Objective To explore the effects of cAMP response element binding protein ( CREB) on taxol-induced cell cycle arrest in HeLa cells .Methods MTT assay was used to determine the optimal concentration and treatment time . PCR method was used to construct the recombinant plasmid pCI neo /CREB( PN) and site-directed mutagenesis recombinant plasmid pCI neo/CREB-M(PM).Cell cycle was assayed by flow cytometry .Expressions of pCREB, CREB, cyclins and CDKs were assayed by Western blotting .Results The effective conditions of taxol treatment on HeLa cells were 0.1μmol/L for 24 hours.After cells were treated with 0.1μmol/L taxol, G2/M phase was arrested in a time-dependent manner , accomplished with the decrease of cyclin A , a significant increase of cyclin B1, D1 and phosphorylated CREB (pCREB) protein expression, whereas, no marked changes were observed in cyclin E , CDK1, CDK2, CDK4 and CREB expressions. However, combinantion of PM and taxol treatment significantly reduced taxol-induced G2/M phase arrest, and reversed the effect of taxol-decreased cyclin A, increased cyclin B1 and D1 expression.Conclusion Tanscription factor CREB-mediated specific cyclins play a pivotal role in taxol-induced G 2/M arrest in HeLa cells .

Objective: To explore the effect of substrate mechanical microenvironment and cell-cell interaction on differentiation of bone marrow mesenchymal stem cells (BMSCs), intrahepatic cellular function and phenotype. Methods: Bone marrow mesenchymal stem cells (BMSCs)-hepatocytes (HCs) and BMSCs-hepatic stellate cells (HSCs) were co-cultured on polyvinyl alcohol (PVA) hydrogel substrates at different stiffness (4.50 ± 0.47 kPa, 19.00 ± 3.51 kPa and 37.00 ± 2.09 kPa) by non-contact co-culture method. Furthermore, the effect of substrate mechanical microenvironment on BMSCs, HCs and HSCs and the activation and proliferation of HCs under different co-cultured condition was studied. A Student's t-test was used to compare the two groups. Results: The expression ofα-smooth muscle actin (α-SMA) and collagenα1- I (Col1A1) in BMSCs and HSCs cultured on its own increased with increase of substrate stiffness. After 72 h, the expression of albumin (ALB) of HCs on three stiff substrates was significantly higher than that of 24 and 48 h. Moreover, the expression of ALB of HCs increased with the increase of substrate stiffness. During the co-culture of BMSCs and HSCs, BMSCs of all three stiffness substrates promoted the expression ofα-SMA, Col1A1 in HSCs, but reduced the expression of PPARγin HSCs cells, thererby promoted the activation of HSCs, with apparent stiffness at 37 kPa. HSCs promoted the expression of ABL in BMSCs at three stiff substrates, but inhibited the expression of alpha-SMA and Col1A1 in BMSCs at 37 kPa, suggesting that co-culture had inhibited the differentiation of BMSCs myofibroblasts, and promoted the differentiation of hepatocyte-like cells, especially at high stiff substrates. In the co-culture of BMSCs and hepatic parenchymal cells, BMSCs had promoted the proliferation of hepatic parenchymal cells at 4.5 kPa. Further, hepatic parenchymal cells had inhibited the expression ofα-SMA in BMSCs, and promoted the expression of Alb, with inhibition of BMSCs differentiation towards myofibroblasts. Conclusion The differentiation of BMSCs affects the substrate mechanical microenvironment, co-culture of HCs and HSCs. Simultaneously, affecting the function of hepatocytes in relation to the mechanical state of the substrates.

This study selected the top five general hospitals in the United States of America, collected the practice information of acupuncturists through the official website, and analyzed the professional department, expertise, professional title, education background, residency, fellowship, board certification, etc. of these acupuncturists to understand the practice situation of acupuncturists in the hospitals. The results of the study showed the practice of acupuncturists in the United States of America is improving. With further localization, locally trained acupuncturists have gradually become the main body of acupuncturists; acupuncture treatment is still mainly for analgesia, but the scope of treatment continues to expand, and departments that accept acupuncturists are gradually increasing. However, the group of licensed acupuncturists is still a minority in the group of acupuncturists in the United States of America. Native American physicians still use acupuncture as an alternative option and neglect to obtain a license. The entry barrier for acupuncturists still needs to be raised.

[This corrects the article DOI: 10.1016/j.apsb.2021.09.017.].

Pulmonary endothelial barrier dysfunction is a hallmark of clinical pulmonary edema and contributes to the development of acute lung injury (ALI). Here we reported that ruscogenin (RUS), an effective steroidal sapogenin of Radix Ophiopogon japonicus, attenuated lipopolysaccharides (LPS)-induced pulmonary endothelial barrier disruption through mediating non-muscle myosin heavy chain IIA (NMMHC IIA)‒Toll-like receptor 4 (TLR4) interactions. By in vivo and in vitro experiments, we observed that RUS administration significantly ameliorated LPS-triggered pulmonary endothelial barrier dysfunction and ALI. Moreover, we identified that RUS directly targeted NMMHC IIA on its N-terminal and head domain by serial affinity chromatography, molecular docking, biolayer interferometry, and microscale thermophoresis analyses. Downregulation of endothelial NMMHC IIA expression in vivo and in vitro abolished the protective effect of RUS. It was also observed that NMMHC IIA was dissociated from TLR4 and then activating TLR4 downstream Src/vascular endothelial cadherin (VE-cadherin) signaling in pulmonary vascular endothelial cells after LPS treatment, which could be restored by RUS. Collectively, these findings provide pharmacological evidence showing that RUS attenuates LPS-induced pulmonary endothelial barrier dysfunction by inhibiting TLR4/Src/VE-cadherin pathway through targeting NMMHC IIA and mediating NMMHC IIA‒TLR4 interactions.