1.Expression and significance of vimentin in different types of chronic rhinosinusitis.
Lili SHI ; Xiang LU ; Zheng LIU ; Yonghua CUI
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2012;26(8):354-359
OBJECTIVE:
To explore the expression and distribution of vimentin in different types of chronic rhinosinusitis and its significance.
METHOD:
There were four groups including control (10 samples), Eos CRSwNP (10 samples), non-Eos CRSwNP (12 samples) and CRSsNP (10 samples). The expression of vimentin in chronic rhinosinusitis were detected by immunohistochemistry technique. The double-immunofluorescence was used to detect the positive staining of both vimentin and E-cadherin, both of which were the marker of epithelial cells.
RESULT:
The positive staining of vimentin were observed both in epithelium and lamina propria. The expression of vimentin were found in myofibroblast, endothelium and other mesenchymal cells. The vimentin positive cells in epithelium were epithelial cells but not mesenchymal cells, as they also expressed E-cadherin.
CONCLUSION
The vimentin positive staining cells distribute in lamina propria and epithelium of both normal nasal mucosa and chronic rhinosinusitis. The positive staining epithelial cells may generate from epithelial-mesenchymal transition. So the vimentin may play an important role in the development of chronic rhinosinusitis.
Adult
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Antigens, CD
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Cadherins
;
metabolism
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Chronic Disease
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Epithelial Cells
;
metabolism
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Female
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Humans
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Male
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Mesoderm
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cytology
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metabolism
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Middle Aged
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Nasal Mucosa
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metabolism
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Sinusitis
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metabolism
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pathology
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Vimentin
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metabolism
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Young Adult
2.Construction of fetal mesenchymal stem cell cDNA subtractive library.
Li YANG ; Dong-Mei WANG ; Liang LI ; Ci-Xian BAI ; Hua CAO ; Ting-Yu LI ; Xue-Tao PEI
Journal of Experimental Hematology 2002;10(2):89-92
UNLABELLEDTo identify differentially expressed genes between fetal mesenchymal stem cell (MSC) and adult MSC, especially specified genes expressed in fetal MSC, a cDNA subtractive library of fetal MSC was constructed using suppression subtractive hybridization (SSH) technique. At first, total RNA was isolated from fetal and adult MSC. Using SMART PCR synthesis method, single-strand and double-strand cDNAs were synthesized. After Rsa I digestion, fetal MSC cDNAs were divided into two groups and ligated to adaptor 1 and adaptor 2 respectively. Results showed that the amplified library contains 890 clones. Analysis of 890 clones with PCR demonstrated that 768 clones were positive. The positive rate is 86.3%. The size of inserted fragments in these positive clones was between 0.2 - 1 kb, with an average of 400 - 600 bp.
CONCLUSIONSSH is a convenient and effective method for screening differentially expressed genes. The constructed cDNA subtractive library of fetal MSC cDNA lays solid foundation for screening and cloning new and specific function related genes of fetal MSC.
Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Fetus ; Gene Library ; Humans ; Mesoderm ; cytology ; metabolism ; Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism
3.Investigation of mesenchymal-epithelial transdifferentiation in the morphogenesis mechanism of embryonic epidermic cells.
Du-yin JIANG ; Xiao-bing FU ; Yu-hua ZHANG ; Zhi-yong SHENG ; Wei CHEN ; Tong-zhu SUN
Chinese Journal of Surgery 2005;43(11):736-739
OBJECTIVETo study the relationship between the morphologic mechanism of human embryonic epidermic cells and mesenchymal-epithelial transformation (MET) and its modulation factor.
METHODSMorphological occurrence of epidermis was detected with histologic methods in earlier period [estimated gestational age (EGA) 6-14 weeks] human embryonic skin samples. At the same time, the characteristic expression and their distribution markers of mesenchymal cells [vimentin and alpha-smooth muscle actin (alpha-SMA)], embryonic specific epidermic protein CK8&18, specific protein of epidermic stem cell CK19, transforming growth factor-beta1) (TGF-beta1) and its receptor (TGFbetaRI) in embryonic epidermis were examined with immunohistochemistry and indirect-immunofluorescent doble-labelling method.
RESULTSDuring EAG 6-8 weeks, ectodermal cells containing Vim+/alpha-SMA(-) were found to transform into epidermal stem cells with CK8&18+/CK19+. In ectodermal cells, protein expression density of TGFbetaRI was moderate (+ +), while positive signal of TGFbeta1 was weak (+/-). After EGA10 weeks, epidermal cells showed typical morphological characteristics.
