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BACKGROUND:Fibronectin type Ⅲ domain-containing protein 5(FNDC5)is a muscle factor that can regulate glucose and lipid metabolism,and exert anti-inflammatory,anti-oxidative effects and improvement in insulin resistance.Moreover,FNDC5 could control various cell pyroptosis. OBJECTIVE:To explore the effect and potential mechanism of FNDC5 on macrophage pyroptosis. METHODS:(1)After completing the construction of the lentivirus virus overexpressing FNDC5 or silencing FNDC5,the THP-1 cells were transfected with the lentivirus vector.The result of transfection was detected by the expression of green fluorescence,qPCR,and western blot assay.(2)Phorbol ester induced THP-1 cells to differentiate into macrophages.Oxidized low-density lipoprotein(ox-LDL)was added to induce the cell pyroptosis model.There were six groups,i.e.,NC group,ox-LDL group,ox-LDL+MOCK1 group,ox-LDL+Ov-FNDC5 group,ox-LDL+MOCK2 group,and ox-LDL+shFNDC5 group.(3)The level of cell pyroptosis was evaluated by Hoechst 33342/propidium iodide fluorescence double staining and lactate dehydrogenase release assay.The expression levels of related molecules in THP-1 cells were analyzed by qPCR and western blot assay.The interleukin-18 and interleukin-1β in cell supernatant were detected by ELISA. RESULTS AND CONCLUSION:Compared with the ox-LDL+MOCK1 group,overexpression of FNDC5 significantly reduced the pyroptosis rate of macrophages and the release levels of lactate dehydrogenase,interleukin-1β and interleukin-18,significantly inhibited the mRNA expression of NF-κB p65,NF-κB p50,NLRP3,ASC,Caspase-1,and GSDMD,and significantly inhibited the protein expression of NF-κB p65,NF-κB p50,NLRP3,ASC,cleaved Caspase-1 and GSDMD-N.Compared with the ox-LDL+MOCK2 group,the silence of FNDC5 showed the opposite result.These findings suggest that FNDC5 attenuates pyroptosis in macrophages by inhibiting the NF-κB/NLRP3 pathway.
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BACKGROUND:At present,bone marrow mesenchymal stem cells are the main seed cells used in cell therapy for the treatment of intervertebral disc degeneration.However,the use of bone marrow mesenchymal stem cells as seed cells for the regeneration of fibrous rings is at risk of heterotopic ossification and teratoma at the repair site.Therefore,it is of great economic and social significance to find a new kind of seed cells for tissue engineering of annulus fibrosus for the treatment of intervertebral disc degeneration. OBJECTIVE:To isolate and purify rat annulus fibrosus-derived stem cells by adherent method combined with fibronectin differential adhesion screening method,and to observe its purification effect and biological characteristics. METHODS:Annulus fibrosus tissue was obtained from a SD rat intervertebral disc.Primary annulus fibrosus-derived stem cells were obtained by the mechanical-enzymatic digestion method.Annulus fibronectin differential adhesion method was used to purify annulus fibrosus-derived stem cells.Morphological changes and proliferation of cells were observed through a microscope.The expression levels of stem cell markers were detected by immunofluorescence technique and qPCR.The screened cells were subjected to multi-lineage cell differentiation and characteristic gene detection. RESULTS AND CONCLUSION:(1)The purified cells grew well,and most of them were angular and star-shaped multi-process cells,which had good proliferation ability.(2)Cells were positive for cell membrane surface antigens CD73,CD90 and CD105,while negative for CD45 and CD34.(3)After specific induction,cells could successfully differentiate into osteoblasts,chondroblasts and lipoblasts.(4)Collagen-I,Runx-2 after osteogenic induction,Collagen Ⅱ,Sox-9 after chondrogenic induction,and PPAR-γ and LPL after lipogenic induction were highly expressed in cells,and the difference was significant compared with that before induction(P<0.05).(5)These findings confirm that the adherent method combined with fibronectin differential adhesion method is effective enough to screen,isolate and purify rat annulus fibrosus-derived stem cells,and has good cell biological properties,good proliferation ability and multiple differentiation potential.
