Journal: Animal Models and Experimental Medicine

Article Title: Baihe Zhimu Decoction ameliorates anxious depression via PI3K / AKT / mTOR pathway‐mediated neuroprotection and inhibition of hippocampal neuronal apoptosis

doi: 10.1002/ame2.70295

Figure Lengend Snippet: BZD‐containing serum protects hippocampal neurons against apoptosis through activation of the PI3K/AKT/mTOR pathway. (A) Hoechst 33342 staining (×200), scale bar: 100 μm. (B) Quantification of apoptosis by Annexin V‐FITC/PI flow cytometry ( n = 3). (C) Representative Western blot bands and quantification of cleaved caspase‐3, Bax, and Bcl‐2 ( n = 4). (D) Representative western blot and quantification of PI3K, AKT, mTOR and their phosphorylated forms ( n = 4). Data are presented as mean ± SD. ## p < 0.01 vs . control group; ** p < 0.01 vs . model group; △ p < 0.05, △△ p < 0.01 vs . BZD group.

Article Snippet: Cells were labeled with 5 μL Annexin V‐FITC and 5 μL PI (E‐CK‐A211, Elabscience) for 15 min at room temperature in the dark.

Techniques: Activation Assay, Staining, Flow Cytometry, Western Blot, Control

Journal: Advanced Science

Article Title: Phosphorylated STYK1 Promotes Gemcitabine Resistance via Autophagy‐Mediated Cancer Stemness in Pancreatic Cancer

doi: 10.1002/advs.202521488

Figure Lengend Snippet: STYK1 promotes gemcitabine resistance in pancreatic cancer cells. (A, B) PANC‐1 cells were treated with different concentrations of gemcitabine for the indicated time periods, and STYK1 mRNA expression was assessed by RT‐qPCR (n = 6). **** p < 0.0001 according to Student's t ‐test. (C) Western blotting analysis of STYK1 expression levels in human pancreatic cancer cell lines PANC‐1 and AsPC‐1, as well as in the gemcitabine‐resistant cell lines PANC‐1/GEM and AsPC‐1/GEM. D) RT‐PCR analysis of STYK1 mRNA levels in PANC‐1, AsPC‐1, PANC‐1/GEM and AsPC‐1/GEM cells. **** p < 0.0001 according to Student's t ‐test. (E) Representative images of IHC staining for STYK1 in gemcitabine‐sensitive and resistant pancreatic cancer PDX tumors tissues. (F–I) Cell viability of PANC‐1/GEM and AsPC‐1/GEM cells with stable STYK1 knockdown, as well as PANC‐1 and AsPC‐1 cells stably overexpressing STYK1, following treatment with different concentrations of gemcitabine for 72 h, as determined by CCK‐8 assay. (J) PANC‐1/GEM cells with stable STYK1 knockdown were treated with gemcitabine (5 µM) for different durations as indicated, and OD values were assessed using the CCK‐8 assay. **** p < 0.0001 according to Student's t ‐test. (K–M) Apoptosis of PANC‐1/GEM and AsPC‐1/GEM cells with stable STYK1 knockdown following gemcitabine treatment (5 µM for PANC‐1/GEM and 40 µM for AsPC‐1/GEM, 24 h), assessed using an Annexin V‐AbFluor 488/PI double staining Apoptosis Detection Kit. Annexin V‐positive cells indicate early apoptosis, whereas PI‐positive cells indicate late apoptosis/necrosis (n = 3). **** p < 0.0001 according to Student's t ‐test. (N) Representative images of resected tumors from nude mice in different groups (n = 5). O) Tumor weights of resected tumors in different groups of mice (n = 5). **** p < 0.0001 according to Student's t ‐test. ** p < 0.01; **** p < 0.0001 according to Student's t ‐test. (P) Tumor volume change curves in different groups of mice (n = 5); Tumor volume was measured every 5 days after cell injection and during gemcitabine treatment (25 mg/kg). **** p < 0.0001 according to Student's t ‐test. (Q) Representative H&E staining and Ki67 immunohistochemical staining images of resected tumor tissues (n = 5). Scale bar: 50 µm. Data were expressed as mean ± SD, * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 according to Student's t ‐test.

Article Snippet: Apoptosis was assessed using an Annexin V‐FITC Apoptosis Detection Kit (Elabscience, E‐CK‐A252).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Analysis, Reverse Transcription Polymerase Chain Reaction, Immunohistochemistry, Knockdown, Stable Transfection, CCK-8 Assay, Double Staining, Injection, Staining, Immunohistochemical staining

Journal: Science Advances

Article Title: Effects of probiotics on joint homeostasis and osteoarthritis: Development of microbe-gut-cartilage axis-on-a-chip

doi: 10.1126/sciadv.aeg8557

Figure Lengend Snippet: ( A ) Timeline of MGCAoC establishment. Caco-2 cells were seeded in the gut channel and cultured under dynamic flow for intestinal epithelium formation over 5 days, which was followed by the seeding of probiotics. BMSC suspension was injected into the microwell array for self-aggregation and chondrogenic differentiation. After 2 days of chondrogenic induction, the cartilage module was connected to the microbe-gut module by adjusting the embedded microvalves to establish the MGCAoC. BB , B. bifidum ; LP , L. plantarum. ( B ) Quantification of apparent permeability of intestinal epithelial monolayers over 7 days [ n = 3 chips, one-way analysis of variance (ANOVA)]. ( C ) Transepithelial resistance (TEER) values of intestinal epithelial monolayers formed in the gut module of MGCAoC over 7 days ( n = 3 chips, one-way ANOVA). d, day. ( D ) Scanning electron microscopy (SEM) image showing the intestinal villus-crypt structures on the established Caco-2 epithelium. Scale bar, 1 μm. ( E ) SEM images of the apical surface of probiotic-seeded intestinal epithelium. The right image shows a magnified view of the area in the white dashed box, where a probiotic community is observed on the mucus. The probiotics were seeded on day 5, and the images were taken 2 days afterward. Scale bars, 1 μm. ( F ) Confocal images showing ZO-1 expression (green) and BB and LP distribution (red) on the intestinal epithelium. ( G ) Confocal images showing the relative positions of the intestinal epithelial layer (blue), the mucus layer (green), and probiotics (red). ( H ) Bright-field images of COs cultured on 96-well plates, CO-on-a-chip (COoC; probiotic-free), and MGCAoC platform. The images were taken on day 14. Scale bar, 200 μm. ( I ) Changes in CO diameters after culturing on 96-well plates, COoC (probiotic-free), and MGCAoC platform for different durations ( n = 10, two-way ANOVA). d, day. ( J ) Terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling (TUNEL) staining of CO sections, with red color indicating apoptotic cells. Scale bars, 200 μm. ( K ) Percentage of TUNEL-positive cells ( n = 5, one-way ANOVA). n.s., not significant.

Article Snippet: The one-step TUNEL apoptosis assay kit (C1090, Beyotime) was used to evaluate cell apoptosis in cryosections of COs.

Techniques: Cell Culture, Probiotics, Suspension, Injection, Permeability, Analysis, Scanning Electron Microscopy, Confocal, Expressing, 96 Well Plates, End Labeling, TUNEL Assay, Staining