Chinese cabbage: an emerging model for functional genomics in leafy vegetable crops

Leafy vegetable crops (LVCs) are consumed worldwide and offer essential nutrients for humans. Unlike model plant species, systematic characterisation of gene function is lacking, although whole-genome sequences (WGSs) are available for various LVCs. Several recent studies in Chinese cabbage have reported high-density mutant populations linking genotype to phenotype, providing blueprints for functional LVC genomics and beyond.     

Understanding chaperone specificity: evidence for a ‘client code’

The interactions of molecular chaperones with clients can be regulated by chaperone post-translational modification (PTMs) collectively known as the ‘chaperone code’. What is less understood is how PTMs on client proteins may impact chaperone–client interactions. In this forum, we discuss the possibility of a ‘client code’.     

Virtual mutation and directional evolution of anti-amoxicillin ScFv antibody for immunoassay of penicillins in milk

In this study, an anti-amoxicillin single chain variable fragment (ScFv) antibody was evolved by directional mutagenesis of a contact amino acid residue based on the analysis of virtual mutation. Comparison with its parental ScFv, the mutant showed highly improved affinity for 11 penicillins with up to 6-folds increased sensitivity. Then, its recognition mechanisms for the 11 drugs were studied by using molecular docking. Results showed that the mutant-penicillins intermolecular forces increased and the total binding energies decreased dramatically, which were responsible for the improvement of antibody sensitivity. The ScFv mutant was used to develop an indirect competitive enzyme linked immunosorbent assay for determination of the 11 drugs in milk. The limits of detection were in the range of 0.2–3.0 ng/mL, the crossreactivities were in the range of 31%–132%, and the recoveries from standards fortified blank milk were in the range of 65.7%–92.4%. This is the first study reporting the directional evolution of a ScFv antibody based on virtual mutation and the use of ScFv antibody for determination of penicillins in foods of animal origin.     

Copeptin,adropin and irisin concentrations in breast milk and plasma of healthy women and those with gestational diabetes mellitus

Copeptin, adropin and irisin are polypeptide hormones implicated in energy homostasis and diabetes. The purposes of this study were (1) to compare the copeptin, adropin and irisin concentrations between colostrum, transitional and mature milk and plasma in lactating women with and without GDM and (2) to compare these values with those from non-lactating women. Venous blood samples were obtained before suckling from 15 healthy lactating women aged 26–30 years, 15 lactating women with GDM aged 26–32 years, and 14 age-matched controls aged 25–31 years. Colostrum, transitional milk and mature milk samples were collected just before suckling. The concentration of copeptin was determined by EIA while the concentrations of adropin and irisin were determined by ELISA. The levels of copeptin, adropin and irisin in the colostrum were significantly higher than those in transitional and mature milk samples from healthy women; also, transitional milk had higher copeptin, adropin and irisin concentrations than mature milk. The amounts of copeptin in the colostrum and transitional milk were significantly higher than in mature milk samples from women with GDM, while the amounts of adropin and irisin were significantly lower. The relative concentrations of copeptin, adropin and irisin in the plasma samples from these groups of women were similar to those in the colostrum, transitional and mature milk samples, but the latter concentrations were higher than those in the plasma. These peptides could influence the regulation of metabolic pathways and the postnatal growth and development of different organs in the newborn.     

Long non-coding RNA 657 suppresses hepatocellular carcinoma cell growth by acting as a molecular sponge of miR-106a-5p to regulate PTEN expression

Previous study has identified the aberrant expression of LINC00657, a long non-coding RNA (lncRNA), in human breast cancer. However, the expression pattern, biological function and underlying mechanism of LINC00657 in human hepatocellular carcinoma (HCC) remain obscure. The expression levels of LINC00657 in HCC tissues and cell lines were determined by quantitative real-time PCR. CCK-8 assay, cell colony formation assay, cell cycle analysis, Transwell assay were performed to determine whether LINC00657 could affect HCC progression. Luciferase reporter assay was used to assess the target of LINC00657. Expressions of the relevant proteins were analyzed by Western blot. Herein, we found that LINC00657 was downregulated in HCC tissue specimens as well as in malignant HCC cell lines. LINC00657 overexpression inhibited the proliferation, migration and invasion of HCC cells, while LINC00657 depletion promoted both cell viability and cell invasion in vitro. We also found that LINC00657 could inhibit tumor growth in vivo. Further experiments demonstrated that down-regulated LINC00657 increased the expression of miR-106a-5p. miR-106a-5p decreased the abundances of PTEN protein, while had no impact on PTEN mRNA. Moreover, we identified that both LINC00657 and PTEN mRNA were targets of miR-106a-5p by using dual-luciferase reporter assay. Our results provide the new evidence supporting the tumor-suppressive role of LINC00657 in HCC, suggesting that LINC00657 might play a role in HCC and can be a novel therapeutic target for treating HCC.     