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{L-End}Objective To investigate the effect and mechanism of low dose metformin in delaying pulmonary fibrosis in silicosis mice. {L-End}Methods The specific pathogen free C57BL/6 male mice were randomly divided into four groups,with six mice in each group. Mice in the silicosis model group and the metformin intervention group were given 20 μL of a mass concentration of 250 g/L silica suspension, and mice in the blank control group and the drug control group were given 20 μL of 0.9% sodium chloride solution, using tracheal exposure method. After 72.0 hours of dust exposure, the mice of drug control group and metformin intervention group were intraperitoneally injected with metformin at a dose of 65 mg/kg body mass, while the mice in the blank control group and the silicosis model group were given 0.9% sodium chloride solution at the same volume, once every other day for 28 days. After the treatment, histopathological change of the lungs was observed, lung organ coefficient was calculated, degree of pulmonary fibrosis was evaluated with Ashcroft score, and mRNA expression of fibronectin (Fn)1 and collagen typeⅠ(COLⅠ) alpha 1 (Col1a1) in lung tissues were detected by real-time fluorescence quantitative polymerase chain reaction. The relative expression of FN and COLⅠ in lung tissues was determined by Western blot. {L-End}Results The results of histopathological examination of the lungs showed that there were no inflammation and fibrosis in the lungs of mice in the blank control group and the drug control group; mice in silicosis model group had inflammation and fibrosis in lung; the degree of lung inflammation and fibrosis was reduced in the mice of metformin intervention group compared with the silicosis model group. The lung organ coefficient, Ashcroft score, the relative expression of Fn1 and Col1a1 mRNA, the relative expression of FN and COLⅠprotein in lung tissues increased in silicosis model group (all P<0.05), compared with those in both blank control group and drug control group. The indexes above decreased of mice in the metformin intervention group than those in the silicosis model group (all P<0.05). {L-End}Conclusion Low-dose metformin can delay the progression of pulmonary fibrosis in silicosis mice. The mechanism may be related to metformin's improving excessive deposition of extracellular matrix induced by silica.
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Aim To investigate the therapeutic effect of Jiangtang Wan (JTW) on mioe with diabetic kidney disease (DKD) and to explore its potential moiecuiar mechanism on ameliorating renal injury and fibrosis. Methods C57BL/6 mice were randomly divided into four groups: the control group, the model group, JTW group (1250 mg • kg
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Objective@#To investigate the mechanism of resveratrol promoting fibronectin type Ⅲ domain-containing 5 (FNDC5) degradation in skeletal muscle of male obese mice．@*Methods@#Six-week-old male C57BL /6 mice were randomly divided into three groups : standard control diet ( SCD) ，high-fat diet ( HFD) and high-fat diet treated with resveratrol (HFD + RES) ．HFD + RES group was intervened with resveratrol via gavage ［400 mg / kg · d) ］ while fed HFD for 20 weeks．The body mass，serum TG，TC，LDL-C and HDL-C levels were detected．The pathological changes in skeletal muscle were detected by HE staining．The expression of FNDC5，SIRT1，SIRT2，LC3， p62，Beclin-1，ATG5，ATG7 was assessed by immunohistochemistry，RT-PCR and Western blot respectively. @*Results@#The body mass ，serum TG ，TC and LDL-C levels increased significantly ，meanwhile HDL-C levels decreased in HFD group．Lipid deposition between skeletal muscle fibers were obvious in HFD group．The immuno- histochemistry results showed that protein expression levels of SIRT1，SIRT2 and LC3 obviously decreased，while the protein levels of FNDC5 and p62 obviously increased．The expression levels of FNDC5 significantly increased， while the gene expression levels of SIRT1，SIRT2，LC3，Atg7 and Beclin-1 obviously decreased．All these responses were attenuated by treatment with RES．@*Conclusion@#RES has obvious effects of lipid-lowering and promoting FNDC5 degradation in skeletal muscle tissues，which may be related with SIRT1 and SIRT2-induced autophagy， thus resulting in degradation of FNDC5 .
