The migration mode transition of cancer cell enhances its invasive capability and the drug resistance,where physical confinement of cell microenvironment has been revealed to induce the mesenchymal-amoeboid transition...The migration mode transition of cancer cell enhances its invasive capability and the drug resistance,where physical confinement of cell microenvironment has been revealed to induce the mesenchymal-amoeboid transition(MAT).However,most existing studies are performed in PDMS microchannels,of which the stiffness is much higher than that of most mammalian tissues.Therefore,the amoeboid migration transition observed in these studies is actually induced by the synergistic effect of matrix stiffness and confinement.Since the stiffness of cell microenvironment has been reported to influence the cell migration in 2D substrate,the decoupling of stiffness and confinement effects is thus in need for elucidating the underlying mechanism of MAT.However,it is technically challenging to construct microchannels with physiologically relevant stiffness and channel size,where existing microchannel platforms with physiological relevance stiffness are all with>10μm channel width.Such size is too wide to mimic the physical confinement that migrating cancer cells confront in vivo,and also larger than the width of PDMS channel,in which the MAT of cancer cell was observed.Therefore,an in vitro cell migration platform,which could mimic both stiffness and confinement of the native physical microenvironment during cancer metastasis,could profoundly contribute to researches on cancer cell migration and cellular mechanotransduction.In this paper,we overcome the limitations of engineering soft materials in microscale by combining the collagen-alginate hydrogel with photolithography.This enables us to improve the accuracy of molded microchannel,and thus successfully construct a 3D microchannel platform,which matches the stiffness and width ranges of native environmental confinement that migrating cancer cells confront in vivo.The stiffness(0.3~20 kPa),confinement(channel width:3.5~14μm)and the adhesion ligand density of the microchannel can be tuned independently.Interestingly,using this platform,we observed that the migration speed of cancer cell is influenced by the synergistic effect of channel stiffness and width,and the increasing stiffness reverses the effect of channel width on the migration speed of cancer cells.In addition,MAT has a strong correlation with the channel stiffness.These findings make us reconsider the widely accepted hypothesis:physical confinement can induce MAT.Actually,this transition can only occur in stiff confined microenvironment not in soft one.For soft microchannels,the compliance of the channel walls could cause little cell/nucleus deformation,and the MAT could not be induced.To further investigate the mechanism of MAT,we developed a computational model to simulate the effect of nucleus deformation on MAT.With the model,we found that deforming the cell nuclear by decreasing the nucleus stiffness will reduce the cellmigration speed.This implies that nuclear stiffness plays an important role in the regulation of cancer migration speed and thus MAT in microchannels.The effect of channel stiffness on MAT and migration speed as observed in our experiment could partially explain previous findings reported in the literature,where the increasing matrix stiffness of tumor microenvironment promotes cancer metastasis.Our observations thus highlight the critical role of cell nuclear deformation not only in MAT,but also in regulating cellular mechanotransduction and cell-ECM interactions.This developed platform is capable of mimicking the native physical microenvironment during metastasis,providing a powerful tool for high-throughput screening applications and investigating the interaction between cancer migration and biophysical microenvironment.展开更多
