5-Hydroxytryptamine 2C(5-HT2C) receptor is one of the major targets of anti-obesity agents, due to its role in regulation of appetite. In the present study, the 70% EtO H extract of the roots of Bupleurum chinense was...5-Hydroxytryptamine 2C(5-HT2C) receptor is one of the major targets of anti-obesity agents, due to its role in regulation of appetite. In the present study, the 70% EtO H extract of the roots of Bupleurum chinense was revealed to have agonistic activity on 5-HT2 C receptor, and the subsequent bioassay-guided isolation led to identification of several saikosaponins as the active constituents with 5-HT2 C receptor agonistic activity in vitro and anti-obesity activity in vivo. The new compound, 22-oxosaikosaponin d(1), was determined by extensive spectroscopic analyses(HR-ESI-MS, IR, and 1D and 2D NMR). The primary structure-activity relationship study suggested that the intramolecular ether bond between C-13 and C-28 and the number of sugars at C-3 position were closely related to the 5-HT2 C receptor agonistic activity. Saikosaponin a(3), the main saponin in B. chinense, showed obviously agonistic activity on 5-HT2 C receptor with an EC50 value of 21.08 ± 0.33 μmol×L^(–1) in vitro and could reduce food intake by 39.1% and 69.2%, and weight gain by 13.6% and 16.4%, respectively, at 3.0 and 6.0 mg×kg^(–1) in vivo. This investigation provided valuable information for the potential use of B. chinense as anti-obesity agent.展开更多
目的建立5-羟色胺2C受体(5-HT_(2C)receptor,5-HT_(2C)R)和增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)标记的活化T细胞核因子2(nuclear factor of activated T cells 2,NFAT2)共表达细胞株。方法人源5-HT_(2C)R质粒...目的建立5-羟色胺2C受体(5-HT_(2C)receptor,5-HT_(2C)R)和增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)标记的活化T细胞核因子2(nuclear factor of activated T cells 2,NFAT2)共表达细胞株。方法人源5-HT_(2C)R质粒转染至U2OS-EGFP-NFAT2细胞,经潮霉素(Hygro)压力筛选到稳定表达5-HT_(2C)R的U2OS-EGFP-NFAT2-5-HT_(2C)R细胞。使用RT-qPCR和Western blot法检测该细胞株中5-HT_(2C)R的mRNA和蛋白表达水平;用核转位功能实验验证U2OS-EGFP-NFAT2-5-HT_(2C)R细胞受体功能的特异性;验证5-HT、LSD、DOM、DOI、赛洛西宾(PSI)和利舒脲(LIS)对5-HT_(2C)R的激活能力。结果筛选得到58号细胞为最强激活的U2OS-EGFP-NFAT2-5-HT_(2C)R单克隆细胞株。RT-qPCR和Western blot结果显示,1~15代内,U2OS-EGFP-NFAT2-5-HT_(2C)R细胞株稳定表达5-HT_(2C)R mRNA和蛋白。1~15代内,Vabicaserin对U2OS-EGFP-NFAT2-5-HT_(2C)R细胞株的激活能力稳定,5-HT_(2C)R特异性拮抗剂SB242084能够拮抗Vabicaserin的作用。5-HT、LIS、PSI能诱导U2OS-EGFP-NFAT2-5-HT_(2C)R细胞部分核转位,而LSD、DOM、DOI没有作用。结论成功构建了共表达5-HT_(2C)R和EGFP-NFAT2的U2OS-EGFP-NFAT2-5-HT_(2C)R细胞,可用于靶向5-HT_(2C)R的高活性小分子化合物筛选。展开更多
oluble solids content(SSC) plays an important role in determining the flavor of tomato fruits. Tomato fruit SSC is transcriptionally regulated via sugar metabolism. Previous studies have predominantly focused on the r...oluble solids content(SSC) plays an important role in determining the flavor of tomato fruits. Tomato fruit SSC is transcriptionally regulated via sugar metabolism. Previous studies have predominantly focused on the role of C2H2-type zinc finger proteins in tomato growth and development. However, the specific regulatory mechanisms of C2H2 in the accumulation of soluble solids in tomato fruits are not fully understood. This study used eight tomato accessions with varying levels of SSC to study the expression of SlC2H2 family genes in red ripe fruits. The levels of SlC2H2-71 expression were found to be significantly reduced in high-SSC accessions compared to low-SSC accessions. Several Slc2h2-71 mutant lines were developed using the CRISPR-Cas9 system, which led to elevated levels of soluble solids, fructose, glucose, malic acid, and citric acid in mature red ripe fruits. However, the sucrose content in the edited Slc2h2-71 mutant lines was generally lower. RNA-seq analysis revealed that fruits from the mutant lines had altered expression of genes related to the sugar and acid metabolic pathways, which was further confirmed by quantitative real-time PCR. Specifically, the expression of SlLIN5 encoding the cell wall invertase(CWIN) was elevated. The yeast one-hybrid(Y1H) assay, 35S::UAS-GUS, dual-luciferase reporter systems and electrophoretic mobility shift assay(EMSA) demonstrated that C2H2-71 regulates tomato sugar metabolism by directly binding to the promoter region of Sl LIN5, culminating in the repression of its transcriptional activity. The activity of acid invertase exhibited a significantly higher level in the SlC2H2-71 knock-out lines compared to the control lines. In summary, the regulation of tomato fruit SSC by C2H2-71 involves the inhibition of SlLIN5 expression.展开更多
