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A New Micropropagation Technology of Tilia amurensis:In VitroMicropropagation of Mature Zygotic Embryos and the Establishment of a PlantRegeneration System
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作者 Shijie Lin Zimo Wang +8 位作者 Hongbo Zhu Conghui Wang Hongfeng Wang Dawei Zhang Tianbing Gou Guangdao Bao Ye Luo Huaijiang He Zhonghui Zhang 《Phyton-International Journal of Experimental Botany》 SCIE 2024年第2期277-289,共13页
Tilia amurensis is an economically valuable broadleaf tree species in Northeast China.The production of highqualityT.amurensis varieties at commercial scales has been greatly limited by the low germination rates.There... Tilia amurensis is an economically valuable broadleaf tree species in Northeast China.The production of highqualityT.amurensis varieties at commercial scales has been greatly limited by the low germination rates.Thereis thus a pressing need to develop an organogenesis protocol for in vitro propagation of T.amurensis to alleviate ashortage of high-quality T.amurensis seedlings.Here,we established a rapid in vitro propagation system forT.amurensis from mature zygotic embryos and analyzed the effects of plant growth regulators and culture mediain different stages.We found that Woody plant medium(WPM)was the optimal primary culture medium formature zygotic embryos.The highest callus induction percentage(68.76%)and number of axillary buds induced(3.2)were obtained in WPM+0.89μmol/L 6-benzyladenine(6-BA)+0.46μmol/L kinetin(KT)+0.25μmol/Lindole-3-butryic acid(IBA)+1.44μmol/L gibberellin A_(3)(GA_(3)).The multiple shoot bud development achievedthe highest percentage(83.32%)in the Murashige and Skoog(MS)+2.22μmol/L 6-BA+0.25μmol/L IBA+1.44μmol/L GA_(3).The rooting percentage(96.70%)was highest in 1/2 MS medium+1.48μmol/L IBA.Thesurvival percentage of transplanting plantlets was 82.22%in soil:vermiculite:perlite(5:3:1).Our study is the firstto establish an effective organogenesis protocol for T.amurensis using mature zygotic embryos. 展开更多
关键词 Tilia amurensis Rupr. mature zygotic embryo MICROPROPAGATION adventitious bud multiplication ROOTING
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Production of marker-free transgenic plants from mature tissues of navel orange using a Cre/loxP site-recombination system
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作者 Yongrui He Lanzhen Xu +7 位作者 Aihong Peng Tiangang Lei Qiang Li Lixiao Yao Guojin Jiang Shanchun Chen Zhengguo Li Xiuping Zou 《Horticultural Plant Journal》 SCIE CAS CSCD 2023年第3期473-480,共8页
Genetic transformation with mature material as the explants could shorten the transgenic period and avoid seed dependence compared with genetic transformation using the epicotyl seedling stem segments as the receptor.... Genetic transformation with mature material as the explants could shorten the transgenic period and avoid seed dependence compared with genetic transformation using the epicotyl seedling stem segments as the receptor. Here, we constructed an Agrobacterium tumefaciensmediated transformation for generation of marker-free transgenic plants from navel orange(Citrus sinensis Osbeck) mature stems using a CreloxP recombination system. To efficiently recover the regenerated buds from mature tissues, five recovery methods were compared: in vitro micrografting of 0.1-0.5(1-2 weeks), > 0.5 cm(3-4 weeks) and > 1 cm long lignified bud and in vitro micrografting of explants with a bud and rooting regenerated bud. The data showed that in vitro micrografting of > 1 cm long regenerated bud with expanded leaves after one month of continuous culture for lignification was the optimal solution for plant recovery from mature tissues. Transgenic plants without selectable marker genes were created from navel orange(Citrus sinensis Osbeck) tissue using a transformation vector PLI-35SPR1aCB containing a Cre/loxP system recombination together with genes encoding the selectable marker isopentenyl transferase(IPT) and an anti-bacterial peptide(PR1aCB).Using IPT positive selection, the transformation efficiency determined by PCR was 0.9%, and in total, 20 transgenic plants were obtained.Southern blotting confirmed further their transgenicity. PCR and sequencing analysis demonstrated that both the Cre and IPT genes had been successfully removed from the transgenic plants(deletion efficiency 100%). Over all, using Cre/loxP system recombination together with the IPT positive selection, marker-free transgenic plants can be recovered efficiently from mature tissues of navel orange(Citrus sinensis Osbeck), which provides a potential method for production of transgenic plants from citrus mature tissue. 展开更多
关键词 Citrus sinensis Osbeck mature stem segment MARKER-FREE TRANSGENE Bud micrografting
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Impact of stage-specific limb function exercises guided by a selfmanagement education model on arteriovenous fistula maturation status 被引量:1
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作者 Yi Li Li-Jun Huang +1 位作者 Jian-Wen Hou Dan-Dan Hu 《World Journal of Clinical Cases》 SCIE 2024年第14期2316-2323,共8页