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【Objective】 To study the removal effect of fibronectin(Fn) from von willebrand factor(vWF) by ion-exchange chromatography through processing human coagulation factor Ⅷ chromatographic washing products, in order to select a method that can effectively reduce Fn without compromising the activity yield. 【Methods】 In a multi-batch process development experiment, Fractogel® EMD TMAE(M) strong anion filler produced by Merck(Germany) was used to conduct chromatography to investigate vWF ristomycins titer (vWF: RCof), vWF recovery, protein content and Fn content. 【Results】 During the development of vWF pilot purification process, the content of Fn in the samples can be effectively reduced by ion-exchange chromatography, with removal rate more than 87%, titer recovery of vWF more than 80%, and no significant change in other quality indexes. 【Conclusion】 The use of ion-exchange chromatography to purify vWF can effectively reduce the content of Fn, which has positive significance for developing new product process and improving the product quality of blood products manufacturers.
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ObjectiveTo investigate the effects of Biling Qutong prescription （BLQT） on serum levels of NOD-like receptor thermal protein domain associated protein 3 （NLRP3）， purinergic ligand-gated ion channel 7 receptor （P2X7R）， fibronectin （FN）， and hepatic steatosis in patients with type 2 diabetes mellitus （T2DM） complicated with gouty arthritis （GA）. MethodSixty-four patients diagnosed with T2DM comorbid with GA and treated at the First Affiliated Hospital of Anhui University of Chinese Medicine from January 2019 to December 2022 were enrolled and randomly divided into a BLQT group （Chinese medicine group， 32 cases） and the ibuprofen group （western medicine group， 32 cases）. Thirty healthy individuals who underwent routine health examinations during the same period were assigned to the control group. The BLQT group and the western medicine group received basic treatment along with BLQT and ibuprofen， respectively. After 8 weeks of continuous treatment， the traditional Chinese medicine syndrome score （TCMSS） of the patients was evaluated before and after treatment. The differences in fasting plasma glucose （FPG）， 2-hour postprandial plasma glucose （2 h PG）， glycated hemoglobin （HbA1c）， fasting insulin （FINS）， homeostasis model assessment of insulin resistance （HOMA-IR）， serum uric acid （SUA）， serum creatinine （SCr）， total cholesterol （TC）， triglyceride （TG）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， controlled attenuation parameter （CAP）， liver stiffness measurement （LSM）， NLRP3， P2X7R， and FN levels before and after treatment were compared. Adverse drug reactions that occurred during treatment were recorded. ResultThe TCMSS for joint redness， swelling， pain， joint burning， yellow urine， and red tongue with yellow and greasy coating， as well as total score were significantly reduced in both the BLQT group and the western medicine group as compared with those before treatment （P<0.05， P<0.01）. The BLQT group also showed a significant reduction in symptom scores such as dry mouth， polyuria， polydipsia， and slippery and rapid pulse （P<0.01）. Compared with the western medicine group after treatment， the BLQT group exhibited a more significant reduction in all symptom scores and total score （P<0.05， P<0.01）. The BLQT group and the western medicine group showed a decrease in FPG， 2 h PG， HbA1c， SCr， SUA， TG， TC， and LDL-C levels （P<0.05， P<0.01） after treatment， and the BLQT group showed decreased HOMA-IR， ALT， AST， and HDL-C levels （P<0.05， P<0.01） compared with those before treatment. When compared with the western medicine group after treatment， the BLQT group showed a more significant reduction in all laboratory parameters except for HDL-C （P<0.05， P<0.01）. Before treatment， NLRP3， P2X7R， and FN levels in both the BLQT group and the western medicine group were higher than those in the control group （P<0.01）. After treatment， NLRP3 and P2X7R levels in both groups significantly decreased （P<0.01）， and FN levels in the BLQT group also decreased significantly （P<0.01）. When compared with the western medicine group after treatment， the BLQT group showed a more significant reduction in NLRP3， P2X7R， and FN levels （P<0.01）. Before treatment， CAP and LSM levels in both the BLQT group and the western medicine group were higher than those in the control group （P<0.01）. After treatment， CAP and LSM levels in both groups decreased （P<0.05， P<0.01）. Compared with the western medicine group after treatment， the BLQT group showed a more significant reduction in CAP and LSM （P<0.01）. The incidence of adverse reactions was 3.13% （1/32） in the BLQT group and 15.63% （5/32） in the western medicine group， with no significant difference. ConclusionBLQT has good efficacy in patients with T2DM complicated with GA， which can significantly alleviate joint redness， swelling， heat， pain， limited mobility， dry mouth， and polydipsia， reduce blood glucose， uric acid， and lipid levels， suppress the high expression of NLRP3， P2X7R， and FN， and improve hepatic steatosis.
