灰飞虱Bursicon基因的克隆、序列分析及在不同发育阶段的表达

Bursicon是通过G蛋白受体调节昆虫表皮硬化及展翅的功能蛋白, 它在昆虫蜕皮后的表皮硬化过程中起着关键作用。为探讨灰飞虱Laodelphax striatellus的 bursicon的功能, 利用RT-PCR和RACE技术克隆获得1 126 bp的bursicon α和761 bp的bursicon β全长序列, 将其分别命名为Lsburs-α和Lsburs-β。生物信息学分析表明： Lsburs-α开放阅读框长483 bp, 编码160个氨基酸, 该蛋白具有2个N-豆蔻酰化位点、 3个酪蛋白激酶Ⅱ磷酸化位点以及2个蛋白激酶C磷酸化位点。Lsburs-β开放阅读框长417 bp, 编码138个氨基酸, 该蛋白具有2个N-豆蔻酰化位点、 3个酪蛋白激酶Ⅱ磷酸化位点以及1个酪氨酸激酶磷酸化位点。qRT-PCR结果表明： Lsburs-α和Lsburs-β在灰飞虱各龄期均有转录表达, 并在若虫期随龄期增加呈上升趋势, 在羽化期达到峰值, 成虫期表达量逐渐降低。结果提示bursicon与灰飞虱蜕皮后的外表皮硬化关系密切。本文结果为深入研究bursicon的功能、受体调节和信号通路等奠定了基础。     

Development of a biosensor assessing SARS-CoV-2 main protease proteolytic activity in living cells for antiviral drugs screening

Highlights:
The biosensor reported in our study can monitor SARS-CoV-2 M^pro activity in living cells instead of in vitro solutions.
The biosensor reported in our study is sensitive and easy to operate.
It is suitable for high-throughput screening.
It has the potential to be used in small animal models.     

Comparison of bioethanol production from cultivated versus wild <Emphasis Type="Italic">Gracilaria verrucosa</Emphasis> and <Emphasis Type="Italic">Gracilaria gigas</Emphasis>

The seaweed genus Gracilaria is a potential candidate for the production of bioethanol due to its high carbohydrate content. Gracilaria is abundant throughout the world and can be found in both wild and cultivated forms. Differences in the ecological factors such as temperature, salinity, and light intensity affecting wild and cultivated specimens may influence the biochemical content of seaweeds, including the carbohydrate content. This study aimed to investigate the proximate composition and potential bioethanol production of wild and cultivated G. gigas and G. verrucosa. Bioethanol was produced using separate hydrolysis fermentation (SHF), employing a combination of enzymatic and acid hydrolysis, followed by fermentation with Saccharomyces cerevisiae ATCC 200062. The highest carbohydrate content was found in wild G. gigas. The highest galactose and glucose contents (20.21 ± 0.32 and 9.70 ± 0.49 g L^?1, respectively), as well as the highest production of bioethanol (3.56 ± 0.02 g L^?1), were also found in wild G. gigas. Thus, we conclude that wild G. gigas is the most promising candidate for bioethanol production. Further research is needed to optimize bioethanol production from wild G. gigas. Domestication of wild G. gigas is a promising challenge for aquaculture to avoid overexploitation of this wild seaweed resource.     

Identification of novel benzimidazole derivatives as inhibitors of leukotriene biosynthesis by virtual screening targeting 5-lipoxygenase-activating protein (FLAP)

Pharmacological suppression of leukotriene biosynthesis by 5-lipoxygenase (5-LO)-activating protein (FLAP) inhibitors is a promising strategy to intervene with inflammatory, allergic and cardiovascular diseases. Virtual screening targeting FLAP based on a combined ligand- and structure-based pharmacophore model led to the identification of 1-(2-chlorobenzyl)-2-(1-(4-isobutylphenyl)ethyl)-1H-benzimidazole (7) as developable candidate. Compound 7 potently suppressed leukotriene formation in intact neutrophils (IC(50)=0.31 μM) but essentially failed to directly inhibit 5-LO suggesting that interaction with FLAP causes inhibition of leukotriene synthesis. For structural optimization, a series of 46 benzimidazole-based derivatives of 7 were synthesized leading to more potent analogues (70-72, 82) with IC(50)=0.12-0.19 μM in intact neutrophils. Together, our results disclose the benzimidazole scaffold bearing an ibuprofen fingerprint as a new chemotype for further development of anti-leukotriene agents.     

Tumor necrosis factor receptor-associated factor family protein 2 is a key mediator of the epidermal growth factor-induced ribosomal S6 kinase 2/cAMP-responsive element-binding protein/Fos protein signaling pathway

TRAF2 has an important function in mediating the TNF-R signaling pathway toward activation of NF-κB and JNKs. Here we reveal a novel function of TRAF2 in the epidermal growth factor (EGF) signaling pathway. Knockdown of TRAF2 blocked EGF-induced AP-1 activity and anchorage- independent cell transformation. Notably, we showed that EGF induces ribosomal S6 kinase 2 (RSK2) ubiquitination, and knocking down TRAF2 suppresses ubiquitination of RSK2 induced by EGF. We also found that TRAF2 affects RSK2 activity through RSK2 ubiquitination. RSK2 plays a critical role in AP-1 activity mediated through CREB and c-Fos, which regulates anchorage-independent cell transformation. In addition, TRAF2 is overexpressed in colon cancer and required for colon cancer development, suggesting that TRAF2 might be a potential molecular target for cancer prevention and treatment.