HIV-1 gains entry into target cells by sequentially interacting with cellular receptors and co-receptors. Both the receptor and co-receptor are recognized by HIV-1 envelope protein gpl20, which plays a key role in the...HIV-1 gains entry into target cells by sequentially interacting with cellular receptors and co-receptors. Both the receptor and co-receptor are recognized by HIV-1 envelope protein gpl20, which plays a key role in the entry process of HIV-1 into cells. The development of new inhibitors is essential since the viral enzyme reverse transcriptase （RT） is one of the first targets of antiretroviral therapy. It has been reported that a variety of natural plants, such as Artemisia rupestris L., have anti-viral pharmacological activity, and they might be the potential inhibitors of RT or V3 loop of gpl20 against HIV-1. RIQRGPGRAFVT1GK （R15K）, the relatively conserved region of V3 loop, can be used for binding research. In this work, we analyzed the interactions between different extracts from Artemisia rupestris L. and R15K by affinity capillary electrophoresis （ACE）. Moreover, we analyzed the interactions between different extracts from Artemisia rupestris L. and RT by capillary zone electrophoresis （CZE）. Our data showed that the chloroform extract ofArtemisia rupestris L. was active among the different plant extracts, which was consistent with previous studies. Taken together, our study provided a rapid screening method to seek anti-HIV ingredients in natural plants＇ extracts.展开更多
Curcumin improves the learning and memory deficits in rats induced by the gp120 V3 loop. The present study cultured rat hippocampal neurons with 1 nM gp120 V3 loop and 1 μM curcumin for 24 hours. The results showed t...Curcumin improves the learning and memory deficits in rats induced by the gp120 V3 loop. The present study cultured rat hippocampal neurons with 1 nM gp120 V3 loop and 1 μM curcumin for 24 hours. The results showed that curcumin inhibited the gp120 V3 loop-induced mitochondrial membrane potential decrease, reduced the mRNA expression of the pro-apoptotic gene caspase-3, and attenuated hippocampal neuronal injury.展开更多
Cervical cancer(CC)is recognized as the most common neoplasm in the female reproductive system worldwide.The lack of chemotherapeutic agents with outstanding effectiveness and safety severely compromises the anti-cipa...Cervical cancer(CC)is recognized as the most common neoplasm in the female reproductive system worldwide.The lack of chemotherapeutic agents with outstanding effectiveness and safety severely compromises the anti-cipated prognosis of patients.Aloperine(ALO)is a natural quinolizidine alkaloid with marked anti-cancer effects on multiple malignancies as well as favorable activity in relieving inflammation,allergies and infection.However,its therapeutic efficacy and underlying mechanism in CC are still unclear.In the current study,MTT assay was employed to evaluate the viability of HeLa cells exposed to ALO to preliminarily estimate the effectiveness of ALO in CC.Then,the effects of ALO on the proliferation and apoptosis of HeLa cells were further investigated by plate colony formation and flow cytometry,respectively,while the migration and invasion of ALO-treated HeLa cells were evaluated using Transwell assay.Moreover,nude mice were subcutaneously inoculated with HeLa cells to demonstrate the anti-CC properties of ALO in vivo.The molecular mechanisms underlying these effects of ALO were evaluated by Western blot and immunohistochemical analysis.This study experimentally demonstrated that ALO inhibited the proliferation of HeLa cells via G2 phase cell cycle arrest.Simultaneously,ALO promoted an increase