Effect of miR-96-5p targeting IRS1 on apoptosis of PC12 cells induced by aluminum maltol / 中华劳动卫生职业病杂志

Objective: To investigate the effect and mechanism of miR-96-5p on apoptosis of PC12 cells induced by maltol aluminum. Methods: In January 2021, PC12 cells at logarithmic growth phase were divided into blank control group and low, medium and high dose group. Cells in each group were treated with 0, 100, 200 and 400 μmol/L maltol aluminum for 24 hours respectively. Cells were collected and cell apoptosis rates were detected by flow cytometry, miR-96-5p and insulin receptor substrate 1 (IRS1) mRNA expressions were detected by qRT-PCR, and the protein expression levels of cysteine protease 3 (Caspase3) 、activated cysteine protease 3 (Cleaved-caspase3) 、IRS1、phosphorylated protein kinase B (p-AKT) and phosphorylated glucose synthesis kinase 3β (p-GSK3β) were detected by western blotting. The target binding relationship between miR-96-5p and IRS1 was detected by double luciferase reporter gene experiment. The miR-96-5p inhibitor cells and negative control cells were constructed after transfecting PC12 cells with miR-96-5p inhibitor for 24 hours. The cells were divided into blank control group, negative control group, aluminum exposure group, aluminum exposure+negative control group, aluminum exposure+miR-96-5p inhibition group, and miR-96-5p inhibition group. After transfecting PC12 cells with miR-96-5p inhibition and IRS1 siRNA for 24 h, the cells were divided into aluminum exposure+miR-96-5p inhibition+negative control group and aluminum exposure+miR-96-5p inhibition+IRS1 inhibition group. The control group was cultured in complete culture medium, and cells in the aluminum exposure group were treated with 200 μmol/L maltol aluminum for 24 hours. Cells in each group were collected and the apoptosis rate, miR-96-5p and IRS1 mRNA expression levels, as well as protein expression levels of Caspase3, Cleaved-caspase3, IRS1, p-AKT, and p-GSK3β were measured. Results: After 24 hours of exposure, compared with blank control group and low-dose group, the apoptosis rates, relative expressions of Caspase3 and Cleaved-caspase3 proteins, and relative expressions of miR-96-5p in the medium and high-dose groups of PC12 cells were significantly increased, while the relative expression levels of IRS1 mRNA, IRS1, p-AKT and p-GSK3β proteins were significantly decreased (P<0.05). Targetscan prediction and double luciferase report experiment both proved that IRS1 was a direct target gene of miR-96-5p. In the transfection experiment, compared with the aluminum exposure group, the apoptosis rate, the relative expressions of Caspase3 and Cleaved-caspase3 proteins, the relative expression of miR-96-5p in the aluminum exposure+miR-96-5p inhibition group were significantly decreased, while the relative expression levels of IRS1 mRNA and IRS1, p-AKT and p-GSK3β proteins were significantly increased (P<0.05). In the IRS1 low expression experiment, compared with the aluminum exposure+miR-96-5p inhibition+negative control group, the apoptosis rate, the relative expressions of Caspase3 and Cleaved-caspase3 proteins in the aluminum exposure+miR-96-5p inhibition+IRS1 inhibition group were significantly increased, while the relative expression levels of IRS1 mRNA and IRS1, p-AKT and p-GSK3β proteins were significantly decreased (P<0.05) . Conclusion: The increased expression of miR-96-5p and the targeted inhibition of IRS1 may be one of the mechanisms of apoptosis of PC12 cells induced by maltol aluminum exposure.

Ultrasonic differential diagnosis of scar pregnancy and lower uterine cavity non-scar pregnancy / 中国医学影像技术

Objective To investigate the value of two-dimensional and three-dimensional ultrasonography in differential diagnosing cesarean scar pregnancy (CSP) and lower uterine cavity non-scar pregnancy.Methods Totally 67 patients with CSP (CSP group)and 29 patients with lower uterine cavity non-scar pregnancy (lower uterine cavity non-scar pregnancy group) were enrolled.Two-dimensional and three-dimensional ultrasonography were performed to observe the gestational sac implantation site,the relationship between gestational sac and cesarean section scar and the main source of blood flow,and the residual muscle thickness of cesarean section scar was measured as well.Logistic regression model was established,and the diagnostic efficacy was evaluated with ROC curve.Results Statistical differences of relationship between gestational sac and scar,the main source of blood flow and residual muscle thickness were found between CSP group and lower uterine cavity non-scar pregnancy group (all P＜0.001).The area under the ROC curve of the Logistic regression model was 0.878 (P＜0.001).Taking prediction accuracy of 0.680 as a cut-off value,the accuracy of this model in predicting was 86.46％,the sensitivity was 89.55％ and the specificity was 79.31％.Taking gestational sac implanted scars and nourish the blood flow from the lower anterior wall of the uterus as the diagnostic criteria for CSP,the Kappa value of two-dimensional and three-dimensional ultrasonograpby in diagnosis CSP and non-scar pregnancy was 0.699 and 0.711,respectively.Conclusion Comprehensive analysis of the relationship between gestational sac and scars,the main source of blood flow and residual muscle thickness with Logistic regression model can improve differential diagnostic efficacy of CSP with lower uterine cavity non-scar pregnancy.

