Objective:Suppression of tumorigenicity 2(ST2),the receptor for interleukin(IL)-33,has a critical role in tumor growth,angiogenesis,metastasis,and immune modulation.The IL-33/ST2 pathway is known to influence the pola...Objective:Suppression of tumorigenicity 2(ST2),the receptor for interleukin(IL)-33,has a critical role in tumor growth,angiogenesis,metastasis,and immune modulation.The IL-33/ST2 pathway is known to influence the polarization and function of macrophages,which is integral to modulating the tumor microenvironment.However,the precise role of IL-33/ST2 in tumors,particularly non-small cell lung cancer(NSCLC),has not been established.Methods:ST2 expression in NSCLC was analysed using a murine model and patient specimens.The effect of the IL-33/ST2 axis on macrophage polarization in NSCLC was determined.Results:Elevated ST2 expression was correlated with aggressive tumor growth.Specifically,ST2 expression on macrophages was associated with lung cancer progression and the absence of ST2 on macrophages was associated with diminished tumor growth.IL-33 promoted polarization of alternatively activated macrophages in an ST2-dependent manner that was mediated via the PI3K/Akt signalling pathway.Moreover,IL-33 inhibited T-cell function by inducing the secretion of transforming growth factorβfrom alternatively activated macrophages.Conclusions:Macrophages expressing ST2 can serve as promising therapeutic targets for NSCLC immunotherapy,highlighting the IL-33/ST2 axis as a potential target for future antitumor strategies.展开更多
In the present study, electrical stimulation to the rat insular cortex induced apnea or respiratory disturbance, reduced amplitude of genioglossal electromyogram, and decreased electromyogram integrals. In addition, a...In the present study, electrical stimulation to the rat insular cortex induced apnea or respiratory disturbance, reduced amplitude of genioglossal electromyogram, and decreased electromyogram integrals. In addition, arterial blood gas analysis showed arterial blood acidosis, reduced pH values, increased alkali reserve negative values, decreased peripheral blood 5-hydroxytryptamine content, and increased 5-hydroxytryptamine expression in cerebellar Purkinje cells. Following lidocaine injection to block the habenular nucleus, abnormalities in breath, genioglossal electromyogram, and blood gas values disappeared, and peripheral blood 5-hydroxytryptamine content returned to levels prior to electric stimulation. However, 5-hydroxytryptamine expression in cerebellar Purkinje cells remained high. The results suggested that 5-hydroxytryptamine expression in Purkinje cells did not correlate with ventilation function involving insular cortex and habenular nucleus.展开更多
Recent studies suggest that PRL-3 is involved in the metastasis of colorectal cancer, but the mechanism concerning that has not been well defined. This article expresses PRL-1, PRL-2, and PRL-3 and the catalytically i...Recent studies suggest that PRL-3 is involved in the metastasis of colorectal cancer, but the mechanism concerning that has not been well defined. This article expresses PRL-1, PRL-2, and PRL-3 and the catalytically inactive mutant forms of those enzymes in SW620 and SW480 cells, two human cell lines derived from non-metastatic cancer and metastatic colorectal cancer, respectively. While the expression of the native forms of PRLs promotes the spreading, adhesion, and proliferation of these cells, the expression of their mutant forms inhibits the earlier-mentioned processes. These data thus provide a cellular mechanism for the role of PRL-3 in tumor metastasis and suggest that all the three PRLs have similar functions.展开更多
Objective 5-azacytidine could induce the differentiation of stem cells into cardiomyocytes （CMs）. The aim of this study was to screen the optimal condition for 5-azacytidine inducing differentiation of human mesench...Objective 5-azacytidine could induce the differentiation of stem cells into cardiomyocytes （CMs）. The aim of this study was to screen the optimal condition for 5-azacytidine inducing differentiation of human mesenchumal stem cells （hMSCs） into CMs, and the effect of 5-azacytidine on adherence, cell vigor and chromosome karyotype of hMSCs. Methods hMSCs were isolated from human bone marrow and cultured in vitro. The phenotypes ofhMSCs were identified by flow cytometric analyses. MTT test was used to investigate the effect of different concentrations of 5-azacytidine on proliferation ofhMSCs. Four weeks after 5-azacytidine induction, semi-quantitative RT-PCR, transmission electron microscopy （TEM）, single-cell action potentials, detection of cardio-enzyme AST and LDH, cell adherence, cell viability and chromosome karyotype test were performed. Results The typical morphological features of hMSCs were fibroblast-like in shape, hMSCs expressed CD44 and CD105,and did not express CD34, CD45 and CD31. The optimal concentration of 5-azacytidine was 10μ mol/L. The shape of hMSCs treated with 5-Azacytidine changed from fusiform to polygon or astrocyte gradually, and passaged cells were evenly arranged as polarity structure. Indueed-hMSCs connected with neighbouring cells, fbrming myotube-like structures 4 weeks later. It was confirmed that induced hMSCs shaped myotubule-like structure and had some of micro-histologic structures of CMs by TEM. RT-PCR showed that induced hMSCs expressed cardiac specific product BNNP and early cardio-myogenesis specific transcription factor NKX2.5mRNA. Besides, induced-MSCs led to the weak action potential and secreted cardio-enzyme AST and LDH. There was no significant difference in cell adherence and viability before and after induction. Both hMSCs and induced-hNSCs kept stable normal diploid nucleus. Conclusion The optimal condition for inducing effect of 5-azacytidine is 10 la mol/L and 24-hour incubation; and under this condition, the adherence, vigor and chromosome karyotype ofhMSCs would not be affected （J Geriatr Cardio12009; 6：182-188）.展开更多
Coronavirus disease 2019(COVID-19)is an acute respiratory infection caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).SARS-CoV-2 infection typically presents with fever and respiratory symptoms,whi...Coronavirus disease 2019(COVID-19)is an acute respiratory infection caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).SARS-CoV-2 infection typically presents with fever and respiratory symptoms,which can progress to severe respiratory distress syndrome and multiple organ failure.In severe cases,these complications may even lead to death.One of the causes of COVID-19 deaths is the cytokine storm caused by an overactive immune response.Therefore,suppressing the overactive immune response may be an effective strategy for treating COVID-19.Mesenchymal stem cells(MSCs)and their derived exosomes(MSCs-Exo)have potent homing abilities,immunomodulatory functions,regenerative repair,and antifibrotic effects,promising an effective tool in treating COVID-19.In this paper,we review the main mechanisms and potential roles of MSCs and MSCs-Exo in treating COVID-19.We also summarize relevant recent clinical trials,including the source of cells,the dosage and the efficacy,and the clinical value and problems in this field,providing more theoretical references for the clinical use of MSCs and MSCs-Exo in the treatment of COVID-19.展开更多