OBJECTIVE Based on the methods of microdialysis,HPLC-MS/MS and gene chip tech.nology,the mechanism of Baicalin and Geniposide(BC/GP) against excitatory amino acid toxicity in ce.rebral ischemia was studied.This will p...OBJECTIVE Based on the methods of microdialysis,HPLC-MS/MS and gene chip tech.nology,the mechanism of Baicalin and Geniposide(BC/GP) against excitatory amino acid toxicity in ce.rebral ischemia was studied.This will provide guidance for the clinical application of BC/GP and the study of excitatory amino acid toxicity in cerebral ischemia.METHODS(1) Microdialysis technique and HPLC-MS/MS was performed to study the pharmacodynamics of BC/GP against cerebral ischemia.(1)18 SD rats with body weight of(280±20) g were randomly divided into control group,treatment groups with BC/CP at low dose,medium dose and high dose(equal to the dosage of crude drugs for 30 mg·kg^(-1),45 mg·kg^(-1) and 60 mg·kg^(-1) respectively).Rats in each group were given intragastric administration for seven days to establish cerebral ischemia model.Then,microdialysis probe was applied to collect cerebrospinal fluid from hippocampus before and after cerebral ischemia.(2) First,we established the HPLC-MS/MS method for measuring drugs and excitatory amino acids.Then we detected the microdi.alysis samples and observed their changes in animals.(2) The mechanism of BC/GP against excitatory toxicity of cerebral ischemia were observed at gene level by chip technique.(1) 16 SD rats with body weight of 240±20 g were randomly divided into sham group,model group,treatment group of BC(60 mg·kg^(-1)),treatment group of GP(60 mg·kg^(-1)) and treatment group of BC/GP(7:3)(60 mg·kg^(-1)).Rats in eachgroup were given intragastric administration for seven days to establish cerebral ischemia model.Then the rats were sacrificed,and the hippocampus were rapidly harvested and stored at-80℃ for further detection.(2) After the quality inspection of the hippocampal,the qualified samples were subjected to detect the levels of neurotransmitter receptor gene in the ischemic of rats by gene chip technology.Finally,the results were analyzed by the method of ΔΔCt.RESULTS(1) Only three compounds includ.ed GP,glutamic acid and aspartic acid were detected in microdialysis samples by HPLC-MS/MS.The concentration of GP increased and lasted for 120 min with a significant dose-dependent after cerebral ischemia.Compared with low dose group,the AUC(0-t),MRT(0-∞),Cmax and t1/2 z in high-dose group showed significant difference(P<0.01).Compared with the model group,the levels of glutamic acid and aspartic acid in the treatment groups decreased significantly,especially in the middle and high dose groups.(2)89 genes in the neurotransmitter receptor gene signaling pathway were detected by gene chip technol.ogy.There were 22 genes with |Fold Regulation| >1.5 in the model group,compared with the sham group.Five of the 22 genes showed statistically significant differences,including Grin2 c(2.9026),Chrna7(-1.5877),and Tacr2(-1.7695).Htr3 a(-1.8172) and Grm6(-2.3527).There were 5 genes with |Fold Regulation|>1.5 in the BC group,compared with the model group,Two of them exhibited statistically significant differences,including Brs3(1.797)and Grin2 c(-1.7979).There were 14 genes with |Fold Reg.ulation| >1.5 in the GP group,compared with the model group.Three of them displayed statistically significant differences,including Hcrtr2(-1.6584),Sctr(-3.8524) and Grin2 c(-4.8408).Compared with model group,the genes of |Fold Regulation| >1.5 in BC/GP(7:3) group are 5,and only one of them showed a significant differences.CONCLUSION(1) After administration of BC and GP,GP can cross the blood-brain barrier and reduce the release of excitatory amino acids in the hippocampus.(2) BC/GP can inhibit the interaction between excitatory amino acids and excitatory amino acid receptors and attenuate the toxicity of excitatory amino acids by down-regulating the expression of glutamic acid receptor Grin2 c gene.(3) BC/GP may exert their brain protection effect by reducing the release of excit.atory amino acids and inhibiting the expression of excitatory amino acid receptors.展开更多
