[目的]探究OR13C2在宫颈癌组织中的表达以及对宫颈癌恶性转移的影响。[方法]通过蛋白免疫印迹和实时的荧光定量PCR检测宫颈癌组织OR13C2表达情况。将宫颈癌细胞分为pcDNA NC组、pcDNA OR13C2组和抑制剂(PI-103)组。CCK-8实验分析宫颈癌...[目的]探究OR13C2在宫颈癌组织中的表达以及对宫颈癌恶性转移的影响。[方法]通过蛋白免疫印迹和实时的荧光定量PCR检测宫颈癌组织OR13C2表达情况。将宫颈癌细胞分为pcDNA NC组、pcDNA OR13C2组和抑制剂(PI-103)组。CCK-8实验分析宫颈癌细胞增殖能力,Transwell实验和划痕实验分析宫颈癌细胞侵袭和迁移能力,流式细胞术检测宫颈癌细胞的凋亡率,蛋白免疫印迹分析OR13C2对PI3K/AKT通路蛋白表达的影响。[结果]OR13C2在宫颈癌组织中低表达,而在宫颈癌旁组织中高表达(0.23±0.02 vs 0.97±0.07;2.01±0.05 vs 4.81±0.13;P<0.05)。OR13C2高表达后能够抑制宫颈癌细胞的增殖、侵袭和迁移能力,促进宫颈癌细胞凋亡。此外,OR13C2高表达能够抑制PI3K/AKT的激活(0.73±0.03 vs 0.19±0.05 vs 0.26±0.08;0.72±0.08 vs 0.23±0.06 vs 0.31±0.03;P<0.05)。[结论]OR13C2在宫颈癌组织中表达减少,上调OR13C2的表达可以抑制宫颈癌HeLa细胞的增殖、侵袭和迁移能力,促进宫颈癌HeLa细胞凋亡,此作用与OR13C2抑制PI3K/AKT通路的激活相关。展开更多
Polypropylene(PP) accounts for approximately 28.0% of the global polyolefin market,valued at $243.4 billion in 2022.Known for its lightweight,chemical resistance,costeffectiveness,high strength and melting point,PP is...Polypropylene(PP) accounts for approximately 28.0% of the global polyolefin market,valued at $243.4 billion in 2022.Known for its lightweight,chemical resistance,costeffectiveness,high strength and melting point,PP is widely used in various applications.Its properties and applications are closely tied to its tacticity.One-dimensional(1D) conventional 13C NMR has been extensively utilized to analyze PP tacticity,but its low sensitivity and longer relaxation time remain drawbacks.Typically,analyzing a single PP sample requires around 9 h of NMR time.Using of a cryogenically cooled 10 mm NMR probe can significantly reduce this measurement time;however,its high cost makes it inaccessible for most NMR laboratories.While the refocused insensitive nuclei enhanced by polarization transfer(RINEPT) technique is well-known for enhancing NMR sensitivity,there are no published studies using 1D 13C RINEPT to quantify PP tacticity.Relaxation agents like chromium(Ⅲ) acetylacetonate(Cr(acac)_(3)) have also been used to reduce relaxation times in polyolefin NMR analyses.Here we introduce a straightforward and easily implementable 1D 13C NMR method for rapid PP tacticity quantification.This method combines Cr(acac)_(3),Bruker's existing RINEPT pulse sequence(ineptrd),and our recently published ~1H decoupling sequence(bi_waltz65_256 pl) to eliminate ~1H decoupling artifacts.It is worth noting that decoupling artifacts are always present.When the signal-to-noise ratio(SNR) is low,these artifacts are obscured by noise.For example,in some two-dimensional(2D) or three-dimensional(3D) NMR spectra,decoupling artifacts are barely visible because of the low SNR.However,when attempting to observe weak signals in ~1D spectra,increasing the sample concentration or the number of scans enhances the SNR,revealing the decoupling artifacts.The decoupling artifacts appeared superimpose with some other weak signals,affecting the measurements of signal intensities.Therefore,improved ~1H-decoupling methods are crucial for such data acquisitio n.This synergy results in a 9.4-to 9.7-fold sensitivity enhancement,equating to an 88-to 94-fold reduction(9.4^(2)≈88,9.7^(2)≈94) in NMR acquisition time compared to conventional 1D 13C NMR experiment with Cr(acac)_3.The time savings are even more substantial compared to experiments without Cr(acac)_3.The faster and quantitative approach is accessible to researchers with or without cryoprobes.Beyond PP,this method can be applied to tacticity measurements of other polyolefins,such as polybutene,polyhexene and polyoctene.展开更多
