Β1 Levels Research Articles

Expression of integrins β1, α4 and cell adhesion molecule ICAM-1 in the presence of sodium deoxyribonucleate with ferrum complex (DNA-Na-Fe) by MT-4 cells transformed by human T-lymphotropic virus type 1 (Retroviridae: Orthoretrovirinae: Deltaretrovirus: Human T-lymphotropic virus type 1)

The important role of integrins (IG) in the initiation and development of cancer processes makes these structures convenient targets for the development of immunomodulatory therapeutic drugs that have an effect directly on these molecules. Among the latter, IG β1, α4 and cell adhesion receptor ICAM-1 (intercellular adhesion molecule 1) are of particular interest. Immunomodulators are capable of changing the IG activity through non-specific mechanisms, which, however, in some cases can cause a decrease in the protective functions of the immune system and health deterioration.The aim of the study was to determine the effect on the levels of cellular expression and the nature of IG metabolism of the drug sodium deoxyribonucleate with ferrum complex, DNA-Na-Fe, which is having been used in the Russian Federation as an immunomodulatory agent, but whose action has not been studied in details so far. We used 2 variants of the neoplastic CD4+ T-lymphocyte cell line transformed with human T-lymphotropic virus type 1 (HTLV-1) of the Retroviridae family, MT-4 (MT-4/1 and MT-4/2). The indicated variants were characterized by different levels of expression of the protein activation markers CD28 and CD38. After cell culture in the presence of 500 μg/ml DNA-Na-Fe, the expression levels of IG β1 (CD29), α4 (CD49d), and ICAM-1 (CD54) were studied by flow cytometry. The cells of the both lines contained many membrane proteins CD29+ (90.4% ± 4.5) and CD54+ (97.9% ± 1.4), while small percentage of cells contained protein CD49d+ (1.9% ± 1.0). No changes in the expression of the studied proteins were observed in the presence of the drug. The levels of IG β1, α4 and ICAM-1 expression may serve as one of the phenotypic characteristics of MT-4 cells. The obtained data are of great importance because the peculiarities of CD4+ T-lymphocytes transformation and their metabolism during HTLV-1 infection have not been sufficiently studied so far. The results of this work may be helpful in determining the pathogenesis of HTLV-1-induced diseases, some types of malignancies, and in searching for new specific pharmacological agents, including molecularly targeted ones. The results of the study will help to expand the existing knowledge on the markers of MT-4 cell line.

Performance of the Polydopamine-Graphene Oxide Composite Substrate in the Osteogenic Differentiation of Mouse Embryonic Stem Cells.

Graphene oxide (GO) is a biocompatible material considered a favorable stem cell culture substrate. In this study, GO was modified with polydopamine (PDA) to facilitate depositing GO onto a tissue culture polystyrene (PT) surface, and the osteogenic performance of the PDA/GO composite in pluripotent embryonic stem cells (ESCs) was investigated. The surface chemistry of the PDA/GO-coated PT surface was analyzed by scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). A high cell viability of ESCs cultured on the PDA/GO composite-coated surface was initially ensured. Then, the osteogenic differentiation of the ESCs in response to the PDA/GO substrate was assessed by alkaline phosphatase (ALP) activity, intracellular calcium levels, matrix mineralization assay, and evaluation of the mRNA and protein levels of osteogenic factors. The culture of ESCs on the PDA/GO substrate presented higher osteogenic potency than that on the uncoated control surface. ESCs cultured on the PDA/GO substrate expressed significantly higher levels of integrin α5 and β1, as well as bone morphogenetic protein receptor (BMPR) types I and II, compared with the control groups. The phosphorylation of extracellular signal-regulated kinase (ERK)1/2, p38, and c-Jun-N-terminal kinase (JNK) mitogen-activated protein kinases (MAPKs) was observed in ESCs culture on the PDA/GO substrate. Moreover, BMP signal transduction by SMAD1/5/8 phosphorylation was increased more in cells on PDA/GO than in the control. The nuclear translocation of SMAD1/5/8 in cells was also processed in response to the PDA/GO substrate. Blocking activation of the integrin α5/β1, MAPK, or SMAD signaling pathways downregulated the PDA/GO-induced osteogenic differentiation of ESCs. These results suggest that the PDA/GO composite stimulates the osteogenic differentiation of ESCs via the integrin α5/β1, MAPK, and BMPR/SMAD signaling pathways.

Conditioned medium of IGF1-induced synovial membrane mesenchymal stem cells increases chondrogenic and chondroprotective markers in chondrocyte inflammation.

