As a crucial regulatory molecule in the context of vascular stenosis, transforming growth factor-β (TGF-β), plays a pivotal role in its initiation and progression. TGF-β, a member of the TGF-β superfamily, can bind to the TGF-β receptor and transduce extracellular to intracellular signals through canonical Smad dependent or noncanonical signaling pathways to regulate cell growth, proliferation, differentiation, and apoptosis. Restenosis remains one of the most challenging problems in cardiac, cerebral, and peripheral vascular disease worldwide. The mechanisms for occurrence and development of restenosis are diverse and complex. The TGF-β pathway exhibits diversity across various cell types. Hence, clarifying the specific roles of TGF-β within different cell types and its precise impact on vascular stenosis provides strategies for future research in the field of stenosis.
Humans ; Transforming Growth Factor beta/metabolism* ; Constriction, Pathologic ; Signal Transduction ; Cell Differentiation ; Vascular Diseases ; Transforming Growth Factors ; Transforming Growth Factor beta1

OBJECTIVETo investigate the expression of E-cadherin, vimentin, β-catenin and transforming growth factor-β1 (TGF-β1) in oral squamous cell carcinomas (OSCC).

METHODSEighty-nine cases of OSCC and 20 cases of normal oral mucosa were collected. Then the 89 cases of OSCC were classified as grade I, II, III. The semiquantitative method was used to calculated the positive intensity and positive rate. The relationship between the OSCC differentiation and the four biomarkers was analyzed.

RESULTSThe median of E-cadherin was 9.00 in the normal tissue, 9.00, 6.00 and 6.00 in OSCC I, II and III, respectively. There was significant difference between the normal group and OSCC group (Z=-4.211, P=0.000). The median of vimentin was 0.00 in the normal tissue, 0.00, 0.00 and 4.00 in OSCC I, II and III, respectively. There was significant difference between the normal group and OSCC group (Z=-3.675, P=0.000). The median of β-catenin was 9.00 in the normal tissue, 3.00, 4.00 and 3.00 in OSCC I, II and III, respectively. There was significant difference between the normal group and OSCC group (Z=-6.300, respectively. There was significant difference between the normal group and OSCC group (Z=-3.329, P=0.000). E-cadherin expression was positively correlated to β-catenin expression (r=0.327, P=0.002), negtively correlated to vimentin expression (r=-0.386, P=0.001) and positively correlated to TGF-β1 expression (r=-0.304, P=0.004). Vimentin expression was positively correlated to TGF-β1 expression (r=0.401, P=0.000).

CONCLUSIONSE-cadherin and β-catenin in OSCC had a down-regulated expression, while the vimentin has an up-regulated expression.

Cadherins ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Differentiation ; Humans ; Mouth Mucosa ; cytology ; metabolism ; Mouth Neoplasms ; metabolism ; pathology ; Neoplasm Proteins ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Transforming Growth Factors ; metabolism ; Vimentin ; metabolism ; beta Catenin ; metabolism

Thymic epithelial cells (TECs) are one of the most important components in thymic microenvironment supporting thymocyte development and maturation. TECs, composed of cortical and medullary TECs, are derived from a common bipotent progenitor, mediating thymocyte positive and negative selections. Multiple levels of signals including intracellular signaling networks and cell-cell interaction are required for TEC development and differentiation. Transcription factors Foxn1 and autoimmune regulator (Aire) are powerful regulators promoting TEC development and differentiation. Crosstalks with thymocytes and other stromal cells for extrinsic signals like RANKL, CD40L, lymphotoxin, fibroblast growth factor (FGF) and Wnt are also definitely required to establish a functional thymic microenvironment. In this review, we will summarize our current understanding about TEC development and differentiation, and its underlying multiple signal pathways.
Cell Communication ; genetics ; Cell Differentiation ; Epithelial Cells ; cytology ; metabolism ; Forkhead Transcription Factors ; genetics ; metabolism ; Humans ; Signal Transduction ; genetics ; Thymocytes ; cytology ; metabolism ; Thymus Gland ; cytology ; growth & development ; Transcription Factors ; genetics ; metabolism

