice. The levels of dopamine (DA) and their metabolites were examined in striatum (STR) by HPLC-EC. The positive expression of tyrosine hydroxylase (TH) was detected by immunohischemical staining and Western blotting. Results: URE dose-dependently increased the cell viability in MPP+-induced SH-SY5Y cells. Quantitative proteomics and bioinformatics results confirmed that HSP90 was an important differentially expressed protein of URE. URE inhibited the expression of HSP90, which further reversed MPP+-induced cell apoptosis and autophagy by increasing the expressions of Bcl-2, Cyclin D1, p-ERK, p-PI3K p85, PI3K p110α, p-AKT, and LC3-I and decreasing cleaved caspase 3, Bax, p-JNK, p-p38, and LC3-II. URE also markedly decreased the apoptotic ratio and elevated mitochondrial transmembrane potential (DΨm). Furthermore, URE treatment ameliorated behavioral impairments, increased the contents of DA and its metabolites and elevated the positive expressions of TH in SN and STR as well as the TH protein. Conclusions: URE possessed the neuroprotective effect in vivo and in vitro, regulated MAPK and PI3K-AKT signal pathways, and inhibited the expression of HSP90. U. rhynchophylla has potentials as therapeutic agent in PD treatment.
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