Inhibition of tyrosinase by gastrodin: An integrated kinetic-computational simulation analysis

We studied the inhibitory effect of gastrodin on tyrosinase using inhibition kinetics and computational simulation. Gastrodin reversibly inhibited tyrosinase in a mixed-type manner with K_i = 123.8 ± 20.2 mM. Time-interval kinetics revealed the inhibition to be a first-order process with mono- and bi-phasic components. Using AutoDock Vina, we calculated a binding energy of ?6.3 kcal/mol for gastrodin and tyrosinase, and we performed a molecular dynamics simulation of the tyrosinase–gastrodin interaction. The simulation results suggested that gastrodin interacts primarily with histidine residues in the active site. A 10-ns molecular dynamics simulation showed that one copper ion in the tyrosinase active site was responsible for the interaction with gastrodin. Our study provides insight into the inhibition of tyrosinase by the hydroxyl groups of gastrodin. A combination of inhibition kinetics and computational calculations may help to confirm the inhibitory action of gastrodin on tyrosinase and define the mechanisms of inhibition.     

Transactivation of Atg4b by C/EBPβ Promotes Autophagy To Facilitate Adipogenesis

Autophagy is a highly conserved self-digestion pathway involved in various physiological and pathophysiological processes. Recent studies have implicated a pivotal role of autophagy in adipocyte differentiation, but the molecular mechanism for its role and how it is regulated during this process are not clear. Here, we show that CCAAT /enhancer-binding protein β (C/EBPβ), an important adipogenic factor, is required for the activation of autophagy during 3T3-L1 adipocyte differentiation. An autophagy-related gene, Atg4b, is identified as a de novo target gene of C/EBPβ and is shown to play an important role in 3T3-L1 adipocyte differentiation. Furthermore, autophagy is required for the degradation of Klf2 and Klf3, two negative regulators of adipocyte differentiation, which is mediated by the adaptor protein p62/SQSTM1. Importantly, the regulation of autophagy by C/EBPβ and the role of autophagy in Klf2/3 degradation and in adipogenesis are further confirmed in mouse models. Our data describe a novel function of C/EBPβ in regulating autophagy and reveal the mechanism of autophagy during adipocyte differentiation. These new insights into the molecular mechanism of adipose tissue development provide a functional pathway with therapeutic potential against obesity and its related metabolic disorders.     

Oral Community Interactions of Filifactor alocis In Vitro

Filifactor alocis is a gram positive anaerobe that is emerging as an important periodontal pathogen. In the oral cavity F. alocis colonizes polymicrobial biofilm communities; however, little is known regarding the nature of the interactions between F. alocis and other oral biofilm bacteria. Here we investigate the community interactions of two strains of F. alocis with Streptococcus gordonii, Fusobacterium nucleatum, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans, organisms with differing pathogenic potential in the oral cavity. In an in vitro community development model, S. gordonii was antagonistic to the accumulation of F. alocis into a dual species community. In contrast, F. nucleatum and the type strain of F. alocis formed a synergistic partnership. Accumulation of a low passage isolate of F. alocis was also enhanced by F. nucleatum. In three species communities of S. gordonii, F. nucleatum and F. alocis, the antagonistic effects of S. gordonii superseded the synergistic effects of F. nucleatum toward F. alocis. The interaction between A. actinomycetemcomitans and F. alocis was strain specific and A. actinomycetemcomitans could either stimulate F. alocis accumulation or have no effect depending on the strain. P. gingivalis and F. alocis formed heterotypic communities with the amount of P. gingivalis greater than in the absence of F. alocis. However, while P. gingivalis benefited from the relationship, levels of F. alocis in the dual species community were lower compared to F. alocis alone. The inhibitory effect of P. gingivalis toward F. alocis was dependent, at least partially, on the presence of the Mfa1 fimbrial subunit. In addition, AI-2 production by P. gingivalis helped maintain levels of F. alocis. Collectively, these results show that the pattern of F. alocis colonization will be dictated by the spatial composition of microbial microenvironments, and that the organism may preferentially accumulate at sites rich in F. nucleatum.     

小麦不同品种和播期对发育阶段的效应

以热时间（thermal time)为尺度研究了小麦不同品种和播期对发育阶段的效应,结果表明,小麦分蘖发生的早晚以生态因子调控为主,基因型差异较小;分薛- 节期为冬性品种（京411）一生中可变性最大的生育阶段,穗分化进入单棱斯的早晚以基因型效应为主,生态因子的影响次之,单棱－二棱期为春化作用的敏感期,冬性品种晚播（3月2日）春化效应可延迟到小花原基分化期之前,小麦物候期与穗发育阶段的对应关系具有一定的可变性,冬性品种较强的春化作用增加了其生态可变叶原基数;春化过程结束前,物候发育及穗发育阶段累计GDD与相应生殖器官原基分化的数的相关性不明显,春性品种（扬麦158）的物候发育及药隔分化期之前的穗发育阶段与各类顶端原基的分化数均具有极显著的正相关关系。     

