HPLC-CAD法测定两性霉素B脂质体中HSPC、DSPG含量分析

摘要 目的：建立高效液相色谱（HPLC）-电喷雾检测器（CAD）法测定两性霉素B脂质体（L-AMB）中氢化大豆磷脂酰胆碱（HSPC）和磷脂二硬酯酰磷脂酰甘油（DSPG）含量的方法。方法：采用Venusil XBP C 8（4.6 mm×250 mm，5 μm）色谱柱，柱温：30℃；以甲醇-水作为流动相；喷雾压力：56.4 psi，喷雾温度：55℃；梯度洗脱，流速1.0 mL?min^-1。结果：L-AMB各磷脂在 5~60 mg?mL^-1内与峰面积呈良好的线性关系。HSPC平均回收率为100.06%，平均RSD为0.42；DSPG平均回收率100.03%，平均RSD为0.55。按HPLC-CAD 色谱条件测定L-AMB中HSPC、 DSPG含量分别为1497.66 mg?L^-1、4710.93 mg?L^-1，且无溶血磷脂检出。采用HPLC-ELSD 法验证，HSPC、 DSPG含量分别为1495.28 mg?L^-1、4712.71 mg?L^-1， 两种方法测定HSPC、 DSPG含量基本一致。结论：HPLC-CAD 测定L-AMB中HSPC、DSPG含量操作简单、准确，可准确测定L-AMB 中脂质体的质量。     

靶向AML突变的CRISPR/Cas9基因敲除文库构建

摘要 目的：构建靶向约200个AML基因突变的sgRNA基因敲除文库,为进一步探索诱发AML的信号通路网络奠定基础。方法：TCGA对200名AML病人进行了全基因组或全外显子组测序,鉴定出约2000个AML相关基因突变,从中选出了约200个突变两次或以上的基因作为靶向基因;接着,从Brie文库中挑选出相应基因的sgRNA序列,每个基因对应4条sgRNA;利用Gibson组装酶连接到慢病毒载体内,得到sgRNA文库;之后,采用pSSA荧光素酶基因报告系统鉴定文库sgRNA的切割活性;对文库进行高通量测序鉴定;用慢病毒包装文库,并测定病毒滴度。结果：1、构建了一个靶向约200个AML突变的sgRNA基因敲除文库;2、pSSA荧光素酶基因报告系统鉴定文库sgRNA具有切割活性;3、鉴定的7个单克隆质粒序列完全正确;4、高通量测序鉴定文库丰度和均一性符合要求;5、用慢病毒包装成病毒文库,测定病毒文库滴度为4.4×10⁷符合后续实验要求。结论：成功构建了靶向约200个基因突变的sgRNA敲除文库,可用于大规模地筛选诱发AML的基因突变,为探索AML发生、发展的分子机制以及药物靶点奠定基础。     

立体定向放疗联合内分泌治疗对转移性激素敏感性前列腺癌患者免疫功能和生活质量的影响

摘要 目的：探讨立体定向放疗（SBRT）联合内分泌治疗对转移性激素敏感性前列腺癌患者生活质量、免疫功能的影响。方法：选取我院2015年2月~2017年2月期间收治的转移性激素敏感性前列腺癌患者100例,根据信封抽签法将患者分为对照组（50例）和放疗组（50例）,对照组给予内分泌治疗,放疗组在对照组的基础上联合SBRT治疗。对比两组前列腺特异性抗原（PSA）进展时间、PSA缓解率、治疗期间不良反应状况、3年生存率、免疫功能（CD3⁺、CD4⁺、CD8⁺、CD4⁺/CD8⁺）和扩展性前列腺癌复合指数量表（EPIC）各项评分。结果：随访3年,对照组有2例失访、放疗组有3例失访,放疗组的PSA进展时间长于对照组（P<0.05）,放疗组的3年生存率高于对照组（P<0.05）。治疗后,两组CD3⁺、CD4⁺/CD8⁺、CD4⁺均下降,但放疗组较对照组升高（P<0.05）,两组治疗后CD8⁺均升高,但放疗组较对照组降低（P<0.05）。治疗后6个月,放疗组性功能、激素功能、泌尿功能、肠道功能领域评分均高于对照组（P<0.05）。两组不良反应总发生率、PSA缓解率组间对比无差异（P>0.05）。结论：SBRT联合内分泌治疗转移性激素敏感性前列腺癌患者,可延长患者PSA进展时间,减轻免疫抑制,提高患者生活质量,同时还可改善患者的预后,患者耐受性良好。     

