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一种新的导致遗传性凝血因子Ⅶ缺陷症的基因突变分析
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作者 王莹宇 张永根 陈文柏 《中国实验血液学杂志》 CAS CSCD 北大核心 2024年第3期857-861,共5页
目的:对一例遗传性凝血因子Ⅶ(FⅦ)缺陷症的患者及其家系进行基因分析,探讨其分子发病机制。方法:使用Sysmex-CS5100全自动血凝分析仪检测先证者及其家系成员(共3代8人)的凝血酶原时间(PT)、活化部分凝血活酶时间、凝血酶时间、D-二聚... 目的:对一例遗传性凝血因子Ⅶ(FⅦ)缺陷症的患者及其家系进行基因分析,探讨其分子发病机制。方法:使用Sysmex-CS5100全自动血凝分析仪检测先证者及其家系成员(共3代8人)的凝血酶原时间(PT)、活化部分凝血活酶时间、凝血酶时间、D-二聚体、纤维蛋白降解产物以及血浆FⅦ的活性(FⅦ:C)水平;PCR法扩增先证者凝血因子Ⅶ基因(F7)所有外显子和侧翼序列,PCR产物纯化后测序,发现突变位点则反向测序给予证实;使用Clustal W软件对突变位点进行保守性分析;应用Mutation Taster和Poly Phen-2在线生物学软件评估突变氨基酸对FⅦ蛋白结构与功能的危害性;运用Swiss软件对突变建模分析。结果:凝血常规检查结果显示,先证者PT单独性延长至42.5 s;FⅦ:C明显降低,仅为2%;同样先证者外婆、母亲和妹妹的FⅦ:C都有轻度降低,分别为49%、51%和42%;父亲各指标均在正常参考范围。基因分析结果显示,先证者F7基因第6号外显子c DNA的646位发生G>A杂合突变,导致FⅦ催化区的156位甘氨酸被替换为丝氨酸(p.Gly156Ser)。F7其他外显子和侧翼序列的测序结果均正常。其外婆、母亲和妹妹均携带c.646G>A杂合突变,父亲为正常野生型。模型构建显示p.Gly156Ser突变使该位点氨基酸极性发生改变并出现侧链,从而使蛋白的不稳定性增加,可能影响所在结构域的催化活性。同时,Mutation Taster和Poly Phen-2两个在线生物信息学软件也高分预测该突变具有致病性。结论:该遗传性凝血因子Ⅶ缺陷症患者FⅦ蛋白p.Gly156Ser错义突变与血浆FⅦ:C水平降低有关。 展开更多
关键词 遗传性凝血因子Ⅶ缺陷症 PCR 基因突变 模型分析
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Effect of Soybean Peptides on in vitro Ruminal Fermentation and Microbial Population 被引量:2
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作者 Wang Li-jun Liu Yan +4 位作者 Liu Shuai Sun Kai-jing Wang Xin-ying zhang Guang-ning zhang yong-gen 《Journal of Northeast Agricultural University(English Edition)》 CAS 2017年第4期40-52,共13页
This study was conducted to investigate the effect of Soybean Peptides(SPs) on ruminal fermentation and microbial diversity in vitro. Four levels of SPs supplements(0, 0.25%, 0.50% and 0.75% dry matter basis) were tes... This study was conducted to investigate the effect of Soybean Peptides(SPs) on ruminal fermentation and microbial diversity in vitro. Four levels of SPs supplements(0, 0.25%, 0.50% and 0.75% dry matter basis) were tested. p H, NH3-N, Neutral Detergent Fiber(NDF), Acid Detergent Fiber(ADF), Dry Matter Digestibility(DMD) and the Total Volatile Fatty Acid(TVFA) were measured at 6, 24, 48 h of in vitro mixed incubation the fluids. Microbial populations were determined at 24 h and Microbial Proteins(MCP) were determined at 24 and 48 h. The gas production after 48 h in vitro incubation presented linear growth(p<0.05) and the highest content was the level of 0.5% SPs(dry matter basis). NH3-N concentration reached the highest concentration with 0.75%(dry matter basis) at 48 h. p H linearly increased(linear, p<0.05) from 6 to 48 h. The digestion of DMD increased with increasing doses of SPs at 24 h. NDF and ADF linearly(p<0.01) decreased by adding SPs. The concentration of TVFA linearly increased(p<0.05) at 6, 24 and 48 h, and reached the highest concentration at the level of 0.75%(dry matter basis). SPs decreased(p<0.05) the molar proportion of acetate and propionate, respectively at 24 and 48 h. Acetat to Propionate ratio(A/P; linear, p<0.05) increased at 48 h, and reached the greatest value at the level