A cathepsin B-like proteinase was purified from eggs of Antheraea pernyi by 5 steps of purification. The molecular mass of the proteinase are estimated to be 47 kDa by using SDS-polyacrylamide gel electro-phoresis. Th...A cathepsin B-like proteinase was purified from eggs of Antheraea pernyi by 5 steps of purification. The molecular mass of the proteinase are estimated to be 47 kDa by using SDS-polyacrylamide gel electro-phoresis. The activity of the purified proteinase were strongly inhibited by E-64 and Leupeptin. The optimum pH is 3.5 as determined by using the bovine hemo-globin as substrate. The activity and quantity of the proteinase during embryo development were studied. The results suggested that the proteolytic activities during embryo development at least partially came from this proteinase.展开更多
The homozygous T3 transgenic lines with sense OsCBL8 gene and antisense OsCBL8 gene obtained by agro-transformation were used to investigate the function of OsCBL8 in rice. Semi-quantitative RT-PCR showed that the exp...The homozygous T3 transgenic lines with sense OsCBL8 gene and antisense OsCBL8 gene obtained by agro-transformation were used to investigate the function of OsCBL8 in rice. Semi-quantitative RT-PCR showed that the expression of OsCBL8 extremely increased in sense transgenic lines, and decreased to some extents in antisense transgenic lines. Such up- and down-regulation of the OsCBL8 gene in these transgenic lines had little effects on main agronomic traits, but significantly decreased the number of filled grains per panicle and seed setting rate in some of transgenic lines. By evaluation of the tolerance to 150 mmol/L NaCl, 20% PEG6000 and low temperature treatments, and relevant physiological indices, 8F12, a sense transgenic line with high salt tolerance, and 8R14, an antisense transgenic line with high drought tolerance, were obtained, which suggests that the OsCBL8 gene is involved in the response of rice to abiotic stresses.展开更多
This study aimed to examine the prognostic factors of luminal B-like breast cancer.Clinical data of 695 luminal B-like breast cancer patients who had been treated in our hospital during the period of past 4.5 years we...This study aimed to examine the prognostic factors of luminal B-like breast cancer.Clinical data of 695 luminal B-like breast cancer patients who had been treated in our hospital during the period of past 4.5 years were collected and analyzed. Estrogen receptor (ER), progesterone receptor (PgR), antigen identified by monoclonal antibody Ki-67 (Ki67) were immunohistochemically detected. Different cutoffs of ER, PgR, and Ki67 were evaluated. Pearson X test was performed to compare categorical parameters. Univariate and multivariate models were used to evaluate predictors of disease free survival (DFS). The results showed that patients who were younger, and had larger tumors, and more positive lymph nodes were more likely to receive neo-adjuvent chemotherapy (NAC). Patients with ER-positive tumors having <10% positive cells received more anthracycline- and taxane-based chemotherapy and less endocrine therapy than those with ER-positive tumors having >10% positive cells (P=0.004 and P=0.007, respectively);however, patients with ER-positive tumors having <10% positive cells experienced more recurrence (P<0.001). PgR expression levels were not associated with therapeutic schedule and DFS. Patients with tumor tissue Ki67 score >30% received more anthracycline- and taxane-based chemotherapy and had worse DFS than those with tumor tissue Ki67 score <30%. Univariate and multivariate analysis showed that clinical T stage, lymph nodes, ER, Ki67, and HER2 status were independent prognostic factors. In conclusion, ER-positive rate <10% and Ki67 score >30%, similar to higher clinical T stage, more metastatic lymph nodes, and HER2 positive status, may indicate a worse prognosis for luminal B-like breast cancer patients. Multi-center prospective trials with larger sample sizes are necessary for the continued perfection of our work.展开更多