Recently, mesenchymal stem cells (MSCs) have been the most explored cells for cell therapy for osteoarthritis (OA) that can be obtained from various sources. Synovial membrane MSCs (SMMSCs) provide best potential for OA therapy, however they are not widely explored. Conditioned medium of SMMSCs (SMMSCs-CM) rich in growth factors and cytokines can inhibit apoptosis and increase chondrocytes cell proliferation. The aim of the present study was to determine growth factors content in SMMSCs-CM as well as the chondrogenic and chondroprotective markers expression in OA model after insulin-like growth factor (IGF)1-induced and non-induced SMMSCs-CM treatments. Chondrocyte cell line (CHON002) was induced by IL1β as OA model (CHON002 with IL1β (IL1β-CHON002)) and treated with SMMSCs-CM with or without IGF1 induction to determine its effectiveness in repairing OA cells model. ELISA was used to assay BMP2, fibroblast growth factor 18 (FGF18) and transforming growth factor (TGF) β1 (TGFβ1) levels in SMMSCs-CM, matrix metalloproteinase (MMP) 13 (MMP13) and a disintegrin and metalloproteinase with thrombospondin motif 4 (ADAMTS4) levels in OA cells model treated with SMMSCs-CM. RT-qPCR analyses were used to investigate the gene expression of SOX9, COL2, and COL10. CM from SMMSCs cultured and induced by IGF1 150 ng/mL was the most effective concentration for increasing the content of growth factor markers of SMMSCs-CM, which had successfully increased negative cartilage hypertrophy markers (SOX9 and COL2) and reduced hypertrophy markers (COL10, MMP13, and ADAMTS4). Preconditioning with IGF1 has better and very significant results in lowering MMP13 and ADAMTS4 levels. The present study supports IGF1 pre-conditioned SMMSCs-CM to develop a new therapeutic approach in OA improvement through its chondrogenic and chondroprotective roles.

Transforming growth factor-β2-mediated mesenchymal transition in lens epithelial cells is repressed in the absence of RAGE

Transforming growth factor-β2 (TGFβ2)-mediated epithelial to mesenchymal transition (EMT) in lens epithelial cells (LECs) has been implicated in fibrosis associated with secondary cataracts. In this study, we investigated whether the receptor for advanced glycation end products (RAGE) plays a role in TGFβ2-mediated EMT in LECs. Unlike in the LECs from wild-type mice, TGFβ2 failed to elicit an EMT response in LECs from RAGE knockout mice. The lack of RAGE also diminished TGFβ2-mediated Smad signaling. In addition, treatment with TGFβ2 increased IL-6 levels in LECs from wild-type mice but not in those from RAGE knockout mice. Treatment of human LECs with the RAGE inhibitor FPS-ZM1 reduced TGFβ2-mediated Smad signaling and the EMT response. Unlike that in wild-type lenses, the removal of fiber cell tissue in RAGE knockout lenses did not result in elevated levels of α-smooth muscle actin (α-SMA), fibronectin (FN), and integrin β1 in capsule-adherent LECs. Taken together, these results suggest that TGFβ2 signaling is intricately linked to RAGE. Targeting RAGE could be explored as a therapeutic strategy against secondary cataracts.

Rac1-mediated sustained β4 integrin level develops reattachment ability of breast cancer cells after anchorage loss.

Previously, we reported that non‐apoptotic cell death was induced in non‐malignant mammary epithelial cells (HMECs) upon loss of anchorage during 48 h incubation in suspension. In this study, we examined HMECs in suspension at an earlier time point and found that most of them lost attachment ability to substrata when replated, although >80% were alive. This suggested that HMECs lost reattachment ability (RA) prior to cell death upon detachment. Concomitant with the loss of RA, a decrease in the levels of β1 and β4 integrin was observed. In sharp contrast, breast cancer cells retained integrin levels, reattached to substrata, and formed colonies after exposure to anchorage loss as efficiently as those maintained under adherent conditions. Such RA of cancer cells is essential for the metastatic process, especially for establishing adhesion contact with ECM in the secondary organ after systemic circulation. Further analysis suggested that sustained levels of β4 integrin, which was mediated by Rac1, was critical for RA after anchorage loss and lung metastasis of breast cancer cells. In the cancer cells, persistent Rac1 activity enhanced escape of β4 integrin from lysosomal degradation depending on actin‐related protein 2/3 and TBC1D2, a GTPase‐activating protein of Rab7 GTPase. Notably, simultaneous high expression of ITGB4 and RAC1 was associated with poor prognosis in patients with breast cancer. Therefore, β4 integrin and Rac1 are attractive therapeutic targets to eliminate RA in cancer cells, thereby preventing the initial step of colonization at the secondary organ during metastasis.

3D magnetic nanocomposite scaffolds enhanced the osteogenic capacities of rat bone mesenchymal stem cells in vitro and in a rat calvarial bone defect model by promoting cell adhesion.

