Neural stem cells(NSCs)are the source of all neurons and glial cells(astrocytes and oligodendrocytes)in the central nervous system.The adult mammalian brain retains NSCs in the subgranular zone of the dentate gyrus in...Neural stem cells(NSCs)are the source of all neurons and glial cells(astrocytes and oligodendrocytes)in the central nervous system.The adult mammalian brain retains NSCs in the subgranular zone of the dentate gyrus in the hippocampus and ventricular subventricular zone lining the lateral ventricle(Olpe and Jessberger,2022).Adult NSCs in rodents are preserved throughout life and continuously produce new neurons that integrate into the pre-existing neuronal network.展开更多
“Last scene of all that ends this strange,eventful history,is second childishness and mere oblivion.I am sans teeth,sans eyes,sans taste,sans everything.”William Shakespeare‘As You Like It'Act 2,Sc.7,l.139Aging...“Last scene of all that ends this strange,eventful history,is second childishness and mere oblivion.I am sans teeth,sans eyes,sans taste,sans everything.”William Shakespeare‘As You Like It'Act 2,Sc.7,l.139Aging of the human brain is characterized by a progressive decline of its functional capacity;this decline however varies widely,and cognitive longevity differs substantially between individuals.展开更多
Background As the global population increases,the demand for protein sources is expected to increase,driving the demand for cell-based cultivated meat.This study aimed to enhance the productivity of cultivated meat th...Background As the global population increases,the demand for protein sources is expected to increase,driving the demand for cell-based cultivated meat.This study aimed to enhance the productivity of cultivated meat through optimization of the cell source and organization process.Results We engineered fibroblasts into myogenic cells via non-viral introduction of the MYOD1 gene,avoiding viral methods for safety.After confirming the stable derivation of myogenic cells,we combined knockout(KO)of MSTN,a negative regulator of myogenesis,with MYOD1-mediated myogenesis to improve cultivated meat production.Primary cells from MSTN KO cattle exhibited enhanced myogenic potential.Additionally,when tested in immortalized fibroblasts,myostatin treatment reduced MYOD1-induced myogenesis in two-dimensional cultures,while MSTN knockout increased it.To achieve muscle-like cell alignment,we employed digital light processing(DLP)-based three-dimensional(3D)bioprinting to organize cells into 3D groove-shaped hydrogels.These bioactive hydrogels supported stable cell proliferation and significantly improved muscle cell alignment.Upon differentiation into myotubes,the cells demonstrated an ordered alignment,particularly the MSTN KO cells,which showed highly efficient differentiation.Conclusions The integration of genetic modification and advanced DLP 3D bioprinting with groove-patterned hydrogels provides an effective strategy for producing high-quality,muscle-aligned cultivated meat.展开更多
Background: Despite considerable advancements in identifying factors contributing to the development of hepatocellular carcinoma(HCC), the pathogenesis of HCC remains unclear. In many cases, HCC is a consequence of pr...Background: Despite considerable advancements in identifying factors contributing to the development of hepatocellular carcinoma(HCC), the pathogenesis of HCC remains unclear. In many cases, HCC is a consequence of prolonged liver fibrosis, resulting in the formation of an intricate premalignant microenvironment. The accumulation of extracellular matrix(ECM) is a hallmark of premalignant microenvironment. Given the critical role of different matrix components in regulating cell phenotype and function, this study aimed to elucidate the interplay between the fibrotic matrix and malignant features in HCC. Methods: Liver tissues from both control(normal) and carbon tetrachloride(CCl_(4))-induced fibrotic rats were decellularized using sodium dodecyl sulfate(SDS) and Triton X-100. The resulting hydrogel from decellularized ECM was processed into micro-particles via the water-in-oil emulsion method. Microparticles were subsequently incorporated into three-dimensional liver biomimetic micro-tissues(MTs) comprising Huh-7 cells, human umbilical vein endothelial cells(HUVECs), and LX-2 cells. The MTs were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS) assay at day 11, immunofluorescence staining, immunoblotting, and spheroid migration assay at day 14 after co-culture. Results: Fibrotic matrix from CCl4-treated rat livers significantly enhanced the growth rate of the MTs and their expression of CCND1 as compared to the normal one. Fibrotic matrix, also induced the expression of epithelial-to-mesenchymal transition(EMT)-associated genes such as TWIST1, ACTA2, MMP9, CDH2, and VIMENTIN in the MTs as compared to the normal matrix. Conversely, the expression of CDH1 and hepatic maturation genes HNF4A, ALB, CYP3A4 was decreased in the MTs when the fibrotic matrix was used. Furthermore, the fibrotic matrix increased the migration of the MTs and their secretion of alpha-fetoprotein. Conclusions: Our findings suggest a regulatory role for the fibrotic matrix in promoting cancerous phenotype, which could potentially accelerate the progression of malignancy in the liver.展开更多
AIM: To evaluate safety and feasibility of autologous bone marrow-enriched CD34+ hematopoietic stem cell Tx through the hepatic artery in patients with decompensated cirrhosis.METHODS: Four patients with decompensated...AIM: To evaluate safety and feasibility of autologous bone marrow-enriched CD34+ hematopoietic stem cell Tx through the hepatic artery in patients with decompensated cirrhosis.METHODS: Four patients with decompensated cirrhosis were included. Approximately 200 mL of the bone marrow of the patients was aspirated, and CD34+ stem cells were selected. Between 3 to 10 million CD34+ cells were isolated. The cells were slowly infused through the hepatic artery of the patients.RESULTS: Patient 1 showed marginal improvement in serum albumin and no significant changes in other test results. In patient 2 prothrombin time was decreased; however, her total bilirubin, serum creatinine, and Model of End-Stage Liver Disease (MELD) score worsened at the end of follow up. In patient 3 there was improvement in serum albumin, porthrombin time (PT), and MELD score. Patient 4 developed radiocontrast nephropathy after the procedure, and progressed to type 1 hepatorenal syndrome and died of liver failure a few days later. Because of the major side effects seen in the last patient, the trial was prematurely stopped.CONCLUSION: Infusion of CD34+ stem cells through the hepatic artery is not safe in decompensated cirrhosis. Radiocontrast nephropathy and hepatorenal syndrome could be major side effects. However, this study doesnot preclude infusion of CD34+ stem cells through other routes.展开更多
Objective To establish the method of isolation, purification, and identification of human amniotic mesenchymal stem cells (hAMSCs). Methods hAMSCs were isolated from human amniotic membrane by trypsin-collagenase dige...Objective To establish the method of isolation, purification, and identification of human amniotic mesenchymal stem cells (hAMSCs). Methods hAMSCs were isolated from human amniotic membrane by trypsin-collagenase digestion, and cultured in Dulbecco's modified Eagle's medinm/F12 medium supplemented with 10% fetal bovine serum. Phenotypic characteristics of these cells were analyzed by means of immunocytochemistry and flow cytometry. Results The cells successfully isolated from human amniotic membrane expressed representative mesenchymal cell surface markers CD44, CD90, and vimentin, but not CD45. Conclusions This study establishes a potential method for isolation of hAMSCs from human amnion, in vitro culture, and identification. The isolated cells show phenotypic characteristics of mesenchymal stem cells.展开更多