CONCLUSIONSAt EGA 6-8 weeks, human embryonic skin epidermal cells began to form through MET, in which the signal pathway mediated by TGFbetaRI might play important roles, but the role of TGFbeta1 need to be further studied.
Cell Differentiation ; physiology ; Epidermis ; cytology ; embryology ; Epithelial Cells ; cytology ; Humans ; In Vitro Techniques ; Mesoderm ; cytology ; Receptors, Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1 ; metabolism
4.Epithelial-mesenchymal transition and human fetal prostate development.
Jian-cheng ZHOU ; Guo-dong ZHU ; Kai-jie WU ; Jin ZENG ; Dong ZHANG ; Yan XUE ; Yu-le CHEN ; Xin-yang WANG ; Da-lin HE
National Journal of Andrology 2011;17(2):146-150
OBJECTIVETo investigate the role and significance of epithelial-mesenchymal transition (EMT) and its transcriptional regulator Twist1 in the development of the human fetal prostate.
METHODSTwenty-five human fetal prostate specimens at various developmental stages (16-39 weeks) were included in this study. EMT markers, such as E-Cadherin, N-Cadherin and Vimentin, and EMT transcriptional regulator Twist1 were determined by immunohistochemistry, and their relationship with the development of the human fetal prostate was analyzed.
RESULTSE-Cadherin was expressed in the fetal prostate epithelium only, while Vimentin, N-Cadherin and Twist1 in both the epithelium and the stroma. The expression of E-Cadherin gradually increased, but those of Vimentin, N-Cadherin and Twist1 gradually decreased with the gestation stages. No significant changes were observed in the staining patterns of Vimentin, N-Cadherin and Twist1 in the stroma during the whole developmental process.
CONCLUSIONEMT is involved in the development of the human fetal prostate, which may promote epithelial cell motility to form prostatic bud tubules in early gestation stages and boost the differentiation of prostate epithelia in later stages.
Cadherins ; metabolism ; Cell Dedifferentiation ; Epithelial Cells ; metabolism ; Epithelial-Mesenchymal Transition ; Fetal Development ; Humans ; Male ; Mesoderm ; metabolism ; Nuclear Proteins ; metabolism ; Prostate ; embryology ; growth & development ; metabolism ; Twist-Related Protein 1 ; metabolism ; Vimentin ; metabolism
5.Relationship between endothelial-to-mesenchymal transition and cardiac fibrosis in acute viral myocarditis.
Jun-yi HUA ; Zhao-cai ZHANG ; Xu-hong JIANG ; Yu-zhou HE ; Peng CHEN
Journal of Zhejiang University. Medical sciences 2012;41(3):298-304
OBJECTIVETo investigate the relationship between endothelial-to-mesenchymal transition (EndMT) and myocardial fibrosis in acute viral myocarditis (VMC).
METHODSTwenty-eight Balb/c mice were randomized into 3 groups: control group (n=8), VMC group(n=10) and intervention group(n=10). Mice in VMC and intervention groups were injected intraperitoneally(i.p) with single dose of coxsackievirus B3, mice in control group were injected with equal amount of viral-free vehicle. In the following day, mice in control and VMC groups were injected i.p with 0.1 ml of saline and intervention group with 0.1 ml of recombinant human bone morphogenetic protein 7(rh-BMP7) at a concentration of 300 μg/kg. The mice hearts were harvested after 7 d, cardiac collagen volume fraction (CVF) was calculated on picrosirius red-stained sections. mRNA and protein expression of TGF-β1, CD31, VE-cadherin, fibroblast special protein 1 (FSP-1) and α-smooth muscle actin (α-SMA) and collagen 1α1 in myocardiac tissues were detected by real-time RT-PCR and Western blot analysis, respectively.
RESULTSCompared to controls, overt fibrosis was presented in necrotic area of myocardium in VMC group. Meanwhile, marked increase of TGF-β1 expression accompanied with EndMT characterized by loss of endothelial phenotype (decreased expression of CD31 and VE-cadherin), gain of mesenchymal proteins (overexpression of FSP-1 and α-SMA) and increased synthesis of collagen was also demonstrated. Both EndMT and cardiac fibrosis were simultaneously reversed by TGF-β1 inhibition.
CONCLUSIONEndMT is involved in cardiac fibrosis in acute viral myocarditis, TGF-β1 might be a main mediator.
Acute Disease ; Animals ; Antigens, CD ; metabolism ; Cadherins ; metabolism ; Collagen ; metabolism ; Coxsackievirus Infections ; metabolism ; pathology ; Disease Models, Animal ; Endothelium ; pathology ; Fibrosis ; Male ; Mesoderm ; pathology ; Mice ; Mice, Inbred BALB C ; Myocarditis ; metabolism ; pathology ; virology ; Myocardium ; metabolism ; pathology ; Transforming Growth Factor beta1 ; metabolism
6.Epithelial to mesenchymal transition in the progression of tubulointerstitial fibrosis.