漏芦醇提取物对小鼠乳腺上皮细胞氧化损伤的缓解作用

摘要 目的：探讨漏芦乙醇提取物（RUEE）对脂多糖（LPS）诱导的小鼠乳腺上皮细胞氧化损伤发挥保护作用。方法：采用体外培养的小鼠乳腺上皮细胞系 HC11 为模型,首先利用MTT法筛选RUEE的最佳作用浓度及检测LPS和RUEE对细胞活力的影响,然后用1 μg?mL^-1 LPS单独处理,20、40、80 μg?mL^-1的RUEE与 1 μg?mL^-1 LPS 共处理 HC11细胞,检测氧化应激相关指标、抗氧化相关基因mRNA和蛋白表达的变化。结果：细胞活力实验确定20、40、80 μg?mL^-1的RUEE作为后续试验的添加浓度。LPS诱导可显著提高HC11细胞内丙二醛（MDA）含量、一氧化氮（NO）水平、一氧化氮合酶（iNOS）活性（P＜0.05）;明显降低超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GPx）、过氧化氢酶（CAT）的活性和总抗氧化能力（T-AOC）（P＜0.05）;显著抑制核因子E2相关因子2（Nrf2）、血红素氧合酶1（HO-1）、NAD(P)H醌氧化还原酶1（NQO1）的mRNA及蛋白表达（P＜0.05）。20、40、80 μg?mL^-1的RUEE预处理后可明显下调LPS诱导的HC11细胞内MDA含量、NO水平及iNOS活性（P＜0.05）;显著提高SOD、GPx、CAT的活性及T-AOC（P＜0.05）;显著上调Nrf2、HO-1、NQO1 mRNA和蛋白表达（P＜0.05）。结论：RUEE可以有效减轻LPS诱导的乳腺上皮细胞氧化损伤,这可能与RUEE能激活乳腺上皮细胞Nrf2进而促进抗氧化基因的表达有关。     

消疬膏对脾胃虚弱证颈部淋巴结核患者CD4+CD25highFoxP3+调节性T淋巴细胞和胃肠激素的影响

摘要 目的：研究消疬膏对脾胃虚弱证颈部淋巴结核患者CD4⁺CD25^highFoxP3⁺调节性T淋巴细胞和胃肠激素的影响。方法：选择南京市中西医结合医院于2020年3月~2022年1月期间收治的颈部淋巴结核患者120例。按照随机数字表法分为对照组（60例）和研究组（60例）。对照组接受常规抗结核方案，研究组接受消疬膏联合常规抗结核方案，两组疗程均为6个月。观察两组治疗前和治疗6个月后的中医证候积分、CD4⁺CD25^highFoxP3⁺调节性T淋巴细胞相关指标和胃肠激素的变化，记录两组治疗期间用药安全性。结果：两组治疗6个月后肿疡大小、肿疡皮色、面色、纳呆、倦怠乏力、便溏、舌质、脉象评分均下降，且研究组低于对照组（P<0.05）。研究组治疗6个月后CD4⁺CD25^highFoxP3⁺调节性T淋巴细胞、白介素-10（IL-10）水平低于对照组，γ-干扰素（IFN-γ）水平高于对照组（P<0.05）。研究组治疗6个月后胆囊收缩素（CCK）、胃泌素（GAS）水平低于对照组，胃动素（MTL）水平高于对照组（P<0.05）。研究组的不良反应总发生率低于对照组（P<0.05）。结论：消疬膏治疗脾胃虚弱证颈部淋巴结核患者，可有效改善患者临床症状，可能与调节CD4⁺CD25^highFoxP3⁺调节性T淋巴细胞和胃肠激素水平有关。     

