VPS4B mutation impairs the osteogenic differentiation of dental follicle cells derived from a patient with dentin dysplasia type I

A splicing mutation in VPS4B can cause dentin dysplasia type I(DD-I),a hereditary autosomal-dominant disorder characterized by rootless teeth,the etiology of which is genetically heterogeneous.In our study,dental follicle cells(DFCs)were isolated and cultured from a patient with DD-I and compared with those from an age-matched,healthy control.In a previous study,this DD-I patient was confirmed to have a loss-of-function splicing mutation in VPS4B(IVS7+46C>G).The results from this study showed that the isolated DFCs were vimentin-positive and CK14-negative,indicating that the isolated cells were derived from the mesenchyme.DFCs harboring the VPS4B mutation had a significantly higher proliferation rate from day 3 to day 8 than control DFCs,indicating that VPS4B is involved in cell proliferation.The cells were then replenished with osteogenic medium to investigate how the VPS4B mutation affected osteogenic differentiation.Induction of osteogenesis,detected by alizarin red and alkaline phosphatase staining in vitro,was decreased in the DFCs from the DD-I patient compared to the control DFCs.Furthermore,we also found that the VPS4B mutation in the DD-I patient downregulated the expression of osteoblast-related genes,such as ALP,BSP,OCN,RUNX2,and their encoded proteins.These outcomes confirmed that the DD-I-associated VPS4B mutation could decrease the capacity of DFCs to differentiate during the mineralization process and may also impair physiological root formation and bone remodeling.This might provide valuable insights and implications for exploring the pathological mechanisms underlying DD-I root development.

Antiviral activity of Basidiomycete mycelia against influenza type A(serotype H1N1) and herpes simplex virus type 2 in cell culture

In this study, we investigated the in vitro antiviral activity of the mycelia of higher mushrooms against influenza virus type A(serotype H1N1) and herpes simplex virus type 2(HSV-2), strain BH. All 10 investigated mushroom species inhibited the reproduction of influenza virus strain A/FM/1/47(H1N1) in MDCK cells reducing the infectious titer by 2.0–6.0 lg ID50. Four species, Pleurotus ostreatus, Fomes fomentarius, Auriporia aurea, and Trametes versicolor, were also determined to be effective against HSV-2 strain BH in RK-13 cells, with similar levels of inhibition as for influenza. For some of the investigated mushroom species—Pleurotus eryngii, Lyophyllum shimeji, and Flammulina velutipes—this is the first report of an anti-influenza effect. This study also reports the first data on the medicinal properties of A. aurea, including anti-influenza and antiherpetic activities. T. versicolor 353 mycelium was found to have a high therapeutic index(324.67), and may be a promising material for the pharmaceutical industry as an anti-influenza and antiherpetic agent with low toxicity. Mycelia with antiviral activity were obtained in our investigation by bioconversion of agricultural wastes(amaranth flour after CO2 extraction), which would reduce the cost of the final product and solve some ecological problems.

Attitude of Military Students and Military Nurses toward Spirituality and Spiritual Care

Military nurses are exposed to various occupational stresses.Spirituality contributes to adaptation to critical situations and stresses.Spiritual care is an essential part of holistic care which,in recent years,has attracted the attention of health-related organizations.The aim of this study was to determine the attitude of military nurses,faculty members,and military nursing students toward spirituality and spiritual care.In a cross-sectional descriptive study,214 military nurses,faculty members,and nursing students of Baqiyatallah University of Medical Sciences in Tehran were selected according to inclusion criteria by simple sampling method and completed the spirituality and spiritual care rating scale.The mean score of attitude toward spirituality and spiritual care in clinical nurses was 69.4(standard deviation[SD]±12.6),in nursing professors 74.6(SD±6.6),and in students 67.5(SD±4/11).The majority of the research community was at a high and desirable level(between 63 and 92,which was the maximum score of the questionnaire).The results of this study showed that military nurses,faculty members,and nursing students of Baqiyatallah University of Medical Sciences in Tehran have a positive attitude toward spirituality.They tend to provide spiritual care.Therefore,it is necessary to provide a suitable platform for providing spiritual care.

A Brief Review of Animal Models for COVID-19

Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)causes an outbreak of corona virus disease 2019(COVID-19)globally.It is to date one of the biggest public health threat in the world.Although studies on this emerging coronavirus have developed rapidly since the COVID-19 case was confirmed,there is much remains unknown about SARS-CoV-2.To better understand the mechanism of cross-species transmission and search for efficient vaccines and therapeutics for COVID-19,animal models are highly required to evaluate countermeasures against emerging COVID-19.An ideal animal model should respond as similar as human be-ings to SARS-CoV-2 infection and show analogical symptoms,reflecting the human beings clinical sign,viral replication,and pathology that can be applied to identify in human beings.Fortunately,several animal models have been developed to study SARS-CoV-2 and aim to find new vaccines,antiviral agents,and Chinese materia medica very shortly.We aim to review the climical presentations of COVID-19 concerning pathologic features in humans and summarize the current animal models'strengths and limitations of use for SARS-CoV-2.