in the percentage of apoptotic HeLa cells by increasing the Bax/Bcl-2 ratio.Additionally,the migration and invasion of HeLa cells were attenuated by ALO treatment,which was considered to result from inhibition of epithelial-to-mesenchymal transition.For molecular mechanisms,the expression and activation of the IL-6-JAK1-STAT3 feedback loop were markedly suppressed by ALO treatment.This study indicated that ALO markedly suppresses the proliferation,migration and invasion and enhances the apoptosis of HeLa cells.In addition,these prominent anti-CC properties of ALO are associated with repression of the IL-6-JAK1-STAT3 feedback loop.展开更多
As a crucial signaling molecule, calcium plays a critical role in many physiological and pathological processes by regulating ion channel activity. Recently, one study resolved the structure of the transient receptor ...As a crucial signaling molecule, calcium plays a critical role in many physiological and pathological processes by regulating ion channel activity. Recently, one study resolved the structure of the transient receptor potential melastatin 2(TRPM2) channel from Nematostella vectensis(nvTRPM2). This identified a calcium-binding site in the S2–S3 loop, while its effect on channel gating remains unclear. Here, we investigated the role of this calcium-binding site in both nvTRPM2 and human TRPM2(hTRPM2) by mutagenesis and patch-clamp recording. Unlike hTRPM2, nvT RPM2 cannot be activated by calcium alone. Moreover, the inactivation rate of nvTRPM2 was decreased as intracellular calcium concentration was increased. In addition, our results showed that the four key residues in the calcium-binding site of S2–S3 loop have similar effects on the gating processes of nvTRPM2 and hTRPM2. Among them, the mutations at negatively charged residues(glutamate and aspartate) substantially decreased the currents of nvT RPM2 and hTRPM2. This suggests that these sites are essential for calcium-dependent channel gating. For the charge-neutralizing residues(glutamine and asparagine) in the calcium-binding site, our data showed that glutamine mutating to alanine or glutamate did not affect the channel activity, but glutamine mutating to lysine caused loss of function. Asparagine mutating to aspartate still remained functional, while asparagine mutating to alanine or lysine led to little channel activity. These results suggest that the side chain of glutamine has a less contribution to channel gating than does asparagine. However, our data indicated that both glutamine mutating to alanine or glutamate and asparagine mutating to aspartate accelerated the channel inactivation rate, suggesting that the calcium-binding site in the S2–S3 loop is important for calcium-dependent channel inactivation. Taken together, our results uncovered the effect of four key residues in the S2–S3 loop of TRPM2 on the TRPM2 gating process.展开更多
Curcumin has been shown to significantly improve spatial memory impairment induced by HIV-1 gp 120 V3 in rats, but the electrophysiological mechanism remains unknown. Using extracellular microelectrode recording techn...Curcumin has been shown to significantly improve spatial memory impairment induced by HIV-1 gp 120 V3 in rats, but the electrophysiological mechanism remains unknown. Using extracellular microelectrode recording techniques, this study confirmed that the gp120 V3 loop could suppress long-term potentiation in the rat hippocampal CA1 region and synaptic plasticity, and that curcumin could antagonize these inhibitory effects. Using a Fura-2/AM calcium ion probe, we found that curcumin resisted the effects of the gp120 V3 loop on hippocampal synaptosomes and decreased Ca2＋ concentration in synaptosomes. This effect of curcumin was identical to nimodipine, suggesting that curcumin improved the inhibitory effects of gpl20 on synaptic plasticity, ameliorated damage caused to the central nervous system, and might be a potential neuroprotective drug.展开更多
