| 2005 |
Heterozygous mutations in TGFBR2 (or TGFBR1) cause a syndrome of cardiovascular, craniofacial, neurocognitive and skeletal abnormalities (Loeys-Dietz syndrome); despite mutant alleles failing to support TGFβ signal propagation in vitro, patient-derived tissues showed increased nuclear phospho-Smad2 and elevated collagen/CTGF expression, indicating paradoxically increased TGFβ signaling in vivo. |
Patient-derived cell culture assays of acute TGFβ signaling kinetics; immunostaining of patient tissues for p-Smad2, collagen, and CTGF; genetic mutation analysis |
Nature genetics |
High |
15731757
|
| 2004 |
Heterozygous mutations in TGFBR2 cause Marfan syndrome type 2 (MFS2); identified mutations lead to loss of TGFβ signaling activity on extracellular matrix formation. |
Chromosomal breakpoint mapping, DNA sequencing of TGFBR2, functional assessment of TGFβ signaling activity and ECM formation in mutation carriers |
Nature genetics |
High |
15235604
|
| 2014 |
Postnatal conditional inactivation of Tgfbr2 in smooth muscle cells causes thoracic aortic thickening, dilation, and dissection; loss of TGFBR2 decreases canonical Smad signaling but unexpectedly increases MAPK signaling, and aortic disease results from perturbed contractile apparatus in medial cells and paracrine growth factor production by adventitial cells; treatment with rapamycin prevents dissection. |
Smooth-muscle-specific Cre/lox conditional knockout in mice; TGFβ serologic neutralization; Western blot for Smad and MAPK signaling; rapamycin treatment; aortic histology |
The Journal of clinical investigation |
High |
24401272
|
| 2007 |
Conditional endothelial deletion of Tgfbr2 (or Alk5) in mice does not affect vessel morphogenesis and does not produce HHT-like vascular malformations, indicating that neither ALK5 nor TGFBR2 is required for ALK1 signaling pertinent to HHT2 pathogenesis. |
Conditional endothelial-specific Cre knockout of Alk1, Alk5, or Tgfbr2 in mice; vascular phenotype analysis; Alk5 inhibitor in zebrafish |
Blood |
High |
17911384
|
| 2010 |
Heterozygous TGFBR2 mutations in patient-derived aortic smooth muscle cells decrease expression of SMC contractile proteins and impair TGFβ1-induced upregulation of contractile genes; patient-derived fibroblasts also fail to differentiate into mature myofibroblasts upon TGFβ1 stimulation, establishing that TGFBR2 mutations impair SMC contractile differentiation. |
Explanted patient aortic SMCs and fibroblasts; TGFβ1 stimulation assays; Western blot and immunostaining for contractile proteins in patient vs. control aortas |
Cardiovascular research |
High |
20628007
|
| 2010 |
Quantitative analysis of nine TGFBR2 patient mutations showed differing effects on protein stability, internalization, Smad phosphorylation, ERK signaling, and transcriptional activity; mutations associated with LDS and MFS2 showed dominant-negative effects; the R460C TAAD-associated mutation retained residual Smad phosphorylation, suggesting that the balance between defective Smad vs. ERK signaling determines phenotypic severity. |
Quantitative cell-based assays for TGFBR2 protein expression, turnover (internalization), Smad phosphorylation, ERK activation, and transcriptional reporter activity for nine patient mutations |
Journal of cell science |
High |
21098638
|
| 2006 |
Cell-autonomous ablation of Tgfbr2 specifically in palatal epithelial cells causes soft palate cleft; mutant MEE cells fail to undergo programmed cell death and instead continue to proliferate and form cysts/epithelial bridges; Irf6 expression is downregulated in Tgfbr2-mutant MEE cells, placing Irf6 downstream of TGFBR2 in palatal epithelial fate determination. |
Cre/lox conditional knockout of Tgfbr2 in palatal epithelial cells; genetic lineage labeling of MEE; immunostaining for Irf6, K14; histological analysis |
