Effect of abnormal activated B cells in patients with ankylosing spondylitis and its molecular mechanism

OBJECTIVE: Ankylosing spondylitis (AS) is a common clinical autoimmune disease, the pathogenesis of which, however, is not yet elucidated. In this study, we aim to explore the effect of B cells in the development and progression of AS and its underlying mechanism.

PATIENTS AND METHODS: B cells were isolated from peripheral blood of AS patients and normal controls. Surface expression of CD40 in B cells was detected by flow cytometry. Expressions of downstream genes in MAPK pathway were detected by Western blot. Moreover, IL-10 expressions in peripheral blood of AS patients and normal controls were detected by ELISA.

RESULTS: No difference was found in the surface expression of CD40 in B cells between AS patients and normal controls. However, CD40 expression was inhibited after B cells in peripheral blood were specifically stimulated by lipopolysaccharide (LPS) in vitro. Abnormal activated B cells, dysregulated p38 expressions and decreased serum expressions of IL-10 were also observed in AS patients.

CONCLUSIONS: Abnormal surface expression of CD40 inn B cells of AS patients may lead to abnormal activation of B cells, thereby interfering the p38 MAPK pathway and reducing the IL-10 secretion.

L'articolo Effect of abnormal activated B cells in patients with ankylosing spondylitis and its molecular mechanism sembra essere il primo su European Review.

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MMP-2 participates in the sclera of guinea pig with form-deprivation myopia via IGF-1/STAT3 pathway

OBJECTIVE: To investigate the expression changes of MMP-2 (matrix metalloproteinases-2) mediated by IGF-1 (insulin-like growth factors-1) STAT3 (signal transducer and activator of transcription 3) pathway in the sclera of the form-deprivation myopia guinea pigs.

MATERIALS AND METHODS: Twenty-four three-week-old guinea pigs were randomly divided into 4 groups: group A (Control), B, C and D. Guinea pigs in group A were sacrificed after 21 days without any special treatment. Guinea pigs in group B were sacrificed 7 days after receiving stitch in the right eye. Guinea pigs in group C were sacrificed 14 days after receiving stitch in the right eye. Guinea pigs in group D were sacrificed 21 days after receiving stitch in the right eye. Eyeball refraction and axial length of guinea pigs were measured before sacrifice. Eyeballs of guinea pigs were enucleated after sacrifice. The expressions of IGF-1, STAT3 and MMP-2 in scleral tissue were detected by Western blot.

RESULTS: Axial length extension and myopia appeared in the right eye of guinea pigs in group B. The expressions of IGF-1, STAT3 and MMP-2 in the sclera significantly increased after 7 days of occlusion compared with that in control group A (p<0.05). In the right eye of group C, the axial prolongation and myopia formation appeared after 14-day occlusion. The expressions of IGF-1, STAT3 and MMP-2 in sclera significantly increased compared with that in group A (p<0.05). In the right eye of group D, the axial extension and myopia formation occurred. IGF-1, STAT3 and MMP-2 in scleral significantly upregulated 21 days after occlusion (p<0.05). Furthermore, at different stages of deprivation, protein expressions of MMP-2 and IGF-1 in sclera were positively correlated (r = 0.962, p<0.01).

CONCLUSIONS: Form-deprivation of guinea pigs lead to increased expressions of IGF-1, STAT3 and MMP-2 in the sclera and myopia of guinea pigs. The expressions of IGF-1, STAT3 and MMP-2 increased progressively over the time of deprivation. Additionally, overexpression of MMP-2 mediated by IGF-1/STAT3 pathway in sclera might promote the formation of myopia.

L'articolo MMP-2 participates in the sclera of guinea pig with form-deprivation myopia via IGF-1/STAT3 pathway sembra essere il primo su European Review.

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Inflammatory cytokines IL-6, IL-10, IL-13, TNF-α and peritoneal fluid flora were associated with infertility in patients with endometriosis

OBJECTIVE: To investigate the correlations of inflammatory factors, interleukin-6 (IL-6), IL-10, IL-13 and tumor necrosis factor-α (TNF-α), and composition of bacterial flora in the peritoneal fluid with infertility in endometriosis patients.

PATIENTS AND METHODS: A total of 55 patients diagnosed with endometriosis and infertility in the Gynecology Clinic of our hospital from June 2014 to July 2017 were selected as observation group, and another 30 non-endometriosis and non-infertility patients were enrolled as control group. The peritoneal fluid was extracted from patients in both groups, and the total white cell count and the percentage of leukocyte subset were determined. The total genome deoxyribonucleic acid (DNA) of microorganisms in peritoneal fluid was extracted, and the composition of microorganisms was analyzed using the Ion Torrent PGM platform (BGI). The levels of IL-6, IL-10, IL-13, and TNF-α in peritoneal fluid were detected via enzyme-linked immunosorbent assay (ELISA). Moreover, the correlations of inflammatory factors in the peritoneal fluid with endometriosis complicated with infertility were analyzed via Logistic regression analysis.