在面向“双碳”目标的废弃物资源化利用进程中,废塑料气化合成气因其富碳氢特性与负碳潜力成为新兴技术焦点。相比于传统焚烧或填埋导致碳资源耗散与二次污染,气化技术将聚烯烃类废弃物定向转化为高纯度合成气,具备低污染、经济效益高...在面向“双碳”目标的废弃物资源化利用进程中,废塑料气化合成气因其富碳氢特性与负碳潜力成为新兴技术焦点。相比于传统焚烧或填埋导致碳资源耗散与二次污染,气化技术将聚烯烃类废弃物定向转化为高纯度合成气,具备低污染、经济效益高、灵活性强等优势,为废弃物高值化利用与绿氢联产提供了关键路径,在实现资源循环和减少环境污染方面有着显著效应。针对常规气化产物气品质不高、催化剂易失活等问题,提出了废塑料分级热解–化学链气化(Staged Chemical Looping Gasification,SCLG)工艺,采用溶胶凝胶法合成的典型钙钛矿LaFeO_(3)载氧体,在固定床反应系统上开展了聚丙烯气化试验研究,考察热解温度、气化温度、LaFeO_(3)(OC)与聚丙烯(PP)质量比对LaFeO_(3)载氧体气化性能的影响机理。该工艺将原料和载氧体分离,能有效避免传统一段式气化工艺中催化剂易被固体残渣、焦炭和焦油污染问题,此外分级反应充分发挥了载氧体的部分氧化能力和还原金属的催化裂解能力,有效提高了合成气的产率和品质。结果表明:热解温度对气化效果的影响较小,而适当提升气化温度效果显著,合适的m(OC):m(PP)能较好地平衡氧化重整和催化裂解两步反应,具备较好的气化效果。在600℃热解温度、850℃气化温度以及m(OC):m(PP)为1:1的反应条件下具有最佳的气化效果,此时合成气产率为.143 mmol/g,碳转化率达82%,CO选择性为80%,并具有较好的循环稳定性。为废塑料资源化及高效稳定气化技术的规模化发展提供了指导。展开更多
本试验根据GenBank中登录的牛副流感病毒3型(BPIV-3)基因序列,利用在线软件Primer Explorer V4Software和Primer Premier 5.0,针对BPIV-3 NP基因序列的保守区设计并筛选了一套环介导逆转录等温核酸扩增(RT-LAMP)引物,建立BPIV-3特异性...本试验根据GenBank中登录的牛副流感病毒3型(BPIV-3)基因序列,利用在线软件Primer Explorer V4Software和Primer Premier 5.0,针对BPIV-3 NP基因序列的保守区设计并筛选了一套环介导逆转录等温核酸扩增(RT-LAMP)引物,建立BPIV-3特异性检测的RT-LAMP方法。在Bst DNA聚合酶作用下,63℃恒温反应1h即可完成扩增过程,扩增产物通过浑浊度比较、凝胶电泳和肉眼可视化进行判定。结果表明,该方法比RT-PCR敏感度更高,最低检出量可达0.069fg/μL。该方法可用于牛副流感病毒3型的实验室检测和临床初步诊断。展开更多
基金National Natural Science Foundation(Grant No.81373372)Specialized Research Fund for the Doctoral Program of Higher Education of China(Grant No.20110001110021 and 20130001110059)
文摘HIV-1 gains entry into target cells by sequentially interacting with cellular receptors and co-receptors. Both the receptor and co-receptor are recognized by HIV-1 envelope protein gpl20, which plays a key role in the entry process of HIV-1 into cells. The development of new inhibitors is essential since the viral enzyme reverse transcriptase （RT） is one of the first targets of antiretroviral therapy. It has been reported that a variety of natural plants, such as Artemisia rupestris L., have anti-viral pharmacological activity, and they might be the potential inhibitors of RT or V3 loop of gpl20 against HIV-1. RIQRGPGRAFVT1GK （R15K）, the relatively conserved region of V3 loop, can be used for binding research. In this work, we analyzed the interactions between different extracts from Artemisia rupestris L. and R15K by affinity capillary electrophoresis （ACE）. Moreover, we analyzed the interactions between different extracts from Artemisia rupestris L. and RT by capillary zone electrophoresis （CZE）. Our data showed that the chloroform extract ofArtemisia rupestris L. was active among the different plant extracts, which was consistent with previous studies. Taken together, our study provided a rapid screening method to seek anti-HIV ingredients in natural plants＇ extracts.
基金supported by the Natural Science Foundation of Guangdong Province, No. 9151040701000, 061050246the Science and Technology Project of Guangdong Province, No. 2010B030700016+1 种基金the Science and Technology Project of Guangzhou, No. 2010Y1-C291the National Natural Science Foundation of China, No. 81171134
文摘Curcumin improves the learning and memory deficits in rats induced by the gp120 V3 loop. The present study cultured rat hippocampal neurons with 1 nM gp120 V3 loop and 1 μM curcumin for 24 hours. The results showed that curcumin inhibited the gp120 V3 loop-induced mitochondrial membrane potential decrease, reduced the mRNA expression of the pro-apoptotic gene caspase-3, and attenuated hippocampal neuronal injury.
