Effect of ion-exchange chromatography on removal of fibronectin in von Willebrand factor / 中国输血杂志

【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.

Effect of Biling Qutong Prescription on NLRP3， P2X7R， FN， and Hepatic Steatosis in Type 2 Diabetes Mellitus Complicated with Gouty Arthritis / 中国实验方剂学杂志

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.

Expansion of cord blood stem cells in fibronectin-coated microfluidic bioreactor

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.

Fibronectin FNDC10 promotes malignant biological behaviors of breast cancer cells by enhancing STAT3 activation / 中国肿瘤生物治疗杂志

@#[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.

Fibronectin-binding Protein F1 (prtF1) Gene Is Highly Distributed in Tetracycline-resistant, Erythromycin-sensitive Streptococcus pyogenes Isolates

@#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.

Bioactivity effects of extracellular matrix proteins on apical papilla cells

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.

Fibronectin as an Early Predictor of Gestational Hypertension/Preeclampsia.

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.

Effect of Tetrandrine on Col-I and FN in TGF-β1-induced MRC-5 Cells / 中国实验方剂学杂志

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.

Effects of integrin av ß3 inhibitor CycIo-RGDfK on epithelial- mesenchymal transition of breast epithelial cells induced by fibronectin / 中国药理学通报

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.

Staphylococcus aureus: generalidades, mecanismos de patogenicidad y colonización celular / Staphylococcus aureus: generalities, mechanisms of pathogenicity and cell colonization

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.

Evaluation of fetal fibronectin rapid bedside test as a predictor of preterm labour: a prospective study

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.

Adhesion molecules facilitate host-pathogen interaction & mediate Mycobacterium tuberculosis pathogenesis

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.

Expression of survivin, fibronectin-1, VEGF and ezrin in thyroid tumors and their relationship with tumor pathological characteristics / 中国肿瘤生物治疗杂志

@# Objective: To investigate the expression of survivin, fibronectin-1, vascular endothelial growth factor (VEGF) and ezrin in thyroid tumors and their relationship with the pathological characteristics of thyroid tumors. Methods: Ninety patients with thyroid tumors admitted to the third affiliated hospital of Zunyi Medical University and the first hospital during Oct. 2016 and Oct. 2018 were selected as the observation group. Seventy-five patients with normal thyroid confirmed by pathology in the same period were selected as the control group. The protein levels of survivin, fibronectin-1, VEGF and ezrin were detected by immunohistochemical method. Results: The positive rates of survivin, fibronectin-1, VEGF and Ezrin in the control group were 2.67%, l4.00%, 1.33% and 1.33%, which were lower than 97.78%, 96.67%, 93.33% and 95.56% in the observation group, respectively (all P<0.05 or P<0.01). The expressions of survivin, fibronectin-1, VEGF and ezrin were significantly correlated with TNM staging, tumor diameter, extrathyroid invasion and lymphatic metastasis (all P<0.05). Conclusion: Survivin, fibronectin-1, VEGF and ezrin proteins are all involved in the occurrence and development of thyroid tumors. The combined detection of these four indicators is of great significance in the diagnosis, treatment and prognosis of thyroid tumors.