Characterization of a naringinase from Aspergillus oryzae 11250 and its application in the debitterization of orange juice

Naringinase plays a rather important role in reducing the bitterness of juice by hydrolyzing naringin. A novel extracellular naringinase was purified from Aspergillus oryzae 11250 cultured in the presence of orange peel. A 26.78-fold purification rate was achieved by salt-induced precipitation, followed by anion-exchange and gel filtration chromatography with 32% recovery and specific activity of 2194.62 units per mg protein (U/mg). The optimum pH and temperature for naringinase activity were 5.0 and 45 °C, respectively. This enzyme was stable at 30 °C for 5 h. The K_m and V_max of naringinase toward naringin determined by Lineweaver-Burk method were 1.60 ± 0.13 mM and 126.21 ± 5.52 μmol/(min mg), respectively. The enzyme activity was inhibited completely by Ag⁺ at 10 mM. Naringinase is capable of hydrolyzing naringin, neohesperidin, and some other glycosides. A supplement of 6 U/mL of this naringinase in citrus juice sufficiently removed naringin to relieve the bitterness of citrus juice. These properties make the enzyme an ideal candidate for commercial application in the debitterization of orange juice.     

降压通络方对大鼠肾小管上皮细胞凋亡的影响

目的:研究降压通络方对大鼠肾小管上皮细胞凋亡的影响。方法:体外培养大鼠肾小管上皮细胞,经过缺血缺氧,将大鼠肾小管细胞随机分为4组:正常组、模型组、降压通络方组、缬沙坦组。采用CCK-8检测细胞增殖情况,免疫组化法检测大鼠肾小管上皮细胞p-AKT蛋白的表达,蛋白免疫印迹法检测大鼠肾小管上皮细胞凋亡相关基因调控蛋白Bcl-2、Bax的表达,对Bcl-2、Bax蛋白的表达水平进行相关性分析。结果:缺血缺氧呈时间依赖性抑制大鼠肾小管上皮细胞活性;免疫组化结果示:p-AKT在正常组呈高表达,模型组低表达,降压通络方组p-AKT表达明显高于模型组(P0.01),缬沙坦组表达低于模型组(P0.05);蛋白免疫印迹法结果示:模型组Bcl-2表达较正常组明显降低(P0.01),降压通络方组和缬沙坦组Bcl-2的表达均高于模型组(P0.05,P0.01);模型组Bax表达较正常组明显升高(P0.01),降压通络方组和缬沙坦组Bax的表达均较模型组降低(P0.05);Bcl-2蛋白与Bax蛋白表达呈负相关性(r=-0.811,P0.01)。结论:降压通络方可抑制大鼠肾小管上皮细胞凋亡,其作用机制可能与上调p-AKT、Bcl-2蛋白,下调Bax蛋白表达有关。     

Phytochromes and flowering: legumes do it another way

Phytochromes have a crucial role in the regulation of flowering in many plants, but the underlying molecular mechanisms vary among species. Recently, Lin et al. described a unique phytochrome A (phyA)-controlled photoperiodic flowering pathway in soybean (Glycine max), revealing a novel mechanism for photoperiodic regulation of flowering.     

PBX1 attenuates 6-OHDA-induced oxidative stress and apoptosis and affects PINK1/PARKIN expression in dopaminergic neurons via FOXA1

Cytotechnology - PBX1 expression has been found to be significantly reduced in nigrostriatal neurons of PD patients, but the effect of PBX1 on ROS and apoptosis in nigrostriatal dopamine neurons is...