Developmental biology |
High |
16780827
|
| 2006 |
Deletion of Tgfbr2 in Col2a-expressing cells disrupts maintenance of boundaries in the axial skeleton; loss of Tgfbr2 expands Pax1/Pax9 expression domains across sclerotome segments and reduces Fibromodulin (intervertebral disc marker) expression, demonstrating that TGFBR2 signaling maintains sclerotome segmental boundaries. |
Col2a-Cre conditional knockout of Tgfbr2 in mice; in situ hybridization and immunostaining for Pax1, Pax9, Msx1, Fmod; histological analysis with PNA staining |
Developmental biology |
High |
16824508
|
| 2009 |
Conditional loss of Tgfbr2 in Prx1Cre-expressing skull mesenchyme causes open skull defects due to reduced mesenchymal cell proliferation and impaired osteoblast differentiation; in isolated Tgfbr2-deleted osteoprogenitor cultures, mRNA levels of Runx2, Osterix, Dlx5, and Msx2 are downregulated, demonstrating that Tgfbr2 regulates osteoblast differentiation independently of proliferation. |
Prx1Cre conditional KO; BrdU proliferation assay; TUNEL apoptosis; immunofluorescence for Runx2/Osterix; Adeno-Cre deletion in cultured osteoprogenitors; RT-PCR |
Developmental biology |
High |
19699732
|
| 2013 |
Deletion of Tgfbr2 in odontoblast/bone mesenchyme (Osx-Cre) disrupts molar root elongation and HERS formation; odontoblast organization and Dspp expression are reduced, while Dmp1 increases; Tgfbr2 in dental mesenchyme indirectly regulates HERS maintenance. |
Osx-Cre conditional KO of Tgfbr2; microCT; immunostaining for K14, Dspp, Dmp1, Nfic; histological analysis |
Developmental biology |
High |
23933490
|
| 2013 |
Tgfbr2 mutant palatal mesenchymal cells (Tgfbr2fl/fl;Wnt1-Cre) spontaneously accumulate lipid droplets due to reduced lipolysis; these cells fail to respond to Sonic Hedgehog-driven proliferation; inhibition of p38 MAPK or telmisartan treatment restores lipid metabolism and cell proliferation, rescuing cleft palate in vivo. |
Cranial neural crest-specific Tgfbr2 conditional KO; lipid droplet staining; lipolysis assay; p38 MAPK inhibitor treatment in vitro and in vivo; rescue of palate fusion |
Human molecular genetics |
High |
23975680
|
| 2014 |
miR-145 directly targets the 3'-UTR of TGFBR2 mRNA in vascular smooth muscle cells; miR-145 overexpression attenuates TGFβ receptor II-dependent extracellular matrix gene expression and fibrosis, while miR-145 knockout mice show increased matrix synthesis and a pronounced fibrotic response to angiotensin II; endothelial cell co-culture induces miR-143/145 expression via Notch signaling. |
miR-145 overexpression/knockout in cell culture and mouse models; luciferase 3'-UTR reporter assay; angiotensin II infusion; ECM gene expression analysis |
Circulation research |
High |
25323858
|
| 2019 |
NG2 glia-derived TGF-β2 signals through TGFBR2 in microglia to maintain brain immune homeostasis; gain- and loss-of-function of NG2 glia-TGF-β2 or microglial TGFBR2 regulate CX3CR1-mediated immune suppression; ablation of NG2 glia or TGFBR2 deficiency in microglia leads to exaggerated neuroinflammation and dopaminergic neuron loss in MPTP model. |
Diphtheria toxin-mediated NG2 glia ablation; RNA sequencing; co-cultures of NG2 glia and microglia; gain/loss-of-function experiments for TGFβ2 and TGFBR2; MPTP mouse model; immunohistochemistry |
BMC medicine |
High |
31727112
|
| 2013 |
miR-21 suppresses TGFBR2 expression by binding its 3'-UTR; androgen receptor (AR) signaling potentiates this effect, creating a positive feedback loop where miR-21 and AR regulate each other's expression; this miR-21/AR axis attenuates TGFβ-mediated Smad2/3 activation, cell growth inhibition, migration inhibition, and apoptosis in prostate cancer cells. |