BACKGROUND The exercise of limb function is the most economical and safe method to promote the maturation of arteriovenous fistula(AVF).However,due to the lack of a uni-fied exercise standard in China,many patients ha... BACKGROUND The exercise of limb function is the most economical and safe method to promote the maturation of arteriovenous fistula(AVF).However,due to the lack of a uni-fied exercise standard in China,many patients have insufficient awareness of the importance of AVF,leading to poor effectiveness of limb function exercise.The self-management education model can effectively promote patients to take pro-active health-related actions.This study focuses on the characteristics of patients during the peri-AVF period and conducts a phased limb function exercise under the guidance of the self-management education model to observe changes in fac-tors such as the maturity of AVF.AIM To assess the impact of stage-specific limb function exercises,directed by a self-management education model,on the maturation status of AVFs.METHODS This study is a randomized controlled trial involving 74 patients with forearm AVFs from the Nephrology Department of a tertiary hospital in Sichuan Province,China.Patients were randomly divided into an observation group and a control group using a random number table method.The observation group underwent tailored stage-specific limb func-tion exercises,informed by a self-management education model which took into account the unique features of AVF at various stages,in conjunction with routine care.Conversely,the control group was given standard limb function exercises along with routine care.The assessment involves the maturity of AVFs post-intervention,post-operative complications,and the self-management level of the fistula in both groups patients.Analyses were conducted using SPSS version 23.0.Count data were represented by frequency and percentage and subjected to chi-square test comparisons.Measurement data adhering to a normal distribution were presented as mean±SD.The independent samples t-test was utilized for inter-group comparisons,while the paired t-test was used for intra-group comparisons.For measurement data not fitting a normal distribution,the median and interquartile range were presented and analyzed using the Wilcoxon rank sum test.RESULTS At the 8-wk postoperative mark,the observation group demonstrated significantly higher scores in AVF symptom recognition,symptom prevention,and self-management compared to the control group(P<0.05).However,the variance in symptom management scores between the observation and control groups lacked statistical signi-ficance(P>0.05).At 4 wk after the operation,the observation group displayed a superior vessel diameter and depth from the skin of the drainage vessels in comparison to the control group(P<0.05).While the observation group did manifest elevated blood flow rates in the drainage vessels relative to the control group,this distinction was not statistically significant(P>0.05).By the 8-wk postoperative interval,the observation group outperformed the control group with notable enhancements in blood flow rates,vessel diameter,and depth from the skin of drainage vessels(P<0.01).Seven days following the procedure,the observation group manifested significantly diminished limb swelling and an overall reduced complication rate in contrast to the control group(P<0.05).The evaluation of infection,thrombosis,embolism,arterial aneurysm stenosis,and incision bleeding showed no notable differences between the two groups(P>0.05).By the 4-wk postoperative juncture,complications between the observation and control groups were statistically indistinguishable(P>0.05).CONCLUSION Stage-specific limb function exercises,under the guidance of a self-management education model,amplify the capacity of AVF patients to discern and prevent symptoms.Additionally,they expedite AVF maturation and miti-gate postoperative limb edema,underscoring their efficacy as a valuable method for the care and upkeep of AVF in hemodialysis patients. 展开更多
关键词 SELF-MANAGEMENT Education model Stage-specific Limb function exercises Arteriovenous fistula maturation status
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Lamellar Bodies Count (LBC) as a Predictor of Fetal Lung Maturity in Preterm Premature Rupture of Membranes Compared to Neonatal Assessment
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作者 Malames Mahmoud Faisal Noha Hamed Rabei +1 位作者 Hoda Ezz El-Arab Abd El-Wahab Abeer Hosny El-Zakkary 《Open Journal of Obstetrics and Gynecology》 2023年第6期1047-1057,共11页
Background: Respiratory distress syndrome (RDS) is a major cause of neonatal morbidity and mortality, affecting approximately 1% of all live births and 10% of all preterm infants. Lamellar bodies represent a storage f... Background: Respiratory distress syndrome (RDS) is a major cause of neonatal morbidity and mortality, affecting approximately 1% of all live births and 10% of all preterm infants. Lamellar bodies represent a storage form of pulmonary surfactant within Type II pneumocytes, secretion of which increases with advancing gestational age, thus enabling prediction of the degree of FLM. Preterm premature rupture of membranes (PPROM) complicates approximately 1/3 of all preterm births. Birth within 1 week is the most likely outcome for any patient with PPROM in the absence of adjunctive treatments. Respiratory distress has been reported to be the most common complication of preterm birth. Sepsis, intraventricular haemorrhage, and necrotizing enterocolitis also are associated with