目的:观察"通督调神针法"与常规针刺治疗急性脑梗死患者的临床疗效差异及其与自噬的关系。方法:将60例急性脑梗死患者分为观察组(30例,2例脱落)与对照组(30例,3例脱落)。两组均采用常规药物治疗。观察组穴取大椎、风池、强间...目的:观察"通督调神针法"与常规针刺治疗急性脑梗死患者的临床疗效差异及其与自噬的关系。方法:将60例急性脑梗死患者分为观察组(30例,2例脱落)与对照组(30例,3例脱落)。两组均采用常规药物治疗。观察组穴取大椎、风池、强间、百会、神庭、印堂、水沟、曲池(患侧)、合谷(患侧)、足三里(患侧),将百会和印堂连接电针仪,30 min后去除电针及非督脉穴位针,继续留督脉穴位针20 min,每隔5 min捻转1次,每次1 min。对照组穴取百会、印堂、曲池(患侧)、外关(患侧)、手三里(患侧)、合谷、足三里、三阴交(患侧)、太溪(患侧)、太冲(患侧),足三里、合谷加电针仪,每次留针30 min。每天治疗1次,连续治疗10 d。观察两组患者治疗前后美国国立卫生院神经功能缺损评分量表(National Institute of Health stroke scale,NIHSS)、简易智能精神状态检查量表(mini-mental state examimation,MMSE)、改良Barthel指数(modified Barthel index,MBI)评分,并评定外周血清中微管相关蛋白1轻链3-Ⅱ(LC3-Ⅱ)和自噬相关蛋白Beclin1表达量的变化及临床疗效。结果:治疗后两组NIHSS评分均低于治疗前,MBI评分、MMSE评分均高于治疗前(均P<0.01),且观察组均优于对照组(均P<0.05)。治疗后两组患者LC3-Ⅱ、Beclin1表达量均高于治疗前(均P<0.01),且观察组均高于对照组(均P<0.05)。治疗后观察组总有效率为92.9%(26/28),优于对照组的70.4%(19/27,P<0.05)。结论:"通督调神针法"减轻急性脑梗死患者临床症状优于常规针刺,可能与自噬体数量增加、自噬活性增强有关。展开更多
Autophagy plays important roles in modulating viral replication and antiviral immune response. Coronavirus infection is associated with the autophagic process, however, little is known about the mechanisms of autophag...Autophagy plays important roles in modulating viral replication and antiviral immune response. Coronavirus infection is associated with the autophagic process, however, little is known about the mechanisms of autophagy induction and its contribution to coronavirus regulation of host innate responses. Here, we show that the membrane-associated papain-like protease PLP2 （PLP2-TM） of coronaviruses acts as a novel autophagy- inducing protein. Intriguingly, PLP2-TM induces incom- plete autophagy process by increasing the accumula- tion of autophagosomes but blocking the fusion of autophagosomes with lysosomes. Furthermore, PLP2- TM interacts with the key autophagy regulators, LC3 and Beclinl, and promotes Beclinl interaction with STING, the key regulator for antiviral IFN signaling. Finally, knockdown of Beclinl partially reverses PLP2-TM＇s inhibitory effect on innate immunity which resulting in decreased coronavirus replication. These results sug- gested that coronavirus papain-like protease induces incomplete autophagy by interacting with Beclinl, which in turn modulates coronavirus replication and antiviral innate immunity.展开更多
Objective： To observe the deregulation of autophagy in diabetic peripheral neuropathy （DPN） and investigate whether Jinmaitong （筋脉通, JMT） alleviates DPN by inducing autophagy. Methods： DPN models were establi...Objective： To observe the deregulation of autophagy in diabetic peripheral neuropathy （DPN） and investigate whether Jinmaitong （筋脉通, JMT） alleviates DPN by inducing autophagy. Methods： DPN models were established by streptozotocin-induced diabetic rats and Schwann cells （SCs） cultured in high glucose medium. The pathological morphology was observed by the improved Bielschowsky＇s nerve fiber axonal staining and the Luxol fast blue-neutral red myelin staining. The ultrastructure was observed by the transmission electron microscopy. Beclinl level was detected by immunohistochemistry and Western blot. The proliferation of cultured SCs was detected by methylthiazolyldiphenyl-tetrazolium bromide. Results： Diabetic peripheral nerve tissues demonstrated pathological morphology and reduced autophagic structure, accompanied with down-regulation of Beclinl. JMT