Single-nucleotide polymorphisms (SNPs) of the IRF6 and TFAP2A in non-syndromic cleft lip with or without cleft palate (NSCLP) in a northern Chinese population

The conserved armadillo repeat (ARM) domain of adenomatous polyposis coli (APC) protein plays an important role in the recognition of its binding partners. In this study, we report the crystal structure of APC-ARM (residues 407-775), which was determined to 2.9 Å resolution. Our structure shows that the seven armadillo repeats of APC-ARM fold together into a compact domain, with Arm2 and Arm5 presenting some deviations from canonical armadillo repeats. There is a positively charged groove on the surface of APC-ARM, which might be the recognition site for APC-binding partners. Comparison of this structure with our previously reported structure of APC (407-751), together with normal mode analysis, reveals that the APC-ARM domain possesses a limited intrinsic flexibility. We propose that this intrinsic flexibility might be an inherent property of ARM domains in general.     

Necessary role for intracellular Ca2+ transients in initiating the apical-basolateral thinning of enveloping layer cells during the early blastula period of zebrafish development

During the early blastula period of zebrafish embryos, the outermost blastomeres begin to undergo a significant thinning in the apical/basolateral dimension to form the first distinct cellular domain of the embryo, the enveloping layer (EVL). During this shape transformation, only the EVL-precursor cells generate a coincidental series of highly restricted Ca(2+) transients. To investigate the role of these localized Ca(2+) transients in this shape-change process, embryos were treated with a Ca(2+) chelator (5,5'-difluoro BAPTA AM; DFB), or the Ca(2+) ionophore (A23187), to downregulate and upregulate the transients, respectively, while the shape-change of the forming EVL cells was measured. DFB was shown to significantly slow, and A23187 to significantly facilitate the shape change of the forming EVL cells. In addition, to investigate the possible involvement of the phosphoinositide and Wnt/Ca(2+) signaling pathways in the Ca(2+) transient generation and/or shape-change processes, embryos were treated with antagonists (thapsigargin, 2-APB and U73122) or an agonist (Wnt-5A) of these pathways. Wnt-5A upregulated the EVL-restricted Ca(2+) transients and facilitated the change in shape of the EVL cells, while 2-APB downregulated the Ca(2+) transients and significantly slowed the cell shape-change process. Furthermore, thapsigargin and U73122 also both inhibited the EVL cell shape-change. We hypothesize, therefore, that the highly localized and coincidental Ca(2+) transients play a necessary role in initiating the shape-change of the EVL cells.     

海洋异养细菌利用溶解有机碳的定量评估

【目的】惰性溶解有机碳(refractory dissolved organic carbon,RDOC)是海洋总有机碳的主体组分,RDOC在深海中可保存数千年,构成了巨大的碳储库,在调节气候变化中有重要作用。但RDOC的定量评估尚未有统一的标准方法。通过测定环境中能被异养细菌利用的溶解有机碳(biodegradable DOC,BDOC)可以反过来评估RDOC的量。本文对BDOC测定中一些关键步骤进行验证,为制定海洋RDOC评估标准奠定基础。【方法】本文评估了3种过滤方式及5种滤膜对DOC测定的影响,并评估了瓶子效应和稀释效应对细菌生长和DOC利用的影响。【结果】研究发现,(1) GF/F滤膜、GF-75滤膜、聚四氟乙烯(PTFE)滤膜(孔径0.2μm)、聚碳酸酯(PC)滤膜(孔径0.2μm)和聚四氟乙烯材质针孔过滤器(HA)(孔径0.2μm) 5种滤膜不会引入DOC污染;抽滤过滤和重力过滤方式过滤效果稳定、无污染,而在线过滤效果不稳定,易污染;(2)不同大小培养体系(30–480 mL;表面积/体积比为:1.64–0.67 cm–1)之间的细菌生长速率和DOC利用量没有显著性差异;(3)培养体系稀释度越高,细菌生长速率越高,对数生长期细菌丰度及DOC利用量越低。【结论】综合考虑,建议BDOC和RDOC测定实验中采用抽滤过滤的方式及不进行稀释的培养体系;常用的滤膜和培养体积对BDOC评估无显著影响。结合研究结果,我们提出了评估海洋RDOC的方法。     