of 0.75%(dry matter basis) at 48 h. The populations of rumen Ruminococcus flavefaciens(R. flavefaciens), Ruminococcusalbus(R. albus), Fibrobacter succinogenes(F. succinogenes), Butyrivibrio fibrisolvens(B. fibrisolvens), Streptococcus bovis(S.bovis), Ruminobacter amylophilus(R. amylophilus) and Succinimonas amylolytica(S. amylolytica) were analyzed based on the total rumen bacterial 16 S ribosomal deoxyribonucleic acid(r DNA). The relative abundance of R. flavefaciens and R. albus increased at 24 h, but the relative abundance of F. succinogenes decreased at this time. The incubation of SPs had no effect on the abundance of S. bovis and R. amylophilus. The relative abundance of B. fibrisolvens and S. amylolytica reached the greatest value(p<0.05) at middle doses of SPs inclusion at 24 h. The value of MCPs linearly increased(p<0.05) at 24 and 48 h. These results showed that SPs could improve in vitro fermentation and nutrient digestion of feed substrates, and had the ability to modulate the ruminal fermentation pattern by regulating the composition of functional rumen microbes. Hence, SPs might be a potential feed additive applied in the diets of ruminants. 展开更多
关键词 Soybean Peptide(SP) ruminal fermentation rumal microbial population DIGESTIBILITY
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Effects of Encapsulated Enzyme and Yeast Products on in Vitro Rumen Fermentation
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作者 zhang Mei-mei Liang Ge-ge +4 位作者 Chen Ming-ming Jin Long Xie Xiao-lai zhang yong-gen Jiao Pei-xin 《Journal of Northeast Agricultural University(English Edition)》 CAS 2022年第2期28-37,共10页
Batch cultures of mixed rumen micro-organisms were conducted to evaluate the effects of encapsulated yeast(+EY)and encapsulated enzyme(+EE)using plant proteins(barley and oats grain)on rumen fermentation in vitro,inve... Batch cultures of mixed rumen micro-organisms were conducted to evaluate the effects of encapsulated yeast(+EY)and encapsulated enzyme(+EE)using plant proteins(barley and oats grain)on rumen fermentation in vitro,investigate the abilities of encapsulated yeast and encapsulated enzyme to prevent rumen digestion in vitro.Treatments of the study were the control,+EY,+EE products(3.33 mg·mL^(-1) of the incubation medium),unencapsulated yeast(-EY)and enzyme(-EE)products(0.17 and 0.17μL·mL^(-1) of the incubation medium,respectively).+EY group increased dry matter disappearance(DMD,P<0.01)and the total volatile fatty acids(TVFA,P<0.01)at 3 h of the incubation compared with the control,regardless of encapsulation of yeast.Gas production(GP)of+EY group was higher(P=0.05,29.94 mL·mL^(-1) organic matter,OM)than that of the control(25.08 mL·g^(-1) OM)at 3 h of the incubation.Supplementation+EY increased DMD(P=0.04,0.394 vs 0.352,respectively)and acetic proportion(P=0.04,52.6 vs 49.8 mol•100 mL^(-1),respectively)at 6 h of the incubation and increased A:P ratio(P<0.01,3.11 and 2.86,respectively)at 24 h of the incubation,as