Molecular Cloning and Functional Characterization of Oxidosqualene Cyclases from Panax vietnamensis

Panax vietnamensis is a valuable medicinal resource with promising preclinical applications. Ginsenosides, which are triterpenoids, are the primary active components in P. vietnamensis. Oxidosqualene cyclases (OSCs) catalyze the formation of the basic skeleton of triterpenes from 2,3-oxidosqualene, which is a crucial step in the biosynthesis of triterpenoids. The OSCs involved in triterpenoid biosynthesis in P. vietnamensis have not yet been characterized. Four OSC genes (PvOSC1–4) were cloned from P. vietnamensis and functionally characterized via heterologous expression in yeast. Transgenic yeast expressing PvOSC1, PvOSC3, and PvOSC4 produced the corresponding products β-amyrin, cycloartenol, and dammarenediol-II, respectively. PvOSC1, PvOSC3, and PvOSC4 are monofunctional OSCs. In this study, we characterized three PvOSC genes, providing a better understanding of the biosynthesis of triterpenoids in P. vietnamensis and the multiple choices of plant OSCs for metabolic engineering in yeast and other hosts.     

摄食水平和食物种类对金鱼生长及氮、磷排泄的影响

本文报道了金鱼的生长率、氨排泄率及磷酸盐排泄率的两个实验的结果。实验A用牛肉作饵料,实验B用配合饲料作饵料。每一实验分6个摄食水平。特定生长率(SGR)与摄食率(R)的关系为:SGR=a十bln(R+1)。实验中两种食物对上式的系数无显著影响。氨排泄率NE与氮摄取率NI的关系为:NE=a+bNI。实验中两种食物对上式的‘b’值无显著影响,但两个实验得出的‘a’值有显著差异。磷酸盐排泄率(PE)与磷摄取率(PI)的关系为:PE=a+bPI。实验中两种食物对上式中的‘b’值无显著影响,但得出的‘a’值有显著差异。若不考虑食物种类的影响,而将两实验数据合并,氮、磷摄取率可分别解释氮、磷排泄率变异的98％和75.7％。     

Dendritic Cells Pulsed with Exosomes in Combination with PD-1 Antibody Increase the Efficacy of Sorafenib in Hepatocellular Carcinoma Model

Advanced hepatocellular carcinoma (HCC) has limited therapeutic options. Immunotherapy is a promising treatment, while sorafenib is a first-line drug-based treatment for advanced HCC. However, the efficacy of sorafenib and immunotherapy in combination, have not been clearly evaluated. Sorafenib treatment has been shown to promote immunosuppression by increasing hypoxia in orthotopic HCC models. Here, we found that sorafenib treatment in mice with orthotopic HCC increased the expression of inhibitor programmed death-ligand 1 (PD-L1) and T-regulatory cells in tumor tissues. We pulsed dendritic cells with exosomes derived from tumor cells (DC-TEX) and found that the number of T-regulatory cells decreased and the number of CD8+T cells increased. However, combining DC-TEX and sorafenib did not prolong survival in these mice. Moreover, we found that the number of PD-1+CD8+T cells significantly increased after DC-TEX treatment. Therefore, we next added PD-1 antibody (PD-1 Ab) to the treatment regimen to block the PD-1/PD-L1 pathway, and found that the exhausted CD8+T cells were restored, without affecting the number of T-regulatory cells. Thus, our data suggest that the combination of DC-TEX and PD-1 Ab enhanced the efficacy of sorafenib, but treatment with either DC-TEX or PD-1 Ab alone, did not.     

世界毁灭性检疫害虫番茄潜叶蛾的生物生态学及危害与控制

番茄潜叶蛾原产于南美洲的秘鲁,严重危害多种茄科作物,是最具毁灭性的世界检疫性害虫,严重发生地块番茄减产80%~100%。该虫主要借助农产品的贸易活动进行远距离传播扩散。截至2017年,番茄潜叶蛾已在全世界的85个国家和地区发生(以及在22个国家和地区疑似发生)。目前,我国尚没有该虫分布,然而其一旦入侵,将对我国的番茄和马铃薯产业构成巨大威胁。从番茄潜叶蛾寄主范围、危害特点及造成的经济损失、生物生态学习性、地理分布和传播扩散途径、防控措施等方面进行概述,以期为该虫的有效防范提供参考。     

Membrane-tethered Notch1 exhibits oncogenic property via activation of EGFR–PI3K–AKT pathway in oral squamous cell carcinoma

Notch proteins are highly conserved cell surface receptors which play essential roles in cellular differentiation, proliferation, and apoptotic events at all stages of development. Recently, NOTCH1 mutations have been extensively observed in oral squamous cell carcinoma (OSCC) and are hinted to be Notch1-inactivating mutations. However, little is known about the biological effect of these reported mutations in OSCC. To mimic the inactivation of Notch1 due to inappropriate mutations and to determine the potential mechanisms, we utilized wild-type Notch1 vectors (Notch1^WT) or mutant Notch1 vectors (Notch1^V1754L) to transfect into OSCC cell lines. Membrane-tethered Notch1 induced by mutation was analyzed by immunofluorescence staining. γ-Secretase inhibitor PF-03084014 was utilized to determine the phenotype in the absence of endogenous Notch1 activation. Here we demonstrated that membrane-tethered Notch1 inactivated the canonical Notch1 signaling and oncogenic phenotypes were identified by promoting cell proliferation and invasion and by inducing epithelial-to-mesenchymal transition in cells. The γ-secretase inhibitor PF-03084014 also showed distinct oncogenic property after treatment. Importantly, both membrane-tethered Notch1 and PF-03084014 inhibitor activated the epidermal growth factor receptor (EGFR)–phosphoinositide 3-kinase (PI3K)–protein kinase B (AKT) signaling pathway, which has been confirmed as an overwhelming modulator in OSCC. This was the first time that we clearly simulated the mutated Notch1 activities and determined the oncogenic phenotypes of membrane-tethered Notch1. Compared with wild-type Notch1, membrane-tethered Notch1 was strongly associated with activated EGFR–PI3K–AKT signaling pathway.