Interactions between thrombin and natural products of Millettia nitita var. hirsutissima using capillary zone electrophoresis

A sensitive and selective high-performance analytical method based on capillary zone electrophoresis (CZE) was developed for investigating interactions between natural products isolated from Millettia nitita var. hirsutissima and thrombin qualitatively and quantitatively for the first time. The results showed that, compared with positive and negative control, the compounds ZYY-5 (genistein-8-C-β-d-apiofuranosyl-(1→6)-O-β-d-glucopyranoside), ZYY-6 (calycosin), ZYY-8 (isoliquiritigenin), ZYY-9 (formononetin), ZYY-12 (gliricidin), ZYY-13 (8-O-methylretusin), FJ-2 (dihydrokaempferol), FJ-3 (biochanin), FJ-5 (afromosin) and XC-2 (hirsutissimiside F) interacted with thrombin, while ZYY-1 (sphaerobioside), ZYY-2 (formononetin-7-O-β-d-apiofuranosyl-(1→6)-O-β-d-glucopyranoside), ZYY-3 (genistein-5-methylether-7-O-α-l-rhamnopyranosyl-(1→6)-O-β-d-glucopyranoside), ZYY-4 (retusin-7,8-O-β-d-diglucopyranoside), ZYY-7 (symplocoside), ZYY-10 (ononin), ZYY-11 (genistin), ZYY-14 (afromosin-7-O-β-d-glucopyranoside), ZYY-15 (lanceolarin), FJ-1 (liquiritigenin), FJ-4 (7,2-dihydroxy,4-methoxyisoflavan) and XC-1 (sphaerobioside) had no binding to thrombin. This indicated that the reported CZE method for the determination of compound–thrombin interactions is powerful, sensitive and fast, and requires less amounts of reagents, and further, it can be employed as a reliable alternative to other methods.     

CADASIL mutant NOTCH3(R90C) decreases the viability of HS683 oligodendrocytes via apoptosis

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the most common hereditary cerebral small vessel disease caused by mutations in NOTCH3. Prevailing models suggest that demyelination occurs secondary to vascular pathology. However, in zebrafish, NOTCH3 is also expressed in mature oligodendrocytes. Thus, we hypothesized that in addition to vascular defects, mutant NOTCH3 may alter glial function in individuals with CADASIL. The aim of this study was to characterize the direct effects of a mutant NOTCH3 protein in HS683 oligodendrocytes. HS683 oligodendrocytes transfected with wild-type NOTCH3, mutant NOTCH3(R90C), and empty control vector were used to study the impact of the NOTCH3(R90C) mutant on its protein hydrolytic processing, cell viability, apoptosis, autophagy, oxidative stress, and the related upstream events using immunoblotting, immunofluorescence, RT-PCR, and flow cytometry. We determined that HS683 oligodendrocytes transfected with mutant NOTCH3(R90C), which is the hotspot mutation site-associated with CADASIL, exhibited aberrant NOTCH3 proteolytic processing. Compared to cells overexpressing wild-type NOTCH3, cells overexpressing NOTCH3(R90C) were less viable and had a higher rate of apoptosis. Immunoblotting revealed that cells transfected with NOTCH3(R90C) had higher levels of intrinsic mitochondrial apoptosis, extrinsic death receptor path-related apoptosis, and autophagy compared with cells transfected with wild-type NOTCH3. This study suggests that in patients with CADASIL, early defects in glia influenced by NOTCH3(R90C) may directly contribute to white matter pathology in addition to secondary vascular defects. This study provides a potential therapeutic target for the future treatment of CADASIL.     

植物中CRISPR/Cas系统的应用研究进展

成簇规律间隔短回文重复—CRISPR(clustered regularly interspaced short palindromic repeat)是近年来新兴的基因组编辑技术,因其操作简便、靶向效率高和应用范围广而备受关注。国内外关于CRISPR/Cas系统在细菌和动物研究中的应用报道较多,但对其在植物研究中的应用报道较少。该文综述了CRISPR/Cas系统的发现、结构、作用机制及其在植物研究中的应用,以期为CRISPR/Cas系统在提高农作物产量及培育抗性品种等方面的应用研究奠定基础。     

