Requirements for human‐induced pluripotent stem cells

‘Requirements for Human‐Induced Pluripotent Stem Cells’ is the first set of guidelines on human‐induced pluripotent stem cells in China, jointly drafted and agreed upon by experts from the Chinese Society for Stem Cell Research. This standard specifies the technical requirements, test methods, and instructions for use, labeling, packaging, storage, transportation, and waste handling for human‐induced pluripotent stem cells, which apply to the production and quality control of human‐induced pluripotent stem cells. It was released by the Chinese Society for Cell Biology on 9 January 2021 and came into effect on 9 April 2021. We hope that the publication of these guidelines will promote institutional establishment, acceptance, and execution of proper protocols and accelerate the international standardization of human‐induced pluripotent stem cells for applications.     

Evidence for two types of Aquilegia ecalcarata and its implications for adaptation to new environments

Spurs have played an important role in the radiation of the genus Aquilegia, but little is known about how the spurless state arose in A. ecalcarata. Here we aim to characterize the genetic divergence within A. ecalcarata and gain insights into the origin of this species. A total of 19 populations from A. ecalcarata and 23 populations from three of its closest relatives (Aquilegia kansuensis, Aquilegia rockii and Aquilegia yabeana) were sampled in this study. We sequenced fifteen nuclear gene fragments across the genome and three chloroplast loci to conduct phylogenetic, PCoA and STRUCTURE analyses.Our analyses indicate that A. ecalcarata may not be monophyletic and can be divided into two distinct lineages (A. ecalcarata I and A. ecalcarata II). A. ecalcarata I is genetically close to A. kansuensis, whereas A. ecalcarata II is close to A. rockii. Isolation-with-migration analysis suggested that historical gene flow was low between A. ecalcarata I and A. rockii, as well as between A. ecalcarata II and A. kansuensis. The two distinct lineages of A. ecalcarata show significant divergence in 13 floral traits and also have distinct distributions. In addition, both A. ecalcarata I and II are adapted to a stony environment that differs from that of their closest relatives, indicating a habitat shift may have driven new adaptations. Our findings enrich the understanding of how floral evolution contributes to species diversification.     

ATG7-enhanced impaired autophagy exacerbates acute pancreatitis by promoting regulated necrosis via the miR-30b-5p/CAMKII pathway

The present study was performed to explore whether and how impaired autophagy could modulate calcium/calmodulin-dependent protein kinase II (CAMKII)-regulated necrosis in the pathogenesis of acute pancreatitis (AP). Wistar rats and AR42J cells were used for AP modeling. When indicated, genetic regulation of CAMKII or ATG7 was performed prior to AP induction. AP-related necrotic injury was positively regulated by the incubation level of CAMKII. ATG7 positively modulated the level of CAMKII and necrosis following AP induction, indicating that there might be a connection between impaired autophagy and CAMKII-regulated necrosis in the pathogenesis of AP. microRNA (miR)-30b-5p was predicted and then verified as the upstream regulator of CAMKII mRNA in our setting of AP. Given that the level of miR-30b-5p was negatively correlated with the incubation levels of ATG7 after AP induction, a rescue experiment was performed and indicated that the miR-30b-5p mimic compromised ATG7 overexpression-induced upregulation of CAMKII-regulated necrosis after AP induction. In conclusion, our results indicate that ATG7-enhanced impaired autophagy exacerbates AP by promoting regulated necrosis via the miR-30b-5p/CAMKII pathway.Subject terms: Cytokines, Acute inflammation     

东海北部滨海湿地铁氧化细菌的分布及其对不同氧气穿透深度的响应

【目的】滨海湿地生态系统位于淡水与海水交互地带,含有高浓度Fe²⁺的地下水渗透到沉积物表层形成的湿地径流和周期性潮汐淹水形成的含氧-缺氧界面有利于铁氧化细菌介导的Fe²⁺的生物氧化过程发生。然而,目前缺乏对滨海湿地生态系统中铁氧化细菌类群的全面评估。【方法】以上海崇明西沙湿地公园及浙江舟山市朱家尖岛东沙沙滩两地共5处滨海湿地沉积物为研究对象,分析沉积物的氧气穿透深度等环境参数,并基于16S rRNA基因扩增子测序技术,全面解析不同滨海湿地生态系统中细菌与铁氧化细菌的群落组成与分布特征。【结果】与崇明西沙湿地相比,朱家尖岛东沙沙滩有更深的氧气穿透深度,达到10 mm以上。非度量多尺度分析(non-metric multidimensional scaling, NMDS)统计结果表明,细菌群落结构主要受到区域位置不同导致的环境条件差异的影响,而铁氧化细菌的群落结构则受到采样的区域位置和沉积物氧气穿透深度的共同影响。崇明西沙湿地和朱家尖岛东沙沙滩的优势细菌为蓝细菌门(Cyanobacteria)、γ-变形菌纲(Gammaproteobacteria)、拟杆菌门(Bacteroidetes)、α-变形菌纲(Alphaproteobacteria)和放线菌门(Actinobacteria);优势铁氧化细菌为嘉利翁氏菌属(Gallionella)、红细菌属(Rhodobacter)、Lepthothrix和Sideroxydans。【结论】通过对崇明西沙湿地和朱家尖岛东沙沙滩沉积物中栖息的铁氧化菌的调查发现,铁氧化细菌的群落组成与湿地沉积物类型导致的氧气穿透深度差异具有密切联系。     