Background:Colorectal adenocarcinoma(COAD)is one of the most common gastrointestinal malignancies.There is a pressing need to recognize reliable biomarkers that can improve diagnostic accuracy,predict prognosis,and se...Background:Colorectal adenocarcinoma(COAD)is one of the most common gastrointestinal malignancies.There is a pressing need to recognize reliable biomarkers that can improve diagnostic accuracy,predict prognosis,and serve as effective molecular targets.Glutathione peroxidase 4(GPX4)is an important antioxidant protein.Evidence demonstrates that abnormal expression of GPX4 is related to cancer initiation and progression.However,the role of GPX4 in COAD remains unclear.Methods:We employed bioinformatics analysis and conducted subsequent validation of biological processes,including cell counting kit-8 assay(CCK-8),colony formation assay,reverse transcription-quantitative polymerase chain reaction(RT-qPCR),5-ethynyl-2′-deoxyuridine assay(EdU),western blot,immunohistochemistry,senescence associatedβ-galactosidase(SA-β-gal)staining and immunofluorescence to explore the expression status,prognostic value and biological function of GPX4 in COAD.Results:Our data revealed that GPX4 mRNA expression was upregulated in COAD tissues and could predict the prognosis in patients with COAD.High GPX4 expression was associated with increased infiltration of malignant cells.We also performed a series of cell experiments confirming that GPX4 knockdown inhibited proliferation and induced cellular senescence,as determined by using CCK-8,colony formation,and EdU assay.In addition,SA-β-gal staining and senescence-associated secretory phenotype(SASP)components,such as P21 and Interleukin-6(IL-6),were increased in GPX4 knockdown cells,while Lamin B1 was decreased.Moreover,we predicted that high expression of GPX4 was related to low immune cell infiltration.Conclusion:This study demonstrates that GPX4 is a potential prognostic biomarker and target gene for COAD.展开更多
Osteoarthritis(OA)is a prevalent degenerative and inflammatory disease posing a significant financial and medical burden on patients and society.Lactic acid,the terminal metabolite of glycolysis,is recognized as a piv...Osteoarthritis(OA)is a prevalent degenerative and inflammatory disease posing a significant financial and medical burden on patients and society.Lactic acid,the terminal metabolite of glycolysis,is recognized as a pivotal signaling molecule governing diverse physiological and pathological processes,particularly in cancer and inflammatory diseases.Emerging evidence suggests that metabolic disorders are closely associated with OA,which may provide a metabolic lens for further exploring its mechanisms.Glycolytic reprogramming is now recognized as a hallmark of OA,leading to the pronounced accumulation of lactic acid within the joint microenvironment.This review synthesizes current evidence to elucidate the role of lactic acid in OA pathogenesis.We summarize the mechanism of glycolytic reprogramming in chondrocytes and macrophages under pathological conditions.Furthermore,we demonstrate that lactic acid exacerbates cartilage degenera-tion while simultaneously promoting inflammation resolution.These dual roles are mediated by extracellular acidification,HCAR1,and lactylation.Given that duality,we suggest that redirecting lactate flux presents considerable potential as a therapeutic approach for the prevention and management of OA.展开更多
Objective:Macrophages are a major component of the tumor microenvironment.M1 macrophages secrete pro-inflammatory factors that inhibit tumor growth and development,whereas tumor-associated macrophages(TAMs)mainly exhi...Objective:Macrophages are a major component of the tumor microenvironment.M1 macrophages secrete pro-inflammatory factors that inhibit tumor growth and development,whereas tumor-associated macrophages(TAMs)mainly exhibit an M2 phenotype.Our previous studies have shown that the interleukin-33/ST2(IL-33/ST2)axis is essential for activation of the M1 phenotype.This study investigates the role of the IL-33/ST2 axis in TAMs,its effects on tumor growth,and whether it participates in the mutual conversion between the M1 and M2 phenotypes.Methods:Bone marrow-derived macrophages were extracted from wildtype,ST2 knockout(ST2-/-),and Il33-overexpressing mice and differentiated with IL-4.The mitochondrial and lysosomal number and location,and the expression of related proteins were used to analyze mitophagy.Oxygen consumption rates and glucose and lactate levels were measured to reveal metabolic changes.Results:The IL-33/ST2 axis was demonstrated to play an important role in the metabolic conversion of macrophages from OXPHOS to glycolysis by altering mitophagy levels.The IL-33/ST2 axis promoted enhanced cell oxidative phosphorylation,thereby further increasing M2 polarization gene expression and ultimately promoting tumor growth(P<0.05)(Figure 4).This metabolic shift was not due to mitochondrial damage,because the mitochondrial membrane potential was not significantly altered by IL-4 stimulation or ST2 knockout;however,it might be associated with the m TOR activity.Conclusions:These results clarify the interaction between the IL-33/ST2 pathway and macrophage polarization,and may pave the way to the development of new cancer immunotherapies targeting the IL-33/ST2 axis.展开更多
Pancreatic cancer(PC)is an aggressive human cancer.Appropriate methods for the diagnosis and treatment of PC have not been found at the genetic level,thus making epigenetics a promising research path in studies of PC....Pancreatic cancer(PC)is an aggressive human cancer.Appropriate methods for the diagnosis and treatment of PC have not been found at the genetic level,thus making epigenetics a promising research path in studies of PC.Histone methylation is one of the most complicated types of epigenetic modifications and has proved crucial in the development of PC.Histone methylation is a reversible process regulated by readers,writers,and erasers.Some writers and erasers can be recognized as potential biomarkers and candidate therapeutic targets in PC because of their unusual expression in PC cells compared with normal pancreatic cells.Based on the impact that writers have on the development of PC,some inhibitors of writers have been developed.However,few inhibitors of erasers have been developed and put to clinical use.Meanwhile,there is not enough research on the reader domains.Therefore,the study of erasers and readers is still a promising area.This review focuses on the regulatory mechanism of histone methylation,and the diagnosis and chemotherapy of PC based on it.The future of epigenetic modification in PC research is also discussed.展开更多
Neurological injury caused by ischemic stroke is a major cause of permanent disability and death. The currently available neuroprotective drugs fail to achieve desired therapeutic efficacy mainly due to short circulat...Neurological injury caused by ischemic stroke is a major cause of permanent disability and death. The currently available neuroprotective drugs fail to achieve desired therapeutic efficacy mainly due to short circulation half-life and poor blood−brain barrier (BBB) permeability. For that, an edaravone-loaded pH/glutathione (pH/GSH) dual-responsive poly(amino acid) nanogel (NG/EDA) was developed to improve the neuroprotection of EDA. The nanogel was triggered by acidic and EDA-induced high-level GSH microenvironments, which enabled the selective and sustained release of EDA at the site of ischemic injury. NG/EDA exhibited a uniform sub-spherical morphology with a mean hydrodynamic diameter of 112.3 ± 8.2 nm. NG/EDA efficiently accumulated at the cerebral ischemic injury site of permanent middle cerebral artery occlusion (pMCAO) mice, showing an efficient BBB crossing feature. Notably, NG/EDA with 50 µM EDA significantly increased neuron survival (29.3%) following oxygen and glucose deprivation by inhibiting ferroptosis. In addition, administering NG/EDA for 7 d significantly reduced infarct volume to 22.2% ± 7.2% and decreased neurobehavioral scores from 9.0 ± 0.6 to 2.0 ± 0.8. Such a pH/GSH dual-responsive nanoplatform might provide a unique and promising modality for neuroprotection in ischemic stroke and other central nervous system diseases.展开更多