prematurity, but these are less common near to term. Objective: To assess the efficacy of the amniotic fluid lamellar body counting from a vaginal pool in predicting fetal lung maturity in women with preterm premature rupture of membranes. Methods: This study was conducted at Ain Shams University Maternity Hospital in the emergency ward from January 2019 to September 2019. It included 106 women with singleton pregnancies, gestational age from 28 - 36 weeks with preterm premature rupture of membranes. This study is designed to assess the efficacy of the amniotic fluid lamellar body counting (LBC) from a vaginal pool in predicting fetal lung maturity in women with preterm premature rupture of membranes. Results: The current study revealed a highly significant increase in the lamellar body count in cases giving birth to neonates without RDS compared to that cases giving birth to neonates with RDS. Also, no statistically significant difference between LBC and age, parity and number of previous miscarriages in the mother was found. Gestational age at delivery was significantly lower among cases with respiratory distress. Steroid administration was significantly less frequent among cases with respiratory distress. However, lamellar bodies had high diagnostic performance in the prediction of respiratory distress. Conclusion: Lamellar body count (LBC) is an effective, safe, easy, and cost-effective method to assess fetal lung maturity (FLM). It does not need a highly equipped laboratory or specially trained personnel, it just needs the conventional blood count analyzer. Measurement of LBC is now replacing the conventional Lecithin/Sphyngomyelin L/S ratio. LBC cut-off value of ≤42.5 × 10<sup>3</sup>/μL can be used safely to decide fetal lung maturity with sensitivity of 95.7% and specificity of 97.6%. 展开更多
关键词 Fetal Lung maturity Lamellar Bodies Count Preterm Premature Rupture of Membranes Respiratory Distress Syndrome
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Studies of the Variability of Primary Metabolites in the Epicarp, Mesocarp and Seed of Dacryodes edulis Fruit at Taste Maturity
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作者 Etou Ossibi Grace Jokael Mbon Nguékou Chrichina +2 位作者 Ongouya Mouekouba Dalcantara Liana Mpika Joseph Attibayeba   《Advances in Biological Chemistry》 CAS 2024年第2期64-72,共9页
The fruits of Dacryodes edulis are rich in biologically active substances, which makes them of great interest in terms of validation. In this study, we targeted the primary metabolites in the epicarp, the mesocarp and... The fruits of Dacryodes edulis are rich in biologically active substances, which makes them of great interest in terms of validation. In this study, we targeted the primary metabolites in the epicarp, the mesocarp and the seed of the fruit of Dacryodes edulis at taste maturity, which were selected for their nutritional quality and its appreciation throughout the Gulf of Guinea area, which is very popular because of its large size, texture and special taste. The evaluation of total carbohydrates, total lipids and soluble proteins in the epicarp, mesocarp and seed of the fruit at taste maturity was made from spectrophotometer measurements. The overall analysis of the results of the present study shows that total carbohydrates, total lipids and proteins accumulate more in the seed with respectively 251.33 ± 1.15 mg/g DM;9.92 ± 0.201 mg/g DM and 55.075 ± 0.024 mg/g DM. Likewise, the results indicate low concentrations of total carbohydrates and total lipids in the epicarp with respectively 245 ± 1 mg/g DM and 4.77 ± 0.047 mg/g DM, on the other hand, it is the mesocarp which presents the lowest content of soluble proteins: 28.075 ± 3.231 mg/g DM. This variation could be linked to the nature of the compartment, more particularly to the storage location. This comparative study could lead to the valorization of the seed of the fruit of Dacryodes edulis for its richness in metabolites and arouse significant interest in nutrition. 展开更多
关键词 Dacryodes edulis FRUIT Taste maturity Epicarp MESOCARP SEED
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Endogenous biosynthesis of docosahexaenoic acid(DHA)regulates fish oocyte maturation by promoting pregnenolone production
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作者 Yi Li Xuehui Li +6 位作者 Ding Ye Ru Zhang Chengjie Liu Mudan He Houpeng Wang Wei Hu Yonghua Sun 《Zoological Research》 SCIE CSCD 2024年第1期176-188,共13页
Omega-3 polyunsaturated fatty acids(n-3 PUFAs),particularly docosahexaenoic acid(22:6n-3,DHA),play crucial roles in the reproductive health of vertebrates,including humans.Nevertheless,the underlying mechanism related... Omega-3 polyunsaturated fatty acids(n-3 PUFAs),particularly docosahexaenoic acid(22:6n-3,DHA),play crucial roles in the reproductive health of vertebrates,including humans.Nevertheless,the underlying mechanism related to this phenomenon remains largely unknown.In this study,we employed two zebrafish genetic models,i.e.,elovl2^(-/-)mutant as an endogenous DHAdeficient model and fat1(omega-3 desaturase encoding gene)transgenic zebrafish as an endogenous DHA-rich model,to investigate the effects of DHA on oocyte maturation and quality.Results show that the elovl2^(-/-)mutants had much lower fecundity and poorer oocyte quality than the wild-type controls,while the fat1 zebrafish had higher fecundity and better oocyte quality than wildtype controls.DHA deficiency in elovl2^(-/-)embryos led to defects in egg activation,poor microtubule stability,and reduced pregnenolone levels.Further study revealed that DHA promoted pregnenolone synthesis by enhancing transcription of cyp11a1,which encodes the cholesterol side-chain cleavage enzyme,thereby stabilizing microtubule assembly during oogenesis.In turn,the hypothalamic-pituitary-gonadal axis was enhanced by DHA.In conclusion,using two unique genetic models,our findings demonstrate that endogenously synthesized DHA promotes oocyte maturation and quality by promoting pregnenolone production via transcriptional regulation of cyp11a1. 展开更多