Several studies have demonstrated that selective serotonin reuptake inhibitor antidepressants can promote neuronal cell proliferation and enhance neuroplasticity both in vitro and in vivo. It is hypothesized that cita...Several studies have demonstrated that selective serotonin reuptake inhibitor antidepressants can promote neuronal cell proliferation and enhance neuroplasticity both in vitro and in vivo. It is hypothesized that citalopram, a selective serotonin reuptake inhibitor, can promote the neuronal differentiation of adult bone marrow mesenchymal stem cells. Citalopram strongly enhanced neuronal characteristics of the cells derived from bone marrow mesenchymal stem cells. The rate of cell death was decreased in citalopram-treated bone marrow mesenchymal stem cells than in control cells in neurobasal medium. In addition, the cumulative population doubling level of the citalopram-treated cells was signiifcantly increased compared to that of control cells. Also BrdU incorporation was elevated in citalopram-treated cells. These ifndings suggest that citalopram can improve the neuronal-like cell differentiation of bone marrow mesenchymal stem cells by increasing cell proliferation and survival while maintaining their neuronal characteristics.展开更多
Despite a vast amount of different methods, protocols and cryoprotective agents(CPA), stem cells are often frozen using standard protocols that have beenoptimized for use with cell lines, rather than with stem cells. ...Despite a vast amount of different methods, protocols and cryoprotective agents(CPA), stem cells are often frozen using standard protocols that have beenoptimized for use with cell lines, rather than with stem cells. Relatively fewcomparative studies have been performed to assess the effects of cryopreservationmethods on these stem cells. Dimethyl sulfoxide (DMSO) has been a key agent forthe development of cryobiology and has been used universally for cryopreservation.However, the use of DMSO has been associated with in vitro and in vivotoxicity and has been shown to affect many cellular processes due to changes inDNA methylation and dysregulation of gene expression. Despite studies showingthat DMSO may affect cell characteristics, DMSO remains the CPA of choice, bothin a research setting and in the clinics. However, numerous alternatives to DMSOhave been shown to hold promise for use as a CPA and include albumin,trehalose, sucrose, ethylene glycol, polyethylene glycol and many more. Here, wewill discuss the use, advantages and disadvantages of these CPAs for cryopreservationof different types of stem cells, including hematopoietic stem cells,mesenchymal stromal/stem cells and induced pluripotent stem cells.展开更多
Extracellular vesicles（EVs）provide a novel mechanism of intercellular communication via the transfer of proteins,lipids,and miR NAs between cells.It is now widely accepted that cargo content of EVs depends on cell t...Extracellular vesicles（EVs）provide a novel mechanism of intercellular communication via the transfer of proteins,lipids,and miR NAs between cells.It is now widely accepted that cargo content of EVs depends on cell type and its physiological state.Accordingly,EVs derived from healthy cells may have a comparable therapeutic potential as cells themselves.展开更多
BACKGROUND The immunosuppressive capacity of mesenchymal stem cells(MSCs)is dependent on the“license”of several proinflammatory factors to express immunosuppressive factors such as programmed cell death 1 ligand 1(P...BACKGROUND The immunosuppressive capacity of mesenchymal stem cells(MSCs)is dependent on the“license”of several proinflammatory factors to express immunosuppressive factors such as programmed cell death 1 ligand 1(PD-L1),which determines the clinical therapeutic efficacy of MSCs for inflammatory or immune diseases.In MSCs,interferon-gamma(IFN-γ)is a key inducer of PD-L1 expression,which is synergistically enhanced by tumor necrosis factor-alpha(TNF-α);however,the underlying mechanism is unclear.AIM To reveal the mechanism of pretreated MSCs express high PD-L1 and explore the application of pretreated MSCs in ulcerative colitis.METHODS We assessed PD-L1 expression in human umbilical-cord-derived MSCs(hUC-MSCs)induced by IFN-γand TNF-α,alone or in combination.Additionally,we performed signal pathway inhibitor experiments as well as RNA interference experiments to elucidate the molecular mechanism by which IFN-γalone or in combination with TNF-αinduces PD-L1 expression.Moreover,we used luciferase reporter gene experiments to verify the binding sites of the transcription factors of each signal transduction pathway to the targeted gene promoters.Finally,we evaluated the immunosuppressive capacity of hUC-MSCs treated with IFN-γand TNF-αin both an in vitro mixed lymphocyte culture assay,and in vivo in mice with dextran sulfate sodium-induced acute colitis.RESULTS Our results suggest that IFN-γinduction alone upregulates PD-L1 expression in hUC-MSCs while TNF-αalone does not,and that the co-induction of IFN-γand TNF-αpromotes higher expression of PD-L1.IFN-γinduces hUCMSCs to express PD-L1,in which IFN-γactivates the JAK/STAT1 signaling pathway,up-regulates the expression of the interferon regulatory factor 1(IRF1)transcription factor,promotes the binding of IRF1 and the PD-L1 gene promoter,and finally promotes PD-L1 mRNA.Although TNF-αalone did not induce PD-L1 expression in hUCMSCs,the addition of TNF-αsignificantly enhanced IFN-γ-induced JAK/STAT1/IRF1 activation.TNF-αupregulated IFN-γreceptor expression through activation of the nuclear factor kappa-B signaling pathway,which significantly enhanced IFN-γsignaling.Finally,co-induced hUC-MSCs have a stronger inhibitory effect on lymphocyte proliferation,and significantly ameliorate weight loss,mucosal damage,inflammatory cell infiltration,and up-regulation of inflammatory factors in colitis mice.CONCLUSION Overall,our results suggest that IFN-γand TNF-αenhance both the immunosuppressive ability of hUC-MSCs and their efficacy in ulcerative colitis by synergistically inducing high expression of PD-L1.展开更多
CD34+cells differentiated from mesenchymal stem cells (MSCs) have a strong biological function in cardiovascular regeneration. However, the molecular mechanisms of and the methods to improve the CD34+ cell differentia...CD34+cells differentiated from mesenchymal stem cells (MSCs) have a strong biological function in cardiovascular regeneration. However, the molecular mechanisms of and the methods to improve the CD34+ cell differentiation from MSCs, especially from human MSCs (hUC-MSCs) are still unclear. In the current study, the effect of CD34 antibody on the CD34+ cell differentiation from human umbilical cord (UC)-derived MSCs (hUC-MSCs) is determined. The results have demonstrated that the expression of cd34 protein is significantly increased in hUC-MSCs treated with CD34 antibody. In addition, the cell proliferation is increased in hUC-MSCs after treatment with CD34 antibody. Moreover, the expression of PI3K, AKT, p-AKT proteins, which are signaling molecules related to stem cell differentiation, is increased by CD34 antibody. The results suggest that CD34 antibody could promote the differentiation of hUC-MSCs into CD34+ cells and PI3K/AKT may be involved in this important process.展开更多