compared to unencapsulation of yeast.Supplementation of enzyme had higher(P≤0.04)GP,DMD and TVFA at 3 and 6 h of the incubation compared with the control,regardless of encapsulation.Moreover,the addition of+EE produced greater GP at 6(P<0.01,92.35 vs 78.21 mL·g^(-1) OM,respectively),12(218.47 vs 159.18 mL·g^(-1) OM)and 24 h(380.97 vs 297.78 mL·g^(-1) OM,respectively)of the incubation,higher DMD(0.347 vs 0.313,respectively)at 3 h of the incubation as compared to-EE group.The study showed that the encapsulation might protect part of yeast and enzyme from releasing to the rumen throughout the digestion in vitro,resulting in higher or no difference of rumen fermentation parameters compared with unencapsulated groups at any incubation times.In comparison with-EY and-EE,the higher rumen fermentation parameters at the early incubation time were observed,which could be attributed to the higher concentration of yeast or enzyme.However,regardless of the encapsulation,the results indicated that both yeast and enzyme only improved the speed rather than the extent of rumen fermentation in vitro. 展开更多
关键词 dry matter digestion encapsulated enzyme encapsulated yeast gas production in vitro volatile fatty acid
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Individual Identification of Dairy Cows Based on Deep Feature Extrac-tion and Matching
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作者 Shen Wei-zheng Sun Jia +4 位作者 Liang Chen Shi Wei Guo Jin-yan zhang Zhe zhang yong-gen 《Journal of Northeast Agricultural University(English Edition)》 CAS 2022年第3期85-96,共12页
Individual identification of dairy cows is the prerequisite for automatic analysis and intelligent perception of dairy cows'behavior.At present,individual identification of dairy cows based on deep convolutional n... Individual identification of dairy cows is the prerequisite for automatic analysis and intelligent perception of dairy cows'behavior.At present,individual identification of dairy cows based on deep convolutional neural network had the disadvantages in prolonged training at the additions of new cows samples.Therefore,a cow individual identification framework was proposed based on deep feature extraction and matching,and the individual identification of dairy cows based on this framework could avoid repeated training.Firstly,the trained convolutional neural network model was used as the feature extractor;secondly,the feature extraction was used to extract features and stored the features into the template feature library to complete the enrollment;finally,the identifies of dairy cows were identified.Based on this framework,when new cows joined the herd,enrollment could be completed quickly.In order to evaluate the application performance of this method in closed-set and open-set individual identification of dairy cows,back images of 524 cows were collected,among which the back images of 150 cows were selected as the training data to train feature extractor.The data of the