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ABSTRACT Background: Hematopoietic stem/progenitor cell transplantation is the main treatment option for hematological malignancies and disorders. One strategy to solve the problem of low stem cell doses used in transplantation is pre-transplant expansion. We hypothesized that using fibronectin-coated microfluidic channels would expand HSPCs and keep self-renewal potential in a three-dimensional environment, compared to the conventional method. We also compared stem cell homing factors expression in microfluidic to conventional cultures. Materials and methods: A microfluidic device was created and characterized by scanning electron microscopy. The CD133+ cells were collected from cord blood and purified. They were subsequently cultured in 24-well plates and microfluidic bioreactor systems using the StemSpan serum-free medium. Eventually, we analyzed cell surface expression levels of the CXCR4 molecule and CXCR4 mRNA expression in CD133+ cells cultured in different systems. Results: The expansion results showed significant improvement in CD133+ cell expansion in the microfluidic system than the conventional method. The median expression of the CXCR4 in the expanded cell was lower in the conventional system than in the microfluidic system. The CXCR4 gene expression up-regulated in the microfluidic system. Conclusion: Utilizing microfluidic systems to expand desired cells effectively is the next step in cell culture. Comparative gene expression profiling provides a glimpse of the effects of culture microenvironments on the genetic program of HSCs grown in different systems.
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Fibronectines , Hémopathies , Cellules souches tumorales , Cellules souches hématopoïétiques , Tumeurs hématologiques , Bioréacteurs , Récepteurs CXCR4 , Sang foetalRésumé
@#[Abstract] Objective: To investigate the effects of fibronectin Ⅲ domain containing protein 10 (FNDC10) on the proliferation, migration and invasion of breast cancer cells, and to primarily explore the mechanism. Methods: TCGA database was used to analyze the expression of FNDC10 in breast cancer tissues. The mRNA level of FNDC10 in normal immortalized breast cells (MCF-10A) and breast cancer cells (MCF-7, MDA-MB-231, BT549, MDA-MB-468, HCC1806, HCC1937) was detected by qPCR. MCF-7 and MDA-MB-231 cells were transfected with FNDC10 siRNA or NC-siRNA for functional experiments. CCK-8 assay was used to detect the effect of FNDC10 on the proliferation of breast cancer cells. Colony forming assay was used to detect the colony forming ability of breast cancer cells. Transwell assay was used to detect the effect of FNDC10 on migration and invasion of breast cancer cells. WB was used to detect the changes of metastasis-related molecules and cell signaling pathways at protein level. Results:The expression of FNDC10 in breast cancer tissues was significantly higher than that in normal tissues (P<0.01), and the expression level of FNDC10 in breast cancer MCF-7 and MDA-MB-231 cells was higher than that in normal breast cells (P<0.01 or P<0.05). Knocking down FNDC10 expression inhibited the proliferation, migration and invasion of breast cancer cells (P<0.01 or P<0.05). The mechanism study showed that knockdown of FNDC10 expression inhibited STAT3 activation in breast cancer cells (P<0.01 or P<0.05) and enhanced the expression of EMT maker E-cadherin (P<0.05), leading to the suppression of EMT progression. Conclusion: FNDC10 promotes proliferation and EMT of breast cancer cells through activating STAT3 signaling pathway, thereby promoting the malignant progression of breast cancer.