关键词 Docosahexaenoic acid Oocyte maturation Oocyte quality PREGNENOLONE MICROTUBULE
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A Proposed Approach for Measuring Maturity Level of Software Delivery
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作者 Osama Hassan Mansour Atef Raslan Nagy Ramadan 《Journal of Software Engineering and Applications》 2024年第5期228-245,共18页
Software delivery is vital for modern organizations, driving innovation and competitiveness. Measuring an organization’s maturity in software delivery is crucial for efficiency and quality. The Capability Maturity Mo... Software delivery is vital for modern organizations, driving innovation and competitiveness. Measuring an organization’s maturity in software delivery is crucial for efficiency and quality. The Capability Maturity Model (CMM) framework provides a roadmap for improvement but assessing an organization’s CMM Level is challenging. This paper offers a quantitative approach tailored to the CMM framework, using Goal-Question-Metric (GQM) frame-works for each key process area (KPA). These frameworks include metrics and questions to compute maturity scores effectively. The study also refines practices into questions for a thorough assessment. The result is an Analysis Matrix that calculates weighted scores and an overall maturity score. This approach helps organizations assess and enhance their software delivery processes systematically, aiming for improved practices and growth. 展开更多
关键词 CMM CMMI Software Delivery maturITY KPAs GQM PRACTICES
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Effect of thermal maturation and organic matter content on oil shale fracturing
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作者 Fatemeh Saberi Mahboubeh Hosseini‑Barzi 《International Journal of Coal Science & Technology》 EI CAS CSCD 2024年第2期16-34,共19页
The Pabdeh Formation represents organic matter enrichment in some oil fields,which can be considered a source rock.This study is based on the Rock–Eval,Iatroscan,and electron microscopy imaging results before and aft... The Pabdeh Formation represents organic matter enrichment in some oil fields,which can be considered a source rock.This study is based on the Rock–Eval,Iatroscan,and electron microscopy imaging results before and after heating the samples.We discovered this immature shale that undergoes burial and diagenesis,in which organic matter is converted into hydro-carbons.Primary migration is the process that transports hydrocarbons in the source rock.We investigated this phenomenon by developing a model that simulates hydrocarbon generation and fluid pressure during kerogen-to-hydrocarbon conversion.Microfractures initially formed at the tip/edge of kerogen and were filled with hydrocarbons,but as catagenesis progressed,the pressure caused by the volume increase of kerogen decreased due to hydrocarbon release.The transformation of solid kerogen into low-density bitumen/oil increased the pressure,leading to the development of damage zones in the source rock.The Pabdeh Formation’s small porethroats hindered effective expulsion,causing an increase in pore fluid pressure inside the initial microfractures.The stress accumulated due to hydrocarbon production,reaching the rock’s fracture strength,further contributed to damage zone development.During the expansion process,microfractures preferentially grew in low-strength pathways such as lithology changes,laminae boundaries,and pre-existing microfractures.When the porous pressure created by each kerogen overlapped,individual microfractures interconnected,forming a network of microfractures within the source rock.This research sheds light on the complex interplay between temperature,hydrocarbon generation,and the development of expulsion fractures in the Pabdeh Formation,providing valuable insights for understanding and optimizing hydrocarbon extraction in similar geological settings. 展开更多
关键词 Oil shale MICROFRACTURE Hydrocarbon generation Organic matter Thermal maturation Primary migration
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Correlation analysis of interstitial maturity and prognosis of colorectal cancer:Meta-analysis
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作者 Zhen-Jun Liu Xu-Wen Zhang +1 位作者 Qi-Qi Liu Shao-Zhao Wang 《World Journal of Gastrointestinal Oncology》 SCIE 2024年第6期2816-2825,共10页