Stem cells are commonly classified based on the developmental stage from which they are isolated, although this has been a source of debate amongst stem cell scientists. A common approach classifies stem cells into th...Stem cells are commonly classified based on the developmental stage from which they are isolated, although this has been a source of debate amongst stem cell scientists. A common approach classifies stem cells into three different groupings: Embryonic Stem Cells (ESCs), Umbilical Cord Stem Cells (UCBSCs) and Adult Stem Cells (ASCs), which include stem cells from bone marrow (BM), fat tissue (FT), engineered induced pluripotent (IP) and peripheral blood (PB). By definition stem cells are progenitor cells capable of self-renewal and differentiation hypothetically “ab infinitum” into more specialized cells and tissue. The main intent of this study was to determine and characterize the different sub-groups of stem cells present within the human PB-SCs that may represent a valid opportunity in the field of clinical regenerative medicine. Stem cells in the isolated mononucleated cells were characterized using a multidisciplinary approach that was based on morphology, the expression of stem cell markers by flowcytometry and fluorescence analysis, RT-PCR and the capacity to self-renew or proliferate and differentiate into specialized cells. This approach was used to identify the expression of hematopoietic, mesenchymal, embryonic and neural stem cell markers. Both isolated adherent and suspension mononucleated cells were able to maintain their stem cell properties during in-vitro culture by holding their capacity for proliferation and differentiation into osteoblast cells, respectively, when exposed to the appropriate induction medium.展开更多
Background:Protein lactylation is a new way for the“metabolic waste”lactic acid to perform novel functions.Nevertheless,our understanding of the contribution of protein lactylation to both tumor progression and ther...Background:Protein lactylation is a new way for the“metabolic waste”lactic acid to perform novel functions.Nevertheless,our understanding of the contribution of protein lactylation to both tumor progression and therapeutic interventions remains imited.The construction of a scoring system for lactylation to predict the prognosis of pancancer patients and to evaluate the tumor immune microenvironment(TIME)would improve our understanding of the clinical significance of lactylation.Methods:Consensus clustering analysis of lactylation-related genes was used to cluster 177 pancreatic adenocarcinoma(PAAD)patients.Subsequently,a scoring system was developed using the least absolute shrinkage and selection operator(LASSO)regression.Internal validation and external validation were both conducted to assess and confirm the predictive accuracy of the scoring system.Finally,leucine rich repeat containing 1(LRRC1),a newly discovered lactylation-related gene,was analyzed in PAAD in vitro.Results:Utilizing the profiles of 332 lactylation-related genes,a total of 177 patients with PAAD were segregated into two distinct groups.LacCluster^(high) patients had a poorer prognosis than LacCluster^(low) patients.Through the differential analysis between the LacCluster^(high) and LacCluster^(low) groups,we identified additional genes associated with lactylation.These genes were then integrated to construct the LacCluster-enhanced system,which enabled more accurate prognosis prediction for patients with PAAD.Then,a lactylation index containing three genes(LacI-3)was constructed using LASSO regression.This was done to enhance the usability of the LacCluster-enhanced system in the clinic.Compared to those in the LacI-3^(high) subgroup,patients in the LacI-3^(low) subgroup exhibited increased expression of immune checkpoint-related genes,more immune cell infiltration,lower tumor mutation burdens,and better prognoses,indicating a“hot tumor”phenotype.Moreover,knocking down the expression of LRRC1,the hub gene in the LacI-3 scoring system,inhibited PAAD cell invasion,migration,and proliferation in vitro.Ultimately,the significance of LacI-3 across cancers was confirmed.Conclusion:Our findings strongly imply that protein lactylation may represent a new approach to diagnosing and treating malignant tumors.展开更多
Alzheimer’s disease(AD)is a major age-related form of dementia with a number of cases exponentially growing,causing enormous social and economic impact on individuals and society.Neuropathological hallmarks of AD,evi...Alzheimer’s disease(AD)is a major age-related form of dementia with a number of cases exponentially growing,causing enormous social and economic impact on individuals and society.Neuropathological hallmarks of AD,evident in postmortem AD brains,include a massive loss of the grey matter in the neocortex,extracellular deposition of amyloid-β(Aβ)in the form of senile plaques and cerebrovascular amyloid angiopathy,and intra-neuronal accumulation of neurofibrillary tangles,formed by hyper-phosphorylated tau protein.展开更多
BACKGROUND ATP sensitive K+(K_(ATP))channels are ubiquitously distributed in various of cells and tissues,including the liver.They play a role in the pathogenesis of myocardial and liver ischemia.AIM To evaluate the r...BACKGROUND ATP sensitive K+(K_(ATP))channels are ubiquitously distributed in various of cells and tissues,including the liver.They play a role in the pathogenesis of myocardial and liver ischemia.AIM To evaluate the radiation-induced changes in the expression of K_(ATP)channel subunits in the mouse liver to understand the potential role of K_(ATP)channels in radiation injury.METHODS Adult C57BL/6 mice were randomly exposed toγ-rays at 0 Gy(control,n=2),0.2 Gy(n=6),1 Gy(n=6),or 5 Gy(n=6).The livers were removed 3 and 24 h after radiation exposure.Hematoxylin and eosin staining was used for morphological observation;immunohistochemical staining was applied to determine the expression of K_(ATP)channel subunits in the liver tissue.RESULTS Compared with the control group,the livers exposed to 0.2 Gyγ-ray showed an initial increase in the expression of Kir6.1 at 3 h,followed by recovery at 24 h after exposure.Exposure to a high dose of 5.0 Gy resulted in decreased expression of Kir6.1 and increased expression of SUR2B at 24 h.However,the expression of Kir6.2,SUR1,or SUR2A had no remarkable changes at 3 and 24 h after exposure to any of these doses.CONCLUSION The expression levels of Kir6.1 and SUR2B in mouse liver changed differently in response to different radiation doses,suggesting a potential role for them in radiation-induced liver injury.展开更多
The Antibiogram of the bacterial isolates from Tuberculosis (TB) patients attending hospitals within Izzi-Abakaliki was evaluated. The bacterial isolates were isolated and identified from the sputum samples according ...The Antibiogram of the bacterial isolates from Tuberculosis (TB) patients attending hospitals within Izzi-Abakaliki was evaluated. The bacterial isolates were isolated and identified from the sputum samples according to microbiological principles while the antibiotics susceptibility test was done by disc diffusion method using Ofloxacin, Pefloxacin, Ciprofloxacin, amoxicillin/clavulanate, Gentamycin, Streptomycin, Cephalosporin, Cotrimoxazole, Nalidixic acid and Ampicillin. Bacteria isolated include 5 E. coli (25%), 3 Streptococcus pyogenes (15%), 2 Streptococcus pneumoniae (10%), 1 Klebsiella spp. (5%), 3 Haemophilus influenza (15%), 2 Pseudomonas (10%), 3 Proteus spp. (15%), 1 Staphylococcus aureus (10%). The result of Antibiogram shows that E. coli was 100% resistant to Amoxicillin/clavulanate and cotrimoxazole, followed by Streptomycin (80%) and 100% susceptible to Pefloxacin with inhibition zone diameter of 18 mm and 18 mm for Ofloxacin (60%). S. pneumoniae and Klebsiella spp. were highly resistant to Amoxicillin/clavulanate (100%), Gentamycin (100%) and Ampicillin (100%) and 100% susceptible to Pefloxacin with inhibition zone 18 mm, Ciprofloxacin (17 mm). S. pyogenes was resistant to streptomycin and Ceporex, with 100% sensitivity to Ofloxacin, Ciprofloxacin and Pefloxacin. Pseudomonas spp. and S. aureus were both 100% resistant to all antibiotics except Ofloxacin, Ciprofloxacin, and Pefloxacin respectively. Proteus spp. was susceptible to Pefloxacin (100%), Ofloxacin (66.7%) and Ciprofloxacin (66.7%) but highly resistant to Streptomycin (100%) and Ampicillin (100%). Haemophilus influenzae were susceptible to Ofloxacin (100%) and Pefloxacin (100%), with high resistance to Amoxicillin/clavulanate (100%) and Ceporex (100%). From the study, Ofloxacin and Pefloxacin are susceptible to all bacteria isolated and are recommended for treatment of the bacterial infection with TB patient.展开更多