OBJECTIVE: To determine the time course and potential mechanism of fibroblast growth factor-1 (FGF-1) in the regulation of adipogenesis.
 METHODS: We cultured human Simpson-Golabi-Behmel syndrome (SGBS) pre-adipocytes with recombinant FGF-1 and harvested cells at various stages prior to and during differentiation; at cell proliferation (D-3), confluence (D0), early (D3), middle (D7) and mature (D14) stages of differentiation. We determined lipid accumulation in mature adipocytes by morphological observation and quantitative measurement of oil red O staining. We also examined the expression of adipogenic genes and related markers involved in the Wnt/β-catenin pathway using quantitative Real-time PCR and Western blot.
 RESULTS: Compared to control SGBS cells, treatment with FGF-1 increased lipid accumulation; induced a sustained increase in the mRNA for peroxisome proliferater-activated receptor γ (PPARγ), glyceraldehyde-3-phosphate dehydrogenase (G3PDH), adiponectin and glucose transporter type 4 (GLUT4); and promoted a sustained decrease in expression of markers of the Wnt/β-catenin pathway, β-catenin and transcription factor 4 (TCF4).
 CONCLUSION The adipogenic effects of FGF-1 are apparent throughout the whole priming and differentiation period in human SGBS pre-adipocytes. Furthermore, our results suggest that FGF-1 
promotes adipogenesis, at least in part, via a sustained decrease in activity of the Wnt/β-catenin pathway.
Adipocytes ; drug effects ; metabolism ; Adipogenesis ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; metabolism ; Cell Differentiation ; Cells, Cultured ; Fibroblast Growth Factor 1 ; pharmacology ; Humans ; Recombinant Proteins ; pharmacology ; Transcription Factor 4 ; Transcription Factors ; metabolism ; Wnt Signaling Pathway ; beta Catenin ; metabolism

The failure rate of dental implantation in patients with well-controlled type 2 diabetes mellitus (T2DM) is higher than that in non-diabetic patients. This due, in part, to the impaired function of bone marrow mesenchymal stem cells (BMSCs) from the jawbone marrow of T2DM patients (DM-BMSCs), limiting implant osseointegration. RNA N6-methyladenine (m6A) is important for BMSC function and diabetes regulation. However, it remains unclear how to best regulate m6A modifications in DM-BMSCs to enhance function. Based on the "m6A site methylation stoichiometry" of m6A single nucleotide arrays, we identified 834 differential m6A-methylated genes in DM-BMSCs compared with normal-BMSCs (N-BMSCs), including 43 and 790 m6A hypermethylated and hypomethylated genes, respectively, and 1 gene containing hyper- and hypomethylated m6A sites. Differential m6A hypermethylated sites were primarily distributed in the coding sequence, while hypomethylated sites were mainly in the 3'-untranslated region. The largest and smallest proportions of m6A-methylated genes were on chromosome 1 and 21, respectively. MazF-PCR and real-time RT-PCR results for the validation of erythrocyte membrane protein band 4.1 like 3, activity-dependent neuroprotector homeobox (ADNP), growth differentiation factor 11 (GDF11), and regulator of G protein signalling 2 agree with m6A single nucleotide array results; ADNP and GDF11 mRNA expression decreased in DM-BMSCs. Furthermore, gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses suggested that most of these genes were enriched in metabolic processes. This study reveals the differential m6A sites of DM-BMSCs compared with N-BMSCs and identifies candidate target genes to enhance BMSC function and improve implantation success in T2DM patients.
Humans ; Bone Marrow/metabolism* ; Bone Morphogenetic Proteins/metabolism* ; Dental Implants/adverse effects* ; Diabetes Mellitus, Type 2/metabolism* ; Growth Differentiation Factors/metabolism* ; Mesenchymal Stem Cells/metabolism* ; RNA/metabolism* ; RNA Processing, Post-Transcriptional