BACKGROUND To investigate the relationship between interstitial maturity and prognosis of colorectal cancer.AIM To examine the correlation between interstitial maturity and the prognosis of colorectal cancer.METHODS T... BACKGROUND To investigate the relationship between interstitial maturity and prognosis of colorectal cancer.AIM To examine the correlation between interstitial maturity and the prognosis of colorectal cancer.METHODS The paper database PubMed,EMBASE,Cochranelibrary,Springerlink,CNKI,and Wanfang database were searched until December 2023."tumor stroma maturity""desmoplastic stroma reaction""desmoplastic reaction""stroma reaction""degree of stroma reaction""stroma classification""stroma density""colorectal cancer""colon cancer""rectal cancer""prognosis"were searched for the search terms.Two system assessors independently screened the literature quality according to the inclusion exclusion criteria,Quality evaluation and data extraction were performed for the included literatures,and meta-analysis was performed for randomized control trials included at using Review Manager 5.2 software.RESULTS Finally,data of 9849 patients with colorectal cancer from 19 cosets in 15 literatures were included,including 4339 patients with mature type(control group),3048 patients with intermediate type(intermediate group)and 2456 patients with immature type(immature group).The results of meta-analysis showed:Relapse-free survival[hazard ratio(HR)=2.66,95%confidence interval(CI):2.30-3.08;P<0.00001],disease-free survival(HR=3.68,95%CI:2.33-5.81;P<0.00001)and overall survival(HR=1.70,95%CI:1.53-1.87;P<0.00001)were significantly lower than those in mature group(control group);relapse-free survival(HR=1.36,95%CI:1.17-1.59;P<0.0001)and disease-free survival rate(HR=1.85,95%CI:1.53-2.24;P<0.0001)was significantly lower than the mature group(control group).CONCLUSION There is the correlation between tumor interstitial maturity and survival prognosis of colorectal cancer,and different degrees of tumor interstitial maturity have a certain impact on the quality of life of colorectal cancer patients. 展开更多
关键词 Colorectal tumor Survival prognosis Tumor interstitial maturity Systematic review META-ANALYSIS
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Debugging Strategies for Parents' Educational Anxiety in the Transition from Kindergarten to Primary School--Based on the Theory of Mature Forces
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作者 WenJie Yu 《教育技术与创新》 2023年第4期59-70,共12页
Due to the rapid growth of the economy and society,the burden of social security has been increasingly escalating,resulting in a continuous rise in parenting stress among certain groups of parents.On one hand,these pa... Due to the rapid growth of the economy and society,the burden of social security has been increasingly escalating,resulting in a continuous rise in parenting stress among certain groups of parents.On one hand,these parents aspire for their young children to grow through play;on the other hand,they are acutely aware that early childhood education constitutes the foundational stage of basic education and serves as the commencement phase of lifelong learning.Thus,they recognize the importance of emphasizing early childhood education.Therefore,preschool education has been given important significance.Parents place particular emphasis on education during this stage.They actively leverage their educational,financial,and social advantages to create favorable educational resources and opportunities for their young children.However,to a certain extent,this exacerbates educational anxiety among a segment of parents.Despite the implementation of various governmental measures,it appears that these initiatives have not fundamentally addressed parental educational anxiety at its root.This study employs interview techniques and literature review methodologies to gain a profound understanding of parental educational anxiety during the transition from early childhood to primary education.By integrating Gesell's Maturation Theory,this research aims to scientifically explicate the developmental patterns of children.Consequently,it endeavors to enable non-professional parents to adopt a"professional"perspective in their approach towards early childhood education. 展开更多
关键词 early childhood connection parental education anxiety mature force theory suggestion strategy
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靶向BCMA CAR-T治疗RRMM的局限性及优化策略
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作者 刘汇洪 王立茹 《中国医学创新》 CAS 2024年第15期183-188,共6页
嵌合抗原受体T细胞(CAR-T)免疫疗法快速发展,为多发性骨髓瘤(MM)治疗带来新的曙光,尤其B细胞成熟抗原(BCMA)作为迄今为止最成功的靶标,靶向BCMA CAR-T疗法可以使MM的症状获得持久且深度的缓解,在复发难治性多发性骨髓瘤(RRMM)治疗中取... 嵌合抗原受体T细胞(CAR-T)免疫疗法快速发展,为多发性骨髓瘤(MM)治疗带来新的曙光,尤其B细胞成熟抗原(BCMA)作为迄今为止最成功的靶标,靶向BCMA CAR-T疗法可以使MM的症状获得持久且深度的缓解,在复发难治性多发性骨髓瘤(RRMM)治疗中取得突破性进展。但由于抗原逃逸、CAR-T衰竭等因素,多数患者仍会进展或复发,靶向BCMA CAR-T治疗后复发或难治患者的后续治疗缺乏标准方案。同时,复杂、昂贵且耗时的个性化CAR-T制造流程也限制其临床疗效的发挥。针对此现状,该综述总结了靶向BCMA CAR-T治疗的局限性及其机制,同时结合MM治疗领域新进展,提出改善进展或RRMM患者结局的潜在优化治疗策略。 展开更多
关键词 B细胞成熟抗原 Car-T疗法 复发难治性多发性骨髓瘤 耐药性
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靶向BCMA的CAR-γδT细胞抗多发性骨髓瘤的疗效
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作者 李盈辉 许依 +2 位作者 张建民 陈慧 何维 《基础医学与临床》 CAS 2024年第6期763-771,共9页