基金This work was supported by the National Natural Science Foundation of China(No.82001850)Shanxi Basic Application Research(No.201901D211491)+2 种基金the Scientific Research Project of Shanxi Health Commission(No.2019038)the Doctoral Research Project of Shanxi Medical University(No.XD1901)the Doctoral Research Project of Shanxi Province(No.SD1901).
文摘Cervical cancer(CC)is recognized as the most common neoplasm in the female reproductive system worldwide.The lack of chemotherapeutic agents with outstanding effectiveness and safety severely compromises the anti-cipated prognosis of patients.Aloperine(ALO)is a natural quinolizidine alkaloid with marked anti-cancer effects on multiple malignancies as well as favorable activity in relieving inflammation,allergies and infection.However,its therapeutic efficacy and underlying mechanism in CC are still unclear.In the current study,MTT assay was employed to evaluate the viability of HeLa cells exposed to ALO to preliminarily estimate the effectiveness of ALO in CC.Then,the effects of ALO on the proliferation and apoptosis of HeLa cells were further investigated by plate colony formation and flow cytometry,respectively,while the migration and invasion of ALO-treated HeLa cells were evaluated using Transwell assay.Moreover,nude mice were subcutaneously inoculated with HeLa cells to demonstrate the anti-CC properties of ALO in vivo.The molecular mechanisms underlying these effects of ALO were evaluated by Western blot and immunohistochemical analysis.This study experimentally demonstrated that ALO inhibited the proliferation of HeLa cells via G2 phase cell cycle arrest.Simultaneously,ALO promoted an increase in the percentage of apoptotic HeLa cells by increasing the Bax/Bcl-2 ratio.Additionally,the migration and invasion of HeLa cells were attenuated by ALO treatment,which was considered to result from inhibition of epithelial-to-mesenchymal transition.For molecular mechanisms,the expression and activation of the IL-6-JAK1-STAT3 feedback loop were markedly suppressed by ALO treatment.This study indicated that ALO markedly suppresses the proliferation,migration and invasion and enhances the apoptosis of HeLa cells.In addition,these prominent anti-CC properties of ALO are associated with repression of the IL-6-JAK1-STAT3 feedback loop.
基金Project supported by the National Natural Science Foundation oX f China(Nos.81371302,81571127,and 31872796)the National Basic Research Program(973)of China(No.2014CB910300)+1 种基金the National Major New Drugs Innovation and Development(No.2018ZX X09711001-004-005)the Zhejiang Provincial Natural Science Foundation of China(Nos.LR16H090001 and LY19B020013)
文摘As a crucial signaling molecule, calcium plays a critical role in many physiological and pathological processes by regulating ion channel activity. Recently, one study resolved the structure of the transient receptor potential melastatin 2(TRPM2) channel from Nematostella vectensis(nvTRPM2). This identified a calcium-binding site in the S2–S3 loop, while its effect on channel gating remains unclear. Here, we investigated the role of this calcium-binding site in both nvTRPM2 and human TRPM2(hTRPM2) by mutagenesis and patch-clamp recording. Unlike hTRPM2, nvT RPM2 cannot be activated by calcium alone. Moreover, the inactivation rate of nvTRPM2 was decreased as intracellular calcium concentration was increased. In addition, our results showed that the four key residues in the calcium-binding site of S2–S3 loop have similar effects on the gating processes of nvTRPM2 and hTRPM2. Among them, the mutations at negatively charged residues(glutamate and aspartate) substantially decreased the currents of nvT RPM2 and hTRPM2. This suggests that these sites are essential for calcium-dependent channel gating. For the charge-neutralizing residues(glutamine and asparagine) in the calcium-binding site, our data showed that glutamine mutating to alanine or glutamate did not affect the channel activity, but glutamine mutating to lysine caused loss of function. Asparagine mutating to aspartate still remained functional, while asparagine mutating to alanine or lysine led to little channel activity. These results suggest that the side chain of glutamine has a less contribution to channel gating than does asparagine. However, our data indicated that both glutamine mutating to alanine or glutamate and asparagine mutating to aspartate accelerated the channel inactivation rate, suggesting that the calcium-binding site in the S2–S3 loop is important for calcium-dependent channel inactivation. Taken together, our results uncovered the effect of four key residues in the S2–S3 loop of TRPM2 on the TRPM2 gating process.