Chinese Medical Journal 2007;120(21):1925-1930
OBJECTIVETo review the mechanisms of epithelial to mesenchymal transition (EMT) and its role in the progression of tubulointerstitial fibrosis.
DATA SOURCESThe data used in this review were obtained mainly from the studies of EMT reported from 2000-2006.
STUDY SELECTIONRelevant articles on studies of EMT in tubulointerstitial fibrosis were selected. Data were mainly extracted from the 45 articles listed in the reference section of this review.
RESULTSThe process of EMT has gained wide recognition as candidate mechanism in progression of chronic fibrotic disorders. New markers were identified and facilitate the observation of EMT. EMT is regulated by many factors through activation of kinase-dependent signaling cascades. Recent findings suggest that EMT is a reversible process, which can be controlled by factors for their epithelial inducing activities.
CONCLUSIONRemarkable progresses of EMT research have been made recently. Preventing or reversing EMT is a promising strategy against renal fibrosis.
Animals ; Connective Tissue Growth Factor ; Disease Progression ; Epithelium ; metabolism ; pathology ; Fibrosis ; Humans ; Immediate-Early Proteins ; metabolism ; Intercellular Signaling Peptides and Proteins ; metabolism ; Mesoderm ; metabolism ; pathology ; Nephritis, Interstitial ; metabolism ; pathology ; Transforming Growth Factors ; metabolism
7.Interaction of Mesenchymal Stem Cells and Osteoblasts for in vitro Osteogenesis.
Hyongbum KIM ; Jong Hun LEE ; Hwal SUH
Yonsei Medical Journal 2003;44(2):187-197
It has recently been reported that bone marrow-derived mesenchymal stem cells (MSCs), which are systemically administrated to different species, undergo site-specific differentiation. This suggests that the tissue specific cells may cause or promote the differentiation of the MSCs toward their cell type via a cell-to-cell interaction that is mediated not only by hormones and cytokines, but also by direct cell-to-cell contact. In this study, in order to assess the possible synergistic interactions for osteogenesis between the two types of cells, the MSCs derived from rabbit bone marrow were co-cultured with rat calvarial osteoblasts in direct cell-to-cell contact in a control medium (CM) and in an osteogenic medium (OM). The cell number, alkaline phosphatase activity, and amount of calcium deposition were assayed in the cultures of MSCs, osteoblasts, and co-cultures of them in either OM or CM for up to 40 days. The cell numbers and the alkaline phosphatase activities in the co-culture were somewhere in between those of the osteoblast cultures and the MSC cultures. The amounts of deposited calcium were lower in the co-culture compared to those of the other cultures. This suggests that there are little synergistic interactions during osteogenesis in vitro between the rat osteoblasts and rabbit MSCs.
Alkaline Phosphatase/metabolism
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Animals
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Calcification, Physiologic
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*Cell Communication
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Cell Count
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Cell Differentiation
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Cell Division
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Female
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Mesoderm/*cytology
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Osteoblasts/*physiology
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*Osteogenesis
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Rabbits
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Stem Cells/*physiology
8.Impact of salvianolic acid-B on TGF-beta1-induced HK-2 epithelial-mesenchymal transition.
Juan ZHOU ; Fei WANG ; Haiying LU ; Yue ZHANG
China Journal of Chinese Materia Medica 2010;35(1):89-93
OBJECTIVETo investigate the effect of salvianolic-acid B (SA-B) on epithelia-mesenchymal transition in human renal proximal tubular cells (HK2), induced by transforming growth factor beta1 (TGF-beta1).
METHODEpithelia-mesenchymal transition (EMT) was induced with TGF-beta1 in HK2 cultured in vitro. Different concentrations (2, 5, 10, 20 microg x L(-1)) and stimulant periods (12, 24, 48 h) were tried to find the perfect condition for EMT. At the same time bone morphogenetic protein-7 (BMP-7, positive control) and the SA-B intervention were given to observe their effect on EMT. Western blot and immunofluorescent microscopy were used to analyze the expression of E-cadherin and alpha-smooth muscle actin (alpha-SMA) in HK2.
RESULTBMP-7 significantly inhibited the down-regulation of E-cadherin and the up-regulation of alpha-SMA induced by TGF-beta1 (P < 0.05), and SA-B significantly inhibited the up-regulation of alpha-SMA expression induced by TGF-beta1 (P < 0.05), but not the down-regulation of E-cadherin induced by TGF-beta1.
