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
Cornelia de Lange syndrome （CdLS; OMIM： 122470） is characterized by distinctive facial features, growthretardation, hirsutism, and upper limb reduction defects. Craniofacial features manifest as synophrys, arched e...Cornelia de Lange syndrome （CdLS; OMIM： 122470） is characterized by distinctive facial features, growthretardation, hirsutism, and upper limb reduction defects. Craniofacial features manifest as synophrys, arched eyebrows, long thick eyelashes, a small upturned nose, small widely-spaced teeth, and microcephaly. The intelligence quotient （IQ） is usually below the normal level. More phenotypes are frequently found, such as cardiac septal defects, gastrointestinal dysfunction, hearing loss, myopia, and cryptorchidism or hypoplastic genitalia.展开更多
目的讨论血清前B细胞克隆增强因子(pre-B cell colony-enhancing factor,PBEF)、Kruppel样转录因子2(kruppel like factor 2,KLF2)、白介素-26(Interleukin-26,IL-26)与重症肺炎(severe pneumonia,SP)患儿病情严重程度及预后转归的关系...目的讨论血清前B细胞克隆增强因子(pre-B cell colony-enhancing factor,PBEF)、Kruppel样转录因子2(kruppel like factor 2,KLF2)、白介素-26(Interleukin-26,IL-26)与重症肺炎(severe pneumonia,SP)患儿病情严重程度及预后转归的关系。方法选择西安市儿童医院2023年3月-2025年3月收治的180例SP患儿作为研究对象,使用ELISA试剂盒分别检测血清中PBEF、KLF2、IL-26的蛋白水平;根据急性生理与慢性健康评分II(acute physiology and chronic health score,APACHEII评分)判定患儿的病情严重程度并分组为非危重症组(n=60)、危重症组(n=75)和极危重症组(n=45);根据SP患儿预后分为预后良好组(n=127)和预后不良组(n=53);血清PBEF、KLF2、IL-26水平与APACHEII评分的相关性采用Pearson法分析;多因素Logistics回归分析影响SP患儿预后转归的因素;ROC曲线分析血清PBEF、KLF2、IL-26水平对SP患儿预后转归的预测价值。结果与非危重症组比较,危重症组、极危重症组APACHEII评分及血清PBEF、IL-26水平升高(P<0.05),血清KLF2水平均低(P<0.05);与危重症组比较,极危重症组APACHEII评分及血清PBEF、IL-26水平升高,血清KLF2水平降低(P<0.05);SP患儿血清PBEF、IL-26水平与APACHEII评分呈正相关,血清KLF2水平与APACHEII评分呈负相关(P<0.05);预后不良组较预后良好组极危重症患者比例、APACHEII评分及血清PBEF、IL-26水平升高,血清KLF2水平降低(P<0.05);血清PBEF、IL-26水平升高,KLF2水平降低是SP患儿发生预后不良的危险因素(P<0.05);血清PBEF、KLF2、IL-26水平单独及联合预测患儿预后不良的AUC分别为0.769、0.820、0.814、0.933,三者联合预测价值更高(Z=4.112、3.893、4.035,P<0.001)。结论伴随SP患儿病情严重程度的增加,血清PBEF、IL-26水平升高,KLF2水平降低,三者可作为预测患儿预后转归的指标。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
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This work was supported by the grants from the Natural Science Foundation of Zhejiang Province （No. Y2090108）, the Qianjiang Talent Program （No. 2010R10063） and the Scientific Research Foundation for the Return Overseas Chinese Scholars, State Education Ministry.
文摘Cornelia de Lange syndrome （CdLS; OMIM： 122470） is characterized by distinctive facial features, growthretardation, hirsutism, and upper limb reduction defects. Craniofacial features manifest as synophrys, arched eyebrows, long thick eyelashes, a small upturned nose, small widely-spaced teeth, and microcephaly. The intelligence quotient （IQ） is usually below the normal level. More phenotypes are frequently found, such as cardiac septal defects, gastrointestinal dysfunction, hearing loss, myopia, and cryptorchidism or hypoplastic genitalia.
文摘目的讨论血清前B细胞克隆增强因子(pre-B cell colony-enhancing factor,PBEF)、Kruppel样转录因子2(kruppel like factor 2,KLF2)、白介素-26(Interleukin-26,IL-26)与重症肺炎(severe pneumonia,SP)患儿病情严重程度及预后转归的关系。方法选择西安市儿童医院2023年3月-2025年3月收治的180例SP患儿作为研究对象,使用ELISA试剂盒分别检测血清中PBEF、KLF2、IL-26的蛋白水平;根据急性生理与慢性健康评分II(acute physiology and chronic health score,APACHEII评分)判定患儿的病情严重程度并分组为非危重症组(n=60)、危重症组(n=75)和极危重症组(n=45);根据SP患儿预后分为预后良好组(n=127)和预后不良组(n=53);血清PBEF、KLF2、IL-26水平与APACHEII评分的相关性采用Pearson法分析;多因素Logistics回归分析影响SP患儿预后转归的因素;ROC曲线分析血清PBEF、KLF2、IL-26水平对SP患儿预后转归的预测价值。结果与非危重症组比较,危重症组、极危重症组APACHEII评分及血清PBEF、IL-26水平升高(P<0.05),血清KLF2水平均低(P<0.05);与危重症组比较,极危重症组APACHEII评分及血清PBEF、IL-26水平升高,血清KLF2水平降低(P<0.05);SP患儿血清PBEF、IL-26水平与APACHEII评分呈正相关,血清KLF2水平与APACHEII评分呈负相关(P<0.05);预后不良组较预后良好组极危重症患者比例、APACHEII评分及血清PBEF、IL-26水平升高,血清KLF2水平降低(P<0.05);血清PBEF、IL-26水平升高,KLF2水平降低是SP患儿发生预后不良的危险因素(P<0.05);血清PBEF、KLF2、IL-26水平单独及联合预测患儿预后不良的AUC分别为0.769、0.820、0.814、0.933,三者联合预测价值更高(Z=4.112、3.893、4.035,P<0.001)。结论伴随SP患儿病情严重程度的增加,血清PBEF、IL-26水平升高,KLF2水平降低,三者可作为预测患儿预后转归的指标。
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.