一次性力竭运动致大鼠骨骼肌氧化应激的机制

目的:观察一次性力竭运动对大鼠骨骼肌氧化应激相关酶表达的影响。方法:雄性SD大鼠40只,分为4组(n=10),分别为对照组(C组)、力竭运动组(E组)、运动+PKC抑制剂组(EC组)、运动+NOX抑制剂组(EA组)。三组运动大鼠进行3 d的跑台适应性运动(5 m/min,1次/日,无坡度),然后休息1 d;EC组于运动前1 d和运动前1 h注射PKC抑制剂白屈菜红碱(5 mg/kg),EA组同期注射NADPH氧化酶抑制剂Apocynin(10 mg/kg),C组和E组注射同等剂量生理盐水;三组运动大鼠进行一次性跑台力竭运动,力竭后取大鼠的跖肌,DCF荧光探针检测活性氧(ROS),Western blot分析NOX2、NOX4、3-NT,免疫沉淀分析PKC、NOX2、NOX4。结果:与C组相比,E组的ROS水平、NOX2和NOX4蛋白表达、PKC-NOX2和PKC-NOX4复合物水平、3-NT生成均显著增加(P＜0.01,P＜ 0.05),EC组、EA组ROS无显著差异(P＞0.05),EC组NOX4蛋白表达显著增加(P＜0.05);与E组相比,EC组和EA组的ROS水平、NOX2和NOX4蛋白表达、PKC-NOX2和PKC-NOX4复合物水平、3-NT生成均显著降低(P＜ 0.01,P＜0.05)。结论:力竭运动诱导骨骼肌NOX2、NOX4蛋白表达增加,PKC通过调控NOX2介导ROS的生成。     

不同密度巨桉人工林林下植物多样性及根际土壤化感物质

为探究巨桉人工林林下植物多样性的影响因素,选择四川省丹棱县4和8年生不同密度(1200、1600和2000株·hm^-2)巨桉人工林为对象,研究林下植物多样性和根际土壤酚类化感物质。结果表明: 共发现植物45种,隶属于33科44属;随林龄增加和林分密度降低,巨桉人工林林下植物种类增加且重要值分布更为均匀;各林分植物生活型以高位芽植物为主。4年生林地灌木Shannon指数和Margalef指数在1600株·hm^-2下显著升高,4年生林地除Margalef指数外其余草本多样性指数及8年生林地草本Pielou指数随林分密度降低均显著升高;灌木Shannon指数及Margalef指数在1200株·hm^-2下8年生林地显著高于4年生林地。根际土壤中鉴定出5种酚类化感物质,4年生林地根际土壤中水杨酸浓度在1600株·hm^-2下显著降低,5,7-二羟基黄酮浓度随林分密度降低而显著降低;水杨酸浓度在8年生林地根际土壤中随林分密度降低显著升高;水杨酸浓度在2000株·hm^-2下4年生林地显著高于8年生林地,在1600株·hm^-2下相反;5,7-二羟基黄酮浓度在1200株·hm^-2下8年生林地显著高于4年生林地。冗余分析(RDA)结果表明,土壤pH、容重、有机质、全磷、全氮及化感物质是林下植物多样性的主要环境影响因子。适度延长轮伐期、调节林分密度以改善林内微环境、缓冲巨桉的化感作用,可促进林下植被发育。     

Rapamycin preserves the primordial follicle pool during cisplatin treatment in vitro and in vivo