To explore the feasibility of mesenchymal stem cells (MSCs) acting as seed cells in tissue engineering, we isolated human bone marrow MSCs and differentiated them into vascular endothelial-like cells (ELCs) in vitro. Bone marrow mononuclear cells (BMSCs) were isolated by the method of percoll density centrifugation, and seeded in Dulbecco Modified Eagle Medium supplemented with 10% fetal bovine serum. MSCs were purified through multiple adherent cultures, and differentiated into ELCs induced by endothelial cell growth medium-2 (EBM-2) medium containing vascular endothelial growth factor (VEGF), human fibroblast growth factor (hFGF), insulin like growth factors 1 (IGF-1), and human epidermal growth factor (hEGF). The relative biologic characteristics of ELCs including cell morphology and phenotype were studied by inverted microscope and flow cytometry. The induced cells were identified by immunofluorescence with CD31 and Von Willebrand factor (vWF). The results showed that the morphology of MSCs was long-spindle and vortex-like growth. After induction of differentiation, the cells were round, and similar to vascular endothelial cells (ECs). Flow cytometric analysis revealed that ELCs expressed ECs specific surface markers of CD31 and vascular endothelial cadherin (VE-cadherin), but not CD133. Immunofluorescence results also confirmed that ELCs expressed CD31 and vWF. The results suggested that ELCs possed similar cell biological characteristics with ECs. In one word, human MSCs derived from bone marrow have the potential to differentiate into ECs in vitro, and show clinical feasibility acting as ideal donor cells of vascular tissue engineering.
Antigens, CD ; metabolism ; Bone Marrow Cells ; Cadherins ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Culture Media ; chemistry ; Endothelial Cells ; cytology ; Epidermal Growth Factor ; pharmacology ; Fibroblast Growth Factors ; pharmacology ; Flow Cytometry ; Humans ; Insulin-Like Growth Factor I ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Tissue Engineering ; Vascular Endothelial Growth Factor A ; pharmacology ; von Willebrand Factor ; metabolism

BACKGROUNDNeural stem cells (NSCs) are a self-renewing and multipotent population of the central nervous system (CNS), which are active during development and maintain homeostasis and tissue integrity throughout life. Microglias are an immune cell population resident in the CNS, which have crucial physiological functions in the developing and adult CNS. This study aimed to investigate that whether microglia co-cultured with NSCs could promote astrogliogenesis from NSCs.

METHODSMicroglia and NSCs were co-cultured in 24-well insert plates. NSCs were plated in the bottom of the well and microglia in the insert. Fluorescent staining, Western blotting and RT-PCR were used to determine the effect of microglia on NSCs differentiation.

RESULTSCo-culture of microglia and NSCs promoted astrogliogenesis from NSCs. Several key genes, such as Notch 1, Notch 2, Notch 3, Hes 5, and NRSF were downregulated, while the critical genes Id1 and Id2 were upregulated. BMP2 and FGF2 were upregulated.

CONCLUSIONMicroglias act as a regulator of NSCs astrogliogenesis.

Animals ; Astrocytes ; cytology ; metabolism ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Blotting, Western ; Bone Morphogenetic Protein 2 ; genetics ; Cell Differentiation ; genetics ; physiology ; Cells, Cultured ; Coculture Techniques ; methods ; Fibroblast Growth Factor 2 ; genetics ; Inhibitor of Differentiation Protein 1 ; genetics ; Inhibitor of Differentiation Protein 2 ; genetics ; Microglia ; cytology ; metabolism ; Microscopy, Fluorescence ; Neural Stem Cells ; cytology ; metabolism ; Rats ; Repressor Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the histogenesis of pulmonary sclerosing hemangioma (PSH).

METHODSTissue microarray and immunohistochemical technique were used to detect the expression of pan-cytokeratin, epithelial membrane antigen(EMA), vimentin, thyroid transcription factor (TTF)-1, napsin A, synaptophysin, chromogranin A, CD56, E-cadherin, β-catenin, CD117, CD68 and transforming growth factor(TGF)-β1 in 49 cases of PSH.

RESULTSImmunohistochemistry revealed that all cuboidal surface cells expressed pan-cytokeratin, EMA, TTF-1 and napsin A. The polygonal cells expressed EMA, TTF-1, napsin A (positive rate 16.3%, 8/49), but not pan-cytokeratin. Both types of cells were negative for synaptophysin, chromogranin A and CD56. Strong positive staining for E-cadherin and β-catenin appeared on the membrane of cuboidal cells in all PSH, with cytoplasm staining for β-catenin as well. The expression levels of these adhesion molecules decreased in the polygonal cells, with the staining localized to the cytoplasm. E-cadherin staining was not detected or was weak. β-catenin staining was not detected on the cell membrane but partially in the cytoplasm. The polygonal cells stained strongly for vimentin, while only a few cuboidal cells were positive. CD117 and CD68 positive inflammatory cells were scattered between the polygonal cells, which was consistent with the distribution of TGF-β1 positive cells.

CONCLUSIONSPSH originates from the primitive respiratory epithelium, and polygonal stromal cells may be derived from epithelial-mesenchymal transformation of the cuboidal cells. TGF-β1 may play an important role in the formation of sclerosing hemangioma.