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@#Introduction: The internalization process of group A streptococci (GAS) into human cells is one of the crucial steps in the pathogenesis of GAS infections, which could also affect their susceptibility responses toward several antibiotics. Currently, data on the distribution of internalization-associated genes and susceptibility patterns are still lacking in Malaysia. This study investigated the distribution of fibronectin-binding protein F1 (prtF1) and streptococcal pyrogenic exotoxin B (speB) genes in GAS isolates with their susceptibility profiles and source of samples. Methods: We used 43 GAS isolates from our previous stock culture and performed antibiotic susceptibility testing by Kirby-Bauer disk diffusion method and interpreted the results according to the established guidelines. We detected virulence (prtF1 and speB) and resistance (ermA, ermB, mefA, tetM and lnuA) genes by PCR method using established primers and protocols. Results: High resistance rates were observed against doxycycline (58.1%) and clindamycin (16.3%). In comparison, 100.0% and 46.5% of GAS isolates carried speB and prtF1 genes, respectively. tetM and lnuA genes were detected in all respective resistant isolates (100% for each). No macrolide resistance genes were detected. Interestingly, prtF1 gene was highly distributed in doxycycline-resistant than doxycycline-sensitive isolates (60.0% versus 27.8%). Conclusions: High resistance rate of GAS toward doxycycline in our study may potentially reflect the uncontrol dissemination of tetM gene among our isolates. The presence of prtF1 gene among this strain would enhance its ability to evade the intracellular action of antibiotics, which may affect the management of GAS diseases. Thus, close monitoring of GAS by molecular methods is required in the future.
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@#ObjectiveTo investigate the value of peroxisome proliferator-activated receptor γ（PPARγ）level in predicting hemorrhagic transformation in patients with non thrombolytic acute cerebral infarction. MethodsA total of 92 patients with non thrombolytic acute cerebral infarction who were treated in our hospital from May 2017 to May 2020 were selected as the research objects，and they were divided into hemorrhagic transformation group（n=14）and non hemorrhagic transformation group（n=78）according to the occurrence of hemorrhagic transformation. The plasma levels of PPARγ and fibronectin（FN）were detected by enzyme-linked immunosorbent assay（ELISA）；the correlation between plasma PPAR-γand blood lipid levels in patients with hemorrhagic transformation of non thrombolytic acute cerebral infarction was analyzed；the predictive value of plasma PPARγ and FN levels on hemorrhagic transformation in patients with non thrombolytic acute cerebral infarction was analyzed；the influencing factors of hemorrhagic transformation in patients with non thrombolytic acute cerebral infarction were analyzed. ResultsThe levels of PPARγ and FN were higher than those in non hemorrhagic transformation group，The area under the curve（AUC）of plasma PPARγ and FN levels in predicting hemorrhagic transformation in patients with non thrombolytic acute cerebral infarction was 0.933 and 0.860，respectively，the specificity was 89.7% and 78.2%，and sensitivity was 85.7% and 78.6%，respectively；the AUC of the combined prediction was 0.952，the specificity was 89.7%，and the sensitivity was 92.9%. PPARγ was an independent risk factor for hemorrhagic transformation in patients with non thrombolytic acute cerebral infarction（P<0.05）. ConclusionPlasma PPARγ is a factor influencing hemorrhagic transformation in patients with non thrombolytic acute cerebral infarction，which may have important predictive value for secondary hemorrhagic transformation.
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Abstract Potent signaling agents stimulate and guide pulp tissue regeneration, especially in endodontic treatment of teeth with incomplete root formation. Objective This study evaluated the bioactive properties of low concentrations of extracellular matrix proteins on human apical papilla cells (hAPCs). Methodology Different concentrations (1, 5, and 10 µg/mL) of fibronectin (FN), laminin (LM), and type I collagen (COL) were applied to the bottom of non-treated wells of sterilized 96-well plates. Non-treated and pre-treated wells were used as negative (NC) and positive (PC) controls. After seeding the hAPCs (5×103 cells/well) on the different substrates, we assessed the following parameters: adhesion, proliferation, spreading, total collagen/type I collagen synthesis and gene expression (ITGA5, ITGAV, COL1A1, COL3A1) (ANOVA/Tukey; α=0.05). Results We observed greater attachment potential for cells on the FN substrate, with the effect depending on concentration. Concentrations of 5 and 10 µg/mL of FN yielded the highest cell proliferation, spreading and collagen synthesis values with 10 µg/mL concentration increasing the ITGA5, ITGAV, and COL1A1 expression compared with PC. LM (5 and 10 µg/mL) showed higher bioactivity values than NC, but those were lower than PC, and COL showed no bioactivity at all. Conclusion We conclude that FN at 10 µg/mL concentration exerted the most intense bioactive effects on hAPCs.