Stroke causes neuronal loss,which ultimately results in persistent neurological dysfunction.Globally,stroke was the third-leading cause of death and disability combined in all ages in 2019,after neonatal disorders and...Stroke causes neuronal loss,which ultimately results in persistent neurological dysfunction.Globally,stroke was the third-leading cause of death and disability combined in all ages in 2019,after neonatal disorders and ischemic heart disease.In that year,there were 12.2 million incident strokes,101 million prevalent strokes,and 143 million disability-adjusted life-years due to stroke.展开更多
Adenomatous polyposis coli(APC)mutations are the most frequently identified genetic alteration in sporadic colorectal cancer(CRC)cases,and a myriad of genetically engineered Apc-mutant CRC mouse models have been devel...Adenomatous polyposis coli(APC)mutations are the most frequently identified genetic alteration in sporadic colorectal cancer(CRC)cases,and a myriad of genetically engineered Apc-mutant CRC mouse models have been developed using various genetic manipulation techniques.The advent of the CRISPR/Cas9 system has revolutionized the field of genetic engineering and facilitated the development of new genetically engineered mouse models.In this study,we aimed to develop a novel Apc knockout allele using the CRISPR/Cas9 system and evaluate the phenotypic effects of this new allele in two different mouse strains.For this purpose,exon 16 of mouse Apc gene was targeted with a single-guide RNA,and the mouse carrying an Apc frameshift mutation at codon 750(^(Δ750))was chosen as the founder.The mutant FVB-Apc^(Δ750)mice were backcrossed with wild-type C57BL/6 mice,and the phenotypic effects of the knockout allele were evaluated in F8-FVB-Apc^(Δ750),F4-B6;FVB-Apc^(Δ750),and F1-B6;FVB-Apc^(Δ750)by a macroscopic and microscopic examination of the gastrointestinal system.The result showed that the mean polyp number was significantly higher in F4-BL6;FVB-Apc^(Δ750)than in F8-FVB-Apc^(Δ750).Intestinal polyposis was more prominent in F4-BL6;FVB-Apc^(Δ750),whereas a higher number of colon polyps than intestinal polyps were observed in F8-FVB-Apc^(Δ750).Additionally,F1-BL6;FVB-Apc^(Δ750)mixed background mice developed gastric polyps that morphologically resembled the pyloric gland adenoma of humans.In conclusion,we developed a novel CRISPR-mediated Apc knockout allele using two mouse strains.We showed that this allele can exert a strainspecific effect on the phenotype of mice and can cause gastric polyp formation.展开更多
Irradiation with X-rays has been widely utilized in the clinical treatment of solid tumors and certain hematopoietic malignancies.However,this method fails to completely distinguish between malignant and normal cells....Irradiation with X-rays has been widely utilized in the clinical treatment of solid tumors and certain hematopoietic malignancies.However,this method fails to completely distinguish between malignant and normal cells.Prolonged or repeated exposure to radiation,whether due to occupational hazards or therapeutical interventions,can cause damage to normal tissues,particularly impacting the hematopoietic system.Therefore,it is important to investigate the effects of total body irradiation on the hematopoietic system of mice and to compare the inhibitory effects of various doses of irradiation on this system.In this study,we primarily employed flow cytometry to analyze mature lineage cells in the peripheral blood,as well as immature hematopoietic stem and progenitor cells(HSPCs)in the bone marrow and spleen.Additionally,we evaluated the multilineage differentiation capacity of HSPCs through colony-forming cell assays.Our results indicated that peripheral B and T cells demonstrated increased sensitivity to irradiation,with significant cell death observed 1-day post-irradiation.Common lymphoid progenitor cells exhibited greater radiotolerance compared to other progenitor cell types,enabling them to maintain a certain population even at elevated doses.Moreover,notable differences were observed between intramedullary and extramedullary hematopoietic stem cells and common lymphoid progenitor cells regarding the extent of damage and recovery rate following irradiation.The multilineage differentiation capacity of HSPCs was also compromised during radiation exposure.In conclusion,different types of mature blood cells,along with immature HSPCs,exhibited varying degrees of sensitivity and tolerance to irradiation,resulting in distinct alterations in cell percentages and numbers.展开更多
Since articular cartilage possesses only a weak capac-ity for repair, its regeneration potential is considered one of the most important challenges for orthopedic surgeons. The treatment options, such as marrow stimul...Since articular cartilage possesses only a weak capac-ity for repair, its regeneration potential is considered one of the most important challenges for orthopedic surgeons. The treatment options, such as marrow stimulation techniques, fail to induce a repair tissue with the same functional and mechanical properties of native hyaline cartilage. Osteochondral transplantation is considered an effective treatment option but is as-sociated with some disadvantages, including donor-site morbidity, tissue supply limitation, unsuitable mechani-cal properties and thickness of the obtained tissue. Although autologous chondrocyte implantation results in reasonable repair, it requires a two-step surgical pro-cedure. Moreover, chondrocytes expanded in culture gradually undergo dedifferentiation, so lose morpho-logical features and specialized functions. In the search for alternative cells, scientists have found mesenchymal stem cells(MSCs) to be an appropriate cellular mate-rial for articular cartilage repair. These cells were origi-nally isolated from bone marrow samples and further investigations have revealed the presence of the cells in many other tissues. Furthermore, chondrogenic dif-ferentiation is an inherent property of MSCs noticedat the time of the cell discovery. MSCs are known to exhibit homing potential to the damaged site at which they differentiate into the tissue cells or secrete a wide spectrum of bioactive factors with regenerative proper-ties. Moreover, these cells possess a considerable im-munomodulatory potential that make them the general donor for therapeutic applications. All of these topics will be discussed in this review.展开更多
基金supported by a Grant-in-Aid for Scientific Research(B)JP21H02808(to TM)JST SPRING JPMJSP2136(to MI)。
文摘Neural stem cells(NSCs)are the source of all neurons and glial cells(astrocytes and oligodendrocytes)in the central nervous system.The adult mammalian brain retains NSCs in the subgranular zone of the dentate gyrus in the hippocampus and ventricular subventricular zone lining the lateral ventricle(Olpe and Jessberger,2022).Adult NSCs in rodents are preserved throughout life and continuously produce new neurons that integrate into the pre-existing neuronal network.