A novel splicing variant of proprotein convertase subtilisin/kexin type 9

Proprotein convertase subtilisin/kexin type 9 (PCSK9) is the most recently identified member of the proprotein convertase family. Genetic and cell biology studies have suggested a critical role of PCSK9 in regulating low-density lipoprotein receptor (LDLR) protein levels and thus modulating plasma LDL cholesterol. Recent data on the molecular basis for PCSK9 action support the model in which PCSK9 is self-cleaved, secreted, and tightly bound to the EGF-A repeat of LDLR extracellular domain. PCSK9 binding to LDLR is essential for the ensuing receptor-mediated endocytosis and is speculated to lock LDLR in a specific conformation that favors degradation in lysosomal compartment instead of recycling back to plasma membrane. We report here a novel human PCSK9 splicing variant, which we named PCSK9sv. PCSK9sv had an in-frame deletion of the eighth exon of 58 amino acids and was expressed in multiple tissues, including liver, small intestine, prostate, uterus, brain, and adipose tissue. Unlike wild-type PCSK9, which is secreted, PCSK9sv expressed in human embryonic kidney HEK293 cells failed to process the prosegment intracellularly and thus was not secreted into the medium. Examination of potential functions revealed that PCSK9sv did not change the LDLR protein levels. Two mutations that have been reported in humans with the associated changes in plasma LDL cholesterol were within exon 8, and thus the expression and function of the two mutants were studied. Both N425S and A443T mutants were processed normally, secreted, and reduced LDLR levels. However, the physiological function of this novel splicing variant of PCSK9 has yet to be determined.     

Effects of radix polygoni multiflori components on tyrosinase activity and melanogenesis

Radix Polygoni multiflori is a herb used effectively to prevent graying and treat skin depigmentation diseases in traditional Chinese medicine but its active ingredients have not been discovered yet. In this investigation, we tested six compounds isolated from Radix Polygoni multiflori, to discover the active component on melanogenesis. Three experiments were performed in the present investigation: mushroom tyrosinase activity, melanin content B16 cell proliferation assay. Among all the six components tested, THSG showed the most potent effects on tyrosinase activation and melanogenesis; it was shown to be a potent tyrosinase activator and a melanogenesis stimulator in this study. On the other hand, we found that gallic acid significantly inhibited tyrosinase and, in addition, anthraquinones were cytotoxic to melanoma cells. They were both harmful to melanogenesis. Therefore, we propose that THSG acts as the active ingredient of Radix Polygoni multiflori on melanogenesis.     

Retrospective analysis of protein kinase C-beta (PKC-β) expression in lymphoid malignancies and its association with survival in diffuse large B-cell lymphomas

Background  

Both mechanistic features and recent correlative findings suggest a potential role for protein kinase C-beta (PKC-β) in tumor pathogenesis, particularly in B-cell malignancies. To evaluate the role of this gene in lymphoid malignancies, we analyzed global gene expression data to quantify PKC-β expression across diagnostic groups and, when possible, determined correlations between PKC-β expression and survival.     

瘤足蕨属(Plagiogyria)植物叶柄和羽片横切面解剖学特征及分类学意义

该文采用光学显微镜,首次对九种瘤足蕨属植物的叶柄和羽片横切面进行了解剖学比较研究。结果表明:九种瘤足蕨属植物在叶柄中部横切面的表皮、基本组织和中柱以及营养叶羽片的表皮、叶肉和主脉等结构特征是相似的,如叶柄中部横切面的表皮不被毛或是鳞片,表皮细胞形状呈类圆形,具有厚壁组织,中柱为周韧型维管束;营养叶羽片横切面的气孔只分布于下表皮,表皮细胞形状呈扁平状,主脉的结构类型属于周韧型维管束。在系统的演化中,瘤足蕨与桫椤科植物有一定的亲缘关系,两者既有相似的特征但也表现出一定的差异,支持瘤足蕨属是一个自然分类的观点。九种瘤足蕨属植物叶柄中部横切面形状有梯形、椭圆形和三角形,叶柄中部横切面维管束的形状成"U"字形、"V"字形或"品"字形;维管束数目1个或3个;维管束结构中的木质部成"八"字形、"U"字形或"一"字形;羽片主脉下侧形成了以三角形、弧形或梯形的突起;叶柄中的木质部形态有两种,即典型的海马形状以及非典型的海马形状,其中典型的海马形状的特点为叶柄的木质部两侧都呈现弯曲成钩的形态,非典型的海马形状特点为2个木质部束的两侧都未出现弯曲成钩的形态或者是其中的一个木质部一端无弯曲成钩的形态,这些形态解剖特征稳定且具类群特异性,为瘤足蕨属植物的分类和系统学研究提供了新的依据。同时,依据其叶柄和羽片横切面解剖叶柄学特征列出了瘤足蕨属植物分种检索表。