Expression of FN1 in gastric cancer and its clinical significance / 医学研究生学报

Objective Fibronectin 1 (FN1) is a glycoprotein involved in cellular adhesion and migration processes. The aim of this study was to investigate the expression and clinicopathological significance of FN1 in gastric cancer and to predict the possible mechanism of FN1. Methods GEO data and TCGA data were downloaded. FN1 expression in gastric cancer and adjacent tissues was analyzed by GSE54129 data, and then verified by GSE29272 and TCGA data. According to the expression profile data and FN1 expression, mRNA expression value was divided into low expression(<-0.475), and medium expression(-0.475~1.036), high expression (>1.036). FN1 expression in gastric cancer and clinicopathological relationship were analyzed. TCGA data and Kaplan Meier were used to analyze the relationship between the expression level of FN1 and the prognosis of gastric cancer patients; while gene concentration analysis (GSEA) was used to predict the related path of FN1. Results TCGA data showed the medium survival time of low, medium, high FN1 expression was respectively 63.5, 55.7, 39.4 months, and the difference between low expression and high expression in survival time was of statistical significance. Kaplan Meier Plotter online data analysis showed the medium survival time of FN1 high expression was shorter than that of low expression(P<0.01), which meant the higher the FN1 expression was, the worse the prognosis was. FN1 expression is an independent prognostic factor (HR=0.480,95% CI:0.336~0.686). High expression of FN1 samples enriched gene sets such as KRAS (FDR=0.052), P53(FDR=0.052), TGF-β(FDR=0.052), cell adhesion(FDR=0.0), extracellular matrix (FDR=0.043)and cytoskeletal protein regulation (FDR=0.052). Conclusion The high expression of FN1 is a poor prognostic factor for gastric cancer and can be used as an effective biomarker for predicting the metastasis and prognosis of gastric cancer. High expression of FN1 leads to abnormalities in KRAS, P53, TGF-β, ECM, cell adhesion and cytoskeletal protein regulatory pathways.

Laboratory detection of blood vessel function in patients with hemorrhagic stroke / 国际检验医学杂志

Objective To investigate the level of vascular endothelial growth factor (VEGF), homocysteine (HCY), fibronectin (Fn) and apolipoprotein E (ApoE) in the serum and cerebrospinal fluid patients with stroke to inquire into patients′endothelial function damage.Methods We chose standard stroke patients in Brain Hospital of Hu′nan Province from August 2015 to August 2016, divided into ischemic stroke (IS) group (52 cases) and hemorrhagic stroke (HS) group (33 cases), and chose patients with the cerebrospinal fluid for diagnosis and treatment and ruled out infections, the nervous system and cardiovascular system diseases as control group (20 cases).The serum VEGF, HCY, Fn, and ApoE levels of the patients on the 1 th, 5 th and14 th day after admission were monitored and compared with those in cerebrospinal fluid expression, NIHSS and Barthel were also performed for assessment of severity and prognosis of HS.Results The serum levels of VEGF, HCY and ApoE in HS group were higher than those in IS group, and the Fn level was lower than that in IS group and control group.At the same time, the Fn levels gradually increased on the 5 th and 14 th day after admission (P＜0.05), close to the control group.Cerebrospinal fluid of VEGF, HCY, Fn, and ApoE levels in HS group were higher than those in the control group, while Fn level was lower, and these indexes were lower in cerebrospinal fluid than those in the serum, and the differences were statistically significant (P＜0.05).In correlation analysis, HCY level of cerebrospinal fluid was positively correlated with ApoE in HS patients (r=0.645, P=0.001), HCY and ApoE levels in HS patients were positively correlated with patients′NIHSS score (r=0.547, P=0.006;r=0.720, P＜0.001), and negatively correlated with Barthel index score (r=-0.703, P＜0.001, r=-0.765, P＜0.001, respectively).Conclusion There were vascular endothelial injury and neuroprotective function decreased in brain tissue of stroke patients.The level of Fn was related to the patient′s prognosis, which was conducive to the differentiation of IS and HS.The cerebrospinal fluid levels of HCY and ApoE can be used to evaluate the prognosis of HS patients.

Effect of IL-6 on fibronectin in bovine trabecular meshwork cells cultured <i>in vitro</i> / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the changes of fibronectin expression in cultured bovine trabecular meshwork cells when cells were stimulated by different concentrations of IL-6 to provide evidence for early diagnosis and new therapy of POAG.<p>METHODS: We identify third-generation bovine trabecular meshwork cells which were got from tissue mass culture method. Then the relative expression of FN gene and protein in cells were detected by Real-Time PCR and Western-blot after 24h stimulation with 0ng/mL, 0.1ng/mL, 0.5ng/mL IL-6.<p>RESULTS: The cultured bovine trabecular cells are coincident with what recorded in the book. Real-time PCR and Western blot showed that the amount of FN mRNA produced by cells was 1.000±0.000, 0.213±0.004, 0.056±0.001, 0.019±0.002 respectively, and the protein expression was 1.167±0.012, 0.662±0.009, 0.238±0.011, 0.061±0.011 respectively. There was a significant difference among four groups(<i>P</i><0.05).<p>CONCLUSION: Cultured bovine trabecular meshwork cells have a negative correlation with the expression of FN protein after being stimulated by exogenous IL-6, and the results are consistent with the actual state of the disease. We speculate that the IL-6 participate in the pathogenesis and progression of POAG by affecting the expression of FN gene and protein and changing the structure of trabecular meshwork.