apparently alleviated the pathological morphology change and increased the autophagy [in vivo, Beclinl integral optical density （IOD） value of the control group 86.6 ± 17.7, DM 43.9± 8.8, JMT 73.3 ± 17.8, P〈0.01 or P〈0.05, in vitro Beclinl IOD value of the glucose group 0.47 ± 0.25 vs the control group 0.88 ± 0.29, P〈0.05]. Consequently, inhibition of autophagy by 3-methyladenine resulted in a time- and concentration- dependent decrease of the proliferation of SCs （P〈0.05, P〈0.01）. Conclusions： Down-regulation of autophagy in SCs might contribute to the pathogenesis of DPN. JMT alleviates diabetic peripheral nerve injury at least in part by inducing autophagy.展开更多
Objective To investigate changes of autophagy after traumatic brain injury （TBI） and its possible role. Methods Rat TBI model was established by controlled cortical injury system. Autophagic double membrane structur...Objective To investigate changes of autophagy after traumatic brain injury （TBI） and its possible role. Methods Rat TBI model was established by controlled cortical injury system. Autophagic double membrane structure was detected by transmission electronic microscope. Microtubule-associated protein 1 light chain 3 （LC3） and Beclin 1 were also used to investigate the activation of autophagy post-TBI. Double labeling with LC3 and caspase-3, or Beclin 1 and Fluoro-Jade, to show the relationship between autophagy and apoptosis or neuron degeneration after TBI. Results An increase of autophagic double membrane structure was observed in early stage （1 h）, and the increase lasted for at least 32 d post-TBI. LC3 and Beclin 1 proteins also began to elevate at 1 h time point post-TBI in neurons, 3 d later in astrocytes, and peaked at about 8 d post-TBI. In both cell types, LC3 and Beclin l maintained at a high level until 32 d post-TBI. Most LC3 and Beclin 1 positive cells were near the side （including hippocampus）, but not in the core of the injury. In addition, in the periphery of the injury site, not all caspase-3 positive （＋） cells merged with LC3 （＋） cells post-TBI; In hippocampal area, almost all Beclin 1 （＋） neurons did not merge with Fluoro-Jade （＋） neurons from 1 h to 48 h post-TBI. Conclusion Autophagy is activated and might protect neurons from degeneration at early stage post-TBI and play a continuous role afterwards in eliminating aberrant cell components.展开更多
文摘目的:观察"通督调神针法"与常规针刺治疗急性脑梗死患者的临床疗效差异及其与自噬的关系。方法:将60例急性脑梗死患者分为观察组(30例,2例脱落)与对照组(30例,3例脱落)。两组均采用常规药物治疗。观察组穴取大椎、风池、强间、百会、神庭、印堂、水沟、曲池(患侧)、合谷(患侧)、足三里(患侧),将百会和印堂连接电针仪,30 min后去除电针及非督脉穴位针,继续留督脉穴位针20 min,每隔5 min捻转1次,每次1 min。对照组穴取百会、印堂、曲池(患侧)、外关(患侧)、手三里(患侧)、合谷、足三里、三阴交(患侧)、太溪(患侧)、太冲(患侧),足三里、合谷加电针仪,每次留针30 min。每天治疗1次,连续治疗10 d。观察两组患者治疗前后美国国立卫生院神经功能缺损评分量表(National Institute of Health stroke scale,NIHSS)、简易智能精神状态检查量表(mini-mental state examimation,MMSE)、改良Barthel指数(modified Barthel index,MBI)评分,并评定外周血清中微管相关蛋白1轻链3-Ⅱ(LC3-Ⅱ)和自噬相关蛋白Beclin1表达量的变化及临床疗效。结果:治疗后两组NIHSS评分均低于治疗前,MBI评分、MMSE评分均高于治疗前(均P<0.01),且观察组均优于对照组(均P<0.05)。治疗后两组患者LC3-Ⅱ、Beclin1表达量均高于治疗前(均P<0.01),且观察组均高于对照组(均P<0.05)。治疗后观察组总有效率为92.9%(26/28),优于对照组的70.4%(19/27,P<0.05)。结论:"通督调神针法"减轻急性脑梗死患者临床症状优于常规针刺,可能与自噬体数量增加、自噬活性增强有关。
基金This research was supported by grants from the National Natural Science Foundation of China （Grant Nos. 81273231,81172799 to Z. C. and 81102478, 81471947 to Y. X.）.