3'-UTR luciferase reporter assay; AR/miR-21 overexpression and knockdown in prostate epithelial cell lines and xenografts; Western blot for p-Smad2/3; cell growth, migration, and apoptosis assays |
Oncogene |
High |
24037531
|
| 2006 |
TGFBR2 frameshift mutations in TGFBR2 exon 3 (polyadenine tract BAT-RII) occur at elevated rates in MLH1-deficient cells; reconstitution of TGFBR2 expression in an MSI colon cancer cell line (HCT116) decreases proliferation and reduces cdk4 expression and kinase activity, establishing that TGFBR2 inactivation drives increased cdk4-dependent proliferation in MSI colorectal cancer. |
TGFBR2 reconstitution in HCT116 MSI cell line; proliferation assays; cdk4 immunostaining and kinase activity assay in primary colon cancers with mutant vs. wild-type BAT-RII |
International journal of cancer |
High |
16108056
|
| 2008 |
MLH1 deficiency produces ~11-fold higher TGFBR2 microsatellite mutation rates compared to MSH6 deficiency; TGFBR2 microsatellite mutates ~3× faster than ACVR2 in both hMLH1 and hMSH6 deficient backgrounds, establishing that TGFBR2 frameshift mutation rate is dependent on specific MMR gene background. |
EGFP frameshift reporter plasmids containing TGFBR2 or ACVR2 coding microsatellites in MMR-deficient cell lines; flow cytometry; DNA sequencing |
PloS one |
High |
18941508
|
| 2019 |
Loss of Tgfbr2 in Kras-mutant lung cells promotes lung squamous cell carcinoma (SCC) formation through a Smad-independent ERK1/2 pathway; low pERK1/2 upregulates SOX2 expression (oncogenic SCC driver), and ERK1/2 suppression cooperates with SMAD4 loss to elevate SOX2; Smad4 loss alone does not drive SCC, placing ERK1/2 downstream of Tgfbr2 as a key suppressor of SOX2-driven SCC. |
Kras × Tgfbr2 conditional KO mouse model; Kras × Smad4 conditional KO comparison; ERK1/2 inhibitor treatment; SOX2 expression analysis; IHC of mouse and human tumors |
Cancer research |
High |
31209059
|
| 2019 |
TGFBR2 reconstitution in MSI colorectal cancer cells alters the protein composition of secreted exosomes; TGFBR2 expression status determines a distinct exosomal proteome and modulates cytokine secretion (IL-4, SCF, PDGF-B) in recipient hepatoma cells. |
Doxycycline-inducible TGFBR2 reconstitution in HCT116-TGFBR2 cells; exosome isolation and mass spectrometry proteomics; nanoparticle tracking; confocal microscopy; Luminex cytokine profiling |
Cell communication and signaling |
High |
28376875
|
| 2016 |
TGFBR2 reconstitution in HCT116 MSI colorectal cancer cells increases LFNG glycosyltransferase expression in a TGFBR2-dependent manner and enhances N-acetyl-d-glucosamine incorporation into Notch1, linking TGFBR2 signaling to Notch1 glycosylation via LFNG. |
Doxycycline-inducible TGFBR2 reconstitution in HCT116; Glyco-Gene Chip array; real-time RT-PCR validation; dual [³H]-GlcNAc/[³⁵S]-methionine metabolic labeling; Notch1 immunoprecipitation |
Cellular signalling |
High |
27156840
|
| 2014 |
Neural-specific deletion of Tgfbr2 (Foxg1-Cre) impairs brain angiogenesis through an altered secretome; VEGFA, IGF1, IGF2, TGFβ, ID1, THBS2, and ADAMTS1 are altered; conditioned medium from Tgfbr2-KO neural cultures causes HUVEC branching and migration defects that are rescued by supplementing VEGFA, FGF2, and IGF1. |
Foxg1-Cre conditional KO of Tgfbr2; conditioned medium transfer to HUVECs; HUVEC branching and migration assays; growth factor supplementation rescue experiments; gene expression analysis |
Human molecular genetics |
High |
24990151
|
| 2023 |
USP33, a deubiquitinating enzyme, interacts with TGFBR2 (identified by mass spectrometry and luciferase complementation), deubiquitinates TGFBR2, prevents its lysosomal degradation, and promotes its accumulation at the cell membrane to sustain TGFβ signaling; TGFβ target gene ZEB1 transcriptionally upregulates USP33, creating a positive feedback loop. |