科学有效地预测水质对于水环境的可持续发展和人类健康具有重要意义,为此以固原市某黄河断面的水质监测数据为研究对象,提出了基于指标客观性的权重赋权(Criteria Importance Though Intercriteria Correlation,CRITIC)法和改进的秃鹰搜...科学有效地预测水质对于水环境的可持续发展和人类健康具有重要意义,为此以固原市某黄河断面的水质监测数据为研究对象,提出了基于指标客观性的权重赋权(Criteria Importance Though Intercriteria Correlation,CRITIC)法和改进的秃鹰搜索(Improved Bald Eagle Search,IBES)算法优化双向长短时记忆网络(Bidirectional Long Short-Term Memory Network,BiLSTM)的组合水质等级预测模型。首先,采用CRITIC法确定各水质指标的权重,加权求和获得一项综合水质指标,从而提出一种改进的水质评价指标体系,以为BiLSTM提供更丰富、更可靠的水质特征信息。其次,在训练过程中引入Logistic映射和莱维飞行策略,并设计交叉共享及准反向搜索策略优化秃鹰搜索(Bald Eagle Search,BES)算法,以提升其种群多样性,增强寻优能力。最后,通过IBES算法迭代寻找BiLSTM的最佳学习率、隐藏层节点数以及正则化系数的超参数组合,进一步提高其预测水平。结果显示:与IBES-BiLSTM、BES-BiLSTM、GA-BiLSTM、PSO-BiLSTM和BiLSTM等模型相比,CRITIC-IBES-BiLSTM模型进行水质等级预测的准确率、精准率、召回率及F_(1)均最高,且具有更好的稳定性。展开更多
文摘目的观察超声引导下星状神经节阻滞(SGB)对乳腺癌患者乳腺癌相关淋巴水肿及术后恢复质量的影响。方法选择2022年10月至2023年10月择期全麻下行乳腺癌改良根治术的女性患者80例,年龄18~64岁,BMI 18.5~25.0 kg/m^(2),ASAⅠ或Ⅱ级。采用随机数字表法将患者随机分为两组:SGB组(S组)和对照组(C组),每组40例。S组术前行超声引导下单次右侧SGB;C组术前仅接受右侧星状神经节超声扫描不进行神经阻滞。记录术后3 d内乳腺癌相关淋巴水肿的发生情况。记录术前1 d和术后1、3 d睡眠时间,术后1、3 d 15项恢复质量评分(QoR-15)及Christensen疲劳评分。记录术后2、8、24 h VAS疼痛评分以及术后24 h内舒芬太尼用量、镇痛泵总按压次数及有效按压次数和补救镇痛例数。记录手术时间、拔管时间、PACU停留时间、术后住院时间、术后腹胀和恶心呕吐的发生情况以及术后神经阻滞相关并发症的发生情况。结果与C组比较,S组术后3 d内乳腺癌相关淋巴水肿发生率明显降低,术后1、3 d睡眠时间明显延长,术后1、3 d QoR-15评分明显升高,Christensen疲劳评分明显降低,术后24 h内舒芬太尼用量、镇疼泵总按压次数及有效按压次数明显减少,术后2、8、24 h VAS疼痛评分明显降低,拔管时间、PACU停留时间和术后住院时间明显缩短,术后腹胀和术后恶心呕吐发生率明显降低(P<0.05)。两组术后24 h内补救镇痛率差异无统计学意义。SGB组无一例发生神经阻滞相关并发症。结论在接受乳腺癌改良根治术的女性患者中术前行超声引导下单次SGB,可以降低乳腺癌相关淋巴水肿的发生率,提高术后恢复质量。
基金financially supported by the National Natural Science Foundation of China ( 11532009, 11602191,21775117)the General Financial Grant from the China Postdoctoral Science Foundation ( 2016M592773)the High Level Returned Overseas Students Foundation ( [2018]642)
文摘The migration mode transition of cancer cell enhances its invasive capability and the drug resistance,where physical confinement of cell microenvironment has been revealed to induce the mesenchymal-amoeboid transition(MAT).However,most existing studies are performed in PDMS microchannels,of which the stiffness is much higher than that of most mammalian tissues.Therefore,the amoeboid migration transition observed in these studies is actually induced by the synergistic effect of matrix stiffness and confinement.Since the stiffness of cell microenvironment has been reported to influence the cell migration in 2D substrate,the decoupling of stiffness and confinement effects is thus in need for elucidating the underlying mechanism of MAT.However,it is technically challenging to construct microchannels with physiologically relevant stiffness and channel size,where existing microchannel platforms with physiological relevance stiffness are all with>10μm channel width.Such size is too wide to mimic the physical confinement that migrating cancer cells confront in vivo,and also larger than the width of PDMS channel,in which the MAT of cancer cell was observed.Therefore,an in vitro cell migration platform,which could mimic both stiffness and confinement of the native physical microenvironment during cancer metastasis,could profoundly contribute to researches on cancer cell migration and cellular mechanotransduction.In this paper,we overcome the limitations of