Neuropathic pain,often featuring allodynia,imposes significant physical and psychological burdens on patients,with limited treatments due to unclear central mechanisms.Addressing this challenge remains a crucial unsol...Neuropathic pain,often featuring allodynia,imposes significant physical and psychological burdens on patients,with limited treatments due to unclear central mechanisms.Addressing this challenge remains a crucial unsolved issue in pain medicine.Our previous study,using protein kinase C gamma(PKCγ)-tdTomato mice,highlights the spinal feedforward inhibitory circuit involving PKCγ neurons in gating neuropathic allodynia.However,the regulatory mechanisms governing this circuit necessitate further elucidation.We used diverse transgenic mice and advanced techniques to uncover the regulatory role of the descending serotonin(5-HT)facilitation system on spinal PKCγ neurons.Our findings revealed that 5-HT neurons from the rostral ventromedial medulla hyperpolarize spinal inhibitory interneurons via 5-HT_(2C) receptors,disinhibiting the feedforward inhibitory circuit involving PKCγ neurons and exacerbating allodynia.Inhibiting spinal 5-HT_(2C) receptors restored the feedforward inhibitory circuit,effectively preventing neuropathic allodynia.These insights offer promising therapeutic targets for neuropathic allodynia management,emphasizing the potential of spinal 5-HT_(2C) receptors as a novel avenue for intervention.展开更多
The novel copper?complex with salicylaldehyde benzoylhydrazone and pyridine ligands, Cu(C14H10N2O2)(C5H5N), has been synthesized and characterized by elemental analysis, IR and thermal analysis. The crystal structure ...The novel copper?complex with salicylaldehyde benzoylhydrazone and pyridine ligands, Cu(C14H10N2O2)(C5H5N), has been synthesized and characterized by elemental analysis, IR and thermal analysis. The crystal structure of the title complex has been determined by single crystal X ray diffraction techniques. The crystal belongs to monoclinic with space group P21/c. The cell parameters are: a=1.6362(9)nm, b=1.7140(9)nm, c=1.2255(7)nm, β=105.168(9)°, V=3.317(3)nm3, Z=8, Dc=1.525g·cm-3, μ(MoKα)=1.334mm-1, F(000)=1560. The structure wasrefined to final R1=0.0376, wR2=0.0909. The copper?ion lies in a distorted square planar environment composed of two oxygen atoms, one nitrogen atom of tridentate acyhydrazone Schiff base ligand and one nitrogen atom of the pyridine ligand. CCDC: 193111.展开更多
SF_6气体由于其优异的电气性能而广泛应用于电力行业,然而,由于其全球变暖潜能值极高,国内外学者在过去的几十年里对SF_6替代气体做了广泛研究,但由于它们都各有缺陷,故很难在实际生产中加以应用。文中以C_5F_(10)O/CO_2混合气体作为主...SF_6气体由于其优异的电气性能而广泛应用于电力行业,然而,由于其全球变暖潜能值极高,国内外学者在过去的几十年里对SF_6替代气体做了广泛研究,但由于它们都各有缺陷,故很难在实际生产中加以应用。文中以C_5F_(10)O/CO_2混合气体作为主要研究对象、以SF_6气体作为对照,研究了混合气体中C_5F_(10)O分压力分别为20 k Pa和40 k Pa、总压力为0.1~0.5 MPa时的工频耐压和雷电冲击(lightning impulse,LI)性能,并分析了其作为SF_6替代气体的可能性。实验结果表明,0.2 MPa的混合气体中混入20 k Pa和40 k Pa的C_5F_(10)O,可分别使混合气体的工频放电电压达到相同气压下SF_6气体的61.89%和81.99%。C_5F_(10)O分压力40 k Pa、总压力0.5 MPa的混合气体正、负极性雷电冲击放电电压分别是0.3 MPa时SF_6气体的88.9%和89.9%。因此,通过增加C_5F_(10)O的含量或提高混合气体的总压力,均可有效提高混合气体的绝缘性能,其中,前者更为有效。展开更多
基金supported by the National Science Foundation of China(No.81573322)the Hundred-Talent Program of CAS+1 种基金the CAS"Light of West China"Programthe Youth Innovation Promotion Association of CAS
