Polarization-Sensitive Sum-Frequency Generation Microscopy of Collagen Fibers

Point-scanning sum-frequency generation (SFG) microscopy enables the generation of images of collagen I fibers in tissues by tuning into specific vibrational resonances of the polypeptide. It is shown that when collagen-rich tissues are visualized near the 2954 cm^–1 stretching vibration of methylene groups, the SFG image contrast is higher compared to the contrast seen in nonresonant second-harmonic generation (SHG) imaging. Polarization and spectrally resolved analysis of the SFG signal as a function of fiber orientation in the CH-stretching range of the vibrational spectrum enabled a comparative characterization of the achiral tensor elements of collagen’s second-order susceptibility. This analysis reveals that selected on-resonance tensor elements are enhanced over other elements, giving rise to a much stronger anisotropy ρ of the signal for SFG (ρ ≈ 15) compared to SHG (ρ ≈ 3). The improved anisotropy of the vibrationally resonant signal contributes to the higher contrast seen in the SFG tissue images

Sphere formation and proliferative capacity of H460 SP and non-SP cells.

<p>A–B, Purified SP and non-SP cells were cultured in serum-free medium in anchorage-independent conditions. The SP cells formed typical floating spheres within 4 days (A) whereas the non-SP cells largely established adherent growth (B). C, The SP cells displayed higher proliferative ability than non-SP cells as determined by CCK-8 kit (<i>P</i><0.05 for all time points, Student <i>t</i>-test).</p

High tumorigenicity in SP cells.

<p>A, Table presentation of the tumorigenic potential of H460 SP and non-SP cells. Three parameters of tumorigenicity, i.e., tumor incidence, latency, and volume (*<i>P</i><0.01) were shown. All animals were terminated (term.) 28 days after implantation. B, The SP cells regenerated larger tumors than corresponding non-SP cells at every cell dose. C, Gross tumor images when tumors were harvested at day 28 after s.c. injection of the SP and non-SP cells into NOD/SCID mice. D, Representative HE-stained photomicrographs of SP and non-SP tumors.</p

SP analysis in cultured H460 lung cancer cells.

<p>A–B, Hoechst staining of H460 cells. Note that although the majority of cells were stained in the nucleus, some cells (indicated by arrows) apparently lacked nuclear Hoechst staining. C–D, SP phenotypes in the absence (C) or presence (D) of verapamil.</p

Cyclopamine inhibits H460 cell proliferation.

<p>A–C, Representative photomicrographs of H460 cells 72 h after treatment with vehicle control (A), Tomatidine (B), or Cyclopamine (C). Original maginifications: ×200. D, Cyclopamine dose-dependently inhibited H460 cell proliferation (<i>P</i><0.05; one-way ANOVA). E, Time course of Cyclopamine inhibition of H460 cells (<i>P</i><0.05, one-way ANOVA). Cyclopamine was used at 20 µmol/L. F, Effects of Cyclopamine on the cell cycle of H460 cells.</p

RT-PCR analysis of ABCG2 and SMO mRNA levels.

<p>A, Representative RT-PCR gel images. M, marker; lane 1, NCI-H460 cells normally cultured in serum-containing medium; lane 2, the H460 spheres; lane 3, purified SP cells; lane 4, purified non-SP cells; lane 5, the Tomatidine control group; lane 6, the Cyclopamine experimental group; lane 7, SP tumors; lane 8, non-SP tumors. B, Quantitative presentation of ABCG2 and SMO mRNA levels as determined by densitometry (<i>P</i><0.001, except ABCG2 mRNA lane 6 vs. lane 8 and SMO mRNA lane 1 vs. lane 5 and lane 4 vs. lane 6).</p

SP re-analysis in various samples.

<p>A–B, SP re-analysis when the SP spheres were disaggregated and dissociated cells cultured in normal serum-containing medium for one week. C–D, SP re-analysis in the SP cell-derived tumors. E–F, SP re-analysis in non-SP cell-derived tumors. A, C, E, without verapamil. B, D, F, with verapamil.</p

C–H/C–H Oxidative Direct Arylation Polycondensation to Isoindigo-Based n‑Type Conjugated Polymers

Two high molecular weight conjugated polymers (CPs), i.e., poly[N,N′-bis(4-tetradecyl­octadecyl)-7,7′-difluoro­isoindigo-alt-2,2′-bithiazole] (P2FI2Tz) and poly[N,N′-bis(4-tetradecyl­octadecyl)-5,5′,7,7′-tetrafluoro­isoindigo-alt-2,2′-bithiazole] (P4FI2Tz), were synthesized via C–H/C–H oxidative direct arylation polycondensation (Oxi-DArP) using PdCl2/CuCl/Cu(OAc)2 as catalytic system and chlorobenzene as solvent. Both polymers are unipolar n-type semiconductors with electron mobility (μe) > 0.2 cm2 V–1 s–1, attributed to their low-lying highest occupied molecular orbitals (HOMOs) and lowest unoccupied molecular orbital (LUMOs), which are −6.01/–3.69 and −6.24/–3.82 eV for P2FI2Tz and P4FI2Tz, respectively. P2FI2Tz was also synthesized via C–Br/C–H direct arylation polycondensation (DArP) and Stille polycondensation. The resultant polymers P2FI2Tz-D and P2FI2Tz-S showed slightly lower μe compared to Oxi-DArP-made polymer (P2FI2Tz-O1) with a similar molecular weight. Notably, Oxi-DArP is much faster and more efficient than DArP and Stille polycondensation. P2FI2Tz with a number-average molecular weight (Mn) above 40 kDa, was obtained in 9 h of Oxi-DArP, significantly shorter than 24 h with DArP and Stille polycondensation. P2FI2Tz with a Mn as high as 191 kDa could be synthesized via Oxi-DArP by extending the polymerization time to 16 h. Our results prove Oxi-DArP to be an efficient protocol to synthesize polymer semiconductors. This is also the first comparative study on three methods, i.e., Oxi-DArP, DArP, and Stille polycondensation

MOESM1 of MicroRNA-125b promotes tumor metastasis through targeting tumor protein 53-induced nuclear protein 1 in patients with non-small-cell lung cancer

Additional file 1: Figure S1. Expressions of miR-125b and TP53INP1 in isolated NSCLC cells were determined by qPCR and analyzed for their negative correlation

Image1_Feiyanning formula modulates the molecular mechanism of osimertinib resistance in lung cancer by regulating the Wnt/β-catenin pathway.TIF

Feiyanning Formula (FYN), a Chinese herbal formula derived from summarized clinical experience, is proven to have anti-tumor effects in lung cancer patients. Osimertinib, a third-generation epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI), can improve progression-free survival and overall survival of patients but drug resistance is inevitable. The current study evaluated the effects of FYN in osimertinib-resistant HCC827OR and PC9OR cells. FYN preferentially inhibited the proliferation and migration of HCC827OR and PC9OR cells. Moreover, FYN and osimertinib exhibited synergistic inhibitory effects on proliferation and migration. Real-time qPCR (RT-qPCR) and western blotting results indicated that FYN downregulated gene and protein levels of GSK3β and SRFS1, which are enriched in the Wnt/β-catenin pathway. Besides, FYN inhibited tumor growth and exhibited synergistic effects with osimertinib in vivo. Collectively, the results suggested that FYN exerted an anti-osimertinib resistance effect via the Wnt/β-catenin pathway.</p