Identification of potential immune-related mechanisms related to the development of multiple myeloma

Background:Although significant advances have been made in the treatment of multiple myeloma(MM),leading to unprecedented response and survival rates among patients,the majority eventually relapse,and a cure remains elusive.This situation is closely related to an incomplete understanding of the immune microenvironment,especially monocytes/macrophages in patients with treatment-naïve MM.The aim of this study was to provide insight into the immune microenvironment,especially monocytes/macrophages,in patients with treatment-naïve MM.Methods:This study used the single-cell RNA sequencing(scRNA-seq)data of both patients with MM and heathy donors to identify immune cells,including natural killer(NK)cells,T cells,dendritic cells(DCs),and monocytes/macrophages.Transcriptomic data and flow cytometry analysis of monocytes/macrophages were used to further examine the effect of monocytes/macrophages in treatment-naïve MM patients.Results:A significant difference was observed between the bone marrow(BM)immune cells of the healthy controls and treatment-naïve MM patients through scRNA-seq.It is noteworthy that,through an scRNA-seq data analysis,this study found that interferon(IFN)-induced NK/T cells,terminally differentiated effector memory(TEMRA)cells,T-helper cells characterized by expression of IFN-stimulated genes(ISG^(+)Th cells),IFN-responding exhausted T cells,mannose receptor C-type 1(MRC1)^(+)DCs,IFN-responding DCs,MHCII^(+)DCs,and immunosuppressive monocytes/macrophages were enriched in patients with treatment-naïve MM.Significantly,transcriptomic data of monocytes/macrophages demonstrated that"don’t eat me"-related genes and IFN-induced genes increase in treatment-naïve MM patients.Furthermore,scRNA-seq,transcriptomic data,and flow cytometry also showed an increased proportion of CD16^(+)monocytes/macrophages and expression level of CD16.Cell-cell communication analysis indicated that monocytes/macrophages,whose related important signaling pathways include migration inhibitory factor(MIF)and interleukin 16(IL-16)signaling pathway,are key players in treatment-naïve MM patients.Conclusions:Our findings provide a comprehensive and in-depth molecular characterization of BM immune cell census in MM patients,especially for monocytes/macrophages.Targeting macrophages may be a novel treatment strategy for patients with MM.

COVID-19 pandemic impact on ocular trauma in a tertiary hospital

AIM:To analyze the impact of the coronavirus disease-2019(COVID-19)pandemic on the presentation and characteristics of patients hospitalized for ocular trauma in a tertiary hospital in China between 2019 and 2020.METHODS:A retrospective case study was designed to collect information on all cases of ocular trauma in a tertiary hospital from 2019 to 2020 and compare differences in inpatients’data(age,sex,admission vision acuity,type of diagnosis,hospital stays,mechanism of injury and location of injury).RESULTS:The total number of patients admitted to the Ophthalmology Department was 883(mean 73.58±11.25patients per month)in 2019 and 714(59.50±17.92patients per month)in 2020.The injury number of in work was also the most within the four types of location in this two year(42.36%in 2019,43.84%in 2020).The mean hospital stays were 12.66d in 2019 and 10.81d in 2020.The highest incidence of ocular trauma was the middleaged(41-65y)groups in 2019 and 2020.The most common cause of ocular trauma was sharp object in 2019(47.34%)and 2020(47.58%).The mechanical ocular trauma reaches98.98%in 2019 and 99.72%in 2020.CONCLUSION:The number of patients with ocular trauma decreased in 2020,but middle-aged(41-65y)are still high incident groups.Mechanical ocular trauma remains the leading cause of hospitalization for ocular trauma patients and the proportion of patients injured at home increases.It is necessary to arouse social attention and the public’s awareness of eye trauma protection should be strengthened during the pandemic.

Development and Application of a Mitochondrial Genetically Encoded Voltage Indicator in Narcosis

Mitochondrial membrane potential(MMP)plays a crucial role in the function of cells and organelles,involving various cellular physiological processes,including energy production,formation of reactive oxygen species(ROS),unfolded protein stress,and cell survival.Currently,there is a lack of genetically encoded fluorescence indicators(GEVIs)for MMP.In our screening of various GEVIs for their potential monitoring MMP,the Accelerated Sensor of Action Potentials(ASAP)demonstrated optimal performance in targeting mitochondria and sensitivity to depolarization in multiple cell types.However,mitochondrial ASAPs also displayed sensitivity to ROS in cardiomyocytes.Therefore,two ASAP mutants resistant to ROS were generated.A double mutant ASAP3-ST exhibited the highest voltage sensitivity but weaker fluorescence.Overall,four GEVIs capable of targeting mitochondria were obtained and named mitochondrial potential indicators 1-4(MPI-1-4).In vivo,fiber photometry experiments utilizing MPI-2 revealed a mitochondrial depolarization during isoflurane-induced narcosis in the M2 cortex.