Stroke is the second leading cause of death and the primary cause of permanent disability worldwide.Ischemic stroke(IS)accounts for 87%of all strokes globally and is characterized by the occlusion of cerebral vasculat...Stroke is the second leading cause of death and the primary cause of permanent disability worldwide.Ischemic stroke(IS)accounts for 87%of all strokes globally and is characterized by the occlusion of cerebral vasculature due to embolic presence.Clinical treatments for IS include enzymatic thrombolysis,mechanical thrombectomy,and neuroprotection.However,these approaches have obvious limitations.First,early vascular recanalization leads to secondary cascade injuries and a high risk of hemorrhagic transformation,resulting in poor clinical outcomes for patients with IS.In addition,neuroprotective agents often fail to achieve satisfactory clinical efficacy due to inadequate drug concentrations and off-target effects[1].Targeted stimuli-responsive nanoformulations for thrombolysis and neuroprotection have been developed to address these limitations in current clinical treatments.These nanoformulations are based on IS-specific thrombus-associated receptors and the pathological microenvironments,showing great promise in treating IS(Figure A).展开更多
Dear editor,Bispecific T-cell engagers(also referred to as BiTEs)are bi/multispecific antibodies that are a clinically used cancer immunotherapy,which enforce formation of an immunological synapse between T-cells and ...Dear editor,Bispecific T-cell engagers(also referred to as BiTEs)are bi/multispecific antibodies that are a clinically used cancer immunotherapy,which enforce formation of an immunological synapse between T-cells and tumour cells by antigenic engagement on the respective cells,resulting in cancer cell apoptosis.1,2 Tumour heterogeneity,however,is a major challenge for identifying the most appropriate cancer target antigen for each patient.Discovery of new tumour targets outweighs new BiTE approval,calling for rapid screening tools to evaluate novel targets and BiTE combinational formats.展开更多
Background Ginsenoside Rg3, the main component isolated from ginseng, inhibits some kinds of tumour growth and angiogenesis. The combination of low dose chemotherapy and antiangiogenesis inhibitors suppresses growth o...Background Ginsenoside Rg3, the main component isolated from ginseng, inhibits some kinds of tumour growth and angiogenesis. The combination of low dose chemotherapy and antiangiogenesis inhibitors suppresses growth of experimental tumours more effectively than conventional therapy. The effect of this combination on ovarian cancer remains to be evaluated. Therefore, we investigated the synergism of ginsenoside Rg3 and cyclophosphamide （CTX） on growth and angiogenesis of human ovarian cancer. Methods Twenty-eight female athymic mice were divided randomly into 4 groups of 7： ginsenoside Rg3, CTX, ginsenoside Rg3 and CTX combination and control, after being transplanted with ovarian cancer cells （SKOV-3）. The mice were given intraperitoneal injection of ginsenoside Rg3 and CTX for the 10 days following inoculation of SKOV-3 cells. The life quality and number of living days of mice were recorded. The size of tumour, tumour inhibitive rate, life elongation rate, proliferating cell nuclear antigen labelling index （PCNALI）, expression of vascular endothelial cell growth factor （VEGF） and microvessel density （MVD） of the tumour tissues were estimated. Results Life quality of mice in ginsenoside Rg3 and combined treatment groups were better and number of living days longer than control. Average tumour weights of each treated group were less than control and there was no significant difference among the treated groups. PCNALI of treated groups was lower than control. The MVD value and VEGF expression in treated groups were significantly lower than control and the MVD values of ginsenoside Rg3 and combined treatment groups were lower than that of CTX group. Conclusions Ginsenoside Rg3 significantly inhibited growth and angiogenesis of ovarian cancer when used alone or combined with CTX. Ginsenoside Rg3 and CTX combination reinforced the antitumour effect each other and improved the living quality and survival time of mice with tumour.展开更多
Background Ginsenosides are main components extracted from ginseng, and ginsenoside Rg3 is one of the most important parts. Ginsenoside Rg3 has been found to inhibit several kinds of tumor growth and metastasis. The p...Background Ginsenosides are main components extracted from ginseng, and ginsenoside Rg3 is one of the most important parts. Ginsenoside Rg3 has been found to inhibit several kinds of tumor growth and metastasis. The present study was undertaken to investigate the effect of ginsenoside Rg3 on human ovarian cancer metastasis and the possible mechanism. Methods The experimental lung metastasis models of ovarian cancer SKOV-3 and the assay of tumor-induced angiogenesis were used to observe the inhibitory effects of Rg3 on tumor metastasis and angiogenesis. The effect of Rg3 on invasive ability of SKOV-3 cells in vitro was detected by Boyden chamber, and immunofluorescence staining was used to recognize the expression of matrix metalloproteinase 9 （MMP-9） in SKOV-3 cells. Results In the experimental lung metastasis models of ovarian cancer, the number of tumor colonies in the lung and vessels oriented toward the tumor mass in each ginsenoside Rg3 group, was lower than that of control group. The invasive ability and MMP-9 expression of SKOV-3 cells decreased significantly after treatment with ginsenoside Rg3. Conclusions Ginsenoside Rg3 can significantly inhibit the metastasis of ovarian cancer. The inhibitory effect is partially due to inhibition of tumor-induced an qioqenesis and decrease of invasive ability and MMP-9 expression of SKOV-3 cells.展开更多
Background Claudin-6 is a protein component of tight junctions and its expression could downregulate the malignant phenotype of breast carcinoma. Here we investigated the mechanisms of claudin-6 induced human MCF-7 br...Background Claudin-6 is a protein component of tight junctions and its expression could downregulate the malignant phenotype of breast carcinoma. Here we investigated the mechanisms of claudin-6 induced human MCF-7 breast cancer cells apoptosis, invasion, and migration. Methods Terminal deoxyribonucleotide transferase-mediated nick-end labeling assay and Annexin-V/PI double stain assay were carried out to evaluate apoptosis. Inhibitors of each pathway were used to inactivate the signaling pathways. The expression of claudin-6 and phosphate p38, Erk 1/2 and Akt protein levels was confirmed by Western blotting analysis. Invasive and migratory traits of claudin-6 expressing cells were determined by Boyden chamber invasion assay and monolayer wound-healing assay. Results Cells with high-level expression of claudin-6 had a higher rate of apoptosis than control cells. Western blotting assay showed that by contrast to control groups, p38 pathways were more activated in claudin-6 expressing cells. However, after inhibitor SB203580 treatment, the activation status could be significantly counteracted. Furthermore, by applying inhibitors to the apoptotic rate, invasive and migratory traits were also recovered in cells with claudin-6 expression. Conclusion Claudin-6 may function through p38 mitogen-activated protein kinase pathway, of which inhibition may reverse claudin-6-induced cell apoptosis, invasion, and migration.展开更多