文摘“Last scene of all that ends this strange,eventful history,is second childishness and mere oblivion.I am sans teeth,sans eyes,sans taste,sans everything.”William Shakespeare‘As You Like It'Act 2,Sc.7,l.139Aging of the human brain is characterized by a progressive decline of its functional capacity;this decline however varies widely,and cognitive longevity differs substantially between individuals.
基金financially supported by the Korea Institute of Planning and Evaluation for Technology in Food,Agriculture and Forestry(IPET-RS-2024–00402320)by the Meterials/Parts Technology Development Pro-gram(1415187291,Development of composite formulation with a sustained release(gene)for the treatment of companion animal sarcopenia)funded By the Ministry of Trade,Industry&Energy(MOTIE,Korea)。
文摘Background As the global population increases,the demand for protein sources is expected to increase,driving the demand for cell-based cultivated meat.This study aimed to enhance the productivity of cultivated meat through optimization of the cell source and organization process.Results We engineered fibroblasts into myogenic cells via non-viral introduction of the MYOD1 gene,avoiding viral methods for safety.After confirming the stable derivation of myogenic cells,we combined knockout(KO)of MSTN,a negative regulator of myogenesis,with MYOD1-mediated myogenesis to improve cultivated meat production.Primary cells from MSTN KO cattle exhibited enhanced myogenic potential.Additionally,when tested in immortalized fibroblasts,myostatin treatment reduced MYOD1-induced myogenesis in two-dimensional cultures,while MSTN knockout increased it.To achieve muscle-like cell alignment,we employed digital light processing(DLP)-based three-dimensional(3D)bioprinting to organize cells into 3D groove-shaped hydrogels.These bioactive hydrogels supported stable cell proliferation and significantly improved muscle cell alignment.Upon differentiation into myotubes,the cells demonstrated an ordered alignment,particularly the MSTN KO cells,which showed highly efficient differentiation.Conclusions The integration of genetic modification and advanced DLP 3D bioprinting with groove-patterned hydrogels provides an effective strategy for producing high-quality,muscle-aligned cultivated meat.
基金financially supported by grants from Royan In-stitute(grant No.400000200)Bahar Tashkhis Teb Co.(BTT,9703,9809,and 9903)。
文摘Background: Despite considerable advancements in identifying factors contributing to the development of hepatocellular carcinoma(HCC), the pathogenesis of HCC remains unclear. In many cases, HCC is a consequence of prolonged liver fibrosis, resulting in the formation of an intricate premalignant microenvironment. The accumulation of extracellular matrix(ECM) is a hallmark of premalignant microenvironment. Given the critical role of different matrix components in regulating cell phenotype and function, this study aimed to elucidate the interplay between the fibrotic matrix and malignant features in HCC. Methods: Liver tissues from both control(normal) and carbon tetrachloride(CCl_(4))-induced fibrotic rats were decellularized using sodium dodecyl sulfate(SDS) and Triton X-100. The resulting hydrogel from decellularized ECM was processed into micro-particles via the water-in-oil emulsion method. Microparticles were subsequently incorporated into three-dimensional liver biomimetic micro-tissues(MTs) comprising Huh-7 cells, human umbilical vein endothelial cells(HUVECs), and LX-2 cells. The MTs were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS) assay at day 11, immunofluorescence staining, immunoblotting, and spheroid migration assay at day 14 after co-culture. Results: Fibrotic matrix from CCl4-treated rat livers significantly enhanced the growth rate of the MTs and their expression of CCND1 as compared to the normal one. Fibrotic matrix, also induced the expression of epithelial-to-mesenchymal transition(EMT)-associated genes such as TWIST1, ACTA2, MMP9, CDH2, and VIMENTIN in the MTs as compared to the normal matrix. Conversely, the expression of CDH1 and hepatic maturation genes HNF4A, ALB, CYP3A4 was decreased in the MTs when the fibrotic matrix was used. Furthermore, the fibrotic matrix increased the migration of the MTs and their secretion of alpha-fetoprotein. Conclusions: Our findings suggest a regulatory role for the fibrotic matrix in promoting cancerous phenotype, which could potentially accelerate the progression of malignancy in the liver.
文摘AIM: To evaluate safety and feasibility of autologous bone marrow-enriched CD34+ hematopoietic stem cell Tx through the hepatic artery in patients with decompensated cirrhosis.METHODS: Four patients with decompensated cirrhosis were included. Approximately 200 mL of the bone marrow of the patients was aspirated, and CD34+ stem cells were selected. Between 3 to 10 million CD34+ cells were isolated. The cells were slowly infused through the hepatic artery of the patients.RESULTS: Patient 1 showed marginal improvement in serum albumin and no significant changes in other test results. In patient 2 prothrombin time was decreased; however, her total bilirubin, serum creatinine, and Model of End-Stage Liver Disease (MELD) score worsened at the end of follow up. In patient 3 there was improvement in serum albumin, porthrombin time (PT), and MELD score. Patient 4 developed radiocontrast nephropathy after the procedure, and progressed to type 1 hepatorenal syndrome and died of liver failure a few days later. Because of the major side effects seen in the last patient, the trial was prematurely stopped.CONCLUSION: Infusion of CD34+ stem cells through the hepatic artery is not safe in decompensated cirrhosis. Radiocontrast nephropathy and hepatorenal syndrome could be major side effects. However, this study doesnot preclude infusion of CD34+ stem cells through other routes.