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Humains , Protéines de la matrice extracellulaire , Fibronectines , Adhérence cellulaire , Cellules cultivées , Laminine , Collagène de type I , Matrice extracellulaireRésumé
【Objective】 To investigate the role of dual-specific phosphatase 6 （DUSP6） in the regulation of human lens epithelial cells （HLECs） epithelial-mesenchymal transition （EMT） and extracellular matrix （ECM） synthesis induced by transforming growth factor β2 （TGF-β2）. 【Methods】 Human lens epithelial B3 （HLE-B3） cells were treated with 10 μg/L of TGF-β2 for 0 h，12 h， 24 h and 48 h. Then we detected the expressions of α-smooth muscle actin （α-SMA） and fibronectin （Fn） by real-time fluorescent quantitative PCR （RT-qPCR） and Western blotting. The overexpression vector of DUSP6 was constructed and transfected into HLE-B3 cell line and divided into three groups： TGF-β2， empty vector （EV）+TGF-β2， and DUSP6+TGF-β2. RT-qPCR and Western blotting were used to detect the effect of overexpression of DUSP6 on the expressions of α-SMA and Fn. Scratch test and Transwell migration test were used to evaluate the effect of DUSP6 overexpression on cell migration. 【Results】 Compared with the control group， the expressions of DUSP6 mRNA and protein decreased after 10 μg/L of TGF-β2 treatment for 24 h （P<0.05）. Compared with EV+TGF-β2 group， DUSP6+TGF-β2 group inhibited the migration of HLE-B3 cells and the expressions of α-SMA and Fn （P<0.05）. 【Conclusion】 DUSP6 could inhibit EMT and ECM synthesis in lens epithelial cells induced by TGF-β2， which might provide a potential therapeutic strategy for posterior capsule opacification.
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Hypertensive disorders complicate 2% to 8% of pregnancies globally. They are one of the leading causes of maternal mortality responsible for 16% of maternal deaths. Preeclampsia is a pregnancy specific syndrome whose pathophysiologic features have not been clearly established, but research during past two decades has suggested that maternal endothelial damage and improper placental development are involved in the genesis of preeclampsia. Fibronectin is known to be a marker of endothelial dysfunction, which occurs in early gestation in women who develop preeclampsia in later gestation and hence levels of fibronectin may vary in first trimester itself in such women. We wanted to examine the usefulness of single biomarker ‘plasma fibronectin’ in screening of pregnant women for gestational hypertension/preeclampsia, study the difference in fibronectin levels in early versus late onset gestational hypertension/preeclampsia and evaluate its levels in preeclampsia with severe features.METHODS200 antenatal women with singleton pregnancy, who were normotensive were enrolled in the study and plasma fibronectin levels were measured at 10-12 weeks of gestation. Women were followed throughout pregnancy and 12 weeks postpartum. Plasma fibronectin levels were compared between normotensive women and women who developed gestational hypertension/preeclampsia.RESULTSThe mean values of plasma fibronectin are significantly higher in group who developed gestational hypertension/preeclampsia compared to group who remained normotensive (167+/-81 vs 114+/-58; p<0.05). The mean value in group with early onset disease as well as preeclampsia with severe features is higher than that of group with late onset disease and preeclampsia without severe features respectively. But the difference is not statistically significant.CONCLUSIONSStudy showed that plasma fibronectin could be used as a marker for early prediction of gestational hypertension/ preeclampsia.
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Aim To investigate the effect of integrin av ß3 inhibitor (Cyclo-RGDfK) on fibronectin (F N) - induced epithelial-mesenchymal transition in human mammary epithelial cells MCF-10A. Methods M C F - 10A cells were cultured in vitro, and the model of epithelial-mesenchymal transition (E M T) induced by FN was established. The expressions of otv integrin, ß3 integrin, epithelial marker E-cadherin and mesenchymal markers N-cadherin and Vimentin were measured by Western blot. The migration ability and invasion ability of cells were detected by Scratch repair experiment and Transwell chamber experiment. Results Western blot results showed that the expressions of ay integrin, ß3 integrin, N-cadherin, and Vimentin significantly were up-regulated and the expression of E-cadherin protein down-regulated after FN treatment for 48 h. After administration of Cyclo-RGDfK, the increased expression of N-cadherin and Vimentin and decreased expression of E-cadherin induced by FN were significantly reversed. Similarly, the results of Scratch repair experiments and Transwell experiments showed that FN significantly enhanced the ability of MCF-10A cells to migrate and invade, and Cyclo-RGDfK significantly inhibited the induction of FN. Conclusions Cyclo-RGDfK can inhibit the process of EMT induced by FN and reduce the invasion and migration of MCF-10A, which may be a potential drug for the treatment of breast cancer metastasis.