Expressions of TGF-β1 and EⅢA-FN after Rat Skeletal Muscle Contusion and Wound Age Estimation / 法医学杂志

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.

Risk assessment for preterm delivery using the fetal fibronectin test associated with the measurement of uterine cervix length in symptomatic pregnant women / Avaliação do risco para parto pré-termo utilizando o teste da fibronectina fetal, associado à medida de comprimento de colo uterino, em gestantes sintomáticas

Abstract Objective To analyze the use of the measurement of uterine cervix length (MUCL) and the fetal fibronectin (fFN) rapid test as predictors of preterm delivery (PTD) in symptomatic pregnant women assisted at the Santa Casa de Misericórdia de Sobral Maternity Hospital. Methods This was a prospective and analytic study involving 53 parturients assisted between September of 2015 and July of 2016; the participants were between 24 and 34 weeks of gestational age (GA) and presented complaints related to preterm labor (PTL) prodromes. Vaginal secretion was collected for fFN testing, and the MUCL was obtained via transvaginal ultrasonography. Results A total of 58.49% of the subjects showed MUCL < 25 mm, and 41.51% were positive in the fFNrapid test.Atotal of 48 patients were followed-up until their delivery date, and 54.17% resulted in PTL. The relative risk (RR) for PTD in patients with MUCL < 25 mm was 1.83 (p = 0.09, 0.99-3.36, 95% confidence interval [CI]), with a mean time before delivery of 2.98 weeks. Based on fFN positive results, the RR was 3.50 (p = 0.002, 1.39- 8.79, 95%CI) and themean time until delivery was 1.94weeks. The RRwas 2.70 (p = 0.002, 1.08-6.72, 95%CI) when both tests were used. The RR of PTD within 48 hours, and 7 and 14 days were, respectively, 1.30 (p = 0.11, 95% CI 1.02-1.67), 1.43 (p = 0.12, 95% CI % 0.99-2.06), and 2.03 (p = 0.008, 95% CI 1.26-3.27), when based on the MUCL, and 1.75 (p = 0.0006, 95% CI 1.20-2.53), 2.88 (p = 0.0001, 95% CI, 1.57-5.31), and 3.57 (p = 0.0002, 95% CI 1.63-7.81) when based on positive fFN results. The RR at 48 hours and 7 and 14 days considering both tests was 1.74 (p = 0.0001, 95% CI 1.14-2.64), 2.22 (p = 0.0001, 95% CI 1.22-4.04), and 2.76 (p = 0.0002, 95% CI 1.27-5.96), respectively. Conclusion In symptomatic pregnant women, we concluded that the MUCL < 25 mm associated with positive fFN rapid test indicate increased the risk for PTD. Further studies with larger sample sizes could contribute in supporting the results presented in the current study.