基金Supported by Science and Technology Program of Shenyang (2009-090063, 2011-F10-222-4-00)
文摘Objective To establish the method of isolation, purification, and identification of human amniotic mesenchymal stem cells (hAMSCs). Methods hAMSCs were isolated from human amniotic membrane by trypsin-collagenase digestion, and cultured in Dulbecco's modified Eagle's medinm/F12 medium supplemented with 10% fetal bovine serum. Phenotypic characteristics of these cells were analyzed by means of immunocytochemistry and flow cytometry. Results The cells successfully isolated from human amniotic membrane expressed representative mesenchymal cell surface markers CD44, CD90, and vimentin, but not CD45. Conclusions This study establishes a potential method for isolation of hAMSCs from human amnion, in vitro culture, and identification. The isolated cells show phenotypic characteristics of mesenchymal stem cells.
基金funded by the Research Center for Science and Technology in Medicine(RCSTiM),Tehran University of Medical Sciences,Tehran(TUMS),Tehran,Iran
文摘Several studies have demonstrated that selective serotonin reuptake inhibitor antidepressants can promote neuronal cell proliferation and enhance neuroplasticity both in vitro and in vivo. It is hypothesized that citalopram, a selective serotonin reuptake inhibitor, can promote the neuronal differentiation of adult bone marrow mesenchymal stem cells. Citalopram strongly enhanced neuronal characteristics of the cells derived from bone marrow mesenchymal stem cells. The rate of cell death was decreased in citalopram-treated bone marrow mesenchymal stem cells than in control cells in neurobasal medium. In addition, the cumulative population doubling level of the citalopram-treated cells was signiifcantly increased compared to that of control cells. Also BrdU incorporation was elevated in citalopram-treated cells. These ifndings suggest that citalopram can improve the neuronal-like cell differentiation of bone marrow mesenchymal stem cells by increasing cell proliferation and survival while maintaining their neuronal characteristics.
基金the Scientific and Technological Research Council of Turkey(TÜBİTAK),No.118S738 and No.219S675.
文摘Despite a vast amount of different methods, protocols and cryoprotective agents(CPA), stem cells are often frozen using standard protocols that have beenoptimized for use with cell lines, rather than with stem cells. Relatively fewcomparative studies have been performed to assess the effects of cryopreservationmethods on these stem cells. Dimethyl sulfoxide (DMSO) has been a key agent forthe development of cryobiology and has been used universally for cryopreservation.However, the use of DMSO has been associated with in vitro and in vivotoxicity and has been shown to affect many cellular processes due to changes inDNA methylation and dysregulation of gene expression. Despite studies showingthat DMSO may affect cell characteristics, DMSO remains the CPA of choice, bothin a research setting and in the clinics. However, numerous alternatives to DMSOhave been shown to hold promise for use as a CPA and include albumin,trehalose, sucrose, ethylene glycol, polyethylene glycol and many more. Here, wewill discuss the use, advantages and disadvantages of these CPAs for cryopreservationof different types of stem cells, including hematopoietic stem cells,mesenchymal stromal/stem cells and induced pluripotent stem cells.
基金supported by National Research Programme,“Healthy ageing”(Grant No.SEN-15090)from Research Council of Lithuania
文摘Extracellular vesicles（EVs）provide a novel mechanism of intercellular communication via the transfer of proteins,lipids,and miR NAs between cells.It is now widely accepted that cargo content of EVs depends on cell type and its physiological state.Accordingly,EVs derived from healthy cells may have a comparable therapeutic potential as cells themselves.
基金Supported by the National Natural Science Foundation of China,No.81871568,No.32100643COVID-19 Infection and Prevention Emergency Special Project of Chongqing Education Commission,No.KYYJ202009.
文摘BACKGROUND The immunosuppressive capacity of mesenchymal stem cells(MSCs)is dependent on the“license”of several proinflammatory factors to express immunosuppressive factors such as programmed cell death 1 ligand 1(PD-L1),which determines the clinical therapeutic efficacy of MSCs for inflammatory or immune diseases.In MSCs,interferon-gamma(IFN-γ)is a key inducer of PD-L1 expression,which is synergistically enhanced by tumor necrosis factor-alpha(TNF-α);however,the underlying mechanism is unclear.AIM To reveal the mechanism of pretreated MSCs express high PD-L1 and explore the application of pretreated MSCs in ulcerative colitis.METHODS We assessed PD-L1 expression in human umbilical-cord-derived MSCs(hUC-MSCs)induced by IFN-γand TNF-α,alone or in combination.Additionally,we performed signal pathway inhibitor experiments as well as RNA interference experiments to elucidate the molecular mechanism by which IFN-γalone or in combination with TNF-αinduces PD-L1 expression.Moreover,we used luciferase reporter gene experiments to verify the binding sites of the transcription factors of each signal transduction pathway to the targeted gene promoters.Finally,we evaluated the immunosuppressive capacity of hUC-MSCs treated with IFN-γand TNF-αin both an in vitro mixed lymphocyte culture assay,and in vivo in mice with dextran sulfate sodium-induced acute colitis.RESULTS Our results suggest that IFN-γinduction alone upregulates PD-L1 expression in hUC-MSCs while TNF-αalone does not,and that the co-induction of IFN-γand TNF-αpromotes higher expression of PD-L1.IFN-γinduces hUCMSCs to express PD-L1,in which IFN-γactivates the JAK/STAT1 signaling pathway,up-regulates the expression of the interferon regulatory factor 1(IRF1)transcription factor,promotes the binding of IRF1 and the PD-L1 gene promoter,and finally promotes PD-L1 mRNA.Although TNF-αalone did not induce PD-L1 expression in hUCMSCs,the addition of TNF-αsignificantly enhanced IFN-γ-induced JAK/STAT1/IRF1 activation.TNF-αupregulated IFN-γreceptor expression through activation of the nuclear factor kappa-B signaling pathway,which significantly enhanced IFN-γsignaling.Finally,co-induced hUC-MSCs have a stronger inhibitory effect on lymphocyte proliferation,and significantly ameliorate weight loss,mucosal damage,inflammatory cell infiltration,and up-regulation of inflammatory factors in colitis mice.CONCLUSION Overall,our results suggest that IFN-γand TNF-αenhance both the immunosuppressive ability of hUC-MSCs and their efficacy in ulcerative colitis by synergistically inducing high expression of PD-L1.
文摘CD34+cells differentiated from mesenchymal stem cells (MSCs) have a strong biological function in cardiovascular regeneration. However, the molecular mechanisms of and the methods to improve the CD34+ cell differentiation from MSCs, especially from human MSCs (hUC-MSCs) are still unclear. In the current study, the effect of CD34 antibody on the CD34+ cell differentiation from human umbilical cord (UC)-derived MSCs (hUC-MSCs) is determined. The results have demonstrated that the expression of cd34 protein is significantly increased in hUC-MSCs treated with CD34 antibody. In addition, the cell proliferation is increased in hUC-MSCs after treatment with CD34 antibody. Moreover, the expression of PI3K, AKT, p-AKT proteins, which are signaling molecules related to stem cell differentiation, is increased by CD34 antibody. The results suggest that CD34 antibody could promote the differentiation of hUC-MSCs into CD34+ cells and PI3K/AKT may be involved in this important process.