基金supported by the National Natural Science Foundation of China,No.81171134 and 81471235a grant from the Introducing Talents of Discipline to Universities,No.B14036+4 种基金a grant from the College Students’Extracurricular Scientific Innovation and Entrepreneurial Activity Research Topic of Jinan University Challenge Cup,No.(2013)27 and (2014)16a grant from the College Students’Extracurricular Scientific Innovation and Entrepreneurial Activity Research Topic of Jinan University in China,No.201410559079,1055914162 and CX14261a grant from the National Basic Research Program of China(973 Program),No.2011CB707501the Science and Technology Foundation of Guangdong Province in China,No.2010B030700016the Natural Science Foundation of Guangdong Province in China,No.2014A030313360
文摘Curcumin has been shown to significantly improve spatial memory impairment induced by HIV-1 gp 120 V3 in rats, but the electrophysiological mechanism remains unknown. Using extracellular microelectrode recording techniques, this study confirmed that the gp120 V3 loop could suppress long-term potentiation in the rat hippocampal CA1 region and synaptic plasticity, and that curcumin could antagonize these inhibitory effects. Using a Fura-2/AM calcium ion probe, we found that curcumin resisted the effects of the gp120 V3 loop on hippocampal synaptosomes and decreased Ca2＋ concentration in synaptosomes. This effect of curcumin was identical to nimodipine, suggesting that curcumin improved the inhibitory effects of gpl20 on synaptic plasticity, ameliorated damage caused to the central nervous system, and might be a potential neuroprotective drug.
文摘在面向“双碳”目标的废弃物资源化利用进程中,废塑料气化合成气因其富碳氢特性与负碳潜力成为新兴技术焦点。相比于传统焚烧或填埋导致碳资源耗散与二次污染,气化技术将聚烯烃类废弃物定向转化为高纯度合成气,具备低污染、经济效益高、灵活性强等优势,为废弃物高值化利用与绿氢联产提供了关键路径,在实现资源循环和减少环境污染方面有着显著效应。针对常规气化产物气品质不高、催化剂易失活等问题,提出了废塑料分级热解–化学链气化(Staged Chemical Looping Gasification,SCLG)工艺,采用溶胶凝胶法合成的典型钙钛矿LaFeO_(3)载氧体,在固定床反应系统上开展了聚丙烯气化试验研究,考察热解温度、气化温度、LaFeO_(3)(OC)与聚丙烯(PP)质量比对LaFeO_(3)载氧体气化性能的影响机理。该工艺将原料和载氧体分离,能有效避免传统一段式气化工艺中催化剂易被固体残渣、焦炭和焦油污染问题,此外分级反应充分发挥了载氧体的部分氧化能力和还原金属的催化裂解能力,有效提高了合成气的产率和品质。结果表明:热解温度对气化效果的影响较小,而适当提升气化温度效果显著,合适的m(OC):m(PP)能较好地平衡氧化重整和催化裂解两步反应,具备较好的气化效果。在600℃热解温度、850℃气化温度以及m(OC):m(PP)为1:1的反应条件下具有最佳的气化效果,此时合成气产率为.143 mmol/g,碳转化率达82%,CO选择性为80%,并具有较好的循环稳定性。为废塑料资源化及高效稳定气化技术的规模化发展提供了指导。
文摘本试验根据GenBank中登录的牛副流感病毒3型(BPIV-3)基因序列,利用在线软件Primer Explorer V4Software和Primer Premier 5.0,针对BPIV-3 NP基因序列的保守区设计并筛选了一套环介导逆转录等温核酸扩增(RT-LAMP)引物,建立BPIV-3特异性检测的RT-LAMP方法。在Bst DNA聚合酶作用下,63℃恒温反应1h即可完成扩增过程,扩增产物通过浑浊度比较、凝胶电泳和肉眼可视化进行判定。结果表明,该方法比RT-PCR敏感度更高,最低检出量可达0.069fg/μL。该方法可用于牛副流感病毒3型的实验室检测和临床初步诊断。