Mechanical stimulation is the key physical factor in cell environment.Mechanotransduction acts as a fundamental regulator of cell behavior,regulating cell proliferation,differentiation,apoptosis,and exhibiting specifi...Mechanical stimulation is the key physical factor in cell environment.Mechanotransduction acts as a fundamental regulator of cell behavior,regulating cell proliferation,differentiation,apoptosis,and exhibiting specific signature alterations during the pathological process.As research continues,the role of epigenetic science in mechanotransduction is attracting attention.However,the molecular mechanism of the synergistic effect between mechanotransduction and epigenetics in physiological and pathological processes has not been clarified.We focus on how histone modifications,as important components of epigenetics,are coordinated with multiple signaling pathways to control cell fate and disease progression.Specifically,we propose that histone modifications can form regulatory feedback loops with signaling pathways,that is,histone modifications can not only serve as downstream regulators of signaling pathways for target gene transcription but also provide feedback to regulate signaling pathways.Mechanotransduction and epigenetic changes could be potential markers and therapeutic targets in clinical practice.展开更多
The five-year survival rate of patients with colorectal cancer(CRC)liver metastasis is less than 30%,and chemotherapy resistance and metastatic microenvironment remodeling are the current treatment bottlenecks.Cancer-...The five-year survival rate of patients with colorectal cancer(CRC)liver metastasis is less than 30%,and chemotherapy resistance and metastatic microenvironment remodeling are the current treatment bottlenecks.Cancer-associated fibroblasts(CAFs)in the tumor microenvironment(TME)form a“CRCs-CAFs crosstalk”with colorectal cancer cells(CRCs)by secreting dense extracellular matrix(ECM),free fatty acids(FFA),and pro-metastatic factors,driving a vicious cycle of drug resistance and metastasis.During liver metastasis,hepatic stellate cells(HSCs)-derived CAFs(HSC-CAFs)promote tumor metastasis by remodeling the pre-metastatic microenvironment.Based on clinical sample RNA sequencing and mouse single-cell sequencing to reveal ECM signal enrichment and CAFs activation characteristics,we innovatively constructed a nano-delivery system using hyaluronic acid-modified MIL-100 nanoparticles(OEMH NPs)co-loaded with oxaliplatin(OXA)and epi-gallocatechin gallate(EGCG).This system can target the CRCs-CAFs crosstalk through CD44 receptor:on the one hand,OEMH NPs can inhibit CAFs activation and reduce ECM deposition,improve drug penetration and downregulate FFA metabolic reprogramming,reverse OXA resistance;on the other hand,OEMH NPs can block the transformation of HSCs to CAFs,down-regulate pro-metastatic factors such as VEGF/IL-11/ANG,induce vascular normalization,and reprogram the pre-metastatic microenvironment.This strategy can simultaneously achieve primary lesion drug sensitization and liver metastasis inhibition,providing a new paradigm for the treatment of advanced colorectal cancer to break through the traditional treatment dilemma through dual reprogramming of metabolism and microenvironment,and has significant clinical translation potential.展开更多
Background Enhanced green fluorescent protein （EGFP） has been an important reporter gene for gene therapy. Human mesenchymal stem cells （hMSCs） are ideal target cells in cell transplantation and tissue engineering...Background Enhanced green fluorescent protein （EGFP） has been an important reporter gene for gene therapy. Human mesenchymal stem cells （hMSCs） are ideal target cells in cell transplantation and tissue engineering. We investigated their biological characteristics and differentiation mediated by PLEGFP-N1 retroviral transduction.Methods hMSCs were isolated from human bone marrow by density gradient fractionation and adherence to plastic flasks. Individual colonies were selected and cultured in tissue dishes. Packaging cells PT67 were transfected by PLEGFP-N1 retroviral vector , and hMSCs were transduced by viral supernatant infection. Meanwhile, hMSCs-EGFP were identified by immune phenotypes and whether it could differentiate into osteoblasts or adipocytes under conditioned media was investigated.Results The rate of stably transduced hMSCs-EGFP was up to 96% after being screened by G418. hMSCs-EGFP exhibited fibroblast-like morphological features. Flow cytometric analyses showed that hMSCs-EGFP were positive for CD73, CD105, CD166, CD90 and CD44, but negative for CD34 and CD45. In addition, it could functionally be induced into osteocytes or adipocytes under conditioned media. These biological features of hMSCs-EGFP were consistent with those of hMSCs.Conclusions hMSCs transduced by PLEGFP-N1 retroviral vector can be used in vivo securely because they can maintain their biological characteristics and differentiation. It is a simple and reliable way to trace the changes of hMSCs in vivo by EGFP during cell transplantation and gene therapy.展开更多
Nonalcoholic fatty liver disease(NAFLD)is a serious threat to public health,but its underlying mechanism remains poorly understood.In screening important genes using Gene Importance Calculator(GIC)we developed previou...Nonalcoholic fatty liver disease(NAFLD)is a serious threat to public health,but its underlying mechanism remains poorly understood.In screening important genes using Gene Importance Calculator(GIC)we developed previously,ribosomal modification protein rimK-like family member A(RIMKLA)was predicted as one essential gene but its functions remained largely unknown.The current study determined the roles of RIMKLA in regulating glucose and lipid metabolism.RIMKLA expression was reduced in livers of human and mouse with NAFLD.Hepatic RIMKLA overexpression ameliorated steatosis and hyperglycemia in obese mice.Hepatocyte-specific RIMKLA knockout aggravated high-fat diet(HFD)-induced dysregulated glucose/lipid metabolism in mice.Mechanistically,RIMKLA is a new protein kinase that phosphorylates betaine-homocysteine S-methyltransferase 1(BHMT1)at threonine 45(Thr45)site.Upon phosphorylation at Thr45 and activation,BHMT1 eliminated homocysteine(Hcy)to inhibit the activity of transcription factor activator protein 1(AP1)and its induction on fatty acid synthase(FASn)and cluster of differentiation 36(CD36)gene transcriptions,concurrently repressing lipid synthesis and uptake in hepatocytes.Thr45 to alanine(T45A)mutation inactivated BHMT1 to abolish RIMKLA’s repression on Hcy level,AP1 activity,FASn/CD36 expressions,and lipid deposition.BHMT1 overexpression rescued the dysregulated lipid metabolism in RIMKLA-deficient hepatocytes.In summary,RIMKLA is a novel protein kinase that phosphorylates BHMT1 at Thr45 to repress lipid synthesis and uptake.Under obese condition,inhibition of RIMKLA impairs BHMT1 activity to promote hepatic lipid deposition.展开更多
基金supported by the National Key R&D Program of China(2023YFC2413100)the National Natural Science Foundation of China(No.81501423)the Fundamental Research Funds for the Central Universities。
文摘Objective:Suppression of tumorigenicity 2(ST2),the receptor for interleukin(IL)-33,has a critical role in tumor growth,angiogenesis,metastasis,and immune modulation.The IL-33/ST2 pathway is known to influence the polarization and function of macrophages,which is integral to modulating the tumor microenvironment.However,the precise role of IL-33/ST2 in tumors,particularly non-small cell lung cancer(NSCLC),has not been established.Methods:ST2 expression in NSCLC was analysed using a murine model and patient specimens.The effect of the IL-33/ST2 axis on macrophage polarization in NSCLC was determined.Results:Elevated ST2 expression was correlated with aggressive tumor growth.Specifically,ST2 expression on macrophages was associated with lung cancer progression and the absence of ST2 on macrophages was associated with diminished tumor growth.IL-33 promoted polarization of alternatively activated macrophages in an ST2-dependent manner that was mediated via the PI3K/Akt signalling pathway.Moreover,IL-33 inhibited T-cell function by inducing the secretion of transforming growth factorβfrom alternatively activated macrophages.Conclusions:Macrophages expressing ST2 can serve as promising therapeutic targets for NSCLC immunotherapy,highlighting the IL-33/ST2 axis as a potential target for future antitumor strategies.