engineering soft materials in microscale by combining the collagen-alginate hydrogel with photolithography.This enables us to improve the accuracy of molded microchannel,and thus successfully construct a 3D microchannel platform,which matches the stiffness and width ranges of native environmental confinement that migrating cancer cells confront in vivo.The stiffness(0.3~20 kPa),confinement(channel width:3.5~14μm)and the adhesion ligand density of the microchannel can be tuned independently.Interestingly,using this platform,we observed that the migration speed of cancer cell is influenced by the synergistic effect of channel stiffness and width,and the increasing stiffness reverses the effect of channel width on the migration speed of cancer cells.In addition,MAT has a strong correlation with the channel stiffness.These findings make us reconsider the widely accepted hypothesis:physical confinement can induce MAT.Actually,this transition can only occur in stiff confined microenvironment not in soft one.For soft microchannels,the compliance of the channel walls could cause little cell/nucleus deformation,and the MAT could not be induced.To further investigate the mechanism of MAT,we developed a computational model to simulate the effect of nucleus deformation on MAT.With the model,we found that deforming the cell nuclear by decreasing the nucleus stiffness will reduce the cellmigration speed.This implies that nuclear stiffness plays an important role in the regulation of cancer migration speed and thus MAT in microchannels.The effect of channel stiffness on MAT and migration speed as observed in our experiment could partially explain previous findings reported in the literature,where the increasing matrix stiffness of tumor microenvironment promotes cancer metastasis.Our observations thus highlight the critical role of cell nuclear deformation not only in MAT,but also in regulating cellular mechanotransduction and cell-ECM interactions.This developed platform is capable of mimicking the native physical microenvironment during metastasis,providing a powerful tool for high-throughput screening applications and investigating the interaction between cancer migration and biophysical microenvironment.
基金supported by National Natural Science Foundation of China(81473385) Shaanxi provincial Natural Science Foundation of China(2017JZ027) Shaanxi Provincial Administration of Traditional Chinese Medicine(13-ZY016)
文摘OBJECTIVE Based on the methods of microdialysis,HPLC-MS/MS and gene chip tech.nology,the mechanism of Baicalin and Geniposide(BC/GP) against excitatory amino acid toxicity in ce.rebral ischemia was studied.This will provide guidance for the clinical application of BC/GP and the study of excitatory amino acid toxicity in cerebral ischemia.METHODS(1) Microdialysis technique and HPLC-MS/MS was performed to study the pharmacodynamics of BC/GP against cerebral ischemia.(1)18 SD rats with body weight of(280±20) g were randomly divided into control group,treatment groups with BC/CP at low dose,medium dose and high dose(equal to the dosage of crude drugs for 30 mg·kg^(-1),45 mg·kg^(-1) and 60 mg·kg^(-1) respectively).Rats in each group were given intragastric administration for seven days to establish cerebral ischemia model.Then,microdialysis probe was applied to collect cerebrospinal fluid from hippocampus before and after cerebral ischemia.(2) First,we established the HPLC-MS/MS method for measuring drugs and excitatory amino acids.Then we detected the microdi.alysis samples and observed their changes in animals.(2) The mechanism of BC/GP against excitatory toxicity of cerebral ischemia were observed at gene level by chip technique.