文摘5-Hydroxytryptamine 2C(5-HT2C) receptor is one of the major targets of anti-obesity agents, due to its role in regulation of appetite. In the present study, the 70% EtO H extract of the roots of Bupleurum chinense was revealed to have agonistic activity on 5-HT2 C receptor, and the subsequent bioassay-guided isolation led to identification of several saikosaponins as the active constituents with 5-HT2 C receptor agonistic activity in vitro and anti-obesity activity in vivo. The new compound, 22-oxosaikosaponin d(1), was determined by extensive spectroscopic analyses(HR-ESI-MS, IR, and 1D and 2D NMR). The primary structure-activity relationship study suggested that the intramolecular ether bond between C-13 and C-28 and the number of sugars at C-3 position were closely related to the 5-HT2 C receptor agonistic activity. Saikosaponin a(3), the main saponin in B. chinense, showed obviously agonistic activity on 5-HT2 C receptor with an EC50 value of 21.08 ± 0.33 μmol×L^(–1) in vitro and could reduce food intake by 39.1% and 69.2%, and weight gain by 13.6% and 16.4%, respectively, at 3.0 and 6.0 mg×kg^(–1) in vivo. This investigation provided valuable information for the potential use of B. chinense as anti-obesity agent.
文摘目的建立5-羟色胺2C受体(5-HT_(2C)receptor,5-HT_(2C)R)和增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)标记的活化T细胞核因子2(nuclear factor of activated T cells 2,NFAT2)共表达细胞株。方法人源5-HT_(2C)R质粒转染至U2OS-EGFP-NFAT2细胞,经潮霉素(Hygro)压力筛选到稳定表达5-HT_(2C)R的U2OS-EGFP-NFAT2-5-HT_(2C)R细胞。使用RT-qPCR和Western blot法检测该细胞株中5-HT_(2C)R的mRNA和蛋白表达水平;用核转位功能实验验证U2OS-EGFP-NFAT2-5-HT_(2C)R细胞受体功能的特异性;验证5-HT、LSD、DOM、DOI、赛洛西宾(PSI)和利舒脲(LIS)对5-HT_(2C)R的激活能力。结果筛选得到58号细胞为最强激活的U2OS-EGFP-NFAT2-5-HT_(2C)R单克隆细胞株。RT-qPCR和Western blot结果显示,1~15代内,U2OS-EGFP-NFAT2-5-HT_(2C)R细胞株稳定表达5-HT_(2C)R mRNA和蛋白。1~15代内,Vabicaserin对U2OS-EGFP-NFAT2-5-HT_(2C)R细胞株的激活能力稳定,5-HT_(2C)R特异性拮抗剂SB242084能够拮抗Vabicaserin的作用。5-HT、LIS、PSI能诱导U2OS-EGFP-NFAT2-5-HT_(2C)R细胞部分核转位,而LSD、DOM、DOI没有作用。结论成功构建了共表达5-HT_(2C)R和EGFP-NFAT2的U2OS-EGFP-NFAT2-5-HT_(2C)R细胞,可用于靶向5-HT_(2C)R的高活性小分子化合物筛选。
基金supported by the grants from the National Key Research & Development Plan of China (2022YFF10030022022YFD1200502)+5 种基金the National Natural Science Foundation of China (32372696)the Wuhan Biological Breeding Major Project, China (2022021302024852)the HZAU–AGIS Cooperation Fund, China (SZYJY2023022)the Funds for High Quality Development of Hubei Seed Industry, China (HBZY2023B004)the Hubei Agriculture Research System, China (2024HBSTX4-06)the Funds of National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops, China (Horti3Y-2024-008)。
文摘oluble solids content(SSC) plays an important role in determining the flavor of tomato fruits. Tomato fruit SSC is transcriptionally regulated via sugar metabolism. Previous studies have predominantly focused on the role of C2H2-type zinc finger proteins in tomato growth and development. However, the specific regulatory mechanisms of C2H2 in the accumulation of soluble solids in tomato fruits are not fully understood. This study used eight tomato accessions with varying levels of SSC to study the expression of SlC2H2 family genes in red ripe fruits. The levels of SlC2H2-71 expression were found to be significantly reduced in high-SSC accessions compared to low-SSC accessions. Several Slc2h2-71 mutant lines were developed using the CRISPR-Cas9 system, which led to elevated levels of soluble solids, fructose, glucose, malic acid, and citric acid in mature red ripe fruits. However, the sucrose content in the edited Slc2h2-71 mutant lines was generally lower. RNA-seq analysis revealed that fruits from the mutant lines had altered expression of genes related to the sugar and acid metabolic pathways, which was further confirmed by quantitative real-time PCR. Specifically, the expression of SlLIN5 encoding the cell wall invertase(CWIN) was elevated. The yeast one-hybrid(Y1H) assay, 35S::UAS-GUS, dual-luciferase reporter systems and electrophoretic mobility shift assay(EMSA) demonstrated that C2H2-71 regulates tomato sugar metabolism by directly binding to the promoter region of Sl LIN5, culminating in the repression of its transcriptional activity. The activity of acid invertase exhibited a significantly higher level in the SlC2H2-71 knock-out lines compared to the control lines. In summary, the regulation of tomato fruit SSC by C2H2-71 involves the inhibition of SlLIN5 expression.