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Objective:To investigate the effect of tetrandrine on transforming growth factor-β1（TGF-β1）stimulated MRC-5 cells. Method:Different concentrations of TGF-β1 (0, 2.5, 5, 10, 20, 40 μg·L-1) were applied to MRC-5 cells. Proliferation toxicity of TGF-β1 to MRC-5 was detected by cell counting kit-8 (CCK-8) method. Detection of alpha smooth muscle actin (α-SMA) and Vimentin's expression levels in MRC-5 by Western blot. Detection of changes of collagen I(Col-I) and fibronectin (FN)'s expression levels in MRC-5 supernatants by enzyme linked immunosorbent assay(ELISA) kit. And the appropriate concentration of TGF-β1 activated MRC-5 cells was screened. The appropriate concentration of TGF-β1 and different concentrations of Tet (0, 2.5, 5, 10, 20, 40 μmol·L-1) were applied to MRC-5 cells, and CCK-8 method was used to screen safe concentration again. Western blot was used to detect changes in α-SMA and Vimentin expression levels in MRC-5 cells, and ELISA method to detect changes in Col-I and FN in MRC-5 cell supernatant. Result:Compared with the blank group, 20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 24 hours (P<0.05), and 10，20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 48 h (P<0.05).When Tet is added for 24 h, the half inhibitory concentration (IC50) value was 14.07 μmol·L-1, and when cultured for 48 h, the IC50 value was 7.51 μmol·L-1. Compared with the blank group, the relative contents of α-SMA, FN and Col-I in the 5 μg·L-1 of TGF-β1 group were obviously increased (P<0.05), and the relative contents of Vimentin were significantly increased (P<0.01), and the relative contents of FN and Col-I, α-SMA and Vimentin in 10 μg·L-1 group were significantly increased (P<0.01). 10 μg·L-1 of TGF-β1 was co-cultured with Tet at different concentrations. Compared with the TGF-β1 group, the relative levels of α-SMA, Vimentin and FN in the 5 μmol·L-1 of Tet group were significantly reduced (P<0.01), and the relative levels of Col-I were obviously reduced (P<0.05). In the Tet 10 μmol·L-1 group, the relative contents of the α-SMA, Vimentin, FN and Col-I were significantly reduced (P<0.01). Conclusion:TGF-β1 can increase the levels of Col-I, FN and other extracellular matrices in MRC-5 cells, and Tet can effectively inhibit the occurrence of this change. It is suggested that Tet may inhibit secreting extracellular matrix of fibroblasts in the formation of pulmonary fibrosis.
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Resumen Staphylococcus aureus se caracteriza por ser la principal causa de bacteriemia nosocomial en el mundo, debido al incremento en la resistencia, a los diferentes factores de patogenicidad y virulencia y la expresión de una gran variedad de proteínas las cuales pertenecen a las moléculas de la matriz adhesiva (MSCRAMM), presentes en la superficie de la bacteria cuya función es la colonización e invasión celular al hospedero y favorecer la formación de biopelícula, El conjunto de estos mecanismos de patogenicidad y virulencia, le permiten a la bacteria persistir en el huésped y en el ambiente, sobreviviendo a factores adversos, al sistema inmune y a los antimicrobianos.
Abstract Staphylococcus aureus is a microorganism characterized by being the main cause of nosocomial bacteremia in different places of the world, due to the different virulence and pathogenicity factors. One of the most important is the biofilm formation, which greatly favors bacterial resistance. For the adhesion of the biofilm to biotic and abiotic surfaces, the microbial surface components recognizing adhesive matrix molecules (MSCRAMM), these proteins play a key role in host cell colonization and invasion by the bacteria.