Mass spectrometry interactome; luciferase complementation assay; ubiquitination assay; lysosome inhibitor experiments; ChIP for ZEB1; Western blot; functional proliferation/migration/invasion assays |
Cell death & disease |
High |
37322017
|
| 2024 |
ITGB5 (integrin β5) promotes TGFBR2 endosomal recycling by acting as a scaffold that interacts with both TGFBR2 and the sorting nexin SNX17, preventing lysosomal degradation of TGFBR2 and maintaining its cell-surface distribution; TGFβ signaling directly upregulates ITGB5, creating a positive feedback loop that enhances TGFβ-mediated EMT and gastric cancer metastasis. |
Co-immunoprecipitation of ITGB5/TGFBR2/SNX17; ITGB5 knockdown in vitro and in vivo; endosomal recycling assay; Western blot for TGFBR2 surface and total levels; feedback loop validation |
Cancer letters |
High |
38729557
|
| 2025 |
PIEZO1 attenuates TGFβ signaling by promoting endocytosis and autophagy of TGFBR2 via Rab GTPase 3C; PIEZO1 knockout in smooth muscle cells of Marfan syndrome mice exacerbates TAA and TGFβ pathway activation by inhibiting TGFBR2 endocytosis/autophagy; pharmacological PIEZO1 activation (Yoda1) prevents TGFβ activation and reverses TAA. |
SMC-specific PIEZO1 conditional KO in Marfan syndrome (Fbn1C1041G/+) mice; Yoda1 pharmacological treatment; Rab3C interaction experiments; endocytosis/autophagy assays; aortic histology and TGFβ pathway analysis |
European heart journal |
Medium |
39585648
|
| 2021 |
The E3 ubiquitin ligase HUWE1 mediates ubiquitination and degradation of TGFBR2 in gastric cancer cells; HUWE1 overexpression promotes GC cell proliferation, migration, and invasion, and these effects are impaired by TGFBR2 overexpression, placing TGFBR2 downstream of HUWE1-mediated ubiquitin-proteasome degradation. |
HUWE1 overexpression/silencing in GC cell lines; ubiquitination assay; Western blot; MTT and Transwell assays; TGFBR2 rescue experiments |
Investigational new drugs |
Medium |
33405091
|
| 2021 |
CRISPR/Cas9-mediated knockout of TGFBR2 in ovarian cancer tumor-infiltrating lymphocytes renders them resistant to TGFβ-mediated immunosuppression, evidenced by absence of SMAD phosphorylation, preserved proinflammatory cytokine secretion, and improved cytotoxicity in the presence of TGFβ. |
CRISPR/Cas9 KO of TGFBR2 in primary patient TILs; Western blot for p-SMAD; cytokine secretion assays; cytotoxicity assay; transcriptome analysis; off-target sequencing |
Journal for immunotherapy of cancer |
High |
35882447
|
| 2019 |
TGFBR2 reconstitution in MSI colorectal cancer cells alters miRNA expression profiles in both cells and secreted extracellular vesicles; specific subsets of miRNAs are regulated in a TGFBR2-dependent manner, including downregulation of miR-381-3p in TGFBR2-deficient cells and EVs. |
Doxycycline-inducible TGFBR2 reconstitution in HCT116-TGFBR2; small RNA sequencing of cell and EV miRNA profiles; isogenic comparison |
International journal of oncology |
Medium |
31432155
|
| 2022 |
The transcription factor GABPA directly activates TGFBR2 transcription (ChIP identified GABPA binding at TGFBR2 locus); GABPA and TGFBR2 phenocopy each other in suppressing renal cell carcinoma proliferation, invasion, and stemness; oncometabolite L-2-HG epigenetically silences GABPA via DNA methylation, disrupting the GABPA-TGFBR2-TGFβ axis. |
siRNA/overexpression of GABPA; RNA sequencing to identify TGFBR2 as GABPA target; ChIP assay for GABPA at TGFBR2 locus; L-2-HG treatment with methylation analysis; xenograft mouse model |
Journal of experimental & clinical cancer research |
High |
35549739
|
| 2021 |