An Or83b-like receptor gene was cloned from antennae of Helicoverpa assulta（Guenée） by using reverse transcription polymerase chain reaction（RT-PCR） and rapid amplification of cDNA ends（RACE）.Sequence analy...An Or83b-like receptor gene was cloned from antennae of Helicoverpa assulta（Guenée） by using reverse transcription polymerase chain reaction（RT-PCR） and rapid amplification of cDNA ends（RACE）.Sequence analysis revealed that the transcript of the Or83b-like receptor gene from H.assulta consists of 1 946 nucleotides and the open reading frame（ORF） encodes a peptide of 473 amino acids with 7 putative transmembrane domains.Alignment analysis suggested that amino acid sequence of Or83b-like receptor from H.assulta shares high identity with other Or83b family receptors and this gene was hence named as HassOr83b.Tissues expression analysis showed that the HassOr83b transcript is clearly observed in the antennae,labial palps and proboscises,but not in bodies,wings and legs.The further development expression analysis suggested HassOr83b is also expressed in several preadult stages,including early-stage larvae,late-stage larvae and pupae,but not in embryos.Locked nucleic acid（LNA）-based in situ hybridization of antennal section indicated that HassOr83b is expressed in a very large number of antennal cells,which suggests that HassOr83b may play a special role in olfaction in H.assulta.展开更多
The complex of calcineurin B-like protein(CBL)and CBL-interacting protein kinase(CIPK)serves as key components in calcium-sensing,orchestrating various signals crucial for plant growth,development,and responses to bio...The complex of calcineurin B-like protein(CBL)and CBL-interacting protein kinase(CIPK)serves as key components in calcium-sensing,orchestrating various signals crucial for plant growth,development,and responses to biotic and abiotic stresses.However,the mechanism underlying the response of this module to cold stress and its role in flower development in wintersweet(Chimonanthus praecox)remains unclear.Through expression pattern analysis,calcium ion(Ca^(2+))concentration assays,correlation analysis,and linear regression analysis,we found that the[Ca^(2+)],along with CpCBL8 and CpCIPK9 expression levels in wintersweet flower buds(FBs),significantly decreased during the initial flowering stage when the chilling requirement reached 570 chill units(CU).Notably,there was a significant positive correlation between[Ca^(2+)]and CpCBL8 expression.Ca^(2+)increased the expression of Cp CBL8 and CpCIPK9 in FBs,causing a significant delay in the flowering of wintersweet.Furthermore,the function of CpCBL8 was studied using heterologous transformation.Overexpression of CpCBL8 significantly delayed flowering time and significantly reduced cold tolerance and altered the expression pattern of endogenous genes related to low-temperature stress and flower development in transgenic Arabidopsis thaliana.Additionally,transcriptome analysis of chilling-induced dormancy breaking and flower bud enlargement revealed that CpCBL8 and CpCIPK9 were negatively regulated by cold,and the expression pattern of endogenous genes related to flower development and cold stress in wintersweet were similar to that of in A.thaliana.Moreover,protein-protein interaction(PPI)analysis revealed that CpCBL8 and CpCIPK9 interacted in the plasma membrane and nucleus.On the basis of these findings,we speculated that the CpCBL8-CpCIPK9 module plays a crucial role in regulating responses to cold stress and flower development in wintersweet.This study elucidated molecular mechanisms through which the downregulation of the Ca^(2+)-induced CpCBL8-CpCIPK9 module results in dormancy breaking and enhances cold tolerance.This study provides valuable insights for the cultivation of new varieties of wintersweet with increased ornamental value and enhanced cold stress tolerance.展开更多