美味猕猴桃原生质体再生植株细胞遗传学研究 Ⅱ.性别性状变异和小孢子发生及其发育命运

对美味猕猴桃同一雌株叶原生质体再生植株进行了形态学、细胞学以及育性特性的比较研究,确认该体细胞无性系性别性状发生变异。其中60％雄性再生植株退化的雌蕊仍保留不同程度的雌性化特征,但雌性全不育;小孢子则能发育成有功能的雄配子体,但有一定的功能缺陷。再生雌株中P1组群性状特征与母株相似;P2组群花发育畸形,导致雌性不育或育性极差。细胞学研究表明,小孢子母细胞减数分裂时染色体异常行为对小孢子发生的影响不能决定其性别类型;雌株类型小孢子败育过程有受基因调控的细胞学特征。认为雌株和雄株小孢子的发育受控于不同的基因体系,具性别的特异性。再生植株性别性状发生变异可能是性别控制基因或染色体发生结构性变异所致。母株染色体上累积的结构性变异与该遗传基础具易变性密切有关。     

自体内皮前体细胞移植对急性心肌梗死大鼠微血管新生及心功能的影响

目的探讨外周血来源的内皮前体细胞自体移植,对大鼠急性心肌梗死后微血管新生与心功能的影响。方法抽取SD大鼠外周动脉血,应用Ficoll密度梯度离心法获取单个核细胞。应用含有VEGF和bFGF的特定培养基体外培养,得到内皮前体细胞;结扎SD大鼠冠状动脉左前降支,建立急性心肌梗死模型;然后将得到的自体内皮前体细胞植入缺血心肌局部区域。对照组动物注入细胞培养液。结果与对照组比较,细胞移植组大鼠心功能明显改善,心肌收缩力显著优于对照组;梗死心肌微血管新生更为明显。结论急性心肌梗死心肌局部移植外周血来源的自体内皮前体细胞,能够促进血管新生;对局部梗死心肌组织结构有一定的保护作用,并可在不同时点不同程度恢复心肌收缩力,显著改善心功能。     

Apple Fruit Diameter and Length Estimation by Using the Thermal and Sunshine Hours Approach and Its Application to the Digital Orchard Management Information System

In apple cultivation, simulation models may be used to monitor fruit size during the growth and development process to predict production levels and to optimize fruit quality. Here, Fuji apples cultivated in spindle-type systems were used as the model crop. Apple size was measured during the growing period at an interval of about 20 days after full bloom, with three weather stations being used to collect orchard temperature and solar radiation data at different sites. Furthermore, a 2-year dataset (2011 and 2012) of apple fruit size measurements were integrated according to the weather station deployment sites, in addition to the top two most important environment factors, thermal and sunshine hours, into the model. The apple fruit diameter and length were simulated using physiological development time (PDT), an indicator that combines important environment factors, such as temperature and photoperiod, as the driving variable. Compared to the model of calendar-based development time (CDT), an indicator counting the days that elapse after full bloom, we confirmed that the PDT model improved the estimation accuracy to within 0.2 cm for fruit diameter and 0.1 cm for fruit length in independent years using a similar data collection method in 2013. The PDT model was implemented to realize a web-based management information system for a digital orchard, and the digital system had been applied in Shandong Province, China since 2013. This system may be used to compute the dynamic curve of apple fruit size based on data obtained from a nearby weather station. This system may provide an important decision support for farmers using the website and short message service to optimize crop production and, hence, economic benefit.     

Aberrant Expression of Osteopontin and E-Cadherin Indicates Radiation Resistance and Poor Prognosis for Patients with Cervical Carcinoma

Radiotherapy is the first-line treatment for all stages of cervical cancer, whether it is used for radical or palliative therapy. However, radioresistance of cervical cancer remains a major therapeutic problem. Consequently, we explored if E-cadherin (a marker of epithelial-mesenchymal transition) and osteopontin could predict radioresistance in patients with locally advanced cervical squamous cell carcinoma (LACSCC). Patients were retrospectively reviewed and 111 patients divided into two groups (radiation-resistant and radiation-sensitive groups) according to progression-free survival (PFS). In pretreated paraffin-embedded tissues, we evaluated E-cadherin and osteopontin expression using immunohistochemical staining. The percentage of patients with high osteopontin but low E-cadherin expression in the radiation-resistant group was significantly higher than those in the radiation-sensitive group (p<0.001). These patients also had a lower 5-year PFS rate (p<0.001). Our research suggests that high osteopontin but low E-cadherin expression can be considered as a negative, independent prognostic factor in patients with LACSCC ([Hazard ratios (95% CI) 6.766 (2.940, 15.572)], p<0.001).