CONCLUSIONSA-B and BMP-7 can inhibit TGF-beta1-induced EMT in HK2. Their common role is to inhibit the up-regulation of alpha-SMA, and the effect of SA-B on the regulation of E-cadherin needs further study to be confirmed.
Benzofurans ; pharmacology ; Bone Morphogenetic Protein 7 ; pharmacology ; Cadherins ; metabolism ; Cell Differentiation ; drug effects ; Cell Line ; Epithelial Cells ; pathology ; Humans ; Mesoderm ; pathology ; Transforming Growth Factor beta1 ; pharmacology
9.Epithelial-Mesenchymal Transitions of Bile Duct Epithelial Cells in Primary Hepatolithiasis.
Lijin ZHAO ; Rigao YANG ; Long CHENG ; Maijian WANG ; Yan JIANG ; Shuguang WANG
Journal of Korean Medical Science 2010;25(7):1066-1070
The purpose of this study was to explore the role of epithelial-mesenchymal transition in the pathogenesis of hepatolithiasis. Thirty-one patients with primary hepatolithiasis were enrolled in this study. Expressions of E-cadherin, alpha-catenin, alpha-SMA, vimentin, S100A4, TGF-beta1 and P-smad2/3 in hepatolithiasis bile duct epithelial cells were examined by immunohistochemistry staining. The results showed that the expressions of the epithelial markers E-cadherin and alpha-catenin were frequently lost in hepatolithiasis (32.3% and 25.9% of cases, respectively), while the mesenchymal markers vimentin, alpha-SMA and S100A4 were found to be present in hepatolithiasis (35.5%, 29.0%, and 32.3% of cases, respectively). The increased mesenchymal marker expression was correlated with decreased epithelial marker expression. The expressions of TGF-beta1 and P-smad2/3 in hepatolithiasis were correlated with the expression of S100A4. These data indicate that TGF-beta1-mediated epithelial-mesenchymal transition might be involved in the formation of hepatolithiasis.
Adult
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*Bile Ducts/cytology/metabolism/pathology
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Biological Markers/*metabolism
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Cell Differentiation/*physiology
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Epithelial Cells/cytology/*physiology
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Epithelium/physiology
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Female
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*Gallstones/metabolism/pathology
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Humans
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Liver Diseases/metabolism/*pathology
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Male
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Mesoderm/cytology/*physiology
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Middle Aged
10.Role of integrin-linked kinase in renal tubular epithelial-mesenchymal transition and the regulatory effect of urokinase on its expression in mice with obstructive nephropathy.
Feng HE ; Lu YU ; Jun-rong TONG ; Zheng-mao LUO ; Qi-zhi ZHU ; Yin WANG ; Jian-lin ZHANG
Journal of Southern Medical University 2009;29(5):909-913
OBJECTIVETo investigate the role of integrin-linked kinase (ILK) on renal tubular epithelial-mesenchymal transition and the regulatory effect of urokinase on LIK expression in mice with obstructive nephropathy.
METHODSNormal male mice were randomly divided into sham-operated group (n=20), unilateral ureteral obstruction (UUO) group (n=28), and UUO with urokinase treatment group (uPA, n=28), and UUO was induced surgically in the latter two groups. The mice were sacrificed on days l, 3, 7 and 14 after the surgery, and renal interstitial fibrosis (RIF) was graded according to the result of Masson staining. The expression of ILK in the renal tissues of the rats was examined by immunofluorescence staining and Western blotting, and the expression of E-cadherin was detected by immunohistochemistry. RT-PCR was used to examine the mRNA expressions of ILK, E-cadherin and alpha-smooth muscle actin (alpha-SMA).
RESULTSThe expressions of ILK mRNA and protein were significantly increased in UUO group, but significantly decreased by treatment with uPA (P<0.05). The expression of alpha-SMA mRNA level was significantly increased, while E-cadherin decreased in mice with UUO on day 3 after the surgery. Treatment with uPA significantly inhibited such effects (P<0.05).
CONCLUSIONILK plays an important role in renal interstitial fibrosis by mediating epithelial-mesenchymal transition. Urokinase attenuates renal tubulointerstitial fibrosis in mice with UUO possibly by inhibiting ILK expression and preventing tubular epithelial-mesenchymal transition.
Animals ; Cell Transdifferentiation ; drug effects ; Epithelial Cells ; metabolism ; pathology ; Fibrosis ; Kidney Tubules ; metabolism ; pathology ; Male ; Mesoderm ; pathology ; Mice ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; physiology ; Ureteral Obstruction ; genetics ; metabolism ; pathology ; Urokinase-Type Plasminogen Activator ; pharmacology