目的构建靶向B细胞成熟抗原(BCMA)的嵌合抗原受体γδT细胞(BCMA CAR-γδT),在体外初步评价其抗多发性骨髓瘤的疗效。方法构建包含BCMA单链抗体序列的三代CAR分子的慢病毒载体,瞬时转染293T细胞,荧光显微镜和Western blot验证外源基因... 目的构建靶向B细胞成熟抗原(BCMA)的嵌合抗原受体γδT细胞(BCMA CAR-γδT),在体外初步评价其抗多发性骨髓瘤的疗效。方法构建包含BCMA单链抗体序列的三代CAR分子的慢病毒载体,瞬时转染293T细胞,荧光显微镜和Western blot验证外源基因的表达情况;包装慢病毒,用流式细胞术测定病毒滴度;分别感染人外周血αβT细胞和γδT细胞,检测感染效率;LDH法检测BCMA CAR-γδT细胞对人多发性骨髓瘤细胞系的体外细胞毒活性,并用Incucyte实时细胞分析系统比较BCMA CAR-γδT细胞与BCMA CAR-T细胞的细胞毒活性差异。结果将BCMA-CAR慢病毒载体转入293T细胞24 h后,荧光显微镜观察到外源ZsGreen的表达;Western blot检测CD3ζ证明BCMA-CAR能顺利表达。慢病毒包装后,收集感染上清浓缩,测得病毒滴度为2.23×10^(8)Tu/mL。用MOI=5感染人外周血αβT细胞与γδT细胞,流式检测结果显示αβT细胞感染效率为59.18%±2.56%,γδT细胞感染效率为48.15%±9.86%。LDH杀伤实验结果显示,CAR-γδT细胞对BCMA高表达的人骨髓瘤细胞系MM1.S、H929和过表达BCMA的K562细胞的细胞毒活性较空载体对照γδT细胞显著增强(P<0.001),而对BCMA表达阴性的细胞系无差异;活细胞时程分析系统结果显示,BCMA CAR-γδT细胞与BCMA CAR-αβT细胞在体外对H929的细胞毒活性均显著优于其空载体对照细胞,且均对BCMA表达阴性的细胞系的细胞毒活性无差异。结论BCMA CAR-γδT细胞在体外显示出明显增强的靶向BCMA的细胞毒活性,有望作为新型同种异体γδT细胞产品,用于多发性骨髓瘤的细胞过继免疫治疗。 展开更多
关键词 B细胞成熟抗原 多发性骨髓瘤 嵌合抗原受体 ΓΔT细胞
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基于BCMA突变体构建BCMA CAR-T细胞体外杀伤功能评价模型
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作者 张小雪 华静涵 +3 位作者 侯睿 刘丹 施明 曹江 《中国肿瘤生物治疗杂志》 CAS CSCD 北大核心 2024年第5期493-500,共8页
目的:为解决野生型B细胞成熟抗原(BCMA)被γ分泌酶切割导致表达不稳定的问题,构建抵抗γ分泌酶切割的BCMA突变体并构建靶细胞,用于评价BCMA CAR-T细胞的杀伤功能。方法:将野生型BCMA的穿膜域替换为人CD8α穿膜域,构建抵抗γ分泌酶切割的... 目的:为解决野生型B细胞成熟抗原(BCMA)被γ分泌酶切割导致表达不稳定的问题,构建抵抗γ分泌酶切割的BCMA突变体并构建靶细胞,用于评价BCMA CAR-T细胞的杀伤功能。方法:将野生型BCMA的穿膜域替换为人CD8α穿膜域,构建抵抗γ分泌酶切割的BCMA突变体(BCMA-CD8αTM),构建过表达该突变体的U266(U266^(BCMA Mut))、K562(K562^(BCMA Mut))、SKOV3(SKOV3^(BCMA Mut))和CHO(CHO^(BCMA Mut))细胞;构建装载NFAT-EGFP报告基因的BCMA CAR Jurkat细胞(BCMA-CAR-Jurkat-Reporter)与U266^(BCMA Mut)细胞共培养,采用FCM检测该细胞中EGFP表达水平以指示NFAT激活水平,荧光素酶法检测BCMA CAR-T细胞对Luciferase标记的K562^(BCMA Mut)细胞的杀伤作用,实时无标记动态细胞分析技术(RTCA)检测BCMA CAR-T细胞对SKOV3^(BCMA Mut)和CHO^(BCMA Mut)细胞的杀伤作用。结果:应用γ分泌酶抑制剂LY411575抑制γ分泌酶活性,显著增强野生型U266细胞表面BCMA表达水平,平均荧光强度上调10倍以上;但撤除抑制剂后BCMA表达水平逐渐降低(P<0.01);BCMA-CD8αTM突变体可抵抗γ分泌酶的切割作用,在U266细胞表面稳定表达(P>0.05);U266细胞及过表达BCMA-CD8αTM的U266细胞与BCMA-CAR-Jurkat-Reporter细胞共培养后都可激活Reporter系统、增强EGFP表达,但该效应在BCMA-CD8αTM过表达的U266细胞中更显著(P<0.01);BCMA-CD8αTM在BCMA表达阴性的K562、SKOV3和CHO 3种靶细胞中成功过表达,且在LY411575处理下该突变体的表达水平仅有小幅度升高;荧光素酶法检测结果显示,不同效靶比下,BCMA CAR-T细胞均可特异、高效杀伤过表达BCMA-CD8αTM的K562细胞;RTCA结果显示,不同效靶比下,BCMA CAR-T细胞均可有效识别、杀伤过表达BCMACD8αTM的SKOV3和CHO细胞,但同等效靶比下的Mock-T细胞无此效应。结论:本实验构建的BCMA-CD8αTM突变体能够抵抗γ分泌酶的切割,在多种靶细胞表面稳定表达,为评价BCMA CAR-T细胞体外杀伤的有效性和特异性提供多种检测手段。 展开更多
关键词 B细胞成熟抗原 Γ分泌酶 CD8α穿膜域 Car-T细胞 杀伤功能评价
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Organic matter and concentrated nitrogen removal by shortcut nitrification and denitrification from mature municipal landfill leachate 被引量:16
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作者 ZHANG Shu-jun PENG Yong-zhen WANG Shu-ying ZHENG Shu-wen GUO Jin 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2007年第6期647-651,共5页
An UASB+Anoxic/Oxic (A/O) system was introduced to treat a mature landfill leachate with low carbon-to-nitrogen ratio and high ammonia concentration. To make the best use of the biodegradable COD in the leaehate, t... An UASB+Anoxic/Oxic (A/O) system was introduced to treat a mature landfill leachate with low carbon-to-nitrogen ratio and high ammonia concentration. To make the best use of the biodegradable COD in the leaehate, the denitrifieation of NOx^--N in the reeireulation effluent from the elarifier was carried out in the UASB. The results showed that most biodegradable organic matters were removed by the denitrifieation in the UASB. The NH4^+-N loading rate (ALR) of A/O reactor and operational temperature was 0.28- 0.60 kg NH4^+-N/(m^3-d) and 17-29℃ during experimental period, respectively. The short-cut nitrification with nitrite accumulation efficiency of 90%-99% was stabilized during the whole experiment. The NH4^+-N removal efficiency varied between 90% and 100%. When ALR was less than 0.45 kg NH4^+-N/(m^3.d), the NH4^+-N removal efficiency was more than 98%. With the influent NH4^+-N of 1200-1800 mg/L, the effluent NH4^+-N was less than 15 mg/L. The shortcut nitrification and denitrifieation can save 40% carbon source, with a highly efficient denitrifieation taking place in the UASB. When the ratio of the feed COD to feed NH4^+-N was only 2-3, the total inorganic nitrogen (TIN) removal efficiency attained 67%-80%. Besides, the sludge samples from A/O reactor were analyzed using FISH. The FISH analysis revealed that ammonia oxidation bacteria (AOB) accounted for 4% of the total eubaeterial population, whereas nitrite oxidation bacteria (NOB) accounted only for 0.2% of the total eubaeterial population. 展开更多
关键词 mature landfill leachate UASB+A/O shortcut nitrification carbon source
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Establishment of a Highly Efficient Regeneration System for the Mature Embryo Culture of Wheat 被引量:19
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作者 YIN Gui-xiang WANG Yan-li +4 位作者 SHE Mao-yun DU Li-pu XU Hui-jun MA Jing-xiu YE Xing-guo 《Agricultural Sciences in China》 CAS CSCD 2011年第1期9-17,共9页
Establishment of a highly efficient regeneration system for the mature embryo of wheat will provide a convenient tool for wheat tissue culture and transformation, thereby facilitating the transformation of foreign gen... Establishment of a highly efficient regeneration system for the mature embryo of wheat will provide a convenient tool for wheat tissue culture and transformation, thereby facilitating the transformation of foreign genes into wheat. By using the mature embryos derived from 20 different wheat lines including Shi 4185, Yumai 66, Lunxuan 987, CB037, Yangmai 6, Xinchun 9, Bobwhite, Han 6172, Zheng 9023, Jimai 20, Ningchun 4, and Jing 411, the effects of some factors including inoculation methods, initiating culture media, organic additives, antioxidants, and auxins on the regeneration from the explants were