基金supported by the National Natural Science Foundation of China, No. 30270502
文摘In the present study, electrical stimulation to the rat insular cortex induced apnea or respiratory disturbance, reduced amplitude of genioglossal electromyogram, and decreased electromyogram integrals. In addition, arterial blood gas analysis showed arterial blood acidosis, reduced pH values, increased alkali reserve negative values, decreased peripheral blood 5-hydroxytryptamine content, and increased 5-hydroxytryptamine expression in cerebellar Purkinje cells. Following lidocaine injection to block the habenular nucleus, abnormalities in breath, genioglossal electromyogram, and blood gas values disappeared, and peripheral blood 5-hydroxytryptamine content returned to levels prior to electric stimulation. However, 5-hydroxytryptamine expression in cerebellar Purkinje cells remained high. The results suggested that 5-hydroxytryptamine expression in Purkinje cells did not correlate with ventilation function involving insular cortex and habenular nucleus.
文摘Recent studies suggest that PRL-3 is involved in the metastasis of colorectal cancer, but the mechanism concerning that has not been well defined. This article expresses PRL-1, PRL-2, and PRL-3 and the catalytically inactive mutant forms of those enzymes in SW620 and SW480 cells, two human cell lines derived from non-metastatic cancer and metastatic colorectal cancer, respectively. While the expression of the native forms of PRLs promotes the spreading, adhesion, and proliferation of these cells, the expression of their mutant forms inhibits the earlier-mentioned processes. These data thus provide a cellular mechanism for the role of PRL-3 in tumor metastasis and suggest that all the three PRLs have similar functions.
文摘Objective 5-azacytidine could induce the differentiation of stem cells into cardiomyocytes （CMs）. The aim of this study was to screen the optimal condition for 5-azacytidine inducing differentiation of human mesenchumal stem cells （hMSCs） into CMs, and the effect of 5-azacytidine on adherence, cell vigor and chromosome karyotype of hMSCs. Methods hMSCs were isolated from human bone marrow and cultured in vitro. The phenotypes ofhMSCs were identified by flow cytometric analyses. MTT test was used to investigate the effect of different concentrations of 5-azacytidine on proliferation ofhMSCs. Four weeks after 5-azacytidine induction, semi-quantitative RT-PCR, transmission electron microscopy （TEM）, single-cell action potentials, detection of cardio-enzyme AST and LDH, cell adherence, cell viability and chromosome karyotype test were performed. Results The typical morphological features of hMSCs were fibroblast-like in shape, hMSCs expressed CD44 and CD105,and did not express CD34, CD45 and CD31. The optimal concentration of 5-azacytidine was 10μ mol/L. The shape of hMSCs treated with 5-Azacytidine changed from fusiform to polygon or astrocyte gradually, and passaged cells were evenly arranged as polarity structure. Indueed-hMSCs connected with neighbouring cells, fbrming myotube-like structures 4 weeks later. It was confirmed that induced hMSCs shaped myotubule-like structure and had some of micro-histologic structures of CMs by TEM. RT-PCR showed that induced hMSCs expressed cardiac specific product BNNP and early cardio-myogenesis specific transcription factor NKX2.5mRNA. Besides, induced-MSCs led to the weak action potential and secreted cardio-enzyme AST and LDH. There was no significant difference in cell adherence and viability before and after induction. Both hMSCs and induced-hNSCs kept stable normal diploid nucleus. Conclusion The optimal condition for inducing effect of 5-azacytidine is 10 la mol/L and 24-hour incubation; and under this condition, the adherence, vigor and chromosome karyotype ofhMSCs would not be affected （J Geriatr Cardio12009; 6：182-188）.
基金Supported by Science and Technology Department Project of Jilin Province,China,No.20230101163JCthe Outstanding Youth Fund Project of Jilin Provincial Department of Education,China,No.JJKH20241324KJ.
文摘Coronavirus disease 2019(COVID-19)is an acute respiratory infection caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).SARS-CoV-2 infection typically presents with fever and respiratory symptoms,which can progress to severe respiratory distress syndrome and multiple organ failure.In severe cases,these complications may even lead to death.One of the causes of COVID-19 deaths is the cytokine storm caused by an overactive immune response.Therefore,suppressing the overactive immune response may be an effective strategy for treating COVID-19.Mesenchymal stem cells(MSCs)and their derived exosomes(MSCs-Exo)have potent homing abilities,immunomodulatory functions,regenerative repair,and antifibrotic effects,promising an effective tool in treating COVID-19.In this paper,we review the main mechanisms and potential roles of MSCs and MSCs-Exo in treating COVID-19.We also summarize relevant recent clinical trials,including the source of cells,the dosage and the efficacy,and the clinical value and problems in this field,providing more theoretical references for the clinical use of MSCs and MSCs-Exo in the treatment of COVID-19.