Magnetic scaffolds incorporated with iron oxide nanoparticles (IONPs) are biocompatible and present excellent osteogenic properties. However, the underlying mechanism is unclear. In this study, 3D-printed poly(lactic-co-glycolic acid) scaffolds were coated with IONPs using layer-by-layer assembly (Fe-scaffold) to prepare magnetic scaffolds. The effects of this modification on osteogenesis were investigated by comparison with untreated scaffolds (Uncoated-scaffold). The results showed that the proliferation of rat bone mesenchymal stem cells (rBMSCs) on the Fe-scaffold was enhanced compared with those on the Uncoated-scaffold (p < 0.05). The alkaline phosphatase activity and expression levels of osteogenic-related genes of cells on the Fe-scaffold were higher than those on the Uncoated-scaffold (p < 0.05). Fe-scaffold was found to promote the cell adhesion compared with Uncoated-scaffold, including increasing the adhered cell number, promoting cell spreading and upregulating the expression levels of adhesion-related genes integrin α1 and β1 and their downstream signaling molecules FAK and ERK1/2 (p < 0.05). Moreover, the amount of new bone formed in rat calvarial defects at 8 weeks decreased in the order: Fe-scaffold > Uncoated-scaffold > Blank-control (samples whose defects were left empty) (p < 0.05). Therefore, 3D magnetic nanocomposite scaffolds enhanced the osteogenic capacities of rBMSCs in vitro and in a rat calvarial bone defect model by promoting cell adhesion. The mechanisms were attributed to the alteration in its hydrophilicity, surface roughness, and chemical composition.

Concomitant inhibition of hedgehog signalling and activation of retinoid receptors abolishes bleomycin-induced lung fibrosis.

Pulmonary fibrosis is a devastating disease with unknown treatment. All-trans retinoic acid (ATRA) attenuates bleomycin-induced lung fibrosis by different mechanistic pathways. However, the role of retinoid receptors in lung fibrosis is still unclear. Forskolin (FSK), a potent inhibitor for the revolutionary hedgehog (Hh) signalling pathway, has a promising antifibrotic effect on other organs such as the liver. This study investigates the interplay between the retinoid receptors modulation and the Hh signalling pathway in bleomycin (BLM)-induced pulmonary fibrosis. Rats were randomised and administrated a single dose of 7.5mg/kg of BLM alone and with ATRA, FSK and both of them. The effects of FSK and ATRA on lung functions, oxidative stress markers (malondialdehyde [MDA], glutathione [GSH], superoxide dismutase [SOD] and catalase [CAT]), retinoid markers (retinoic acid receptors [RAR] and rexinoid X receptors [RXR]) and Hh signalling markers (patched homolog 1 [Ptch-1], Smoothened [Smo] and glioblastoma-2 [Gli-2]) were assessed. In single therapies, ATRA and FSK ameliorated BLM-induced lung fibrosis. On the contrary, a combination of both drugs synergistically reversed the effect of BLM-induced lung fibrosis, as indicated by the enhancement of lung functions and the decrease of the α-smooth muscle actin (α-SMA) expression and collagen deposition. Additionally, FSK and ATRA ameliorated oxidative stress and inflammation, reduced transforming growth factor β1 (TGF-β1) levels and reversed the effect of BLM on the mRNA expression of Ptch-1, Smo and Gli-2. FSK inhibited the Hh pathway and also activated protein kinase A (PKA) that is, in part, involved in phosphorylation of RAR/RXR heterodimer (a key step in retinoid receptor activation). The present results suggest that a combination of FSK and ATRA has a promising therapeutic value for lung fibrosis management.

PRR11 induces filopodia formation and promotes cell motility via recruiting ARP2/3 complex in non-small cell lung cancer cells

Filopodia, a finger-like structure and actin-rich plasma-membrane protrusion at the leading edge of the cell, has important roles in cell motility. However, the mechanisms of filopodia generation are not well-understood via the actin-related protein 2/3 (ARP2/3) complex in Non-Small Cell Lung Cancer (NSCLC) cells. We previously have demonstrated that PRR11 associates with the ARP2/3 complex to regulate cytoskeleton-nucleoskeleton assembly and chromatin remodeling. In this study, we further demonstrate that PRR11 involves in filopodia formation, focal adhesion turnover and cell motility through ARP2/3 complex. Cell phenotype assays revealed that the silencing of PRR11 increased cellular size and inhibited cell motility in NSCLC cells. Mechanistically, PRR11 recruited and co-localized with Arp2 at the membrane protrusion to promote filopodia formation but not lamellipodia formation. Notably, PRR11 mutant deletion of the proline-rich region 2 (amino acid residues 185–200) abrogated the effect of filopodia formation. In addition, PRR11-depletion inhibited filopodial actin filaments assembly and increased the level of active integrin β1 in the cell surface, whereas reduced the phosphorylation level of focal adhesion kinase (FAKY397) to repress focal adhesion turnover and cell motility in NSCLC cells. Taken together, our findings indicate that PRR11 has critical roles in controlling filopodia formation, focal adhesion turnover and cell motility by recruiting ARP2/3 complex, thus dysregualted expression of PRR11 potentially facilitates tumor metastasis in NSCLC cells.