evaluated. The results indicated that the scraping embryo culture was better than the whole embryo culture, the Aa medium was better than the SD2 medium and dicamba was better than 2,4-D in increasing the regeneration frequency. An Adi medium was established in this study by adding silver nitrate, cysteine, ascorbic acid, dicamba, glutamine into the Aa medium at the concentration of 4,40, 100, 2, and 5 mg L^-1, respectively. By using the Adi medium and the scraping technique, the regeneration frequencies of the mature embryos of CB037, Lunxuan 987, Hart 6172, Yangmai 6, Bobwhite, Zheng 9023, Shi 4 185, and Jimai 20 became 85.6, 60,1, 46.0, 42.1,42.0, 34.0, 33.0, and 32.0%, respectively, which were about 5-8 times higher than that obtained from the conventional culture mediums and techniques. This novel regeneration system could be helpful in wheat transformation. 展开更多
关键词 WHEAT mature embryo culture scraping treatment Adi medium highly efficient regeneration
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Dicamba and Sugar Effects on Callus Induction and Plant Regeneration from Mature Embryo Culture of Wheat 被引量:11
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作者 REN Jiang-ping,WANG Xin-guo and YIN Jun National Engineering Research Center for Wheat/Henan Agricultural University,Zhengzhou 450002,P.R.China 《Agricultural Sciences in China》 CSCD 2010年第1期31-37,共7页
To establish a highly efficient plant regeneration system for wheat genetic transformation, the effects of three different concentrations of dicamba and two different sugar types on callus induction and plant regenera... To establish a highly efficient plant regeneration system for wheat genetic transformation, the effects of three different concentrations of dicamba and two different sugar types on callus induction and plant regeneration from mature embryo cultures were evaluated. Callus induction and plant regeneration were obtained from mature embryos of two commercial cultivars Zhoumai 18 and Yumai 34 (Triticum aestivum L.) cultured on L3 basal medium. The results showed that the efficiency of mature embryo culture was significantly influenced by the genotypes, sugar types and dicamba concentrations. 4 mg L^-1 dicamba proved the best effective for inducing embryogenic callus and also gave the highest proportion of plants regenerated across the two cultivars. Substitution of maltose by sucrose significantly improved the plant regeneration efficiency in both cultivars. There was a significant interaction between genotype-by-sugar types, and sugar types-bydicamba concentrations. Overall, Zhoumai 18 gave the highest frequency of plant regeneration (82.65%) when dicamba concentration was 4.0 mg L^-1 and with sucrose in initial callus induction. These results will facilitate genetic transformation work with elite wheat. 展开更多
关键词 WHEAT mature embryo tissue culture plant regeneration
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Species diversity,population structure and regeneration of woody species in fallows and mature stands of tropical woodlands of southeast Angola 被引量:5
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作者 Francisco M.P.Goncalves Rasmus Revermann +2 位作者 Manuel J.Cachissapa Amandio L.Gomes Marcos P.M.Aidar 《Journal of Forestry Research》 SCIE CAS CSCD 2018年第6期1569-1579,共11页
This study in the dry tropical woodlands of SE Angola in Cuando-Cubango Province assessed the diversity and composition of woody species in fallows compared to those in mature woodlands. We assessed the population str... This study in the dry tropical woodlands of SE Angola in Cuando-Cubango Province assessed the diversity and composition of woody species in fallows compared to those in mature woodlands. We assessed the population structure of the most harvested woody species by calculating size class distribution and evaluated their regeneration potential based on the density of saplings. The vegetation was surveyed in 20 plots of 20 × 50 m(1000 m2). In each plot, we measured the diameter at breast height(DBH) of all woody species with DBH ≥ 5 cm. The saplings were counted, identified and recorded; 718 individuals, corresponding to 34 species in 32 genera and 15 families were recorded. The size class distribution of target woody species showed three different patterns in fallows and mature woodlands. In general,most of the smaller diameter classes had more individuals than the larger ones did, showing that the regeneration may take place. However, in some diameter classes, the absence of larger stems indicated selective logging of larger trees. Few saplings were recorded in the fallows or mature woodlands; fire frequency and intensity is probably the main obstacle for seed germination and seedling survival rates in the studied area. 展开更多
关键词 Baikiaea woodlands Fallows mature woodlands REGENERATION Woody species
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Study on Plant Regeneration of Wheat Mature Embryos Under Endosperm-Supported Culture 被引量:11
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作者 CHEN Jun-ying YUE Run-qing XU Hai-xian CHEN Xin-jian 《Agricultural Sciences in China》 CAS CSCD 2006年第8期572-578,共7页