2型糖尿病并发肺结核患者PCT、HMGB1、CD4+/CD8+比值与继发肺部感染的关系

摘要 目的：探讨2型糖尿病并发肺结核患者降钙素原（PCT）、高迁移率族蛋白1（HMGB1）、CD4⁺/CD8⁺比值与继发肺部感染的关系。方法：选择2019年1月至2022年6月四川大学华西医院呼吸与危重症医学科收治的97例2型糖尿病并发肺结核患者,根据入院治疗时是否继发肺部感染分为肺部感染组（53例）及非肺部感染组（44例）。检测两组血清PCT、HMGB1水平以及外周血CD4⁺/CD8⁺比值。单因素和多因素Logistic回归分析2型糖尿病并发肺结核患者继发肺部感染的因素。受试者工作特征（ROC）曲线分析PCT、HMGB1和CD4⁺/CD8⁺比值预测2型糖尿病并发肺结核患者继发肺部感染的价值。结果：肺部感染组血清PCT、HMGB1水平高于非肺部感染组（P＜0.05）,外周血CD4⁺/CD8⁺比值低于非肺部感染组（P＜0.05）。糖化血红蛋白及血清PCT、HMGB1水平升高是2型糖尿病并发肺结核患者继发肺部感染的危险因素（P＜0.05）,高CD4⁺/CD8⁺比值是保护因素（P＜0.05）。PCT、HMGB1、CD4⁺/CD8⁺比值预测2型糖尿病并发肺结核患者继发肺部感染的曲线下面积为0.719、0.761、0.738,联合PCT、HMGB1和CD4⁺/CD8⁺比值预测的曲线下面积为0.878,高于各指标单独预测。结论：2型糖尿病并发肺结核患者血清PCT、HMGB1水平增高,外周血CD4⁺/CD8⁺比值降低,均与继发肺部感染有关,PCT、HMGB1联合CD4⁺/CD8⁺比值可辅助预测2型糖尿病并发肺结核患者继发肺部感染的风险。     

脓毒症患者外周血T淋巴细胞PD-1表达变化及胸腺肽α-1的免疫调理作用研究

摘要 目的：探讨脓毒症患者外周血T淋巴细胞程序性细胞死亡受体1（PD-1）表达特点,分析胸腺肽?琢-1治疗对患者免疫功能的影响。方法：选择2018年3月至2020年6月我院重症医学科收治的140例脓毒症患者（脓毒症组）和同期于我院进行体检的95例健康志愿者（对照组）,根据急性生理与慢性健康评估Ⅱ（APACHE Ⅱ）、序贯器官衰竭评估（SOFA）评分结果将脓毒症患者分为APACHE Ⅱ 0~10分组（51例）、11~20分组（62例）和＞20分组（27例）;SOFA评分0~5分组（48例）、6~10分组（60例）和＞10分组（32例）。检测外周血CD4⁺T细胞上PD-1表达、CD8⁺T细胞上PD-1表达,比较组间差异性。Pearson秩相关性分析外周血CD4⁺T细胞上PD-1表达、CD8⁺T细胞上PD-1表达与APACHE Ⅱ、SOFA评分相关性。根据治疗方法将脓毒症患者分为A组（60例）和B组（80例）,A组给予常规综合治疗和乌司他丁治疗,B组在A组的基础上联合胸腺肽α-1治疗,比较两组治疗前后外周血T淋巴细胞（CD3⁺、CD4⁺、CD8⁺）、NK细胞（CD3-CD16⁺CD56⁺）差异。结果：脓毒症组外周血CD4⁺T细胞上PD-1表达、CD8⁺T细胞上PD-1表达高于对照组（P＜0.001）,外周血CD4⁺T细胞上PD-1表达、CD8⁺T细胞上PD-1表达随APACHE Ⅱ、SOFA评分的增加而增高,各组间差异显著（P＜0.05）。Pearson秩相关分析结果显示外周血CD4⁺T细胞上PD-1表达、CD8+T细胞上PD-1表达与APACHE Ⅱ评分、SOFA评分呈正相关（r=0.569、0.475;0.653、0.509,P均＜0.05）。B组治疗后CD3⁺、CD4⁺、CD3-CD16⁺CD56⁺高于A组（P＜0.05）,CD8⁺低于A组（P＜0.05）。结论：脓毒症患者外周血CD4⁺、CD8⁺T细胞上PD-1表达均增高,其表达与病情严重程度密切相关。给予胸腺肽α-1治疗可改善患者免疫功能。     