Diagnostic Role of Transforming Growth Factor Beta Level in Necrotizing Enterocolitis in Extremely Low Birthweight Infants

Necrotizing enterocolitis (NEC) continues to be a leading cause of morbidity and mortality in premature infants.Premature infants may be at risk of NEC due to developmental deficiency of transforming growth factor Beta-1 level in the intestine, which is further accentuated during NEC. No significant differences in the incidence of NEC are noticed between male and female patients. TGF- β1 is an important marker for diagnosis and prognosis of necrotizing enterocolitis in extremely low birthweight infants and the aim of this work was to evaluate the diagnostic and prognostic value of transforming growth factor Beta-1 (TGF- β1) Level On necrotizing enterocolitis for greatly low birthweight babies. This might have been an instance - regulated study that incorporated 50 greatly low birthweight newely destined babies diagnosed Likewise necrotizing enterocolitis clinically and arranged as stated by altered Bell’s arranging criteria and 25 greatly low birthweight newely conceived babies without nec clinched alongside nicu of from june 2017 should june 2018. We established that the mean conception weight clinched alongside nec aggregation might have been 1. 61 ± 0. 35 (kg) Furthermore in control gathering might have been 1. 65 ± 0. 35 ( kg ) with male predominance. TGF- β1 level ≤ 864. 3 at onset about manifestations might altogether ( p = 0. 004 ) foresee nec Also TGF- β1 level ≥ 964. 7 after 7 times from claiming onset for manifestations could essentially ( p = 0. 011 ) anticipate seriousness of nec.

Endoplasmic reticulum oxidoreductase 1 alpha modulates prostate cancer hallmarks.

BackgroundTherapies available for late stage prostate cancer (PCa) patients are limited and mostly palliative. The necessary development of unexplored therapeutic options relies on a deeper knowledge of molecular mechanisms leading to cancer progression. Redox signals are known to modulate the intensity and duration of oncogenic circuits; cues originating from the endoplasmic reticulum (ER) and downstream exocytic organelles are relevant in secretory tumors, including PCa. Ero 1α is a master regulator of redox homeostasis and oxidative folding.MethodsWe assessed Ero 1α mRNA expression by bioinformatic analysis of three public datasets and protein expression levels in PCa cell lines representing different degrees of tumor progression and different human prostate specimens. Transient Ero 1α knockdown was achieved by RNA interference (siRNA). Consequences of Ero 1α downregulation were monitored by PCa proliferation, migration and invasion properties.ResultsEro 1α mRNA and protein levels are upregulated in PCa cell lines compared to non-tumorigenic cells (P=0.0273). Ero 1α expression increases with the grade of malignancy, reaching the highest level in the androgen resistant PC3. In patients’ samples from 3 datasets, Ero 1α mRNA expression correlates with pathological Gleason scores. Ero 1α knockdown inhibits proliferation (P=0.0081), migration (P=0.0085) and invasion (P=0.0007) of PC3 cells and alters the levels of integrin β1 (P=0.0024).ConclusionsResults indicate that Ero 1α levels correlate with PCa aggressiveness; Ero 1α silencing inhibits key steps over the PCa metastatic process. Therefore, Ero 1α has the potential to be exploited as a novel biomarker and a therapeutic target in PCa.

Clinical value of serum Ape1/Ref-1 combined with TGF-β1 monitoring in predicting the occurrence of radiation pneumonitis (RP) in non-small cell lung cancer patients.

The aim of the present study was to explore the predictive value of serum apurinic/apyrimidinic endonuclease 1/redox factor-1 (Ape1/Ref-1) combined with transforming growth factor β1 (TGF-β1) levels in the occurrence of radiation pneumonitis (RP) in patients with non-small cell lung cancer (NSCLS). Eighty-one patients with NSCLS who were admitted from August 2017 to July 2019 were enrolled in the present study. All patients were treated with concurrent radiotherapy and chemotherapy. Serum Ape1/Ref-1 and TGF-β1 levels were measured before treatment and 12 weeks after treatment. Patients with radiation-induced lung injury were assessed and divided into the RP group (lung injury ≥2) and non-RP (NRP) group (grade <2). The levels of serum Ape1/Ref-1 and TGF-β1 before and after treatment between the 2 groups were compared. The relationship between clinical characteristics, serum Ape1/Ref-1, TGF-β1 levels, and the occurrence of RP were then analyzed, and the relationship between serum Ape1/Ref-1, TGF-β1 levels, and their predictive value for the occurrence of RP was also assessed. The incidence of RP in 81 patients was 30.86%. After treatment, the serum Ape1/Ref-1 and TGF-β1 levels of the 2 groups were significantly higher than those before treatment (P<0.05). Furthermore, after treatment, the levels of serum Ape1/Ref-1 and TGF-β1 in the RP group were significantly higher than those of the NRP group (P<0.05). Multivariate logistic regression analysis showed that V20, Ape1/Ref-1, and TGF-β1 were associated with the occurrence of RP (P<0.05). The levels of serum Ape1/Ref-1 were positively correlated with TGF-β1 (r=0.734, P<0.05). Finally, the area under the curve of RP occurrence, which was predicted by the levels of serum Ape1/Ref-1, TGF-β1, and the combination of both were 0.779, 0.69, and 0.842, respectively. The occurrence of RP in NSCLS patients is closely related to the levels of serum Ape1/Ref-1 and TGF-β1, and the combination of both has important predictive values for the occurrence of RP.