文摘Background:Colorectal adenocarcinoma(COAD)is one of the most common gastrointestinal malignancies.There is a pressing need to recognize reliable biomarkers that can improve diagnostic accuracy,predict prognosis,and serve as effective molecular targets.Glutathione peroxidase 4(GPX4)is an important antioxidant protein.Evidence demonstrates that abnormal expression of GPX4 is related to cancer initiation and progression.However,the role of GPX4 in COAD remains unclear.Methods:We employed bioinformatics analysis and conducted subsequent validation of biological processes,including cell counting kit-8 assay(CCK-8),colony formation assay,reverse transcription-quantitative polymerase chain reaction(RT-qPCR),5-ethynyl-2′-deoxyuridine assay(EdU),western blot,immunohistochemistry,senescence associatedβ-galactosidase(SA-β-gal)staining and immunofluorescence to explore the expression status,prognostic value and biological function of GPX4 in COAD.Results:Our data revealed that GPX4 mRNA expression was upregulated in COAD tissues and could predict the prognosis in patients with COAD.High GPX4 expression was associated with increased infiltration of malignant cells.We also performed a series of cell experiments confirming that GPX4 knockdown inhibited proliferation and induced cellular senescence,as determined by using CCK-8,colony formation,and EdU assay.In addition,SA-β-gal staining and senescence-associated secretory phenotype(SASP)components,such as P21 and Interleukin-6(IL-6),were increased in GPX4 knockdown cells,while Lamin B1 was decreased.Moreover,we predicted that high expression of GPX4 was related to low immune cell infiltration.Conclusion:This study demonstrates that GPX4 is a potential prognostic biomarker and target gene for COAD.
基金supported by the National Natural Science Foundation of China(Nos.82271425,81870974)the Jilin Province Health Science and Technology Capacity Enhancement Program(No.2025WS-KA017).
文摘Osteoarthritis(OA)is a prevalent degenerative and inflammatory disease posing a significant financial and medical burden on patients and society.Lactic acid,the terminal metabolite of glycolysis,is recognized as a pivotal signaling molecule governing diverse physiological and pathological processes,particularly in cancer and inflammatory diseases.Emerging evidence suggests that metabolic disorders are closely associated with OA,which may provide a metabolic lens for further exploring its mechanisms.Glycolytic reprogramming is now recognized as a hallmark of OA,leading to the pronounced accumulation of lactic acid within the joint microenvironment.This review synthesizes current evidence to elucidate the role of lactic acid in OA pathogenesis.We summarize the mechanism of glycolytic reprogramming in chondrocytes and macrophages under pathological conditions.Furthermore,we demonstrate that lactic acid exacerbates cartilage degenera-tion while simultaneously promoting inflammation resolution.These dual roles are mediated by extracellular acidification,HCAR1,and lactylation.Given that duality,we suggest that redirecting lactate flux presents considerable potential as a therapeutic approach for the prevention and management of OA.
基金supported by grants from the National Natural Science Foundation of China(Grant Nos.81672948 and 81772794)Jilin Provincial Research Foundation for Health Technology Innovation(Grant No.2019J009)+1 种基金Jilin Provincial Research Foundation for the Development of Science and Technology Projects(Grant Nos.20191004004TC and 20190103095JH)Jilin Provincial Industrial Innovation Project(Grant No.2018C052-7)。
文摘Objective:Macrophages are a major component of the tumor microenvironment.M1 macrophages secrete pro-inflammatory factors that inhibit tumor growth and development,whereas tumor-associated macrophages(TAMs)mainly exhibit an M2 phenotype.Our previous studies have shown that the interleukin-33/ST2(IL-33/ST2)axis is essential for activation of the M1 phenotype.This study investigates the role of the IL-33/ST2 axis in TAMs,its effects on tumor growth,and whether it participates in the mutual conversion between the M1 and M2 phenotypes.Methods:Bone marrow-derived macrophages were extracted from wildtype,ST2 knockout(ST2-/-),and Il33-overexpressing mice and differentiated with IL-4.The mitochondrial and lysosomal number and location,and the expression of related proteins were used to analyze mitophagy.Oxygen consumption rates and glucose and lactate levels were measured to reveal metabolic changes.Results:The IL-33/ST2 axis was demonstrated to play an important role in the metabolic conversion of macrophages from OXPHOS to glycolysis by altering mitophagy levels.The IL-33/ST2 axis promoted enhanced cell oxidative phosphorylation,thereby further increasing M2 polarization gene expression and ultimately promoting tumor growth(P<0.05)(Figure 4).This metabolic shift was not due to mitochondrial damage,because the mitochondrial membrane potential was not significantly altered by IL-4 stimulation or ST2 knockout;however,it might be associated with the m TOR activity.Conclusions:These results clarify the interaction between the IL-33/ST2 pathway and macrophage polarization,and may pave the way to the development of new cancer immunotherapies targeting the IL-33/ST2 axis.
文摘Pancreatic cancer(PC)is an aggressive human cancer.Appropriate methods for the diagnosis and treatment of PC have not been found at the genetic level,thus making epigenetics a promising research path in studies of PC.Histone methylation is one of the most complicated types of epigenetic modifications and has proved crucial in the development of PC.Histone methylation is a reversible process regulated by readers,writers,and erasers.Some writers and erasers can be recognized as potential biomarkers and candidate therapeutic targets in PC because of their unusual expression in PC cells compared with normal pancreatic cells.Based on the impact that writers have on the development of PC,some inhibitors of writers have been developed.However,few inhibitors of erasers have been developed and put to clinical use.Meanwhile,there is not enough research on the reader domains.Therefore,the study of erasers and readers is still a promising area.This review focuses on the regulatory mechanism of histone methylation,and the diagnosis and chemotherapy of PC based on it.The future of epigenetic modification in PC research is also discussed.
基金supported by the National Natural Science Foundation of China(Grant No.U23A20591,52203201,52173149,and 81971174)the Youth Talents Promotion Project of Jilin Province(Grant No.202019)+1 种基金the Science and Technology Development Program of Jilin Province(Grant No.20210101114JC)Research Cooperation Platform Project of Sino-Japanese Friendship Hospital of Jilin University and Basic Medical School of Jilin University(Grant No.KYXZ2022JC04).
文摘Neurological injury caused by ischemic stroke is a major cause of permanent disability and death. The currently available neuroprotective drugs fail to achieve desired therapeutic efficacy mainly due to short circulation half-life and poor blood−brain barrier (BBB) permeability. For that, an edaravone-loaded pH/glutathione (pH/GSH) dual-responsive poly(amino acid) nanogel (NG/EDA) was developed to improve the neuroprotection of EDA. The nanogel was triggered by acidic and EDA-induced high-level GSH microenvironments, which enabled the selective and sustained release of EDA at the site of ischemic injury. NG/EDA exhibited a uniform sub-spherical morphology with a mean hydrodynamic diameter of 112.3 ± 8.2 nm. NG/EDA efficiently accumulated at the cerebral ischemic injury site of permanent middle cerebral artery occlusion (pMCAO) mice, showing an efficient BBB crossing feature. Notably, NG/EDA with 50 µM EDA significantly increased neuron survival (29.3%) following oxygen and glucose deprivation by inhibiting ferroptosis. In addition, administering NG/EDA for 7 d significantly reduced infarct volume to 22.2% ± 7.2% and decreased neurobehavioral scores from 9.0 ± 0.6 to 2.0 ± 0.8. Such a pH/GSH dual-responsive nanoplatform might provide a unique and promising modality for neuroprotection in ischemic stroke and other central nervous system diseases.