remaining 374 cows were used to generate the template data set and the data to be identified.The experiment results showed that in the closed-set individual identification of dairy cows,the highest identification accuracy of top-1 was 99.73%,the highest identification accuracy from top-2 to top-5 was 100%,and the identification time of a single cow was 0.601 s,this method was verified to be effective.In the open-set individual identification of dairy cows,the recall was 90.38%,and the accuracy was 89.46%.When false accept rate(FAR)=0.05,true accept rate(TAR)=84.07%,this method was verified that the application had certain research value in open-set individual identification of dairy cows,which provided a certain idea for the application of individual identification in the field of intelligent animal husbandry. 展开更多
关键词 cow individual identification convolutional neural networks deep feature extraction feature matching
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黄芪多糖通过Nrf2/SLC7A11/GPX4信号通路诱导卵巢癌细胞铁死亡
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作者 张永跟 颜小飞 +5 位作者 刘锋 郝文哲 蔡玥 刘莹 刘兰林 李学军 《中国中药杂志》 CAS CSCD 北大核心 2024年第23期6459-6467,共9页
该研究主要探讨中药黄芪多糖(Astragalus polysaccharides, APS)调控核转录因子E2相关因2(Nrf2)/溶质载体家族7成员11(SLC7A11)/谷胱甘肽过氧化物酶4(GPX4)信号通路,诱导卵巢癌细胞(Caov-3和SKOV3细胞)铁死亡的作用机制。将Caov-3和SKOV... 该研究主要探讨中药黄芪多糖(Astragalus polysaccharides, APS)调控核转录因子E2相关因2(Nrf2)/溶质载体家族7成员11(SLC7A11)/谷胱甘肽过氧化物酶4(GPX4)信号通路,诱导卵巢癌细胞(Caov-3和SKOV3细胞)铁死亡的作用机制。将Caov-3和SKOV3细胞分为对照(Vehicle)组、APS组、谷胱甘肽过氧化物酶4抑制剂(RSL3)组、APS+RSL3组,干预48 h后观察各组细胞的活性和形态。cell counting kit-8(CCK-8)法检测细胞半数抑制浓度(IC_(50));克隆形成实验和EdU实验检测细胞增殖情况;生化试剂检测Caov-3细胞内脂质活性氧(L-ROS)、丙二醛(MDA)、二价铁离子(Fe^(2+))、谷胱甘肽(GSH)水平;透射电镜观察Caov-3细胞线粒体的形态变化;通过生物信息学分析筛选APS作用卵巢癌细胞潜在的目标基因;采用蛋白免疫印迹法(Western blot)和实时荧光定量PCR(RT-PCR)检测Nrf2、SLC7A11和GPX4蛋白和mRNA的表达水平。结果显示,APS能有效抑制卵巢癌细胞活性与增殖,显著升高L-ROS、MDA、Fe^(2+)水平(P<0.001),同时显著降低GSH水平(P<0.001);电镜下观察到Caov-3细胞线粒体明显变小,双层膜密度明显增加,线粒体嵴基本消失,外层线粒体膜破裂,APS+RSL3效果更加明显;生物信息学分析筛选出APS作用卵巢癌细胞潜在目标基因分别是Nrf2、SLC7A11、GPX4;APS能降低Nrf2、SLC7A11、GPX4蛋白和mRNA的表达(P<0.01),APS+RSL3降低效果更明显(P<0.001)。结果表明,APS可诱导卵巢癌细胞铁死亡,其机制可能与APS通过调节Nrf2/SLC7A11/GPX4信号通路,诱导卵巢癌细胞铁死亡相关,为APS治疗卵巢癌提供了实验基础。 展开更多
关键词 黄芪多糖 Nrf2/SLC7A11/GPX4信号通路 卵巢癌细胞 铁死亡
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通督调神针法对PSD患者COR的影响及临床研究 被引量:3
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作者 颜小飞 张永跟 张闻东 《医学信息》 2018年第15期137-139,142,共4页
目的通过通督调神针法对脑卒中后抑郁患者进行针刺治疗,观察血浆皮质醇水平的变化及临床疗效。方法选取2015年12月~2016年11月我院60例PSD患者,按数字随机法分为治疗组和对照组,每组30例。治疗组行通督调神针刺治疗,对照组行盐酸氟西汀... 目的通过通督调神针法对脑卒中后抑郁患者进行针刺治疗,观察血浆皮质醇水平的变化及临床疗效。方法选取2015年12月~2016年11月我院60例PSD患者,按数字随机法分为治疗组和对照组,每组30例。治疗组行通督调神针刺治疗,对照组行盐酸氟西汀胶囊药物治疗,治疗4周后,采用临床神经功能缺损程度评分表评定神经功能,汉密尔顿抑郁量表24项评定抑郁状态,Barthe I指数评定日常活动能力,以及应用放射免疫法测定COR的含量浓度,观察两组对PSD患者治疗的疗效。结果治疗组总有效率93.10%,对照组总有效率90.00%,两组对比差异无统计学意义(P>0.05);两组在NIHSS、HAMD、ADL量表的改善方面,治疗组评分改善情况优于对照组,差异有统计学意义(P<0.05);两组COR含量比较方面,两组差异无统计学意义(P>0.05)。结论通督调神针法治疗PSD可显著改善患者的临床症状和生活质量,值得临床上进一步深入研究。 展开更多
关键词 通督调神针法 脑卒中后抑郁 血浆皮质醇
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