钙调磷酸酶B蛋白(calcineurin B like protein, CBL)互作蛋白激酶(CBL-interacting protein kinase,CIPK)是一类Ser/Thr蛋白激酶,在植物生长发育和逆境胁迫响应过程中发挥重要作用。CIPK可自磷酸化或靶向磷酸化其他信号因子以响应生物...钙调磷酸酶B蛋白(calcineurin B like protein, CBL)互作蛋白激酶(CBL-interacting protein kinase,CIPK)是一类Ser/Thr蛋白激酶,在植物生长发育和逆境胁迫响应过程中发挥重要作用。CIPK可自磷酸化或靶向磷酸化其他信号因子以响应生物和非生物逆境胁迫,也可参与脱落酸(abscisic acid,ABA)、赤霉素(gibberellic acid, GA)、乙烯(ethylene, ETH)、水杨酸(salicylic acid, SA)等植物激素信号通路来调节植物生长发育过程。此外,CIPK通过与CBL蛋白特异性互作形成复合物对盐碱、干旱、极端低温及重金属等逆境胁迫做出应答反应。本文对植物CIPKs的发现、结构、分类、调控机制及其在逆境胁迫响应中的作用等方面研究成果进行综述,并对其未来研究方向进行展望,为农作物抗逆性遗传改良和生物育种提供了参考。展开更多
背景与目的:弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)的生发中心B细胞样(germinal center B-cell-like,GCB)亚型和非GCB(non-GCB)亚型在患者预后和治疗上存在差异,但目前依赖有创病理学检查。本研究基于多参数MRI构建...背景与目的:弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)的生发中心B细胞样(germinal center B-cell-like,GCB)亚型和非GCB(non-GCB)亚型在患者预后和治疗上存在差异,但目前依赖有创病理学检查。本研究基于多参数MRI构建影像组学和深度学习模型,旨在于术前无创性区分这两种亚型。方法:本研究回顾性分析2013年3月—2024年12月在复旦大学附属华山医院及外院经病理学检查确诊的DLBCL患者。使用多参数MRI扫描数据,结合4种影像组学机器学习[支持向量机(support vector machine,SVM)、逻辑回归(logistic regression,LR)、高斯过程(Gaussian process,GP)和朴素贝叶斯(Naive Bayes,NB)]和3种深度学习[密集连接卷积网络121(densely-connected convolutional networks 121,DenseNet121)、残差网络101(residual network 101,ResNet101)和高效网络B5(Efficient Net-b5)]建立DLBCL亚型分类模型。此外,两名经验不同的放射科医师在盲法下基于MRI图像独立分类DLBCL。模型和医师的诊断性能均通过接收者操作特征曲线下面积(area under the curve,AUC)、准确度(accuracy,ACC)和F1分数(F1-score,F1)等指标进行量化评估,以衡量其区分GCB和non-GCB亚型的能力。本研究经复旦大学附属华山医院伦理委员会批准(KY2024-663),所有患者均知情同意。结果:本研究共纳入173例患者(GCB型55例,non-GCB型118例)。影像组学和深度学习方法能有效地区分DLBCL亚型。其中,GP影像组学模型(基于T1-CE+T2-FLAIR+ADC序列)和DenseNet121深度学习模型(基于T1-CE+T2-FLAIR+ADC序列)表现最佳,在内部验证集上分别取得优异性能(GP:AUC=0.900,ACC=0.896,F1=0.840;DenseNet121:AUC=0.846,ACC=0.854,F1=0.774),并在外部验证集上保持稳健。并且,最优AI模型的分类效能优于经验丰富的放射科医师(医师最高AUC=0.678)。结论:基于多参数MRI特征的影像组学与深度学习模型可有效地鉴别DLBCL的GCB与non-GCB亚型。其中,GP与DenseNet121模型在处理复杂图像数据、特别是融合多序列特征组进行亚型分类时,呈现出优异的性能。展开更多
文摘A cathepsin B-like proteinase was purified from eggs of Antheraea pernyi by 5 steps of purification. The molecular mass of the proteinase are estimated to be 47 kDa by using SDS-polyacrylamide gel electro-phoresis. The activity of the purified proteinase were strongly inhibited by E-64 and Leupeptin. The optimum pH is 3.5 as determined by using the bovine hemo-globin as substrate. The activity and quantity of the proteinase during embryo development were studied. The results suggested that the proteolytic activities during embryo development at least partially came from this proteinase.
基金supported by the Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT)Ph.D. the Program Foundation of Ministry of Education of China (Grant No.20020307036)+1 种基金the Natural Science Foundation of China (Grant Nos.30771329 and 30800677)the Natural Science Foundation of Zhejiang Province,China (Grant No.Y3080359)
文摘The homozygous T3 transgenic lines with sense OsCBL8 gene and antisense OsCBL8 gene obtained by agro-transformation were used to investigate the function of OsCBL8 in rice. Semi-quantitative RT-PCR showed that the expression of OsCBL8 extremely increased in sense transgenic lines, and decreased to some extents in antisense transgenic lines. Such up- and down-regulation of the OsCBL8 gene in these transgenic lines had little effects on main agronomic traits, but significantly decreased the number of filled grains per panicle and seed setting rate in some of transgenic lines. By evaluation of the tolerance to 150 mmol/L NaCl, 20% PEG6000 and low temperature treatments, and relevant physiological indices, 8F12, a sense transgenic line with high salt tolerance, and 8R14, an antisense transgenic line with high drought tolerance, were obtained, which suggests that the OsCBL8 gene is involved in the response of rice to abiotic stresses.