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Staphylococcus aureus , Bactériémie , Facteurs de virulence , Système immunitaire , Anti-infectieuxRésumé
Background: Preterm birth defined as birth before 37 weeks of gestation is a significant public health issue. Identification of patients at risk of preterm labour while ruling out those who are not is a fundamental but challenging goal for clinicians. This study was done to evaluate bed side dipstick test for detecting fetal fibronectin in cervico-vaginal secretions as a predictor of preterm delivery in symptomatic and asymptomatic high risk group.Methods: This was a hospital based prospective, double blinded study. We enrolled 100 pregnant women presenting with or without symptoms of preterm delivery, between 20 and 35 weeks of gestation. A rapid bed side dipstick test was performed to detect FFN in cervico-vaginal secretions of all enrolled women (symptomatic and asymptomatic high risk women) and results were evaluated for prediction of preterm labour. Qualitative data were analyzed by using Chi-square and Fisher’s exact test and quantitative data were analyzed by using unpaired Student’s t test and Mann-Whitney test. P value < 0.05 was considered significant.Results: In symptomatic group sensitivity, specificity, PPV and NPV of FFN test in predicting delivery within 48 hours, 7days 14days and preterm delivery was 100%, 63.2%, 46.2%, 100%; 100%, 72.7%, 65.4%, 100%; 100%, 75%, 69.2%, 100%; 80%, 76%, 76.9%, 79.2% respectively. In asymptomatic high risk group, sensitivity, specificity, PPV and NPV of FFN test in predicting preterm delivery (<37weeks) was 0%, 87.5%, 0%, 77.8%.Conclusions: The high negative predictive value may be of value in avoiding unnecessary interventions with potentially hazardous medications and identifying symptomatic women who are not in true labour and also allaying anxiety of asymptomatic high risk women.
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Most of the microorganisms display adhesion molecules on their surface which help them to bind and interact with the host cell during infection. Adhesion molecules help mycobacteria to colonize and invade immune system of the host, and also trigger immune response explicated by the host against the infection. Hence, understanding the signalling pathways illustrated by these molecules to enhance our knowledge on mycobacterial survival and persistence inside the host cell is required. Hence, this review was focussed on the role of adhesion molecules and their receptor molecules. The various mechanisms adopted by adhesion molecules to bind with the specific receptors on the host cell and their role in invasion and persistence of mycobacterium inside the host cell are explained.
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Objective To study the expressions of transforming growth factor-β1 （TGF-β1） and EⅢA-fibronectin （EⅢA-FN） at different time points of antemortem injury, antemortem injury postmortem expression and postmortem injury and to explore their application value in wound age estimation. Methods A model of rat skeletal muscle contusion was established. The rats were randomly divided into normal control group （n=5）, antemortem contusion group （n=40）, antemortem contusion postmortem expression group （n=110） and postmortem injury group （n=25）. The expressions of TGF-β1 and EⅢA-FN after rat skeletal muscles antemortem contusion were detected with immunohistochemical staining. Expression changes of TGF-β1 and EⅢA-FN mRNA in each group were analyzed with real-time fluorescence quantitative PCR. Results Immunohistochemical staining results showed that a large number of polymorphonuclear leukocyte, mononuclear cells and fibroblastic cells showed a strong expression of TGF-β1 in wounded zones 12 h-14 d after antemortem contusion. EⅢA-FN was mainly distributed in the extracellular matrix, 3 to 7 d post-traumatic. Real-time fluorescence quantitative PCR results showed that TGF-β1 and EⅢA-FN mRNA in antemortem injury group reached the peak at 3 and 5 d post-traumatic respectively. The expressions of TGF-β1 and EⅢA-FN mRNA in antemortem contusion postmortem expression group peaked at 6 h and 12 h postmortem. The expression of TGF-β1 and EⅢA-FN mRNA in postmortem injury group 0.5-12 h postmortem was significantly lower than those of the normal control group and the antemortem contusion group. Conclusion TGF-β1 and EⅢA-FN might become a reference index for skeletal muscle wound age estimation.