(1) 16 SD rats with body weight of 240±20 g were randomly divided into sham group,model group,treatment group of BC(60 mg·kg^(-1)),treatment group of GP(60 mg·kg^(-1)) and treatment group of BC/GP(7:3)(60 mg·kg^(-1)).Rats in eachgroup were given intragastric administration for seven days to establish cerebral ischemia model.Then the rats were sacrificed,and the hippocampus were rapidly harvested and stored at-80℃ for further detection.(2) After the quality inspection of the hippocampal,the qualified samples were subjected to detect the levels of neurotransmitter receptor gene in the ischemic of rats by gene chip technology.Finally,the results were analyzed by the method of ΔΔCt.RESULTS(1) Only three compounds includ.ed GP,glutamic acid and aspartic acid were detected in microdialysis samples by HPLC-MS/MS.The concentration of GP increased and lasted for 120 min with a significant dose-dependent after cerebral ischemia.Compared with low dose group,the AUC(0-t),MRT(0-∞),Cmax and t1/2 z in high-dose group showed significant difference(P<0.01).Compared with the model group,the levels of glutamic acid and aspartic acid in the treatment groups decreased significantly,especially in the middle and high dose groups.(2)89 genes in the neurotransmitter receptor gene signaling pathway were detected by gene chip technol.ogy.There were 22 genes with |Fold Regulation| >1.5 in the model group,compared with the sham group.Five of the 22 genes showed statistically significant differences,including Grin2 c(2.9026),Chrna7(-1.5877),and Tacr2(-1.7695).Htr3 a(-1.8172) and Grm6(-2.3527).There were 5 genes with |Fold Regulation|>1.5 in the BC group,compared with the model group,Two of them exhibited statistically significant differences,including Brs3(1.797)and Grin2 c(-1.7979).There were 14 genes with |Fold Reg.ulation| >1.5 in the GP group,compared with the model group.Three of them displayed statistically significant differences,including Hcrtr2(-1.6584),Sctr(-3.8524) and Grin2 c(-4.8408).Compared with model group,the genes of |Fold Regulation| >1.5 in BC/GP(7:3) group are 5,and only one of them showed a significant differences.CONCLUSION(1) After administration of BC and GP,GP can cross the blood-brain barrier and reduce the release of excitatory amino acids in the hippocampus.(2) BC/GP can inhibit the interaction between excitatory amino acids and excitatory amino acid receptors and attenuate the toxicity of excitatory amino acids by down-regulating the expression of glutamic acid receptor Grin2 c gene.(3) BC/GP may exert their brain protection effect by reducing the release of excit.atory amino acids and inhibiting the expression of excitatory amino acid receptors.
文摘科学有效地预测水质对于水环境的可持续发展和人类健康具有重要意义,为此以固原市某黄河断面的水质监测数据为研究对象,提出了基于指标客观性的权重赋权(Criteria Importance Though Intercriteria Correlation,CRITIC)法和改进的秃鹰搜索(Improved Bald Eagle Search,IBES)算法优化双向长短时记忆网络(Bidirectional Long Short-Term Memory Network,BiLSTM)的组合水质等级预测模型。首先,采用CRITIC法确定各水质指标的权重,加权求和获得一项综合水质指标,从而提出一种改进的水质评价指标体系,以为BiLSTM提供更丰富、更可靠的水质特征信息。其次,在训练过程中引入Logistic映射和莱维飞行策略,并设计交叉共享及准反向搜索策略优化秃鹰搜索(Bald Eagle Search,BES)算法,以提升其种群多样性,增强寻优能力。最后,通过IBES算法迭代寻找BiLSTM的最佳学习率、隐藏层节点数以及正则化系数的超参数组合,进一步提高其预测水平。结果显示:与IBES-BiLSTM、BES-BiLSTM、GA-BiLSTM、PSO-BiLSTM和BiLSTM等模型相比,CRITIC-IBES-BiLSTM模型进行水质等级预测的准确率、精准率、召回率及F_(1)均最高,且具有更好的稳定性。
文摘目的比较超声引导下腹横筋膜平面(TFP)阻滞与腹横肌平面(TAP)阻滞在剖宫产术后镇痛中的效果。方法择期蛛网膜下腔阻滞下行剖宫产术产妇60例,年龄20~35岁,体重50~75 kg,ASAⅠ或Ⅱ级,采用随机数字表法分为两组:腹横筋膜平面阻滞组(TFP组)和腹横肌平面阻滞组(TAP组),每组29例。术毕TFP组行超声引导下双侧腹横筋膜平面阻滞,TAP组行超声引导下双侧腹横肌平面阻滞,两组均每侧注射0.375%罗哌卡因1.25 mg/kg。术后均行曲马多PCIA,若VAS疼痛评分≥4分,肌肉注射曲马多100 mg行补救镇痛。记录术后6、12、24、36、48 h PCIA中曲马多累积用量;记录术后48 h内曲马多补救镇痛情况;记录镇痛期间恶心呕吐、局麻药中毒、呼吸抑制、穿刺部位血肿、穿刺部位感染、腹膜刺穿造成腹腔内注射等不良反应的发生情况。结果与TAP组比较,TFP组术后6、12、24、36、48 h PCIA中曲马多累积用量明显减少(P<0.05),术后48 h内曲马多补救镇痛率明显降低(P<0.05)。两组镇痛期间恶心呕吐发生率差异统计学意义。两组均无其他不良反应发生。结论与超声引导下腹横肌平面阻滞比较,腹横筋膜平面阻滞可减少剖宫产术后阿片类药物用量,镇痛效果更佳。