文摘Stroke is the second leading cause of death and the primary cause of permanent disability worldwide.Ischemic stroke(IS)accounts for 87%of all strokes globally and is characterized by the occlusion of cerebral vasculature due to embolic presence.Clinical treatments for IS include enzymatic thrombolysis,mechanical thrombectomy,and neuroprotection.However,these approaches have obvious limitations.First,early vascular recanalization leads to secondary cascade injuries and a high risk of hemorrhagic transformation,resulting in poor clinical outcomes for patients with IS.In addition,neuroprotective agents often fail to achieve satisfactory clinical efficacy due to inadequate drug concentrations and off-target effects[1].Targeted stimuli-responsive nanoformulations for thrombolysis and neuroprotection have been developed to address these limitations in current clinical treatments.These nanoformulations are based on IS-specific thrombus-associated receptors and the pathological microenvironments,showing great promise in treating IS(Figure A).
基金supported by the Novo Nordisk Foundation Grant,CEMBID(Center for Multifunctional Biomolecular Drug Design,Grant Number:NNF17OC0028070)the National Key Research and Development Programme(2024 YFA0918600)+1 种基金Venus Project from China-Japan Union Hospital of Jilin University(Funding No.:XHQMX20242)the Science and Technology Development Plan Project of Jilin Province(Grant No.:20230402040GH).
文摘Dear editor,Bispecific T-cell engagers(also referred to as BiTEs)are bi/multispecific antibodies that are a clinically used cancer immunotherapy,which enforce formation of an immunological synapse between T-cells and tumour cells by antigenic engagement on the respective cells,resulting in cancer cell apoptosis.1,2 Tumour heterogeneity,however,is a major challenge for identifying the most appropriate cancer target antigen for each patient.Discovery of new tumour targets outweighs new BiTE approval,calling for rapid screening tools to evaluate novel targets and BiTE combinational formats.
文摘Background Ginsenoside Rg3, the main component isolated from ginseng, inhibits some kinds of tumour growth and angiogenesis. The combination of low dose chemotherapy and antiangiogenesis inhibitors suppresses growth of experimental tumours more effectively than conventional therapy. The effect of this combination on ovarian cancer remains to be evaluated. Therefore, we investigated the synergism of ginsenoside Rg3 and cyclophosphamide （CTX） on growth and angiogenesis of human ovarian cancer. Methods Twenty-eight female athymic mice were divided randomly into 4 groups of 7： ginsenoside Rg3, CTX, ginsenoside Rg3 and CTX combination and control, after being transplanted with ovarian cancer cells （SKOV-3）. The mice were given intraperitoneal injection of ginsenoside Rg3 and CTX for the 10 days following inoculation of SKOV-3 cells. The life quality and number of living days of mice were recorded. The size of tumour, tumour inhibitive rate, life elongation rate, proliferating cell nuclear antigen labelling index （PCNALI）, expression of vascular endothelial cell growth factor （VEGF） and microvessel density （MVD） of the tumour tissues were estimated. Results Life quality of mice in ginsenoside Rg3 and combined treatment groups were better and number of living days longer than control. Average tumour weights of each treated group were less than control and there was no significant difference among the treated groups. PCNALI of treated groups was lower than control. The MVD value and VEGF expression in treated groups were significantly lower than control and the MVD values of ginsenoside Rg3 and combined treatment groups were lower than that of CTX group. Conclusions Ginsenoside Rg3 significantly inhibited growth and angiogenesis of ovarian cancer when used alone or combined with CTX. Ginsenoside Rg3 and CTX combination reinforced the antitumour effect each other and improved the living quality and survival time of mice with tumour.
文摘Background Ginsenosides are main components extracted from ginseng, and ginsenoside Rg3 is one of the most important parts. Ginsenoside Rg3 has been found to inhibit several kinds of tumor growth and metastasis. The present study was undertaken to investigate the effect of ginsenoside Rg3 on human ovarian cancer metastasis and the possible mechanism. Methods The experimental lung metastasis models of ovarian cancer SKOV-3 and the assay of tumor-induced angiogenesis were used to observe the inhibitory effects of Rg3 on tumor metastasis and angiogenesis. The effect of Rg3 on invasive ability of SKOV-3 cells in vitro was detected by Boyden chamber, and immunofluorescence staining was used to recognize the expression of matrix metalloproteinase 9 （MMP-9） in SKOV-3 cells. Results In the experimental lung metastasis models of ovarian cancer, the number of tumor colonies in the lung and vessels oriented toward the tumor mass in each ginsenoside Rg3 group, was lower than that of control group. The invasive ability and MMP-9 expression of SKOV-3 cells decreased significantly after treatment with ginsenoside Rg3. Conclusions Ginsenoside Rg3 can significantly inhibit the metastasis of ovarian cancer. The inhibitory effect is partially due to inhibition of tumor-induced an qioqenesis and decrease of invasive ability and MMP-9 expression of SKOV-3 cells.
基金This research was supported by grants from the National Natural Science Foundation of China （No. 81172499） and Sciende and Technology Development Plan of the Office of Science and Technology Project in Jilin Province （No. 201115113）.
文摘Background Claudin-6 is a protein component of tight junctions and its expression could downregulate the malignant phenotype of breast carcinoma. Here we investigated the mechanisms of claudin-6 induced human MCF-7 breast cancer cells apoptosis, invasion, and migration. Methods Terminal deoxyribonucleotide transferase-mediated nick-end labeling assay and Annexin-V/PI double stain assay were carried out to evaluate apoptosis. Inhibitors of each pathway were used to inactivate the signaling pathways. The expression of claudin-6 and phosphate p38, Erk 1/2 and Akt protein levels was confirmed by Western blotting analysis. Invasive and migratory traits of claudin-6 expressing cells were determined by Boyden chamber invasion assay and monolayer wound-healing assay. Results Cells with high-level expression of claudin-6 had a higher rate of apoptosis than control cells. Western blotting assay showed that by contrast to control groups, p38 pathways were more activated in claudin-6 expressing cells. However, after inhibitor SB203580 treatment, the activation status could be significantly counteracted. Furthermore, by applying inhibitors to the apoptotic rate, invasive and migratory traits were also recovered in cells with claudin-6 expression. Conclusion Claudin-6 may function through p38 mitogen-activated protein kinase pathway, of which inhibition may reverse claudin-6-induced cell apoptosis, invasion, and migration.