文摘Stem cells are commonly classified based on the developmental stage from which they are isolated, although this has been a source of debate amongst stem cell scientists. A common approach classifies stem cells into three different groupings: Embryonic Stem Cells (ESCs), Umbilical Cord Stem Cells (UCBSCs) and Adult Stem Cells (ASCs), which include stem cells from bone marrow (BM), fat tissue (FT), engineered induced pluripotent (IP) and peripheral blood (PB). By definition stem cells are progenitor cells capable of self-renewal and differentiation hypothetically “ab infinitum” into more specialized cells and tissue. The main intent of this study was to determine and characterize the different sub-groups of stem cells present within the human PB-SCs that may represent a valid opportunity in the field of clinical regenerative medicine. Stem cells in the isolated mononucleated cells were characterized using a multidisciplinary approach that was based on morphology, the expression of stem cell markers by flowcytometry and fluorescence analysis, RT-PCR and the capacity to self-renew or proliferate and differentiate into specialized cells. This approach was used to identify the expression of hematopoietic, mesenchymal, embryonic and neural stem cell markers. Both isolated adherent and suspension mononucleated cells were able to maintain their stem cell properties during in-vitro culture by holding their capacity for proliferation and differentiation into osteoblast cells, respectively, when exposed to the appropriate induction medium.
基金supported by the National Key Research and Development Program of China(Grant Number 2022YFA1205003)Major Research Projects of the National Natural Science Foundation of China(Grant Number 92059204)+1 种基金General Research Projects of the National Natural Science Foundation of China(Grant Number 82273419)Major Projects of Technological Innovation and Application Development Foundation in Chongqing(Grant Number CSTB2022TIAD-STX0012).
文摘Background:Protein lactylation is a new way for the“metabolic waste”lactic acid to perform novel functions.Nevertheless,our understanding of the contribution of protein lactylation to both tumor progression and therapeutic interventions remains imited.The construction of a scoring system for lactylation to predict the prognosis of pancancer patients and to evaluate the tumor immune microenvironment(TIME)would improve our understanding of the clinical significance of lactylation.Methods:Consensus clustering analysis of lactylation-related genes was used to cluster 177 pancreatic adenocarcinoma(PAAD)patients.Subsequently,a scoring system was developed using the least absolute shrinkage and selection operator(LASSO)regression.Internal validation and external validation were both conducted to assess and confirm the predictive accuracy of the scoring system.Finally,leucine rich repeat containing 1(LRRC1),a newly discovered lactylation-related gene,was analyzed in PAAD in vitro.Results:Utilizing the profiles of 332 lactylation-related genes,a total of 177 patients with PAAD were segregated into two distinct groups.LacCluster^(high) patients had a poorer prognosis than LacCluster^(low) patients.Through the differential analysis between the LacCluster^(high) and LacCluster^(low) groups,we identified additional genes associated with lactylation.These genes were then integrated to construct the LacCluster-enhanced system,which enabled more accurate prognosis prediction for patients with PAAD.Then,a lactylation index containing three genes(LacI-3)was constructed using LASSO regression.This was done to enhance the usability of the LacCluster-enhanced system in the clinic.Compared to those in the LacI-3^(high) subgroup,patients in the LacI-3^(low) subgroup exhibited increased expression of immune checkpoint-related genes,more immune cell infiltration,lower tumor mutation burdens,and better prognoses,indicating a“hot tumor”phenotype.Moreover,knocking down the expression of LRRC1,the hub gene in the LacI-3 scoring system,inhibited PAAD cell invasion,migration,and proliferation in vitro.Ultimately,the significance of LacI-3 across cancers was confirmed.Conclusion:Our findings strongly imply that protein lactylation may represent a new approach to diagnosing and treating malignant tumors.
基金the following financial support grant FAR-2019 to DL from The Universita del Piemonte Orientale。
文摘Alzheimer’s disease(AD)is a major age-related form of dementia with a number of cases exponentially growing,causing enormous social and economic impact on individuals and society.Neuropathological hallmarks of AD,evident in postmortem AD brains,include a massive loss of the grey matter in the neocortex,extracellular deposition of amyloid-β(Aβ)in the form of senile plaques and cerebrovascular amyloid angiopathy,and intra-neuronal accumulation of neurofibrillary tangles,formed by hyper-phosphorylated tau protein.
基金Supported by the Program of the Network-type Joint Usage/Research Center for Radiation Disaster Medical Science of Hiroshima University,Nagasaki University.
文摘BACKGROUND ATP sensitive K+(K_(ATP))channels are ubiquitously distributed in various of cells and tissues,including the liver.They play a role in the pathogenesis of myocardial and liver ischemia.AIM To evaluate the radiation-induced changes in the expression of K_(ATP)channel subunits in the mouse liver to understand the potential role of K_(ATP)channels in radiation injury.METHODS Adult C57BL/6 mice were randomly exposed toγ-rays at 0 Gy(control,n=2),0.2 Gy(n=6),1 Gy(n=6),or 5 Gy(n=6).The livers were removed 3 and 24 h after radiation exposure.Hematoxylin and eosin staining was used for morphological observation;immunohistochemical staining was applied to determine the expression of K_(ATP)channel subunits in the liver tissue.RESULTS Compared with the control group,the livers exposed to 0.2 Gyγ-ray showed an initial increase in the expression of Kir6.1 at 3 h,followed by recovery at 24 h after exposure.Exposure to a high dose of 5.0 Gy resulted in decreased expression of Kir6.1 and increased expression of SUR2B at 24 h.However,the expression of Kir6.2,SUR1,or SUR2A had no remarkable changes at 3 and 24 h after exposure to any of these doses.CONCLUSION The expression levels of Kir6.1 and SUR2B in mouse liver changed differently in response to different radiation doses,suggesting a potential role for them in radiation-induced liver injury.