RESULTS: The total white cell count, monocytes, neutrophils, eosinophils and basophils in endometriosis patients complicated with infertility were significantly higher than those in control group (p<0.05). Results of ELISA showed that the levels of IL-6, IL-10, IL-13, and TNF-α in peritoneal fluid of endometriosis patients complicated with infertility were significantly higher than those in control group (p<0.05). In peritoneal fluid of patients in both groups, Proteobacteria and Firmicutes were mainly dominated, followed by Actinobacillus, Bacteroidetes, Fusobacteria and Tenericutes, and there was no significant difference in the Eumycota between the two groups (p>0.05). Logistic regression analysis results showed that there were significant correlations of inflammatory factors (IL-6, IL-10, IL-13, and TNF-α) with endometriosis complicated with infertility.

CONCLUSIONS: There are many kinds of Eumycota in the peritoneal fluid of endometriosis patients complicated with infertility, but they are not the main pathogenic factors. Inflammatory factors (IL-6, IL-10, IL-13, and TNF-α) can be used as important reference indexes for the diagnosis of endometriosis complicated with infertility.

L'articolo Inflammatory cytokines IL-6, IL-10, IL-13, TNF-α and peritoneal fluid flora were associated with infertility in patients with endometriosis sembra essere il primo su European Review.

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MEG3 is involved in the development of glaucoma through promoting the autophagy of retinal ganglion cells

OBJECTIVE: In this study, we aimed at investigating whether MEG3 may be involved in the pathogenesis of glaucoma by regulating the autophagy of retinal ganglion cells (RGCs).

MATERIALS AND METHODS: We used qRT-PCR to detect the expression of MEG3 in RGC-5s cell line under high hydrostatic pressure. RGC-5s were transfected with a lentiviral vector to achieve MEG3 overexpression or knockdown. The influence of overexpression or inhibition of MEG3 on cell proliferation and apoptosis was observed using CCK-8 test and flow cytometry. After overexpression of MEG3 and/or knockdown of MEG3 or Beclin-1, detection of the expressions of autophagy-related and apoptosis-related proteins was performed using Western blot.

RESULTS: MEG3 expression level increased in RGC-5 cells under high hydrostatic pressure, while exogenously decreased MEG3 expression can reverse the impact of the high pressure on RGC-5 cells. Additionally, overexpression of MEG3 can improve Atg3 expression, promote cell apoptosis, inhibit cell proliferation, and enhance autophagy levels. Meanwhile, knockdown of Beclin-1 up-regulated Bcl-2 level.

CONCLUSIONS: Upregulation of MEG3 is involved in the pathogenesis of glaucoma through promoting apoptosis of retinal ganglion cells, the mechanism of which may be related to the enhanced autophagy levels.

L'articolo MEG3 is involved in the development of glaucoma through promoting the autophagy of retinal ganglion cells sembra essere il primo su European Review.

https://ift.tt/2Id7SJg

Correlations of SOX9 expression with serum IGF1 and inflammatory cytokines IL-1α and IL-6 in skin lesions of patients with acne

OBJECTIVE: To study the correlations of sex determining region Y-box 9 (SOX9) expression with serum type-1 insulin-like growth factor (IGF-1), interleukin-1α (IL-1α), and interleukin-6 (IL-6) in skin lesion tissues of patients with acne.

PATIENTS AND METHODS: Six patients with acne who were treated for the first time in our outpatient clinic from June 2017 to July 2017 were selected as observation group, and 6 normal subjects were selected as control group. The expression of SOX9 was detected by immunohistochemistry. The protein expressions of IGF-1, IL-1α, and IL-6 were detected by enzyme-linked immunosorbent assay (ELISA). SOX9 was detected by quantitative polymerase chain reaction (qPCR).

RESULTS: Compared with that in control group, the expression of SOX9 in observation group was significantly increased (p < 0.05). Compared with those in control group, the expressions of IGF-1, IL-1α and IL-6 in observation group were significantly increased (p < 0.05). Compared with that in control group, the mRNA expression of SOX9 in observation group was significantly increased (p < 0.05). SOX9 was positively correlated with IGF-1, IL-1α and IL-6.

CONCLUSIONS: The expressions of SOX9, IGF-1, IL-1α, and IL-6 in skin lesion tissues of patients with acne are increased, and SOX9 is positively correlated with IGF-1, IL-1α, and IL-6 and can be used as a target for the treatment of acne inflammation.