Copper-Free Click Chemistry-Mediated Assembly of Single Quantum Dot Nanosensor for Accurately Monitoring Locus-Specific m^(6)A in Cancer Cells

N6-methyladenosine(m^(6)A)plays an important role in embryogenesis,nuclear export,transcription splicing,and protein translation control.Herein,we demonstrate a copper-free click chemistry-mediated assembly of single quantum dot(QD)nanosensor for accurately monitoring locus-specific m^(6)A in cancer cells.The m^(6)A-sensitive endoribonuclease MazF can digest the unmethylated A-RNA,and the intact m^(6)A-RNA then hybridizes with DNA probes a and b to produce a sandwich hybrid,initiating the click chemistry to generate probe a–b ligation product via first tandem ligation detection reaction(LDR)cycle.Subsequently,DNA probes c and d can hybridize with the probe a–b ligation product to generate the probe c–d ligation product via second LDR cycle.Both LDR cycles can be repeated through denaturation and annealing reaction to generate abundant biotin-/fluorophore-modified probe c–d ligation products that can easily assemble on the QD surface to induce distinct fluorescence resonance energy transfer(FRET)between QD and Cy5.This assay can be homogenously performed without the involvement of copper catalyst,m^(6)A-specific antibody,radioactive labeling,ligase enzyme,enzymatic reverse transcription,and next-generation sequencing.Moreover,it can discriminate even 0.01% m^(6)A level in complex samples and accurately measure cellular m^(6)A-RNA expression,providing a promising avenue for clinical diagnostics and biomedical research.

Seed amplification assay of nasal swab extracts for accurate and non-invasive molecular diagnosis of neurodegenerative diseases

Nasal swabs are non-invasive testing methods for detecting diseases by collecting samples from the nasal cavity or nasopharynx.Dysosmia is regarded as an early sign of coronavirus disease 2019(COVID-19),and nasal swabs are the gold standard for the detection.By nasal swabs,severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)nucleic acids can be cyclically amplified and detected using real-time reverse transcriptase-polymerase chain reaction after sampling.Similarly,olfactory dysfunction precedes the onset of typical clinical manifestations by several years in prion diseases and other neurodegenerative diseases,such as Alzheimer’s disease,Parkinson’s disease,dementia with Lewy bodies,and multiple system atrophy.In neurodegenerative diseases,nasal swab tests are currently being explored using seed amplification assay(SAA)of pathogenic misfolded proteins,such as prion,α-synuclein,and tau.These misfolded proteins can serve as templates for the conformational change of other copies from the native form into the same misfolded form in a prion-like manner.SAA for misfolded prion-like proteins from nasal swab extracts has been developed,conceptually analogous to PCR,showing high sensitivity and specificity for molecular diagnosis of degenerative diseases even in the prodromal stage.Cyclic amplification assay of nasal swab extracts is an attractive and feasible method for accurate and non-invasive detection of trace amount of pathogenic substances for screening and diagnosis of neurodegenerative disease.

Mkrn2 deficiency induces teratozoospermia and male infertility through p53/PERP-mediated apoptosis in testis

The apoptosis that occurs in the immature testis under physiological conditions is necessary for male germ cell development,whereas improper activation of apoptosis can impair spermatogenesis and cause defects in reproduction.We previously demonstrated that in mice,the makorin-2(Mkrn 2)gene is expressed exclusively in the testis and its deletion leads to male infertility.To understand the potential molecular mechanism,in this study,we found that levels of apoptosis in the testis were abnormally high in the absence of Mkrn 2.To identify specific gene(s)involved,we performed digital gene expression profiling(DGE)and pathway analysis via gene set enrichment analysis(GSEA)and the Kyoto Encyclopedia of Genes and Genomes(KEGG)database,and we found that MKRN2 inhibits p53 apoptosis effector related to PMP22(PERP)expression and that levels of the protein in sperm samples have an inverse correlation with infertility levels.GSEA additionally indicated that PERP is a negative regulator of spermatogenesis and that its ectopic expression induces male infertility.Further,Gene Expression Omnibus(GEO)dataset analysis showed that p53,upstream of PERP,was upregulated in oligoasthenoteratozoospermia(OAT).These observations suggest that Mkrn 2 is crucial for protecting germ cells from excessive apoptosis and implicate Mkrn 2-based suppression of the p53/PERP signaling pathway in spermatogenesis and male fertility.