Resumo Objetivo Analisar a utilização da medida do comprimento do colo uterino (MCCU), e do teste da fibronectina fetal (FNf) como preditores do trabalho de parto pré-termo (PPT), em gestantes sintomáticas, atendidas na Maternidade da Santa Casa de Misericórdia de Sobral. Métodos Foi realizado umestudo prospectivo e analítico, envolvendo 53 parturientes atendidas no período de setembro de 2015 a julho de 2016, com idade gestacional (IG) entre 24 e 34 semanas que tiveram queixas relacionadas a pródromos de trabalho de parto prematuro (TPP), sendo realizada coleta de secreção vaginal para FNf e MCCU por via ultrassonográfica transvaginal. Resultados Um total de 58,49% das pacientes tinham MCCU < 25 mm, e 41,51% tiveram teste rápido de fFN positivo. Foi feito o acompanhamento de 48 pacientes, com 54,17% de PPTs. O risco relativo (RR) para PPT com MCCU < 25 mm foi de 1,83 (p = 0,09, 0,99-3,36, intervalo de confiança [IC] 95%), com média de tempo até o parto de 2,98 semanas. Para fFN, o RR foi de 3.50 (p = 0.002, 1.39-8.79, IC 95%) e a média até o parto foi de 1,94 semanas. Quando os dois testes forampositivos, o RR foi de 2,70 (1,08-6,72). Para a MCCU, o RR para PPT em 48 horas, 7 e 14 dias foram 1,30 (p = 0.11, 95% IC 1.02-1.67), 1,43 (p = 0.12, 95% CI % 0.99-2.06) e 2,03 (p = 0.008, 95% IC 1.26-3.27), respectivamente. Para FNf, em 48 horas, 7 e 14 dias foi de 1,75 (p = 0.0006, 95% IC 1.20-2.53, 2,88 (p = 0.0001, 95% IC, 1.57-5.31) e 3,57 (p = 0.0002, 95% IC 1.63-7.81) respectivamente. Com os dois testes, o RR em 48 horas, 7 e 14 dias foi 1,74 (p = 0.0001, 95%IC 1.14-2.64), 2,22 (p = 0.0001, 95% IC 1.22-4.04) e 2,76 (p = 0.0002, 95% IC 1.27-5.96) respectivamente. Conclusão Em mulheres grávidas sintomáticas, concluímos que a MCCU < 25 mm e o teste rápido de FNf positivo indicam aumento do risco de PPT. Outros estudos com tamanhos de amostra maiores podem contribuir para apoiar os resultados apresentados no presente estudo.

Parto pretérmino: causas y medidas de prevención

Preterm birth is the final common pathway of a series of different physiopathological processes, so it is considered a syndrome. Spontaneous preterm birth represents two thirds of preterm deliveries. Prevention can be primary or secondary, and is basically focused on prediction, which is currently done by risk calculators that combine medical history, cervical length and fetal fibronectin. Vaginal progesterone reduces the rate of spontaneous preterm birth to 35-40%, if used from week 16 through week 34.

El parto pretérmino es la vía final común de una serie de procesos fisiopatológicos diferentes, por lo que constituye un síndrome. El parto pretérmino espontáneo representa las dos terceras partes del parto pretérmino. Su prevención puede ser primaria o secundaria, y se centra básicamente en la predicción, la que actualmente se realiza mediante calculadoras de riesgo que combinan factores de historia clínica, longitud cervical y fibronectina fetal. La progesterona vaginal reduce la tasa de parto pretérmino espontáneo en 35 a 40%, si se administra desde las 16 hasta las 34 semanas.

The Effect of Fibronectin-Immobilized Microgrooved Titanium Substrata on Cell Proliferation and Expression of Genes and Proteins in Human Gingival Fibroblasts

BACKGROUND: We aimed to determine the effect of fibronectin (FN)-immobilized microgrooved titanium (Ti) on human gingival fibroblast proliferation, gene expression and protein expression. METHODS: Photolithography was used to fabricate the microgrooved Ti, and amine funtionalization (silanization) was used for FN immobilization on titanium surfaces. Cell proliferation, gene expression and protein expression were analyzed, followed by multiple regression analysis for determining the influential factors on cell proliferation. RESULTS: FN-immobilized microgrooved Ti significantly enhanced the fibroblast proliferation in various timelines of culture, among which a burst of fivefold increase is induced at 96 h of culture compared to that on the control smooth Ti. We suggest a presence of the synergistic promotion effect of microgrooves and FN immobilization on fibroblast proliferation. Through a series of analyses on the expression of various genes and proteins involved in cell adhesion and proliferation, cyclin-dependent kinase 6, cyclin D1, integrin α5, oncogene c-Src, osteonectin, paxillin and talin-2 were determined as influential factors on promoting fibroblast proliferation induced by FN-immobilized microgrooved Ti. CONCLUSION: FN-immobilized microgrooved Ti can act as an effective surface for enhancing fibroblast proliferation, and can be used for promoting soft tissue response on the connective tissue attachment zone of biomaterial surfaces.