L'articolo Correlations of SOX9 expression with serum IGF1 and inflammatory cytokines IL-1α and IL-6 in skin lesions of patients with acne sembra essere il primo su European Review.

https://ift.tt/2I84GTp

MiR-133a inhibits fracture healing via targeting RUNX2/BMP2

OBJECTIVE: To analyze the mechanism of miR-133a in inhibiting fracture healing through regulating runt-related transcription factor 2 (RUNX2) signaling pathway.

PATIENTS AND METHODS: A total of 80 patients with fracture admitted to our hospital from January 2016 to January 2017 were divided into 2 groups according to nonunion fracture or healing fracture: nonunion fracture group (n = 40) and control group (n= 40). After admission, patients underwent the surgery, respectively, and the bone tissues were taken for stand-by application. The expression level of bone morphogenetic protein 2 (BMP2) was detected using the immunohistochemical method, the expression level of RUNX2 protein was detected by Western blotting, and the expression level of micro ribonucleic acid (miR)-133a was detected by quantitative polymerase chain reaction (qPCR). Moreover, the bioinformatics method was used to predict the target gene of miR-133a, and the luciferase reporter gene was used to detect the binding of miR-133a to RUNX2.

RESULTS: Compared with those in control group, the expression of BMP2 and the relative expressions of RUNX2 protein and miR-133a were significantly decreased; the differences were statistically significant (p < 0.05). Pearson correlation analysis showed that miR-133a was negatively correlated with RUNX2. After overexpression of miR-133a, the expression level of RUNX2 was decreased, but it was increased significantly after interference in miR-133a. Besides, it was found in dual-luciferase reporter assay that miR-133a bound to RUNX2.

CONCLUSIONS: MiR-133a inhibits the bone formation through inhibiting the RUNX2/BMP2 signaling pathway, thereby negatively regulating the fracture healing.

L'articolo MiR-133a inhibits fracture healing via targeting RUNX2/BMP2 sembra essere il primo su European Review.

https://ift.tt/2IebZoC

BAY-11-7082 induces apoptosis of multiple myeloma U266 cells through inhibiting NF-κB pathway

OBJECTIVE: To study the effects of BAY-11-7082 on proliferation and apoptosis of U266 cells and its mechanism of action.

MATERIALS AND METHODS: Multiple myelomas U266 cells were cultured and divided into control group and gradient-concentration BAY-11-7082 groups (1 μmol/L, 2 μmol/L, 4 μmol/L and 8 μmol/L). Cells in BAY-11-7082 groups were treated with drugs in different concentrations for 4 h, while those in control group were added with an equal volume of solvent. The cell viability was detected via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, and lactate dehydrogenase (LDH) release assay was used to detect the cytotoxicity. Furthermore, cells in low-concentration and high-concentration BAY-11-7082 groups were compared with those in control group. The cell proliferation level was evaluated via cell cycle assay, the interleukin-6 (IL-6) level in cells was detected via enzyme-linked immunosorbent assay (ELISA), and the β-catenin protein expression was detected via Western blotting. Moreover, flow cytometry and Hoechst staining were performed to detect the number of apoptotic cells, and the apoptosis level was detected via caspase3 activity and apoptosis-related protein expression. Finally, the levels of p65, p50 and inhibitor kappa B kinase β (IKKβ) were detected via polymerase chain reaction (PCR), and the expressions and changes of phosphorylated (p)-p65 and p-IKKβ were detected via Western blotting.

RESULTS: BAY-11-7082 could reduce the U266 cell viability and increase the cytotoxic effect. Based on gradient concentration, 2 μmol/L was selected as the low concentration, while 4 μmol/L was selected as the high concentration. Compared with those in control group, the number of cells in the S and G2/M phases in drug administration groups was significantly decreased, but that in the G0/G1 phase was significantly increased. Besides, the secretion of IL-6 in cells in drug administration groups was significantly decreased compared with that in control group. The β-catenin protein expression was decreased in drug administration groups, and there was also a difference between high concentration group and low concentration group. Flow cytometry and Hoechst staining showed that the proportion of apoptotic cells in drug administration groups was significantly increased. Western blotting and detection of caspase3 activity revealed that the expression and activation of apoptosis-related protein were increased in drug administration groups. It was found in the detection of nuclear factor-κB (NF-κB) pathway that the NF-κB pathway was inhibited in drug administration groups, and there was also a statistically significant difference between high concentration group and low concentration group.

CONCLUSIONS: BAY-11-7082 inhibits the proliferation and induces the apoptosis of U266 cells through inhibiting NF-κB pathway.

L'articolo BAY-11-7082 induces apoptosis of multiple myeloma U266 cells through inhibiting NF-κB pathway sembra essere il primo su European Review.

https://ift.tt/2I6QJVH