SOX4 directly activates TGFBR2 transcription; SOX4 and the SWI/SNF ATPase SMARCA4 form a complex that maintains open chromatin at TGFBR2 regulatory regions; TGFBR2 is required to mediate SOX4-dependent PI3K/Akt signaling in triple-negative breast cancer. |
Genomic/proteomic analyses; ChIP for SOX4/SMARCA4 at TGFBR2 locus; chromatin accessibility assay (ATAC-seq equivalent); TGFBR2 knockdown rescue of SOX4-driven PI3K signaling; Co-IP of SOX4/SMARCA4 complex |
NPJ breast cancer |
High |
33837205
|
| 2017 |
YAP-1 promotes TGFBR2 transcription by binding directly to the TGFBR2 promoter through TEAD transcription factors in T cells, thereby promoting Treg differentiation and immunosuppression in hepatocellular carcinoma. |
Bioinformatics identification of YAP-1/TEAD binding sites in TGFBR2 promoter; luciferase reporter assay; ChIP/promoter binding assay; overexpression of YAP-1 in T cells; flow cytometry for Treg markers |
Cellular physiology and biochemistry |
Medium |
28472799
|
| 2020 |
YAP/TAZ represses TGFBR2 both post-transcriptionally through the miR-106b-25 cluster and transcriptionally via EZH2 (identified as a YAP/TAZ transcriptional target); YAP/TAZ and EZH2 cooperate to repress TGFBR2 in NSCLC. |
miRNA target validation; EZH2 as YAP/TAZ target gene analysis; reporter assays; Western blot; correlation in clinical NSCLC samples |
Cancer letters |
Medium |
33296708
|
| 2021 |
MYOCD localizes to the TGFBR2 promoter region and recruits PRMT5/MEP50 complex to epigenetically silence TGFBR2 transcription; loss of MYOCD leads to TGFBR2 hyperactivation, and NSCLC cells deficient in MYOCD are particularly sensitive to TGFBR kinase inhibitors. |
ChIP showing MYOCD at TGFBR2 promoter; PRMT5/MEP50 co-recruitment assay; TGFBR2 expression analysis with MYOCD manipulation; sensitivity assays with TGFBRi |
Theranostics |
Medium |
33995678
|
| 2019 |
TGFβ signaling through SMAD2/3 transcriptionally induces miR-520e expression (confirmed by ChIP-PCR), which in turn targets and downregulates TGFBR2, constituting a TGFβ-SMAD-miR-520e-TGFBR2 negative-feedback loop in NSCLC cells. |
ChIP-PCR for SMAD2/3 at miR-520e promoter; miR-520e overexpression/inhibition; luciferase 3'-UTR reporter for TGFBR2; Western blot; clinical correlation in NSCLC samples |
Carcinogenesis |
Medium |
30475986
|
| 2019 |
GDF15-induced apoptosis, caspase-9/3 activation, and ERK1/2/p38 MAPK inhibition in A549 cells depends on TGFBR2 expression; TGFBR2 silencing blocks all GDF15-induced apoptotic and MAPK effects, placing TGFBR2 as a required mediator of GDF15 signaling in this cell context. |
GDF15 overexpression (full and mature forms); siRNA knockdown of TGFBR2; flow cytometry for annexin V/PI; caspase activity assays; ERK1/2 and p38 phosphorylation by Western blot |
Cell biochemistry and function |
Medium |
31172564
|
| 2023 |
NRP1 (neuropilin-1) interacts with TGFBR2 at endothelial adherens junctions (Co-IP), reduces plasma membrane localization of TGFBR2, and suppresses TGFβ signaling; NRP1 knockdown increases TGFBR2 membrane abundance and TGFβ-driven inflammation. |
Co-IP of NRP1 with TGFBR2; NRP1 endothelial-specific KO mice; surface TGFBR2 localization analysis; inflammatory marker expression; atherosclerosis model |
Science signaling |
High |
37220183
|
| 2019 |
A super-enhancer (marked by H3K27Ac) at the TGFBR2 locus controls its transcription in pancreatic cancer cells; CRISPR deletion of this super-enhancer (sgTGFBR2-SEΔ) reduces TGFBR2 expression, impairs TGFβ-induced p-SMAD2/3 signaling, and attenuates TGFβ-induced migration and EMT. |
H3K27Ac ChIP-seq; BRD4 inhibitor (JQ1) treatment; CRISPR sgRNA-mediated super-enhancer deletion; p-SMAD2/3 Western blot; migration and EMT assays |
Cellular signalling |
Medium |
31730895
|
| 2021 |