Лабораторна оцінка рівнів трансформуючого фактору росту-β1 у хворих на ішемічну хворобу серця у поєднанні з цукровим діабетом 2-го типу

The purpose of the study was to examine the levels of transforming growth factor-β1 in the serum of patients with coronary heart disease in combination with type 2 diabetes and without it. Material and methods. We conducted a survey of 65 patients (25 men, 40 women) aged from 36 to 69 years (mean age was (59±3.5) years). All patients were diagnosed with coronary heart disease in the form of stable angina pectoris I-II functional classes. The group of examined patients included 33 patients with concomitant type 2 diabetes mellitus (mild form was in 15 people; moderate was in 18 people) and 32 patients without diabetes mellitus. The scope of the survey covered the generally accepted methods of clinical, laboratory and instrumental examination. The group of patients with coronary heart disease with type 2 diabetes had 14 (43%) men and 19 (57%) women (mean age was (62±2.6) years. Heart failure of stage I-II A (I-II functional classes) was diagnosed in 22 (68%) patients. The duration of coronary heart disease was from 3 to 15 years, the duration of type 2 diabetes lasted from 3 to 14. We detected hypertension in 19 (57%) patients, it was within 1-2 degrees (according to the criteria of the Ukrainian Association of Cardiologists, 2008). In the group of patients with coronary heart disease without diabetes there were 11 men (34%), 21 women (66%) (mean age was (57.0±2.4) years). Hypertension within 1-2 degrees was detected in 15 (46%) patients. Heart failure of I-II A stages (I-II functional classes) was diagnosed in 15 (46%) patients. The control group consisted of 15 practically healthy individuals who were representative by sex and age of patients from the study group and who did not have diseases of the cardiovascular system and endocrinopathies. The level of transforming growth factor-β1 in the blood serum was determined using sets of standard test systems "TGF-β1 ELISA" produced by company "DRG Instruments" (Germany). The level of native transforming growth factor-β1 in the serum was determined by solid-phase enzyme-linked immunosorbent assay. Results and discussion. Groups of patients with coronary heart disease with type 2 diabetes and without it could be compared by age, sex, duration and severity of coronary heart disease, the frequency of concomitant hypertension. At the same time, among patients with coronary heart disease with type 2 diabetes there was a higher frequency of heart failure. The results showed that probable increase in serum transforming growth factor-β1 levels in patients with coronary heart disease was more pronounced when combining coronary heart disease with type 2 diabetes. There was a significant increase in serum of transforming growth factor -β1 levels in patients with coronary heart disease both with type 2 diabetes and without it with a longer course of coronary heart disease and were severer, in patients with coronary heart disease with type 2 diabetes was with a longer course of diabetes. Analysis of the nature of changes in the levels of transforming growth factor-β1 in the serum of the examined patients with coronary heart disease with type 2 diabetes and without it, depending on gender, did not reveal any significant differences. The results of the study also indicated that in patients with coronary heart disease with type 2 diabetes and without it for all duration of coronary heart disease, the levels of this indicator in the serum were probably (p &lt;0.05) higher than those in the control group. However, in patients with a significant duration of coronary heart disease (5-10 years and over 10 years) serum levels of transforming growth factor-β1 were probably (p &lt;0.05) higher than in patients with a duration of coronary heart disease less than 5 years, and in the presence of and in the absence of type 2 diabetes. At the same time, for all periods of coronary heart disease, the levels of transforming growth factor-β1 in patients with type 2 diabetes were probably higher than in patients without diabetes. Conclusion. A probable increase in the levels of transforming growth factor -β1 in the serum of patients with coronary heart disease, which was more pronounced when combining coronary heart disease with type 2 diabetes. There was a significant increase in the levels of transforming growth factor-β1 in the serum of patients with coronary heart disease both with type 2 diabetes and without it with a longer course of coronary heart disease and its severe degree, in patients with coronary heart disease with type 2 diabetes especially with a longer course of diabetes. In order to increase the informativeness of assessing the risk of cardiovascular complications and the nature of coronary heart disease in patients with type 2 diabetes, the survey should include determination of serum levels of potent profibrogenic factor like transforming growth factor-β1

Fortification of biscuit with sidr leaf and flaxseedmitigates immunosuppression and nephrotoxicity induced by cyclosporine A.