文摘The Antibiogram of the bacterial isolates from Tuberculosis (TB) patients attending hospitals within Izzi-Abakaliki was evaluated. The bacterial isolates were isolated and identified from the sputum samples according to microbiological principles while the antibiotics susceptibility test was done by disc diffusion method using Ofloxacin, Pefloxacin, Ciprofloxacin, amoxicillin/clavulanate, Gentamycin, Streptomycin, Cephalosporin, Cotrimoxazole, Nalidixic acid and Ampicillin. Bacteria isolated include 5 E. coli (25%), 3 Streptococcus pyogenes (15%), 2 Streptococcus pneumoniae (10%), 1 Klebsiella spp. (5%), 3 Haemophilus influenza (15%), 2 Pseudomonas (10%), 3 Proteus spp. (15%), 1 Staphylococcus aureus (10%). The result of Antibiogram shows that E. coli was 100% resistant to Amoxicillin/clavulanate and cotrimoxazole, followed by Streptomycin (80%) and 100% susceptible to Pefloxacin with inhibition zone diameter of 18 mm and 18 mm for Ofloxacin (60%). S. pneumoniae and Klebsiella spp. were highly resistant to Amoxicillin/clavulanate (100%), Gentamycin (100%) and Ampicillin (100%) and 100% susceptible to Pefloxacin with inhibition zone 18 mm, Ciprofloxacin (17 mm). S. pyogenes was resistant to streptomycin and Ceporex, with 100% sensitivity to Ofloxacin, Ciprofloxacin and Pefloxacin. Pseudomonas spp. and S. aureus were both 100% resistant to all antibiotics except Ofloxacin, Ciprofloxacin, and Pefloxacin respectively. Proteus spp. was susceptible to Pefloxacin (100%), Ofloxacin (66.7%) and Ciprofloxacin (66.7%) but highly resistant to Streptomycin (100%) and Ampicillin (100%). Haemophilus influenzae were susceptible to Ofloxacin (100%) and Pefloxacin (100%), with high resistance to Amoxicillin/clavulanate (100%) and Ceporex (100%). From the study, Ofloxacin and Pefloxacin are susceptible to all bacteria isolated and are recommended for treatment of the bacterial infection with TB patient.
基金supported by JSPS KAKENHI Grant Number JP24K18622(to TI)JSPS KAKENHI Grant Number JP23K18451(to TM)。
文摘Stroke causes neuronal loss,which ultimately results in persistent neurological dysfunction.Globally,stroke was the third-leading cause of death and disability combined in all ages in 2019,after neonatal disorders and ischemic heart disease.In that year,there were 12.2 million incident strokes,101 million prevalent strokes,and 143 million disability-adjusted life-years due to stroke.
基金The Scientific and Technological Research Council of Turkey 1001 program,Grant/Award Number:SBAG-215S926。
文摘Adenomatous polyposis coli(APC)mutations are the most frequently identified genetic alteration in sporadic colorectal cancer(CRC)cases,and a myriad of genetically engineered Apc-mutant CRC mouse models have been developed using various genetic manipulation techniques.The advent of the CRISPR/Cas9 system has revolutionized the field of genetic engineering and facilitated the development of new genetically engineered mouse models.In this study,we aimed to develop a novel Apc knockout allele using the CRISPR/Cas9 system and evaluate the phenotypic effects of this new allele in two different mouse strains.For this purpose,exon 16 of mouse Apc gene was targeted with a single-guide RNA,and the mouse carrying an Apc frameshift mutation at codon 750(^(Δ750))was chosen as the founder.The mutant FVB-Apc^(Δ750)mice were backcrossed with wild-type C57BL/6 mice,and the phenotypic effects of the knockout allele were evaluated in F8-FVB-Apc^(Δ750),F4-B6;FVB-Apc^(Δ750),and F1-B6;FVB-Apc^(Δ750)by a macroscopic and microscopic examination of the gastrointestinal system.The result showed that the mean polyp number was significantly higher in F4-BL6;FVB-Apc^(Δ750)than in F8-FVB-Apc^(Δ750).Intestinal polyposis was more prominent in F4-BL6;FVB-Apc^(Δ750),whereas a higher number of colon polyps than intestinal polyps were observed in F8-FVB-Apc^(Δ750).Additionally,F1-BL6;FVB-Apc^(Δ750)mixed background mice developed gastric polyps that morphologically resembled the pyloric gland adenoma of humans.In conclusion,we developed a novel CRISPR-mediated Apc knockout allele using two mouse strains.We showed that this allele can exert a strainspecific effect on the phenotype of mice and can cause gastric polyp formation.
基金supported by grants from the CAMS Innovation Fund for Medical Science(2021-I2M-1-019 to Y.G.and W.Z.)the National Natural Science Foundation of China(92068204 to Y.G.,82370120 to Y.L.,and 82200126 to W.Z.)+1 种基金Young Scientific and Technological Talents(Level Two)in Tianjin(QN20230216 to Y.L.)a SKLEH-Pilot Research Grant。
文摘Irradiation with X-rays has been widely utilized in the clinical treatment of solid tumors and certain hematopoietic malignancies.However,this method fails to completely distinguish between malignant and normal cells.Prolonged or repeated exposure to radiation,whether due to occupational hazards or therapeutical interventions,can cause damage to normal tissues,particularly impacting the hematopoietic system.Therefore,it is important to investigate the effects of total body irradiation on the hematopoietic system of mice and to compare the inhibitory effects of various doses of irradiation on this system.In this study,we primarily employed flow cytometry to analyze mature lineage cells in the peripheral blood,as well as immature hematopoietic stem and progenitor cells(HSPCs)in the bone marrow and spleen.Additionally,we evaluated the multilineage differentiation capacity of HSPCs through colony-forming cell assays.Our results indicated that peripheral B and T cells demonstrated increased sensitivity to irradiation,with significant cell death observed 1-day post-irradiation.Common lymphoid progenitor cells exhibited greater radiotolerance compared to other progenitor cell types,enabling them to maintain a certain population even at elevated doses.Moreover,notable differences were observed between intramedullary and extramedullary hematopoietic stem cells and common lymphoid progenitor cells regarding the extent of damage and recovery rate following irradiation.The multilineage differentiation capacity of HSPCs was also compromised during radiation exposure.In conclusion,different types of mature blood cells,along with immature HSPCs,exhibited varying degrees of sensitivity and tolerance to irradiation,resulting in distinct alterations in cell percentages and numbers.
文摘Since articular cartilage possesses only a weak capac-ity for repair, its regeneration potential is considered one of the most important challenges for orthopedic surgeons. The treatment options, such as marrow stimulation techniques, fail to induce a repair tissue with the same functional and mechanical properties of native hyaline cartilage. Osteochondral transplantation is considered an effective treatment option but is as-sociated with some disadvantages, including donor-site morbidity, tissue supply limitation, unsuitable mechani-cal properties and thickness of the obtained tissue. Although autologous chondrocyte implantation results in reasonable repair, it requires a two-step surgical pro-cedure. Moreover, chondrocytes expanded in culture gradually undergo dedifferentiation, so lose morpho-logical features and specialized functions. In the search for alternative cells, scientists have found mesenchymal stem cells(MSCs) to be an appropriate cellular mate-rial for articular cartilage repair. These cells were origi-nally isolated from bone marrow samples and further investigations have revealed the presence of the cells in many other tissues. Furthermore, chondrogenic dif-ferentiation is an inherent property of MSCs noticedat the time of the cell discovery. MSCs are known to exhibit homing potential to the damaged site at which they differentiate into the tissue cells or secrete a wide spectrum of bioactive factors with regenerative proper-ties. Moreover, these cells possess a considerable im-munomodulatory potential that make them the general donor for therapeutic applications. All of these topics will be discussed in this review.
