Cell growth rate.

<p>Cell growth in each group was detected using the MTT assay. Compared with the NC group (controls transfected by 1 irrelevant interference sequence), the growth rate of ICBD-1 cells was significantly decreased after Gab1 siRNA interference or VEGFR-2 siRNA interference, **<i>P</i><0.01. Compared with the un-transfected group, there was no significant change in the growth rate of ICBD-1 cells within the NC group.</p

Changes in PI3K/Akt pathway activity and MMP-9 expression after RNA interference.

<p>A: Compared with the NC group (controls transfected by 1 irrelevant interference sequence), significantly decreased expression of p-Gab1, but no obvious change in total Gab1 expression in ICDB-1 cells were observed after VEGFR-2 siRNA interference, **<i>P</i><0.01. Compared with the un-transfected group, there were no significant changes in Gab1 and p-Gab1 expression in ICBD-1 cells within the NC group. B: Compared with the NC group, there were no significant changes in VEGFR-2 and p-VEGFR-2 expression in ICBD-1 cells after Gab1 siRNA interference. C: Compared with the NC group, significantly decreased expression of p-Akt, but no obvious change in total Akt expression in ICDB-1 cells were observed after Gab1 siRNA interference, **<i>P</i><0.01. Compared with the un-transfected group, there were no significant changes in Akt and p-Akt expression in ICBD-1 cells within the NC group. D: Compared with the NC group, significantly decreased p-Akt expression in ICDB-1 cells was observed after VEGFR-2 siRNA interference, **<i>P</i><0.01. E: Compared with the NC group, significantly decreased MMP-9 mRNA level in ICDB-1 cells was observed after Gab1 siRNA or VEGFR-2 siRNA interference, **<i>P</i><0.01. Compared with the un-transfected group, there was no significant change in MMP-9 level in ICBD-1 cells within the NC group. F: Compared with the NC group, significantly decreased expression of MMP-9 in ICDB-1 cells was observed after Gab1 siRNA interference, **<i>P</i><0.01. G: Compared with the NC group, significantly decreased expression of MMP-9 in ICDB-1 cells was observed after VEGFR-2 siRNA interference, **<i>P</i><0.01.</p

Expression of E-cadherin (red) and α-SMA (green) in Huh-7 cells observed by immunofluorescence.

<p>α-SMA was detected in the cytoplasm; E-cadherin was found in the plasmalemma and the cytoplasm. HSP70/HSP70-PCs treatment reduced the expressions of E-cadherin and promoted the expression of α-SMA.</p

HSP70/HSP70-PCs affect the protein levels of E-cadherin , α-SMA (A), total-p38, phosphor-p38 (C) and mRNA (B) expression of E-cadherin and α-SMA.

<p>24h after induction, cells were harvested for western blotting and real-time RT-PCR. Data are presented as means ± SD from three independent experiments (normalized to GADPH expression).</p

The mRNA and protein expression of E-cadherin, p-p38, HSP70 and α-SMA in tumors and the correlation between tumor-free survival and expression of HSP70, α-SMA and p-p38.

<p>(A) The mRNA expressions of E-cadherin, α-SMA and HSP70 and p38 (normalized to GADPH expression). (B) Photomicrographs of well-differentiated hepatic cancer (top panel) and poorly differentiated hepatic cancer. p-p38, HSP70 and α-SMA were detected in the cytoplasm; E-cadherin was detected in the plasmalemma. Original magnification, 200´. (C) The protein expressions of α-SMA, HSP70, p38 and p-p38 (normalized to GAPDH expression). Kaplan-Meier tumor-free survival curves for hepatic cancer patients showing that the median tumor-free survival time of patients correlated with HSP70 (D), α-SMA (E) and p-p38 (F) expression.</p

DataSheet_1_Mannose-modified erythrocyte membrane-encapsulated chitovanic nanoparticles as a DNA vaccine carrier against reticuloendothelial tissue hyperplasia virus.doc

IntroductionThe erythrocyte membranes used in nanovaccines include high membrane stability, long circulation life, adaptability and extremely good bio compatibility. Nanoparticles encapsulated by erythrocyte membranes are widely used as ideal drug delivery vehicles because of their high drug loading, long circulation time, and excellent biocompatibility. The mannose modification of delivery materials can help target mannose receptors (MRs) to deliver antigens to antigen-presenting cells (APCs).MethodsIn this study, the antigen gene gp90 of avian reticuloendotheliosis virus (REV) was encapsulated with carboxymethyl chitosan (CS) to obtain CSgp90 nanoparticles, which were coated with mannose-modied fowl erythrocyte membranes to yield CS-gp90@M-M nanoparticles. The physicochemical characterization and immune response of the CS-gp90@M-M nanoparticles were investigated in vitro and in vivo.ResultsCS-gp90@M-M nanoparticles were rapidly phagocytized in vitro by macrophages to induce the production of cytokines and nitric oxide. In vivo, CS-gp90@M-M nanoparticles increased cytokine levels, the CD4+/8+ ratio, REV-specific antibodies in the peripheral blood of chicks, and the mRNA levels of immune-related genes in the spleen and bursa of immunized chicks. CS-gp90@M-M nanoparticles could be targeted to lymphoid organs to prolong the retention time of the nanoparticles at the injection site and lymphatic organs, leading to a strong, sustained immune response. Moreover, the CS-gp90@M-M nano-vaccine showed a lasting immunoprotective effect and improved the body weight of chicks after the challenge.ConclusionOverall, CS-gp90@M-M nanoparticles can be used in vaccine designs as an effective delivery carrier with immune response-enhancing effects.</p

DataSheet_1_Precipitation variation: a key factor regulating plant diversity in semi-arid livestock grazing lands.docx

Livestock presence impacts plant biodiversity (species richness) in grassland ecosystems, yet extent and direction of grazing impacts on biodiversity vary greatly across inter-annual periods. In this study, an 8-year (2014-2021) grazing gradient experiment with sheep was conducted in a semi-arid grassland to investigate the impact of grazing under different precipitation variability on biodiversity. The results suggest no direct impact of grazing on species richness in semi-arid Stipa grassland. However, increased grazing indirectly enhanced species richness by elevating community dominance (increasing the sheltering effect of Stipa grass). Importantly, intensified grazing also regulates excessive community biomass resulting from increased inter-annual wetness (SPEI), amplifying the positive influence of annual humidity index on species richness. Lastly, we emphasize that, in water-constrained grassland ecosystems, intra-annual precipitation variability (PCI) was the most crucial factor driving species richness. Therefore, the water-heat synchrony during the growing season may alleviate physiological constraints on plants, significantly enhancing species richness as a result of multifactorial interactions. Our study provides strong evidence for how to regulate grazing intensity to increase biodiversity under future variable climate patterns. We suggest adapting grazing intensity according to local climate variability to achieve grassland biodiversity conservation.</p

Additional file 1: of Autophagy promotes metastasis and glycolysis by upregulating MCT1 expression and Wnt/β-catenin signaling pathway activation in hepatocellular carcinoma cells

Primers used for Quantitative real-time PCR. (DOCX 67 kb