马尾松幼苗生长及生理特性对铝胁迫的响应

以马尾松幼苗为试验材料,采用水培法研究铝胁迫（Al³⁺浓度为0、0.2、0.4、0.8、1.6 mmol·L^-1）对马尾松幼苗生长及其针叶中叶绿素、渗透调节物质（可溶性糖、可溶性蛋白、脯氨酸）、丙二醛（MDA）和超氧化物歧化酶（SOD）、过氧化物酶（POD）等保护酶活性的影响,为揭示马尾松铝毒害生理机制及提高马尾松的耐铝能力提供理论依据。结果显示：当Al³⁺处理浓度为0.2 mmol·L^-1时对马尾松株高和基径生长的影响较小,但对马尾松根系生长有一定的促进作用;Al³⁺处理浓度大于0.2 mmol·L^-1时对马尾松株高、基径和根长的生长均会产生一定的抑制作用,且这种抑制作用随着Al³⁺浓度的增大而增强。马尾松针叶中叶绿素含量和SOD、POD活性均随着Al³⁺处理浓度的增加呈先上升后下降的趋势;Al³⁺处理浓度大于0.2 mmol·L^-1时马尾松针叶中可溶性糖、可溶性蛋白、脯氨酸等渗透调节物质均呈上升趋势,且随着Al³⁺浓度的升高而增大;Al³⁺处理浓度大于0.2 mmol·L^-1时马尾松针叶中MDA含量也呈上升趋势,且随着Al³⁺浓度的增大而升高,说明大于0.2 mmol·L^-1的Al³⁺处理可导致马尾松膜脂产生氧化。研究表明,马尾松幼苗具有一定的耐铝能力,在铝胁迫生境下可通过提高自身SOD和POD等保护酶的合成和主动积累脯氨酸、蛋白质和可溶性糖等渗透物质,产生适应性生理响应以维持自身的生理平衡来降低铝毒害作用。     

ALPL缺陷加速高脂诱导小鼠肝内脂肪沉积

摘要 目的：明确肝/骨/肾型碱性磷酸酶基因（liver/bone/kidney alkaline phosphatase,ALPL）在高脂诱导肝内脂肪沉积的作用。方法：利用野生型（WT）和ALPL敲除小鼠（ALPL^+/-）,给予高脂饮食8周诱导脂肪肝模型,检测小鼠肝内脂肪沉积和血清中葡糖糖、甘油三脂及胆固醇含量,并应用RT-PCR、Western blotting和免疫荧光染色检测肝组织中脂肪酸生成和转运相关基因表达变化。结果：ALPL^+/-组在正常饮食条件下较WT组肝内脂肪沉积无明显变化,而血清中葡萄糖和胆固醇含量增加;高脂条件下,敲除ALPL小鼠肝内脂肪沉积明显增加,且伴随血清中甘油三脂含量增加。RT-PCR和Western blotting结果显示,在高脂诱导下ALPL^+/-小鼠肝组织中关键脂肪酸生成基因ACC1、ACC2和 PPAR-γ,及脂肪酸生成基因LPL表达明显增加。此外,免疫荧光染色结果显示高脂诱导下敲除ALPL小鼠肝组织中的PPAR-γ阳性肝细胞明显增加。结论：ALPL敲除促进肝内脂肪酸生成和转运,加速高脂诱导小鼠肝内脂肪沉积,为阐明脂肪肝病变分子机制提供理论支持。     

2017-2019年某某医院ICU患者痰液病原菌分布及耐药性分析

摘要 目的：探究医院ICU患者痰液病原菌分布,并就其耐药性进行分析,以期为临床经验性选择抗生素治疗提供必要的细菌学依据。方法：选择2017年1月至2019年12月于北京中医医院顺义医院入住ICU的84例患者为研究对象,采集其痰液标本,并实施细菌学培养和药敏学试验。结果：（1）84例患者合计提取菌株91种,其中G^-杆菌45种,占比49.45 %,G⁺菌36种,占比39.56 %,真菌10种,占比10.99 %;（2）分析显示亚胺培南对G^-杆菌的敏感性较好,万古霉素对G⁺菌的敏感性较好,尚未发现耐万古霉素的肠球菌和葡萄球菌出现。结论：ICU患者痰液病原菌类型较多,其中以G^-杆菌为主,病原菌呈现多重耐药性,亚胺培南对G^-杆菌敏感性较好,万古霉素对G⁺菌敏感性较好。     

A porphyrin‐based fluorescence method for zinc determination in commercial propolis extracts without sample pretreatment

The quantification of zinc in over‐the–counter drugs as commercial propolis extracts by molecular fluorescence technique using meso ‐tetrakis(4‐carboxyphenyl)porphyrin (H₂TCPP⁴) was developed for the first time. The calibration curve is linear from 6.60 to 100 nmol L^?1 of Zn²⁺. The detection and quantification limits were 6.22 nmol L^?1 and 19.0 nmol L^?1, respectively. The reproducibility and repeatability calculated as the percentage variation of slopes of seven calibration curves were 6.75% and 4.61%, respectively. Commercial propolis extract samples from four Brazilian states were analyzed and the results (0.329–0.797 mg/100 mL) obtained with this method are in good agreement with that obtained with the Atomic Absorption Spectroscopy (AAS) technique. The method is simple, fast, of low cost and allows the analysis of the samples without pretreatment. Moreover the major advantage is that Zn‐porphyrin complex presents fluorescent characteristic promoting the selectivity and sensitivity of the method.     