Adolescent ; Adult ; Aged ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Aspartic Acid Endopeptidases ; metabolism ; Cadherins ; metabolism ; Female ; Humans ; Immunohistochemistry ; Keratins ; metabolism ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Nuclear Proteins ; metabolism ; Pneumonectomy ; Proto-Oncogene Proteins c-kit ; metabolism ; Pulmonary Sclerosing Hemangioma ; metabolism ; pathology ; surgery ; Thyroid Nuclear Factor 1 ; Transcription Factors ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Vimentin ; metabolism ; Young Adult ; beta Catenin ; metabolism

Naturally occurred CD4(+)CD25(+) regulatory T cells derived from thymus. It plays an important role in self-tolerance and allograft-tolerance through cell-contact dependent mechanism. This review described the advances of study on the probable regulatory factors of the naturally occurring regulatory T cells, such as Foxp3, IL-2, TGF-beta(1), dendritic cells and CTLA-4. As a marker of Treg, the expression of Foxp3 could be used to identify regulatory T cells. The combination of interferon 2 and IL-2Ralpha would activate Treg and promote its proliferation through the phosphorylation of STAT5. TGF-beta(1) on the cell surface may influence the function of Treg, while the secretion type of TGF-beta may promote the proliferation of Treg. Dendritic cells can positively or negatively regulate Treg, which depends on the signal transduction pathway. CTLA-4 expressed on the surface of Treg might bind to the B7 molecule on the DC, effective cell or Treg itself directly or indirectly regulate Treg.
Animals ; CD4 Antigens ; metabolism ; Cell Differentiation ; immunology ; Cells, Cultured ; Dendritic Cells ; immunology ; Forkhead Transcription Factors ; physiology ; Humans ; Interleukin-2 ; physiology ; Interleukin-2 Receptor alpha Subunit ; metabolism ; T-Lymphocytes, Regulatory ; cytology ; immunology ; metabolism ; Thymus Gland ; cytology ; Transforming Growth Factor beta ; physiology

Neural progenitor cells (NPs) have shown several promising benefits for the treatment of neurological disorders. To evaluate the therapeutic potential of human neural progenitor cells (hNPs) in amyotrophic lateral sclerosis (ALS), we transplanted hNPs or growth factor (GF)-expressing hNPs into the central nervous system (CNS) of mutant Cu/Zn superoxide dismutase (SOD(1G93A)) transgenic mice. The hNPs were engineered to express brain-derived neurotrophic factor (BDNF), insulin-like growth factor-1 (IGF-1), VEGF, neurotrophin-3 (NT-3), or glial cell-derived neurotrophic factor (GDNF), respectively, by adenoviral vector and GDNF by lentiviral vector before transplantation. Donor-derived cells engrafted and migrated into the spinal cord or brain of ALS mice and differentiated into neurons, oligodendrocytes, or glutamate transporter-1 (GLT1)-expressing astrocytes while some cells retained immature markers. Transplantation of GDNF- or IGF-1-expressing hNPs attenuated the loss of motor neurons and induced trophic changes in motor neurons of the spinal cord. However, improvement in motor performance and extension of lifespan were not observed in all hNP transplantation groups compared to vehicle-injected controls. Moreover, the lifespan of GDNF-expressing hNP recipient mice by lentiviral vector was shortened compared to controls, which was largely due to the decreased survival times of female animals. These results imply that although implanted hNPs differentiate into GLT1-expressing astrocytes and secrete GFs, which maintain dying motor neurons, inadequate trophic support could be harmful and there is sexual dimorphism in response to GDNF delivery in ALS mice. Therefore, additional therapeutic approaches may be required for full functional recovery.
Adenoviridae/genetics ; Amyotrophic Lateral Sclerosis/metabolism/mortality/*therapy ; Animals ; Astrocytes/metabolism ; Brain/*embryology ; Cell Differentiation ; Disease Models, Animal ; Excitatory Amino Acid Transporter 2/metabolism ; Female ; Fetal Stem Cells/*metabolism ; Genetic Vectors ; Humans ; Immunoenzyme Techniques ; Male ; Mice ; Mice, Transgenic ; Motor Neurons/*physiology ; Nerve Growth Factors/*metabolism ; *Stem Cell Transplantation ; Superoxide Dismutase/genetics ; Transfection ; Vascular Endothelial Growth Factor A/genetics/metabolism