文摘This study aimed to examine the prognostic factors of luminal B-like breast cancer.Clinical data of 695 luminal B-like breast cancer patients who had been treated in our hospital during the period of past 4.5 years were collected and analyzed. Estrogen receptor (ER), progesterone receptor (PgR), antigen identified by monoclonal antibody Ki-67 (Ki67) were immunohistochemically detected. Different cutoffs of ER, PgR, and Ki67 were evaluated. Pearson X test was performed to compare categorical parameters. Univariate and multivariate models were used to evaluate predictors of disease free survival (DFS). The results showed that patients who were younger, and had larger tumors, and more positive lymph nodes were more likely to receive neo-adjuvent chemotherapy (NAC). Patients with ER-positive tumors having <10% positive cells received more anthracycline- and taxane-based chemotherapy and less endocrine therapy than those with ER-positive tumors having >10% positive cells (P=0.004 and P=0.007, respectively);however, patients with ER-positive tumors having <10% positive cells experienced more recurrence (P<0.001). PgR expression levels were not associated with therapeutic schedule and DFS. Patients with tumor tissue Ki67 score >30% received more anthracycline- and taxane-based chemotherapy and had worse DFS than those with tumor tissue Ki67 score <30%. Univariate and multivariate analysis showed that clinical T stage, lymph nodes, ER, Ki67, and HER2 status were independent prognostic factors. In conclusion, ER-positive rate <10% and Ki67 score >30%, similar to higher clinical T stage, more metastatic lymph nodes, and HER2 positive status, may indicate a worse prognosis for luminal B-like breast cancer patients. Multi-center prospective trials with larger sample sizes are necessary for the continued perfection of our work.
文摘Sox(SRY-related HMG-box)基因编码一类重要的转录因子,其产物都具有一个HMG基序保守区。之前我们在中华绒螯蟹分离鉴定了一个只在性腺中特异表达转录本Es Sox21b-like,为了验证EsSOX21b-like蛋白是否也在中华绒螯蟹性腺中特异表达,本研究原核表达了EsSOX21b-like蛋白,即将EsSox21b-like基因序列克隆到pGEX-2T表达载体,构建重组表达质粒pGEX-2T-Es Sox21b-like,转化大肠杆菌BL21,IPTG诱导融合表达,经SDS-PAGE分析表明,融合蛋白主要以包涵体形式存在,分子量约为54 k D。利用Ni柱亲和纯化融合蛋白后免疫家兔,制备了多克隆抗体,Western blotting检测表明该抗体能特异地识别性腺组织中EsSOX21b-like蛋白,而在其他组织未检测到其表达,目的蛋白分子量约为28 k D,在精巢组织中的表达量比在卵巢中高,该结果暗示Es SOX21b-like可能在性腺发育过程中起重要调控作用。
基金supported by the Excellent Youth Foundation of Henan Province of China (074100510013)the Science and Technology Innovation Foundation of Henan Agricultural University of China (2007-CX-014)
文摘An Or83b-like receptor gene was cloned from antennae of Helicoverpa assulta（Guenée） by using reverse transcription polymerase chain reaction（RT-PCR） and rapid amplification of cDNA ends（RACE）.Sequence analysis revealed that the transcript of the Or83b-like receptor gene from H.assulta consists of 1 946 nucleotides and the open reading frame（ORF） encodes a peptide of 473 amino acids with 7 putative transmembrane domains.Alignment analysis suggested that amino acid sequence of Or83b-like receptor from H.assulta shares high identity with other Or83b family receptors and this gene was hence named as HassOr83b.Tissues expression analysis showed that the HassOr83b transcript is clearly observed in the antennae,labial palps and proboscises,but not in bodies,wings and legs.The further development expression analysis suggested HassOr83b is also expressed in several preadult stages,including early-stage larvae,late-stage larvae and pupae,but not in embryos.Locked nucleic acid（LNA）-based in situ hybridization of antennal section indicated that HassOr83b is expressed in a very large number of antennal cells,which suggests that HassOr83b may play a special role in olfaction in H.assulta.