文摘Autophagy plays important roles in modulating viral replication and antiviral immune response. Coronavirus infection is associated with the autophagic process, however, little is known about the mechanisms of autophagy induction and its contribution to coronavirus regulation of host innate responses. Here, we show that the membrane-associated papain-like protease PLP2 （PLP2-TM） of coronaviruses acts as a novel autophagy- inducing protein. Intriguingly, PLP2-TM induces incom- plete autophagy process by increasing the accumula- tion of autophagosomes but blocking the fusion of autophagosomes with lysosomes. Furthermore, PLP2- TM interacts with the key autophagy regulators, LC3 and Beclinl, and promotes Beclinl interaction with STING, the key regulator for antiviral IFN signaling. Finally, knockdown of Beclinl partially reverses PLP2-TM＇s inhibitory effect on innate immunity which resulting in decreased coronavirus replication. These results sug- gested that coronavirus papain-like protease induces incomplete autophagy by interacting with Beclinl, which in turn modulates coronavirus replication and antiviral innate immunity.
基金Supported by the Natural Science Foundation of China(No.30572438)the Beijing Science Nuture Foundation(No.7132189)+1 种基金Science Foundation of Peking Union Medical College Hospital(No.2013-098)Youth Foundation of Peking Union Medical College Hospital
文摘Objective： To observe the deregulation of autophagy in diabetic peripheral neuropathy （DPN） and investigate whether Jinmaitong （筋脉通, JMT） alleviates DPN by inducing autophagy. Methods： DPN models were established by streptozotocin-induced diabetic rats and Schwann cells （SCs） cultured in high glucose medium. The pathological morphology was observed by the improved Bielschowsky＇s nerve fiber axonal staining and the Luxol fast blue-neutral red myelin staining. The ultrastructure was observed by the transmission electron microscopy. Beclinl level was detected by immunohistochemistry and Western blot. The proliferation of cultured SCs was detected by methylthiazolyldiphenyl-tetrazolium bromide. Results： Diabetic peripheral nerve tissues demonstrated pathological morphology and reduced autophagic structure, accompanied with down-regulation of Beclinl. JMT apparently alleviated the pathological morphology change and increased the autophagy [in vivo, Beclinl integral optical density （IOD） value of the control group 86.6 ± 17.7, DM 43.9± 8.8, JMT 73.3 ± 17.8, P〈0.01 or P〈0.05, in vitro Beclinl IOD value of the glucose group 0.47 ± 0.25 vs the control group 0.88 ± 0.29, P〈0.05]. Consequently, inhibition of autophagy by 3-methyladenine resulted in a time- and concentration- dependent decrease of the proliferation of SCs （P〈0.05, P〈0.01）. Conclusions： Down-regulation of autophagy in SCs might contribute to the pathogenesis of DPN. JMT alleviates diabetic peripheral nerve injury at least in part by inducing autophagy.
基金This work was supported by the National Natural Science Foundation of China （No. 30571909） the Youth Teacher Foundation of Jiangsu Province （No. BU134701）+1 种基金 Medical Development Foundation of Soochow University （No. EE 134615） We also thank Jin-Sheng YANG for his help in polishing the English language.
文摘Objective To investigate changes of autophagy after traumatic brain injury （TBI） and its possible role. Methods Rat TBI model was established by controlled cortical injury system. Autophagic double membrane structure was detected by transmission electronic microscope. Microtubule-associated protein 1 light chain 3 （LC3） and Beclin 1 were also used to investigate the activation of autophagy post-TBI. Double labeling with LC3 and caspase-3, or Beclin 1 and Fluoro-Jade, to show the relationship between autophagy and apoptosis or neuron degeneration after TBI. Results An increase of autophagic double membrane structure was observed in early stage （1 h）, and the increase lasted for at least 32 d post-TBI. LC3 and Beclin 1 proteins also began to elevate at 1 h time point post-TBI in neurons, 3 d later in astrocytes, and peaked at about 8 d post-TBI. In both cell types, LC3 and Beclin l maintained at a high level until 32 d post-TBI. Most LC3 and Beclin 1 positive cells were near the side （including hippocampus）, but not in the core of the injury. In addition, in the periphery of the injury site, not all caspase-3 positive （＋） cells merged with LC3 （＋） cells post-TBI; In hippocampal area, almost all Beclin 1 （＋） neurons did not merge with Fluoro-Jade （＋） neurons from 1 h to 48 h post-TBI. Conclusion Autophagy is activated and might protect neurons from degeneration at early stage post-TBI and play a continuous role afterwards in eliminating aberrant cell components.