To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars (Triticum aestivum L. cv.) were cultured with or without endosperm to ... To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars (Triticum aestivum L. cv.) were cultured with or without endosperm to test their efficiency of callus induction and plant regeneration. When embryos were cultured together with endosperm (endosperm-supported culture, ES), the percentage of callus induction was significantly lower than that when embryos were cultured in the absence of endosperm (non-endosperm-supported culture, NES). This pattern was evident in most genotypes, regardless of whether 2 or 8 mg L^-1 2,4-D was added in the NES culture. However, in ES culture, more induced calli were differentiated into distinct green spots and they further developed into plantlets. Thus, more plants were regenerated in ES culture than in the NES treatment. Most of the eight tested genotypes showed a significant difference in callus induction rate and plantlet regeneration in both ES and NES cultures. In addition, the enzymatic activity of oxalate oxidase in the callus of ES culture condition was obviously higher than that in the callus of NES culture condition, suggesting that the activity of oxalate oxidase may be a parameter for selection of calli with potential for plantlet regeneration. These results indicate that wheat mature embryos are valuable explants for highly efficient callus induction and plant regeneration, if proper treatment and medium are used. 展开更多
关键词 mature wheat embryo tissue culture endosperm-supported plant regeneration oxalate oxidase activity
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Evolution and organic geochemical significance of bicyclic sesquiterpanes in pyrolysis simulation experiments on immature organic-rich mudstone 被引量:3
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作者 Gang Yan Yao-Hui Xu +2 位作者 Yan Liu Peng-Hai Tang Wei-Bin Liu 《Petroleum Science》 SCIE CAS CSCD 2019年第3期502-512,共11页
Sesquiterpanes are ubiquitous components of crude oils and ancient sediments.Liquid saturated hydrocarbons from simulated pyrolysis experiments on immature organic-rich mudstone collected from the Lower Cretaceous Hes... Sesquiterpanes are ubiquitous components of crude oils and ancient sediments.Liquid saturated hydrocarbons from simulated pyrolysis experiments on immature organic-rich mudstone collected from the Lower Cretaceous Hesigewula Sag were analyzed by gas chromatography-mass spectrometry(GC-MS).C14 bicyclic sesquiterpanes,namely,8β(H)-drimane,8β(H)-homodrimane,and 8 a(H)-homodrimane were detected and identified on basis of their diagnostic fragment ions(m/z123,179,193,and 207),and previously published mass spectra data,and these bicyclic sesquiterpanes presented relatively regular characteristics in their thermal evolution.The ratios 8β(H)-drimane/8β(H)-homodrimane,8β(H)-homodrimane/8 a(H)-homodrimane,and 8β(H)-drimane/8 a(H)-homodrimane all show a clear upward trend with increasing temperature below the temperature turning point.Thus,all these ratios can be used as evolution indexes of source rocks in the immature-lowmaturity stage.However,the last two ratios may be more suitable than the first ratio as valid parameters for measuring the extent of thermal evolution of organic matter in the immature-low-maturity stage because their change amplitude with increasing temperature is more obvious. 展开更多
关键词 Immature-low-maturity stage Simulated PYROLYSIS experiment BICYCLIC sesquiterpanes Thermal EVOLUTION maturITY indicators
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The Comparison in Tissue Culture Ability of Mature Embryo in Different Cultivars of Rice 被引量:6
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作者 YAN Li-na LI Xia WU Dan 《Agricultural Sciences in China》 CSCD 2010年第6期840-846,共7页
In order to study the regeneration technology of mature embryos in different rice varieties,nine japonica,nine indica and eleven hybrid rice varieties of two line or three line or superiority combinations were selecte... In order to study the regeneration technology of mature embryos in different rice varieties,nine japonica,nine indica and eleven hybrid rice varieties of two line or three line or superiority combinations were selected as explants to study the callus induction,differentiation and regeneration rates on different media.The higher callus induction (61.7-89.2%) was observed in japonica rice,when cytokinin was added at lower concentration (0.3 mg L-1 6-BA) in M8 basal medium,supplemented with 30 g L-1 sucrose,8 g L-1 agar and 2 mg L-1 2,4-D.Further,the addition of two cytokinins (2 mg L-1 6-BA,0.5 mg L-1 KT) and 1 mg L-1 NAA in the M8 basal supplemented medium resulted in 9.1-100% of the callus induction in indica rice.The percent callus induction in hybrid rice varieties was 40-86.3% when addition of 1 mg L-1 6-BA and 1 mg L-1 KT was added,and the cytokinins was required by the japonica and indica rice varieties in the M8 basal supplemented medium.It was observed that when the 0.5 mg L-1 2,4-D and 1 mg L-1 6-BA were added in japonica rice,and 0.2 mg L-1 2,4-D and 0.5 mg L-1 6-BA were added in indica and hybrid rice in the MS different media,the regeneration rates were 9.2-59.5%,3.6-87.5% and 17.2-43.2% for japonica,indica and hybrid rice,respectively.Thus,the regeneration technology with higher output is established in the mature embryos of similar rice varieties. 展开更多
关键词 JAPONICA INDICA hybrid rice mature embryos HORMONE tissue culture ability
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