The focus of consumers in healthy food turned to the possible health benefits of particular foods and food ingredients. This study aimed to evaluate the newly fortified biscuits supplemented with sidr leaves and flaxseed and to highlight their nutritional quality and health benefits against cyclosporine A-induced dexterous effects. Sidr leaves (SL), and flaxseed (FS) were used in the preparation of fortified biscuits. Proximate analysis and sensoryevaluation were carried out on thebiscuits. In in vivo study, 15 male albino mice were used for each group. Groups were divided into control, CsA, SL, FS, and SL+FS-treated groups. Hematological analysis, kidney function tests, oxidative stress, and anti-oxidant status were estimated. Flow cytometry was utilized to detect apoptosis and autophagy levels. The enzyme-linked immunosorbent assay (ELISA) was used for detection of interleukin-2 (IL-2), interferon-γ (IFN-γ), and transforming growth factor β1 (TGF-β1) levels. The composition of biscuits complemented by SL and FS demonstrated significant improvement in the nutritional value represented by the increase in overall protein, crude fat, crude fiber, ash, and carbohydrate contents. Treatment with SL and FS restored the disturbance in hematological, kidney function, oxidative, and antioxidant biomarkers. CsA-induced apoptotic and autophagic renal cell death was suppressed. Cytokines and pro-inflammatory markers were ameliorated. The use of SL and FS in dietary products can be recommended as a functional food. Moreover, they showed renal-protective, antioxidant, anti-inflammatory, and immune-enhancing activities. PRACTICAL APPLICATIONS: Sidr leaves (SL) and flaxseed (FS) were used in the preparation of fortified biscuits. The composition of biscuits complemented by SL and FS demonstrated a significant improvement in the nutritional values represented by the increase in overall protein, crude fat, crude fiber, ash, and carbohydrate contents. SL and FS showed a potential therapeutic activity in reversing CsA-induced dexterous side effects by acting as an antioxidant, antiapoptotic, antiautophagic, anti-inflammatory, renal-protective, and immune-enhancing agents. The use of sidr leaves and flaxseed in dietary products can be recommended as a functional food. Supplementation of SL and/or FS to the diet is recommended to ensure a good health. Moreover, introducing awareness for the patients utilizing CsA to use SL and FS in their diets.

Preparation of Nano Hydroxyapatite/Polyamide Composite and Its Application in Lumbar Disc Herniation Operation

Carbon fiber (CFR) was used to strengthen the mechanical properties of hydroxyapatite/polyamide (ha/pa) to synthesize ternary composites with different mass fraction. At the same time, its mechanical and material characterization were compared with that of ha/pa without CFR. CFR-ha/pa was used as the substrate for the culture of mouse bone marrow mesenchymal stem cells (C3H10T1/2). The cell morphology and adhesion properties of materials were analyzed by immuno light staining of cytoskeleton and adhesion protein, and FESEM. The interbody fusion cage based on the material was used in the treatment of lumbar disc herniation. According to the X-ray film and the corresponding scoring standards (JOA, ODI and VAS), the therapeutic effect of the interbody fusion cage was evaluated. The results showed that there were cavities on the surface of CFR-ha/pa composites. With the increase of the mass fraction of CFR in CFR-ha/pa composites, the compressive strength (from 115 MPa to 218 Mpa), the bending strength (from 85 MPa to 140 MPa) and other mechanical indexes of the composite increased obviously. After 24 h of cell inoculation, the substrate of CFR-ha/pa group was covered by layered, well spread and fused cells, and there was dense cell adhesion around it, that was to say, the material and cells showed good adhesion performance. At the same time, the levels of Integrin α1 and Integrin β1 significantly increased in CFR-ha/pa group (P &lt; 0.05). The VAS and ODI scores of polyether ether ketone (Peek) interbody fusion cage group and CFR-ha/pa interbody fusion cage group were lower than those before operation (P &lt; 0.05). The therapeutic effect of CFR-ha/pa interbody fusion cage is better than Peek interbody fusion cage.

Identification of Na+/K+-ATPase α/β isoforms in Rhinella marina tissues by RNAseq and a molecular docking approach at the protein level to evaluate α isoform affinities for bufadienolides

Na+/K+-ATPase (NKA) function is inhibited by Bufadienolides (BD), a group of cardiotonic steroids (CTS) primarily produced by anurans of the Bufonidae family, such as Rhinella marina. This study characterized the presence of α and β NKA subunit isoforms in R. marina via RNAseq in four tissues: oocytes, skin, heart, and skeletal muscle. Transcripts encoding three α-like isoforms (α1, α2, α3) and three β-like isoforms (β1, β2, β4) were identified. The amino acid sequence of α1-like isoform shared 99.4% identity with the α1 isoform previously published for R. marina. Sequences for α2, α3, and β4 from R. marina were previously unavailable. The first extracellular loop in the α2-like isoform in R. marina showed similar substitutions to those found in their susceptible homologues in other taxa (L/Q111T and S119T); in contrast, this same loop in α3-like isoform showed similar substitutions (Q111L and G120R) to those reported for toad-eating animals such as snakes, which suggests relatively lower affinity for CTS. Docking results showed that all three α-like isoforms identified in R. marina transcriptomes have low affinity to CTS compared to the susceptible α1 isoform of Sus scrofa (pig), with α1-like isoform being the most resistant. The tissue-specific RNAseq results showed the following expression of NKA α-like and β-like subunit isoforms: Oocytes expressed α1 and β1; skin α1, β1, and low levels of β2; heart α1, α3, and β1; skeletal muscle α1, β4, with low levels of α2, α3, and β1. R. marina could be used as an important model for future structural, functional and pharmacological studies of NKA and its isoforms.