文摘[目的]探究OR13C2在宫颈癌组织中的表达以及对宫颈癌恶性转移的影响。[方法]通过蛋白免疫印迹和实时的荧光定量PCR检测宫颈癌组织OR13C2表达情况。将宫颈癌细胞分为pcDNA NC组、pcDNA OR13C2组和抑制剂(PI-103)组。CCK-8实验分析宫颈癌细胞增殖能力,Transwell实验和划痕实验分析宫颈癌细胞侵袭和迁移能力,流式细胞术检测宫颈癌细胞的凋亡率,蛋白免疫印迹分析OR13C2对PI3K/AKT通路蛋白表达的影响。[结果]OR13C2在宫颈癌组织中低表达,而在宫颈癌旁组织中高表达(0.23±0.02 vs 0.97±0.07;2.01±0.05 vs 4.81±0.13;P<0.05)。OR13C2高表达后能够抑制宫颈癌细胞的增殖、侵袭和迁移能力,促进宫颈癌细胞凋亡。此外,OR13C2高表达能够抑制PI3K/AKT的激活(0.73±0.03 vs 0.19±0.05 vs 0.26±0.08;0.72±0.08 vs 0.23±0.06 vs 0.31±0.03;P<0.05)。[结论]OR13C2在宫颈癌组织中表达减少,上调OR13C2的表达可以抑制宫颈癌HeLa细胞的增殖、侵袭和迁移能力,促进宫颈癌HeLa细胞凋亡,此作用与OR13C2抑制PI3K/AKT通路的激活相关。
文摘Polypropylene(PP) accounts for approximately 28.0% of the global polyolefin market,valued at $243.4 billion in 2022.Known for its lightweight,chemical resistance,costeffectiveness,high strength and melting point,PP is widely used in various applications.Its properties and applications are closely tied to its tacticity.One-dimensional(1D) conventional 13C NMR has been extensively utilized to analyze PP tacticity,but its low sensitivity and longer relaxation time remain drawbacks.Typically,analyzing a single PP sample requires around 9 h of NMR time.Using of a cryogenically cooled 10 mm NMR probe can significantly reduce this measurement time;however,its high cost makes it inaccessible for most NMR laboratories.While the refocused insensitive nuclei enhanced by polarization transfer(RINEPT) technique is well-known for enhancing NMR sensitivity,there are no published studies using 1D 13C RINEPT to quantify PP tacticity.Relaxation agents like chromium(Ⅲ) acetylacetonate(Cr(acac)_(3)) have also been used to reduce relaxation times in polyolefin NMR analyses.Here we introduce a straightforward and easily implementable 1D 13C NMR method for rapid PP tacticity quantification.This method combines Cr(acac)_(3),Bruker's existing RINEPT pulse sequence(ineptrd),and our recently published ~1H decoupling sequence(bi_waltz65_256 pl) to eliminate ~1H decoupling artifacts.It is worth noting that decoupling artifacts are always present.When the signal-to-noise ratio(SNR) is low,these artifacts are obscured by noise.For example,in some two-dimensional(2D) or three-dimensional(3D) NMR spectra,decoupling artifacts are barely visible because of the low SNR.However,when attempting to observe weak signals in ~1D spectra,increasing the sample concentration or the number of scans enhances the SNR,revealing the decoupling artifacts.The decoupling artifacts appeared superimpose with some other weak signals,affecting the measurements of signal intensities.Therefore,improved ~1H-decoupling methods are crucial for such data acquisitio n.This synergy results in a 9.4-to 9.7-fold sensitivity enhancement,equating to an 88-to 94-fold reduction(9.4^(2)≈88,9.7^(2)≈94) in NMR acquisition time compared to conventional 1D 13C NMR experiment with Cr(acac)_3.The time savings are even more substantial compared to experiments without Cr(acac)_3.The faster and quantitative approach is accessible to researchers with or without cryoprobes.Beyond PP,this method can be applied to tacticity measurements of other polyolefins,such as polybutene,polyhexene and polyoctene.