Enzalutamide decreases TGFBR2 protein expression in osteoblasts through PTH1R-mediated endocytosis; PTH1R blockade rescues enzalutamide-mediated TGFBR2 reduction and restores enzalutamide sensitivity in prostate cancer cells co-cultured with osteoblasts. |
Co-culture of PCa cells with osteoblasts; PTH1R inhibitor rescue; Western blot for TGFBR2 protein; bone metastasis mouse models; clinical data analysis |
Cancer letters |
Medium |
34752846
|
| 2022 |
TGFBR2 reconstitution in MSI/MMR-deficient VCMsh2/Tgfbr2 mice modulates gut microbiota composition; combined loss of Msh2 and Tgfbr2 in intestinal epithelium synergistically promotes inflammation-associated colorectal cancer, and gut microbiota strongly modulates tumor incidence. |
Villin-Cre dual conditional KO of Msh2 and Tgfbr2; intestinal tumor analysis; mutational spectra; gene expression profiling; microbiota compositional analysis |
Cellular and molecular gastroenterology and hepatology |
Medium |
35688320
|
| 2016 |
Tgfbr2 deletion in Osx-Cre-expressing immature osteoblasts causes postnatal reduction in bone volume due to decreased osteoblast proliferation (PCNA reduction) and impaired maturation (Bglap/osteocalcin mRNA reduced) without altering Runx2 mRNA or apoptosis. |
Osx-Cre conditional KO; microCT; histomorphometry; qRT-PCR for Runx2, Bglap, PCNA, Bax/Bcl2; osteoblast and osteoclast enumeration |
Bone |
Medium |
28043895
|
| 2015 |
Odontoblast-specific loss of Tgfbr2 disrupts odontoblast polarity and matrix secretion after mantle dentin formation, reduces alkaline phosphatase and Dsp expression, increases Dmp1, and causes ectopic fibrous matrix calcification leading to pulpal obliteration. |
Odontoblast-specific Cre conditional KO; immunohistochemistry for ALP, Dsp, Dmp1, collagen I, biglycan; microCT |
Journal of dental research |
Medium |
25818583
|
| 2019 |
A TGFBR2/SMAD2/DNMT1/miR-145 negative regulatory loop operates in LPS-induced sepsis: miR-145 directly targets TGFBR2 (confirmed by luciferase reporter and RIP); activated SMAD2 binds the DNMT1 promoter (ChIP), upregulating DNMT1 which methylates the miR-145 promoter, suppressing miR-145 and further increasing TGFBR2 expression. |
Luciferase reporter; ChIP for SMAD2 at DNMT1 promoter; RIP assay; miR-145 promoter methylation; TGFBR2 knockdown; LPS sepsis mouse model |
Biomedicine & pharmacotherapy |
Medium |
30784922
|
| 2020 |
TGFBR2 interacts with PDGFR in GBM cells (Co-immunoprecipitation); concurrent TGFBR2 and PDGFR expression attenuates PDGFRi toxicity and maintains downstream PDGFR signaling under inhibitor treatment, contributing to PDGFRi resistance. |
Co-immunoprecipitation of TGFBR2 with PDGFR in GBM cells; PDGFRi toxicity assays; PDGFR downstream signaling Western blot |
American journal of cancer research |
Medium |
29888103
|
| 2020 |
Hypoxia-induced EZH2 promotes H3K27me3 at the TGFBR2 promoter, causing hypermethylation and epigenetic silencing of TGFBR2 in prostate cancer; separately, hypoxia upregulates miR-93 which also targets and suppresses TGFBR2. |
EZH2 siRNA/inhibitor experiments; methylation-specific PCR; ChIP for H3K27me3 at TGFBR2 promoter; luciferase 3'-UTR reporter for miR-93/TGFBR2; demethylation agent rescue |
Journal of experimental & clinical cancer research |
Medium |
29699590
|
| 2017 |
Combined mutation of Apc, Kras, and Tgfbr2 deletion in intestinal epithelial cells cooperates to drive colorectal cancer metastasis; Tgfbr2 deletion combined with Kras mutation induces EMT-like morphology and lymph vessel intravasation; liver metastasis requires Kras combined with either Trp53 or Tgfbr2 deletion, with highest metastasis in the Apc/Kras/Tgfbr2 combination. |
Intestinal epithelial conditional KO mouse models with combinations of Apc, Kras, Tgfbr2, Trp53, Fbxw7; tumor-derived organoid splenic transplantation metastasis assay; RNA sequencing |
Cancer research |
High |
29282223
|