基金the Department of Science and Technology of Jilin Province,China(No.20230101163JC and No.20210509003RQ)。
文摘Mechanical stimulation is the key physical factor in cell environment.Mechanotransduction acts as a fundamental regulator of cell behavior,regulating cell proliferation,differentiation,apoptosis,and exhibiting specific signature alterations during the pathological process.As research continues,the role of epigenetic science in mechanotransduction is attracting attention.However,the molecular mechanism of the synergistic effect between mechanotransduction and epigenetics in physiological and pathological processes has not been clarified.We focus on how histone modifications,as important components of epigenetics,are coordinated with multiple signaling pathways to control cell fate and disease progression.Specifically,we propose that histone modifications can form regulatory feedback loops with signaling pathways,that is,histone modifications can not only serve as downstream regulators of signaling pathways for target gene transcription but also provide feedback to regulate signaling pathways.Mechanotransduction and epigenetic changes could be potential markers and therapeutic targets in clinical practice.
基金First Hospital of Jilin University and Changchun Institute of Applied Chemistry,Joint Laboratory of Medicine and Polymers Project(2022YYGFZJC005)the National Natural Science Foundation of China(52073278,52373161)+2 种基金the“Medical Science+X”Cross-innovation Team of the Norman Bethune Health Science of Jilin University(2022JBGS10)the Education Department of Jilin Province(JJKH20231205KJ)the Jilin Province Science and Technology Development Program(20230101045JC).
文摘The five-year survival rate of patients with colorectal cancer(CRC)liver metastasis is less than 30%,and chemotherapy resistance and metastatic microenvironment remodeling are the current treatment bottlenecks.Cancer-associated fibroblasts(CAFs)in the tumor microenvironment(TME)form a“CRCs-CAFs crosstalk”with colorectal cancer cells(CRCs)by secreting dense extracellular matrix(ECM),free fatty acids(FFA),and pro-metastatic factors,driving a vicious cycle of drug resistance and metastasis.During liver metastasis,hepatic stellate cells(HSCs)-derived CAFs(HSC-CAFs)promote tumor metastasis by remodeling the pre-metastatic microenvironment.Based on clinical sample RNA sequencing and mouse single-cell sequencing to reveal ECM signal enrichment and CAFs activation characteristics,we innovatively constructed a nano-delivery system using hyaluronic acid-modified MIL-100 nanoparticles(OEMH NPs)co-loaded with oxaliplatin(OXA)and epi-gallocatechin gallate(EGCG).This system can target the CRCs-CAFs crosstalk through CD44 receptor:on the one hand,OEMH NPs can inhibit CAFs activation and reduce ECM deposition,improve drug penetration and downregulate FFA metabolic reprogramming,reverse OXA resistance;on the other hand,OEMH NPs can block the transformation of HSCs to CAFs,down-regulate pro-metastatic factors such as VEGF/IL-11/ANG,induce vascular normalization,and reprogram the pre-metastatic microenvironment.This strategy can simultaneously achieve primary lesion drug sensitization and liver metastasis inhibition,providing a new paradigm for the treatment of advanced colorectal cancer to break through the traditional treatment dilemma through dual reprogramming of metabolism and microenvironment,and has significant clinical translation potential.
基金This study was supported by a grant from the National 863 Program( No.2004AA205020 ), Doctoral Foundation of Ministry ofEducation (No.20020183064),and Scientific Foundation for YoungTeachers of Jilin University (No.419070100050).
文摘Background Enhanced green fluorescent protein （EGFP） has been an important reporter gene for gene therapy. Human mesenchymal stem cells （hMSCs） are ideal target cells in cell transplantation and tissue engineering. We investigated their biological characteristics and differentiation mediated by PLEGFP-N1 retroviral transduction.Methods hMSCs were isolated from human bone marrow by density gradient fractionation and adherence to plastic flasks. Individual colonies were selected and cultured in tissue dishes. Packaging cells PT67 were transfected by PLEGFP-N1 retroviral vector , and hMSCs were transduced by viral supernatant infection. Meanwhile, hMSCs-EGFP were identified by immune phenotypes and whether it could differentiate into osteoblasts or adipocytes under conditioned media was investigated.Results The rate of stably transduced hMSCs-EGFP was up to 96% after being screened by G418. hMSCs-EGFP exhibited fibroblast-like morphological features. Flow cytometric analyses showed that hMSCs-EGFP were positive for CD73, CD105, CD166, CD90 and CD44, but negative for CD34 and CD45. In addition, it could functionally be induced into osteocytes or adipocytes under conditioned media. These biological features of hMSCs-EGFP were consistent with those of hMSCs.Conclusions hMSCs transduced by PLEGFP-N1 retroviral vector can be used in vivo securely because they can maintain their biological characteristics and differentiation. It is a simple and reliable way to trace the changes of hMSCs in vivo by EGFP during cell transplantation and gene therapy.
基金supported by National Natural Science Foundation of China(82300957/82230024/82025008/82070844)Beijing Natural Science Foundation(7212123).
文摘Nonalcoholic fatty liver disease(NAFLD)is a serious threat to public health,but its underlying mechanism remains poorly understood.In screening important genes using Gene Importance Calculator(GIC)we developed previously,ribosomal modification protein rimK-like family member A(RIMKLA)was predicted as one essential gene but its functions remained largely unknown.The current study determined the roles of RIMKLA in regulating glucose and lipid metabolism.RIMKLA expression was reduced in livers of human and mouse with NAFLD.Hepatic RIMKLA overexpression ameliorated steatosis and hyperglycemia in obese mice.Hepatocyte-specific RIMKLA knockout aggravated high-fat diet(HFD)-induced dysregulated glucose/lipid metabolism in mice.Mechanistically,RIMKLA is a new protein kinase that phosphorylates betaine-homocysteine S-methyltransferase 1(BHMT1)at threonine 45(Thr45)site.Upon phosphorylation at Thr45 and activation,BHMT1 eliminated homocysteine(Hcy)to inhibit the activity of transcription factor activator protein 1(AP1)and its induction on fatty acid synthase(FASn)and cluster of differentiation 36(CD36)gene transcriptions,concurrently repressing lipid synthesis and uptake in hepatocytes.Thr45 to alanine(T45A)mutation inactivated BHMT1 to abolish RIMKLA’s repression on Hcy level,AP1 activity,FASn/CD36 expressions,and lipid deposition.BHMT1 overexpression rescued the dysregulated lipid metabolism in RIMKLA-deficient hepatocytes.In summary,RIMKLA is a novel protein kinase that phosphorylates BHMT1 at Thr45 to repress lipid synthesis and uptake.Under obese condition,inhibition of RIMKLA impairs BHMT1 activity to promote hepatic lipid deposition.