Rapamycin has been proven to effectively inhibit the activation of primordial follicles while cisplatin‐induced the loss of primordial follicles due to the over‐activation of the primordial follicle stockpile. Whether rapamycin could inhibit the loss of primordial follicles induced by cisplatin is still unknown. The ovaries of neonatal Sprague Dawley rats were cultured in vitro in different doses of rapamycin (0.08, 0.16, and 0.32 μg/ml) and cisplatin (0.1, 0.4, and 0.8 μg/ml). The immature BALB/c mice were administered cisplatin with or without rapamycin by intraperitoneal injection. Ovaries were collected to analyze the histomorphology, the messenger RNA (mRNA) expression of anti‐Mullerian hormone (AMH), growth differentiation factor 9 (GDF9), and bone morphogenetic protein 15 (BMP15) and the expression of key proteins of mammalian target of rapamycin (mTOR) pathway. Growing follicle counts of ovaries cultured in vitro in the R0.16 and R0.32 groups were decreased and the ratio of growing to primordial follicles was also decreased in a dose‐dependent manner. In the C0.8 group, growing follicles were decreased compared with the other groups while the ratio was substantially increased in the C0.4 and C0.8 group. Co‐treatment attenuated primordial follicle loss and reduced the upregulated ratio induced by cisplatin. Ovarian follicle dynamics in vivo was consistent with the in vitro results. Primordial follicles counts were statistically increased and the ratio was reduced in the rapamycin group compared with the control group. Primordial follicle counts were dramatically reduced in the cisplatin group whereas co‐treatment with rapamycin slightly recovered its counts. There was no obvious difference in the number of growing follicles between the cisplatin group and other groups. The ratio was significantly increased in cisplatin‐treated mice whereas decreased in the co‐treatment group. The apoptosis rate of antral follicles in cisplatin‐treated mice was higher than the other groups while the apoptosis rate was decreased in the co‐treatment group in vivo. Compared with the control and rapamycin group, the mRNA expression of AMH, GDF9, and BMP15 were downregulated in the cisplatin group. The co‐treatment group recovered the mRNA expression of BMP15. In addition, the expression of key protein of mTOR pathway rpS6 and its phosphorylated forms were increased in the cisplatin‐treated group while co‐treatment decreased their expression. Rapamycin attenuated the loss of primordial follicles induced by cisplatin through the inhibitory effect of rapamycin on the mTOR pathway. These results suggest that rapamycin may be an effective drug for the protection of ovarian function during chemotherapy.     

Abundant fungi adapt to broader environmental gradients than rare fungi in agricultural fields

Soil communities are intricately linked to ecosystem functioning, and a predictive understanding of how communities assemble in response to environmental change is of great ecological importance. Little is known about the assembly processes governing abundant and rare fungal communities across agro‐ecosystems, particularly with regard to their environmental adaptation. By considering abundant and rare taxa, we tested the environmental thresholds and phylogenetic signals for ecological preferences of fungal communities across complex environmental gradients to reflect their environmental adaptation, and explored the factors influencing their assembly based on the large‐scale soil survey in agricultural fields across eastern China. We found that the abundant taxa exhibited remarkably broader response thresholds and stronger phylogenetic signals for the ecological preferences across environmental gradients compared to the rare taxa. Neutral processes played a key role in shaping the abundant subcommunity compared to the rare subcommunity. Null model analysis revealed that the abundant subcommunity was less clustered phylogenetically and governed primarily by dispersal limitation, while homogeneous selection was the major assembly process in the rare subcommunity. Soil available sulfur was the major factor mediating the balance between stochastic and deterministic processes of both the abundant and rare subcommunities, as indicated by an increase in stochasticity with higher available sulfur concentration. Based on macroecological spatial scale datasets, our study revealed the potential broader environmental adaptation of abundant fungal taxa compared to rare fungal taxa, and identified the factors mediating their distinct community assembly processes in agricultural fields. These results contribute to our understanding of the mechanisms underlying the generation and maintenance of fungal diversity in response to global environmental change.