典型油气藏区域环境因素差异对甲烷氧化菌丰度与群落结构分布的影响

【目的】甲烷氧化菌(methane-oxidizing bacteria, MOB)是油气微生物勘探中重要的指标微生物,其丰度与群落结构分布受到地理位置和多种环境因素的影响。本研究以采集的7个典型油气藏区域的土壤样品为研究对象,探究油气藏区域环境因素差异对甲烷氧化菌丰度与群落结构分布的影响。【方法】对采集自7个典型油气藏区域土壤样品进行pmoA基因实时荧光定量PCR (real-time fluorescence quantitative PCR, qPCR)、细菌16S rRNA基因测序和甲烷氧化菌功能基因pmoA测序,结合环境因子,比较甲烷氧化菌丰度差异,分析环境因子对甲烷氧化菌分布的影响。【结果】土壤样品的理化性质测定发现江汉盆地样品的含水量最高,约22.8%,硝态氮平均含量最高位于玉北油田,达到31.96 μg/g干重土壤,春光油田出现最高的SO₄^2- (6 425.0 mg/g干重土壤)及Cl^- (1 617.0 mg/g干重土壤)浓度。qPCR分析发现pmoA基因丰度仅为土壤样本总细菌丰度的0.77%,表明甲烷氧化菌绝对数量较少。16S rRNA基因测序发现type I型甲基八叠球菌属(Methylosarcina)、甲基嗜热菌属(Methylocaldum)、甲基球菌属(Methylococcus)与type II型的甲基胞囊菌属(Methylocystis) 4种主要的甲烷氧化菌,甲烷氧化菌中的优势菌属的相对丰度极低,最高仅为0.124%。甲烷氧化菌功能基因pmoA测序发现type II型的甲基胞囊菌属、甲基弯曲菌属(Methylosinus)为主要优势pmoA基因型。环境因子与甲烷氧化菌丰度相关性分析表明甲烷氧化菌绝对丰度与铵态氮、pH、颗粒大小、硫酸根和氯离子显著相关,而环境因子与甲烷氧化菌的相对丰度的相关性分析仅发现与颗粒大小、总氮、总磷、金属离子(Al、Fe、K、Ca、Mg、Mn、Zn和Cu)显著相关。甲烷氧化菌pmoA功能基因组成判别相关分析(discriminant correlation analysis, DCA)表明含水量(土壤湿度)、pH值、<2 μm的土壤颗粒、总氮及钙离子对甲烷氧化菌pmoA功能基因组成影响具有显著性。【结论】本研究通过对来自7个典型油气藏区域的土壤样品的甲烷氧化菌的绝对丰度与相对丰度进行分析,发现基于不同的测量方法,甲烷氧化菌的丰度存在较大差异,16S rRNA基因测序无法较为全面地反映土壤样本中优势甲烷氧化菌的群落结构,不同地理位置的甲烷氧化菌指示群落存在差异,在全国尺度下油气区及背景点均无特有的甲烷氧化菌指示群落。对环境因子与甲烷氧化菌相关性分析发现,基于不同的甲烷氧化菌丰度指标,环境因子的显著影响具有较大差别。     

Autophagy-associated circRNA circATG7 facilitates autophagy and promotes pancreatic cancer progression

Dysregulation of autophagy and circular RNAs (circRNAs) are involved in the pancreatic cancer (PC) progression. However, the regulatory network between circRNAs, autophagy, and PC progression remains unknown. Herein, we demonstrated that autophagy-associated circRNA circ-autophagy related 7 (circATG7) was elevated in PC tissues compared to adjacent tissues, and in PC cells treated with EBSS and hypoxia. circATG7 expression was positively associated with tumor diameter and lymph node invasion in patients with PC. circATG7 overexpression promoted PC cell proliferation, mobility, and autophagy in vitro, while circATG7 knockdown induced the opposite effects. ATG7 inhibition attenuated the effects of circATG7 on the biological functions of PC cells. CircATG7 is located in the cell cytoplasm and nucleus. Cytoplasmic circATG7 sponged miR-766-5p and decreased its expression, and increased the expression of ATG7, a target gene of miR-766-5p. Nuclear circATG7 acted as a scaffold to increase the interaction between the human antigen R protein and ATG7 mRNA and enhanced ATG mRNA stability. Furthermore, we demonstrated that circATG7 regulates PC cell proliferation and metastasis in vivo via ATG7-dependent autophagy. In conclusion, our results demonstrated that circATG7 accelerates PC progression via miR-766-5p/ATG7 and that HUR/ATG7 depends on autophagic flux. Thus, circATG7 may be a potential therapeutic target for PC.Subject terms: Pancreatic cancer, Pancreatic cancer     