Inhibitory effect of compound W3D on LPS-induced release of inflammatory mediators of RAW264.7 cells through TLR4-MyD88-NF-κB signaling pathway / 中国药理学通报

Aim To investigate the effect of the novel benzoxazolone derivative 4-( 5′-dimethylamino )-naph-thalenesulfonyl-2 ( 3H )-benzoxazolone ( W3D ) on TLR4-MyD88-NF-κB signaling pathway in LPS-in-duced RAW264.7 cells. Methods The cell viability was detected by MTT assay, and the contents of TNF-α, IL-6, IL-1β and COX-2 in the cell supernatant were analyzed using ELISA methods. The protein ex-pression of IL-6, TLR4, MyD88, p-IRAK4 and NF-κB were investigated by western blot analysis, and the mRNA expressions of TLR4, MyD88 and IL-6 were an-alyzed by RT-PCR. Results W3D could obviously in-hibit the production of TNF-α, IL-6 and IL-1β in LPS- induced RAW264.7 cell supernatant, but it had no effect on the release of COX-2. Compared with the model group, the expressions of TLR4, MyD88 and IL-6 were decreased significantly in a dose dependent manner. Meanwhile, the expressions of p-IRAK4 and nucleus of NF-κB were decrease in W3D treated group compared with the model group. Conclusion The no-vel compound W3D could inhibit the release of the in-flammatory mediators through the regulation of TLR4-MyD88-NF-κB signaling pathway.

End-binding protein 1 stimulates paclitaxel sensitivity in breast cancer by promoting its actions toward microtubule assembly and stability

Paclitaxel is a microtubule-targeting agent widely used for the treatment of many solid tumors. However, patients show variable sensitivity to this drug, and effective diagnostic tests predicting drug sensitivity remain to be investigated. Herein, we show that the expression of end-binding protein 1 (EB1), a regulator of microtubule dynamics involved in multiple cellular activities, in breast tumor tissues correlates with the pathological response of tumors to paclitaxel-based chemotherapy. In vitro cell proliferation assays reveal that EB1 stimulates paclitaxel sensitivity in breast cancer cell lines. Our data further demonstrate that EB1 increases the activity of paclitaxel to cause mitotic arrest and apoptosis in cancer cells. In addition, microtubule binding affinity analysis and polymerization/depolymerization assays show that EB1 enhances paclitaxel binding to microtubules and stimulates the ability of paclitaxel to promote microtubule assembly and stabilization. These findings thus reveal EB1 as a critical regulator of paclitaxel sensitivity and have important implications in breast cancer chemotherapy.

Diagnosis and treatment of capillary leak syndrome in children / 中国小儿急救医学

Capillary leak syndrome (CLS) is a common and life-threatening disorder in children.The common pathogenies are sepsis,severe trauma,shock,asphyxia,extracorporeal circulation,ischemical reperfusion injury and so on.It is characterized by shock resulting from leakage of plasma,which is refected by accompanying hemoconcentration,hypoalbuminemia and edema,sometimes followed by multiple organ dysfunction syndrome.Clinicians should consider the diagnosis in patients with unexplained edema and hypotension.To know the pathophysiology and clinical course will be benefit to proper medicine choosen.

Risk factors for capillary leak syndrome in neonates / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the risk factors for capillary leak syndrome (CLS) in neonates.</p><p><b>METHODS</b>The clinical data of 52 neonates with CLS (case group) were retrospectively reviewed. Fifty hospitalized neonates without CLS were used as the control group. The possible factors for the development of CLS were identified by univariate analysis. The independent risk factors for CLS were determined by multivariate logistic regression analysis.</p><p><b>RESULTS</b>The univariate analysis showed that the incidences of hyperglycemia, respiratory distress syndrome (RDS), sepsis and cold injury syndrome in the case group were significantly higher than those in the control group (P<0.05). The logistic regression analysis showed that sepsis (OR=5.004, P=0.001), RDS (OR=3.880, P=0.013) and cold injury syndrome (OR=3.207, P=0.023) were the independent risk factors for the development of CLS.</p><p><b>CONCLUSIONS</b>RDS, sepsis and cold injury syndrome are independent risk factors for CLS in neonates. Hyperglycemia may be associated with the development of CLS.</p>

Suppression of allergic airway inflammation in a mouse model by Der p2 recombined BCG-induced adaptive CD4~+CD25~+ Treg / 中国病理生理杂志

AIM:The bacillus calmette-guerin(BCG) vaccine is the most widely used Th1-inducing vaccine.In recent years,some studies argued that mycobacterium vaccae can be used as adjuvant to induce regulatory T cells(Treg) and then suppress asthmatic airway inflammation.We previously have engineered recombined BCG that expressed Der p2 of house dust mites(Der p2 rBCG) on the cell wall.The aim of this study is to investigate the immune regulatory mechanisms of Der p2 rBCG.METHODS:Mice were vaccined with PS,BCG or rBCG.The relative proportion and the absolute numbers of related Tregs in spleen cells were analyzed.The suppressive activity of Der p2 rBCG-induced CD4+CD25+ T cells was detected both in vitro and in vivo.RESULTS:(1) Der p2 rBCG induced a CD4+CD25+Foxp3+ T cell subtype.(2) Der p2 rBCG-induced CD4+CD25+ T cells suppressed the proliferation of Th2 effector cells in vitro in an antigen-specific way.(3) Der p2 rBCG-induced CD4+CD25+ T cells mediated Der p2 specific suppression of airway allergy in vivo.CONCLUSION:Der p2 rBCG induces a CD4+CD25+Foxp3+ T cell subtype,which suppresses inflammation in allergic airway in a mouse model.