基金supported by the National Natural Science Foundation of China(81971058,82371226,82101295,82301398)the National Funded Postdoctoral Researcher Program(GZC20233585)The Boost Plan of Xijing Hospital(XJZT24QN25,XJZT25CX22).
文摘Neuropathic pain,often featuring allodynia,imposes significant physical and psychological burdens on patients,with limited treatments due to unclear central mechanisms.Addressing this challenge remains a crucial unsolved issue in pain medicine.Our previous study,using protein kinase C gamma(PKCγ)-tdTomato mice,highlights the spinal feedforward inhibitory circuit involving PKCγ neurons in gating neuropathic allodynia.However,the regulatory mechanisms governing this circuit necessitate further elucidation.We used diverse transgenic mice and advanced techniques to uncover the regulatory role of the descending serotonin(5-HT)facilitation system on spinal PKCγ neurons.Our findings revealed that 5-HT neurons from the rostral ventromedial medulla hyperpolarize spinal inhibitory interneurons via 5-HT_(2C) receptors,disinhibiting the feedforward inhibitory circuit involving PKCγ neurons and exacerbating allodynia.Inhibiting spinal 5-HT_(2C) receptors restored the feedforward inhibitory circuit,effectively preventing neuropathic allodynia.These insights offer promising therapeutic targets for neuropathic allodynia management,emphasizing the potential of spinal 5-HT_(2C) receptors as a novel avenue for intervention.
文摘The novel copper?complex with salicylaldehyde benzoylhydrazone and pyridine ligands, Cu(C14H10N2O2)(C5H5N), has been synthesized and characterized by elemental analysis, IR and thermal analysis. The crystal structure of the title complex has been determined by single crystal X ray diffraction techniques. The crystal belongs to monoclinic with space group P21/c. The cell parameters are: a=1.6362(9)nm, b=1.7140(9)nm, c=1.2255(7)nm, β=105.168(9)°, V=3.317(3)nm3, Z=8, Dc=1.525g·cm-3, μ(MoKα)=1.334mm-1, F(000)=1560. The structure wasrefined to final R1=0.0376, wR2=0.0909. The copper?ion lies in a distorted square planar environment composed of two oxygen atoms, one nitrogen atom of tridentate acyhydrazone Schiff base ligand and one nitrogen atom of the pyridine ligand. CCDC: 193111.
文摘SF_6气体由于其优异的电气性能而广泛应用于电力行业,然而,由于其全球变暖潜能值极高,国内外学者在过去的几十年里对SF_6替代气体做了广泛研究,但由于它们都各有缺陷,故很难在实际生产中加以应用。文中以C_5F_(10)O/CO_2混合气体作为主要研究对象、以SF_6气体作为对照,研究了混合气体中C_5F_(10)O分压力分别为20 k Pa和40 k Pa、总压力为0.1~0.5 MPa时的工频耐压和雷电冲击(lightning impulse,LI)性能,并分析了其作为SF_6替代气体的可能性。实验结果表明,0.2 MPa的混合气体中混入20 k Pa和40 k Pa的C_5F_(10)O,可分别使混合气体的工频放电电压达到相同气压下SF_6气体的61.89%和81.99%。C_5F_(10)O分压力40 k Pa、总压力0.5 MPa的混合气体正、负极性雷电冲击放电电压分别是0.3 MPa时SF_6气体的88.9%和89.9%。因此,通过增加C_5F_(10)O的含量或提高混合气体的总压力,均可有效提高混合气体的绝缘性能,其中,前者更为有效。