基金funded by the Natural Science Foundation of Chongqing(CSTB2023NSCQ-MSX0236)Fundamental Research Funds for the Central Universities(SWU-XDJH202308)Earmarked Funds for the China Agriculture Research System(CARS-26)。
文摘The complex of calcineurin B-like protein(CBL)and CBL-interacting protein kinase(CIPK)serves as key components in calcium-sensing,orchestrating various signals crucial for plant growth,development,and responses to biotic and abiotic stresses.However,the mechanism underlying the response of this module to cold stress and its role in flower development in wintersweet(Chimonanthus praecox)remains unclear.Through expression pattern analysis,calcium ion(Ca^(2+))concentration assays,correlation analysis,and linear regression analysis,we found that the[Ca^(2+)],along with CpCBL8 and CpCIPK9 expression levels in wintersweet flower buds(FBs),significantly decreased during the initial flowering stage when the chilling requirement reached 570 chill units(CU).Notably,there was a significant positive correlation between[Ca^(2+)]and CpCBL8 expression.Ca^(2+)increased the expression of Cp CBL8 and CpCIPK9 in FBs,causing a significant delay in the flowering of wintersweet.Furthermore,the function of CpCBL8 was studied using heterologous transformation.Overexpression of CpCBL8 significantly delayed flowering time and significantly reduced cold tolerance and altered the expression pattern of endogenous genes related to low-temperature stress and flower development in transgenic Arabidopsis thaliana.Additionally,transcriptome analysis of chilling-induced dormancy breaking and flower bud enlargement revealed that CpCBL8 and CpCIPK9 were negatively regulated by cold,and the expression pattern of endogenous genes related to flower development and cold stress in wintersweet were similar to that of in A.thaliana.Moreover,protein-protein interaction(PPI)analysis revealed that CpCBL8 and CpCIPK9 interacted in the plasma membrane and nucleus.On the basis of these findings,we speculated that the CpCBL8-CpCIPK9 module plays a crucial role in regulating responses to cold stress and flower development in wintersweet.This study elucidated molecular mechanisms through which the downregulation of the Ca^(2+)-induced CpCBL8-CpCIPK9 module results in dormancy breaking and enhances cold tolerance.This study provides valuable insights for the cultivation of new varieties of wintersweet with increased ornamental value and enhanced cold stress tolerance.
文摘钙调磷酸酶B蛋白(calcineurin B like protein, CBL)互作蛋白激酶(CBL-interacting protein kinase,CIPK)是一类Ser/Thr蛋白激酶,在植物生长发育和逆境胁迫响应过程中发挥重要作用。CIPK可自磷酸化或靶向磷酸化其他信号因子以响应生物和非生物逆境胁迫,也可参与脱落酸(abscisic acid,ABA)、赤霉素(gibberellic acid, GA)、乙烯(ethylene, ETH)、水杨酸(salicylic acid, SA)等植物激素信号通路来调节植物生长发育过程。此外,CIPK通过与CBL蛋白特异性互作形成复合物对盐碱、干旱、极端低温及重金属等逆境胁迫做出应答反应。本文对植物CIPKs的发现、结构、分类、调控机制及其在逆境胁迫响应中的作用等方面研究成果进行综述,并对其未来研究方向进行展望,为农作物抗逆性遗传改良和生物育种提供了参考。
文摘背景与目的:弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)的生发中心B细胞样(germinal center B-cell-like,GCB)亚型和非GCB(non-GCB)亚型在患者预后和治疗上存在差异,但目前依赖有创病理学检查。本研究基于多参数MRI构建影像组学和深度学习模型,旨在于术前无创性区分这两种亚型。方法:本研究回顾性分析2013年3月—2024年12月在复旦大学附属华山医院及外院经病理学检查确诊的DLBCL患者。使用多参数MRI扫描数据,结合4种影像组学机器学习[支持向量机(support vector machine,SVM)、逻辑回归(logistic regression,LR)、高斯过程(Gaussian process,GP)和朴素贝叶斯(Naive Bayes,NB)]和3种深度学习[密集连接卷积网络121(densely-connected convolutional networks 121,DenseNet121)、残差网络101(residual network 101,ResNet101)和高效网络B5(Efficient Net-b5)]建立DLBCL亚型分类模型。此外,两名经验不同的放射科医师在盲法下基于MRI图像独立分类DLBCL。模型和医师的诊断性能均通过接收者操作特征曲线下面积(area under the curve,AUC)、准确度(accuracy,ACC)和F1分数(F1-score,F1)等指标进行量化评估,以衡量其区分GCB和non-GCB亚型的能力。本研究经复旦大学附属华山医院伦理委员会批准(KY2024-663),所有患者均知情同意。结果:本研究共纳入173例患者(GCB型55例,non-GCB型118例)。影像组学和深度学习方法能有效地区分DLBCL亚型。其中,GP影像组学模型(基于T1-CE+T2-FLAIR+ADC序列)和DenseNet121深度学习模型(基于T1-CE+T2-FLAIR+ADC序列)表现最佳,在内部验证集上分别取得优异性能(GP:AUC=0.900,ACC=0.896,F1=0.840;DenseNet121:AUC=0.846,ACC=0.854,F1=0.774),并在外部验证集上保持稳健。并且,最优AI模型的分类效能优于经验丰富的放射科医师(医师最高AUC=0.678)。结论:基于多参数MRI特征的影像组学与深度学习模型可有效地鉴别DLBCL的GCB与non-GCB亚型。其中,GP与DenseNet121模型在处理复杂图像数据、特别是融合多序列特征组进行亚型分类时,呈现出优异的性能。