Expansion and CD2/CD3/CD28 stimulation enhance Th2 cytokine secretion of human invariant NKT cells with retained anti-tumor cytotoxicity

Background aimsKey obstacles in human iNKT cell translational research and immunotherapy include the lack of robust protocols for dependable expansion of human iNKT cells and the paucity of data on phenotypes in post-expanded cells.MethodsWe delineate expansion methods using interleukin (IL)-2, IL-7 and allogeneic feeder cells and anti-CD2/CD3/CD28 stimulation by which to dependably augment Th2 polarization and direct cytotoxicity of human peripheral blood CD3⁺Vα24⁺Vβ11⁺ iNKT cells.ResultsGene and protein expression profiling demonstrated augmented Th2 cytokine secretion (IL-4, IL-5, IL-13) in expanded iNKT cells stimulated with anti-CD2/CD3/CD28 antibodies. Cytotoxic effector molecules including granzyme B were increased in expanded iNKT cells after CD2/CD3/CD28 stimulation. Direct cytotoxicity assays using unstimulated expanded iNKT cell effectors revealed α-galactosyl ceramide (α-GalCer)-dependent killing of the T-ALL cell line Jurkat. Moreover, CD2/CD3/CD28 stimulation of expanded iNKT cells augmented their (α-GalCer-independent) killing of Jurkat cells. Co-culture of expanded iNKT cells with stimulated responder cells confirmed contact-dependent inhibition of activated CD4⁺ and CD8⁺ responder T cells.DiscussionThese data establish a robust protocol to expand and novel pathways to enhance Th2 cytokine secretion and direct cytotoxicity in human iNKT cells, findings with direct implications for autoimmunity, vaccine augmentation and anti-infective immunity, cancer immunotherapy and transplantation.     

A New Synthesis of (E,E)-8, 10-Dodecadien-1-ol,Sex Pheromone of Codling Moth

A methanol-utilizing bacterium, which produced red to pink pigments and assimilated methanol via icl^- serine pathway, was isolated from soil and tentatively designated as Pseudomonas MS 31. This bacterium produced L-serine when glycine was added to the growth medium at the late exponential phase of growth. The cells showed high L-serine degradation activity. Chelating agents and some metal ions, which inhibited L-serine degradation, stimulated the L-serine accumulation. In the presence of 0.1 ? 1 mM EDTA, o-phenanthroline, 8-hydroxyquinoline, α,α'-dipyridyl, cobalt sulfate or nickel sulfate, this bacterium produced 0.7?2.1mg L-serine from 4mg glycine per ml culture.     

Influence of temperature, light and nutrients on the growth rates of the macroalga Gracilaria domingensis in synthetic seawater using experimental design

In the present study, the daily relative growth rates (DRGR, in percent per day) of the red macroalga Gracilaria domingensis in synthetic seawater was investigated for the combined influence of five factors, i.e., light (L), temperature (T), nitrate (N), phosphate (P), and molybdate (M), using a statistical design method. The ranges of the experimental cultivation conditions were T, 18–26°C; L, 74–162?μmol photons m^?2?s^?1; N, 40–80?μmol?L^?1; P, 8–16?μmol?L^?1; and M, 1–5?nmol?L^?1. The optimal conditions, which resulted in a maximum growth rate of ≥6.4% d^?1 from 7 to 10?days of cultivation, were determined by analysis of variance (ANOVA) multivariate factorial analysis (with a 2⁵ full factorial design) to be L, 74?μmol photons m^?2?s^?1; T, 26°C; N, 80?μmol?L^?1; P, 8?μmol?L^?1; and M, 1?nmol?L^?1. In additional, these growth rate values are close to the growth rate values in natural medium (von Stosch medium), i.e., 6.5–7.0% d^?1. The results analyzed by the ANOVA indicate that the factors N and T are highly significant linear terms, X _L, (α?=?0.05). On the other hand, the only significant quadratic term (X _Q) was that for L. Statistically significant interactions between two different factors were found between T vs. L and N vs. T. Finally, a two-way (linear/quadratic interaction) model provided a quite reasonable correlation between the experimental and predicted DRGR values (R _adjusted ² ?=?0.9540).     