Moderate mechanical stimulation rescues degenerative annulus fibrosus by suppressing caveolin-1 mediated pro-inflammatory signaling pathway

Mechanical loading can induce or antagonize the extracellular matrix (ECM) synthesis, proliferation, migration, and inflammatory responses of annulus fibrosus cells (AFCs), depending on the loading mode and level. Caveolin-1 (Cav1), the core protein of caveolae, plays an important role in cellular mechanotransduction and inflammatory responses. In the present study, we presented that AFCs demonstrated different behaviors when subjected to cyclic tensile strain (CTS) for 24 h at a magnitude of 0%, 2%, 5% and 12%, respectively. It was found that 5% CTS had positive effects on cell proliferation, migration and anabolism, while 12% CTS had the opposite effects. Besides, cells exposed to interleukin-1β stimulus exhibited an increase expression in inflammatory genes, and the expression of these genes decreased after exposure to moderate mechanical loading with 5% CTS. In addition, 5% CTS decreased the level of Cav1 and integrin β1 and exhibited anti-inflammatory effects. Moreover, the expression of integrin β1 and p-p65 increased in AFCs transfected with Cav1 plasmids. In vivo results revealed that moderate mechanical stimulation could recover the water content and morphology of the discs. In conclusion, moderate mechanical stimulation restrained Cav1-mediated signaling pathway and exhibited anti-inflammatory effects on AFCs. Together with in vivo results, this study expounds the underlying molecular mechanisms on the effect of moderate mechanical stimulation on intervertebral discs (IVDs) and may provide a new therapeutic strategy for the treatment of IVD degeneration.

Abstract 13385: Protective Effect of Beta3-Adrenergic Receptor Deficiency on Diabetic Cardiomyopathy: Reversal of Type 2 Diabetes-Induced Contrast Changes of Cardiac SERCA 2a and Inducible Nitric Oxide Synthase

Background: We have shown that diabetes (DM)-induced cardiac dysfunction and β-adrenergic desensitization were prevented in β 3 -adrenergic receptor (AR) knockout (β 3 KO) mice. However, the molecular mechanism is unclear. We hypothesize that reversal of type 2 (T2) DM-induced alterations of cardiac SR Ca 2+ -ATPase (SERCA2a) and inducible nitric oxide synthase (iNOS) by β 3 KO may play a key role for the protective effect. Methods: LV myocyte protein levels of β 1 - and β 3 -AR, SERCA2a and iNOS, myocyte contractile and [Ca 2+ ] i transient ([Ca 2+ ] iT ) responses to isoproterenol (ISO, 10 -8 M) were compared in 4 groups (n=6) female mice: 2 Vehicle controls wild-type (CWT) and Cβ 3 KO; and 2 T2DM of T2WT and T2β 3 KO.T2 was induced by fed high-fat diet (HFD) for 14 weeks (W), but after HFD for 4 W receiving streptozotocin (STZ, 40 mg/kg/day, i.p. for 5 days). Results: Mice-treated with HFD plus STZ had high blood glucose (~350 to 380 mg/dl). Versus CWT, T2WT myocytes had significantly decreased protein levels of SERCA2a (0.18 vs 0.29), and β 1 -AR (0.35 vs 0.49), but increased iNOS (0.25 vs 0.15) and β 3 -AR (0.25 vs 0.14) with reduced cell contraction (dL/dt max ) (72.7 vs 140.9 μm/s), relaxation (dR/dt max ) (50.9 vs 117.3 μm/s) and [Ca 2+ ] iT (0.16 vs 0.22). Moreover, in T2WT myocytes, ISO caused significantly less increases in dL/dt max (37% vs 59%), dR/dt max (30% vs 53%) and [Ca 2+ ] iT (19% vs 30%). Versus CWT, T2β 3 KO did not alter basal myocyte contraction and relaxation and response to ISO stimulation, but had significantly increased protein levels of SERCA2a (0.38 vs 0.29). T2β 3 KO prevented DM-caused increase in iNOS (T2β 3 KO: 0.16 vs CWT: 0.15) and decrease in β 1 -AR (0.46 vs 0.49). Versus T2β 3 KO, Cβ 3 KO mice had similar alterations. Importantly, in contrast to T2WT, in T2β 3 KO myocytes, the increased SERCA2a and reduced iNOS correlated with normal basal cell contraction and relaxation with preserved ISO-stimulated inotropic response. ISO caused similar increases in dL/dt max (60% vs 59%), dR/dt max (51% vs 53%) and [Ca 2+ ] iT (32% vs 30%). Conclusions: β 3 KO prevents T2DM-caused downregulation of cardiac β 1 -ARs and reverses increased iNOS and decreased SERCA2a, leading to the preservation of myocyte function, [Ca 2+ ] iT , and β-adrenergic reserve. Thus, β 3 -AR may provide a new target for DCM.