输尿管软镜结合输尿管硬镜治疗复杂上尿路结石的效果评价及对预后的影响

目的 探讨输尿管软镜协同输尿管硬镜治疗复杂上尿路结石的临床效果.方法 选择我院收治的复杂上尿路结石患者96例,按入院顺序分为对照组和观察组,每组48例.对照组采取输尿管硬镜钬激光碎石术治疗,观察组采取输尿管软镜协同输尿管硬镜钬激光碎石术治疗.比较两组患者1次性碎石成功率、结石彻底清除率、炎症反应情况以及生活质量、住院时...     

High glucose represses the proliferation of tendon fibroblasts by inhibiting autophagy activation in tendon injury

Diabetic foot ulcer (DFU) is a kind of common and disabling complication of Diabetes Mellitus (DM). Emerging studies have demonstrated that tendon fibroblasts play a crucial role in remodeling phase of wound healing. However, little is known about the mechanism underlying high glucose (HG)-induced decrease in tendon fibroblasts viability. In the present study, the rat models of DFU were established, and collagen deposition, autophagy activation and cell apoptosis in tendon tissues were assessed using Hematoxylin–Eosin (HE) staining, immunohistochemistry (IHC), and TdT-mediated dUTP Nick-End Labeling (TUNEL) assay, respectively. Tendon fibroblasts were isolated from Achilles tendon of the both limbs, and the effect of HG on autophagy activation in tendon fibroblasts was assessed using Western blot analysis, Cell Counting Kit-8 (CCK-8) assay, and flow cytometry. We found that cell apoptosis was increased significantly and autophagy activation was decreased in foot tendon tissues of DFU rats compared with normal tissues. The role of HG in regulating tendon fibroblasts viability was then investigated in vitro, and data showed that HG repressed cell viability and increased cell apoptosis. Furthermore, HG treatment reduced LC3-II expression and increased p62 expression, indicating that HG repressed autophagy activation of tendon fibroblasts. The autophagy activator rapamycin reversed the effect. More importantly, rapamycin alleviated the suppressive role of HG in tendon fibroblasts viability. Taken together, our data demonstrate that HG represses tendon fibroblasts proliferation by inhibiting autophagy activation in tendon injury.     

Hsc70-Auxilin复合物的拉伸分子动力学模拟

Hsc70与auxilin蛋白组成的系统是Hsp70/Hsp40分子伴侣系统家族的一员,在热休克反应中发挥重要作用。本文为得出auxilin蛋白J结构域的关键氨基酸,首先采用由二硫键交联的Hsc70 R171C与auxilin D876C的复合物结晶结构作为初始模型,进行分子动力学模拟,通过比较平衡后的结合部位发现,将形成二硫键的氨基酸突变为原来的氨基酸结构在结合位点上与生化结果较为相近,之后利用此结构通过拉伸动力学模拟分析了auxilin蛋白J结构域与Hsc70的ATPase功能域的解离过程,并探讨了Hsc70与auxilin蛋白之间的相互作用力。结果表明位于HPD loop上的His874,Asp876,Thr879,螺旋Ⅲ上的Glu884,Asn895,Asp896,Ser899,Glu902,Asn903为关键氨基酸,这些数据符合之前核磁共振实验证实的T抗原J结构域的HPD基序和螺旋Ⅲ与Hsc70的ATPase功能域之间的相互作用。     

多球壳菌素介导线粒体功能调控酵母细胞衰老

多球壳菌素是前期鉴定出的一种抗衰老活性小分子,为了较全面理解多球壳菌素的抗衰机制,本研究利用模式生物芽殖酵母为材料,借助DNA Microarray技术、生物信息学手段并结合相关功能实验分析了多球壳菌素对对数期芽殖酵母细胞基因表达谱、基因本体聚类及相关信号通路的影响。结果显示,多球壳菌素对细胞转录组产生了显著影响,共导致1 648个基因的差异表达(FDR0.05,Fold Change1.5),其中843个基因显著上调,805个基因显著下调。进一步对基因本体聚类、信号通路分析及功能实验验证显示,线粒体相关功能和信号通路是多球壳菌素作用的一个主要靶点,本研究对多球壳菌素的进一步开发利用奠定了重要的理论基础。