Surfactant enhanced l-α-glycerylphosphorylcholine production from phosphatidylcholine using phospholipase A1 in the aqueous phase

Abstract

Enzymatic production of L-α-glycerylphosphorylcholine (L-α-GPC) is difficult due to the limited solubility of phosphatidylcholine (PC) in the aqueous phase. Surfactants can be used to improve the solubility and the dispersibility of non-polar chemicals in the aqueous media. In this study, various surfactants were investigated to improve L-α-GPC enzymatic production using phospholipase A₁ (PLA₁) in the aqueous phase. The results showed that Tween 20 was the most effective surfactant for enhancing L-α-GPC concentration. With 20?g^.L^?1 of Tween 20, the optimal conditions of PC hydrolysis were determined to be enzyme loading of 0.64?g^.L^?1 and substrate concentration of 60?g^.L^?1 at 45?°C for 1?h. In addition, the fed-batch catalytic process of PC was conducted to avoid substrate inhibition and increase product accumulation, resulting in 112.56?g^.L^?1 of L-α-GPC from 360.00?g^.L^?1 PC with yield of 91.36% within 3?h.     

Physiological responses,tolerance efficiency,and phytoextraction potential of Hylotelephium spectabile (Boreau) H. Ohba under Cd stress in hydroponic condition

Abstract

A sand hydroponic experiment with different concentrations of 0, 5, 10, 20, 40?mg L^?1 Cd was used to study the growth and physiological response of Hylotelephium spectabile (Boreau) H. Ohba. and its phytoextraction potential for Cd. The results showed that total plant biomass under 5?mg L^?1 Cd treatment was slightly affected. The content of malondialdehyde (MDA) in leaf exposed to Cd was higher, and the POD and CAT activity exhibited a positive response to the low level of Cd addition (5?mg·L^?1). The photosynthesis pigments were slightly inhibited, and the ultrastructure of chloroplast remained intact after treatment with 10?mg L^?1 Cd. The maximum leaf Cd content (603?mg·kg^?1) was found in 5?mg L^?1 Cd treatment, then decreased with the Cd level increased. The maximum Cd content in the shoots far exceeds the threshold level (100?mg kg^?1) for a Cd-hyperaccumulator plant with the value of translocation factor (TF_shoot/root) for Cd reaching up to 5.62. In conclusion, H. spectabile showed normal growth and physiological response and high shoot Cd accumulation under 5?mg L^?1 Cd stress, which made it to be a good candidate for phytoextraction of low-level Cd polluted environment.     

基于CRISPR/Cas9技术构建CELF6基因敲除细胞系并探讨CELF6与p53基因的关系

目的:构建CELF6基因敲除细胞系并探讨CELF6与p53基因之间的关系。方法:CRISPR/Cas9双载体慢病毒系统通过两个慢病毒载体分别向细胞中导入Cas9蛋白和sg RNA序列表达框,从而实现对CELF6基因的敲除。通过Surveyor(错配酶法)检测sg RNA活性并利用Western blot(蛋白质免疫印迹)检测CELF6的敲除效率。进一步利用CCK-8试剂盒检测敲除CELF6和过表达CELF6对细胞增殖的影响。利用公开可用的癌症基因组图谱(TCGA)数据库分析CELF6在多种肿瘤组织中的表达情况。结果:CELF6基因敲除的HCT116细胞系成功构建。Western blot检测发现CELF6基因敲除的细胞系中p53、p-RB蛋白的表达水平显著下调,而CELF6过表达的细胞中p53、p-RB蛋白的表达水平明显上调。CCK-8实验结果显示敲除CELF6基因后,细胞增殖活性显著增强;而过表达CELF6后,细胞的增殖活性受到明显抑制。生物信息学分析发现CELF6在结肠癌、胶质母细胞瘤、子宫内膜癌、肾嫌色细胞癌、乳腺癌、甲状腺癌等肿瘤组织中显著低表达。结论:推测CELF6是一种新的潜在肿瘤抑制基因,其可能在肿瘤的发生、发展过程中发挥重要作用。     

A Positive Feedback Loop between ATOH7 and a Notch Effector Regulates Cell-Cycle Progression and Neurogenesis in the Retina

1. Download : Download full-size image

Highlights? Atoh7 regulates the conversion of progenitor cells to newborn neurons ? Atoh7 is required to activate HES5.3 before the penultimate mitosis ? HES5.3 lengthens the cell cycle ? HES5.3 is required for Atoh7-expressing cells to differentiate into neurons