Pharmaceutical Evaluation of Honokiol and Magnolol on Apoptosis and Migration Inhibition in Human Bladder Cancer Cells.

Among herbal medicines, magnolia bark extract, particularly its components honokiol (Hono) and magnolol (Mag), has been widely documented to have antineoplastic properties. The present study aimed to evaluate the synergism of Hono and Mag in bladder cancer therapy both in vitro and in vivo. Treatment with Mag alone at concentrations up to 80 μM failed to have an antiproliferative effect. In contrast, the combination of Hono and Mag at 40 μM decreased viability, caused cell cycle arrest and enhanced the proportion of Annexin V/7AAD-positive cells. Moreover, Mag with Hono at 40 μM induced caspase 3-dependent apoptosis and autophagy. Neither Hono nor Mag alone had an anti-migratory effect on bladder cancer cells. In contrast, Hono and Mag at 20 μM inhibited the motility of TSGH8301 and T24 cells in wound-healing and Transwell assays. The above phenomena were further confirmed by decreased phosphorylated focal adhesion kinase (p-FAK), p-paxillin, integrin β1, and integrin β3 protein levels. In a nude mouse xenograft model, Mag/Hono administration preferentially retarded T24 tumor progression, which was consistent with the results of cellular experiments. Current findings suggest Hono and Mag treatment as a potential anticancer therapy for both low- and high-grade urothelial carcinoma.

Vaspin deficiency failed to promote the proliferation of BMSCs in osteoarthritis.

The aim of this study was to estimate the possible role of vaspin in the proliferation of bone mesenchymal stem cells (BMSCs) and its molecular mechanisms in the bone marrow microenvironment of osteoarthritis (OA). This study included 15 non-obese elderly patients with severe knee OA and 15 non-obese controls with femoral neck fracture. Patients all underwent hip or knee arthroplasty surgery to restore joint shape and function. Bone marrow samples were taken during surgery to estimate vaspin and transforming growth factor (TGF)-β1 levels by enzyme-linked immunosorbent assay and to observe the effect of vaspin on BMSCs proliferation by Cell Counting Kit-8. Real-time polymerase chain reaction and western blot were performed to evaluate the effect of vaspin on the genes and proteins of Akt involved in the PI3K/AKT signaling pathway. Bone marrow vaspin levels were significantly lower in OA patients compared to controls (P=.03). Furthermore, we found a significant correlation between vaspin and TGF-β1 concentrations in bone marrow (r=.60, P<.01). In addition, the in vitro studies indicated the proliferation of BMSCs was significantly promoted when the vaspin treatment concentration was 150ng/mL (P<.01). Meanwhile, we found that the Akt messenger RNA and pAkt protein levels in BMSCs were increased after vaspin treatment (P<.05). The findings of this study suggest there was abnormal expression of vaspin in OA bone marrow microenvironment, and vaspin likely had a mediator role in the proliferation of BMSCs, which may work by promoting the activation of the PI3K/AKT signaling pathway.

Effect of Wenxia Changfu Formula Combined With Cisplatin Reversing Non-Small Cell Lung Cancer Cell Adhesion-Mediated Drug Resistance.

Non-small cell lung cancer (NSCLC), the major form of primary lung cancer, is a common cause of cancer-related death worldwide. Cell adhesion-mediated drug resistance (CAM-DR), a form of chemotherapy resistance, has been reported to confer resistance to various chemotherapeutic agents. Integrin β1 signaling plays an important role in CAM-DR and has been proposed as a potential target for NSCLC. Wenxia Changfu Formula (WCF) is a Traditional Chinese Compound Prescription for the intervention treatment of NSCLC combined with cisplatin (DDP). This study aims to investigate the effect and mechanism of WCF combined with DDP in reversing CAM-DR. Firstly, the chemical profile of WCF was characterized by UPLC/Q-TOF-MS analysis. A total of 237 compounds with mzCloud Best Match of greater than 70 were identified by using the online database mzCloud. Secondly, we established A549 three-dimensional(3D) cells cultured in vitro and nude mice xenografts models of the A549 cell line with Integrin β1 overexpression. In vitro, the cell viability, migration and adhesion were measured though MTT Assay, Wound Healing Assay and Cell Adhesion Assay, the Integrin β1 expression of the A549 cells was assessed through immunocytochemistry; in vitro, the transplanted tumor morphology and the colocalization of Integrin β1 and its ligands were tested by HE staining and immunofluorescence. As a result, we found that the combination effectively reduced cell viability, suppressed migration and adhesion, and downregulated the protein level of Integrin β1 in three-dimensional cultured A549 cells. And the combination of WCF with DDP significantly inhibited tumor growth, increased organelle vacuolations and decreased colocalization of Integrin β1 and its ligands including fibulin-2 and laminin. Taken together, our results confirm that the combination of WCF with DDP could reverse the lung cancer CAM-DR through the Integrin β1 signaling pathway.