Comparative metabolic activity related to flavonoid synthesis in leaves and flowers of Chrysanthemum morifolium in response to K deficiency

K limitation could decrease the flavonoids content in many Chinese traditional herbs. These results may be caused by the influence of K deficiency on the synthesis pathway of flavonoids. In this paper, we aim to study the influence of K deficiency on secondary metabolites and activities of phenylalanine ammonia lyase (PAL), 4-coumarate coenzyme A ligase (4CL) and cinnamate 4-hydroxylase (C4H) enzymes in the process of flavonoid synthesis in Chrysanthemum morifolium Ramat. The results show that K deficiency decreased the flavonoid and chlorogenic acid contents slightly in flower of C. morifolium while the same effect was obtained on C4H and PAL. Total flavonoids content increased with the content of cinnamic acid and p-coumaric acid in the plant under K deficiency, respectively. The regression equations between content of flavonoids and cinnamic acid/ p-coumaric acid should be expressed in term of linear effects (R²?=?0.9809, P?<?0.001 for cinnamic acid and R²?=?0.9929, P?<?0.0001 for p-coumaric acid, respectively). But the effect of phenylalanine on flavonoids should be expressed in term of quadratic pattern (R²?=?0.9375, P?<?0.05). There were two kinds principal components from chorismate to coumaryl CoA synthesis process, principal 1 was the substrate (phenylalanine, cinnamic acid, p-coumaric acid and PAL) and principal 2 was the enzyme (4CL, C4H), the principal 1 was the domination principal in both K deficient (88.36% and 10.57%) and sufficient treatments (88.17% and 9.64%), however, the factor loading (correlation coefficient of measured targets and the corresponding principal component) in principal 1 was opposite under different K application.     

An improved intracellular staining protocol for efficient detection of nuclear proteins in YFP-expressing cells

Intracellular staining is a widely used flow cytometry (FCM)-based technique to detect the expression of cytoslio nucleic antigens. However, intracellular staining of cells expressing cytosolic fluorescent protein (FP) markers was proven to be problematic as significant loss of the FP-signal was routinely observed. Using splenocytes harvested from mice constitutively expressing the enhanced yellow fluorescent proteins (YFP) as a model, we modified the widely used intracellular staining protocol and successfully achieved simultaneous detection of both the nuclear proteins and YFP in T-regulatory cells. The improved protocol can be used to perform antibody-based intracellular characterization of FP-labeled target cells, while maintaining their fluorescent reporter signals for easy tracing and identification.     

Dysfunction of myocardial sarcoplasmic reticulum in rats with myocardial calcification

We investigated the relationship between cardiac dysfunction and Ca2+ transport in the myocardial sarcoplasmic reticulum (SR) during the pathogenesis of cardiovascular calcification in rats. The possible mechanism of SR dysfunction was explored by detecting the alteration of the nitric oxide/nitric oxide synthase (NO/NOS) pathway in the SR. Using the vitamin D plus nicotine (VDN treatment for 2 week and 6 week) experimental model of cardiac calcification, cardiac function and sarcoplasmic reticulum function were measured. Inhibition of cardiac functions in vivo (peak rate of contraction and peak rate of relaxation, P < 0.05 or P < 0.01) were observed in all calcification groups, simultaneously, Ca2+ release and uptake in the SR as well as the Ca2+ release channel and Ca2+ pump activity were inhibited. Myocardial Ca2+ concentration and cardiac and SR dysfunction were inversely related (P < 0.05). The specific NO/NOS pathway (NO production, NOS activity and nNOS expression in the SR) was upregulated in the SR and associated with calcification (both 2- and 6 week VDN groups). These results indicate that cardiac dysfunction associated with myocardial calcification might be mediated by SR dysfunction, which may result from an impaired SR-specific NO/NOS pathway.     

神经迁移因子在血管系统中的表达与功能

神经迁移因子是近10年来在发育神经生物学中的研究热点,主要由ephrin、neuropilin、Slit和netrin四大家族成员构成,其主要功能是吸引或排斥神经元轴突的迁移,在神经系统中发挥着重要作用。现在,越来越多的实验证据表明：神经迁移因子的作用不仅仅局限在神经系统发育过程中,在血管发生或新生血管形成中同样具有不可替代的功能。     

Tumor regression induced by intratumor therapy with a disabled infectious single cycle (DISC) herpes simplex virus (HSV) vector, DISC/HSV/murine granulocyte-macrophage colony-stimulating factor, correlates with antigen-specific adaptive immunity

Direct intratumor injection of a disabled infectious single cycle HSV-2 virus encoding the murine GM-CSF gene (DISC/mGM-CSF) into established murine colon carcinoma CT26 tumors induced a significant delay in tumor growth and complete tumor regression in up to 70% of animals. Pre-existing immunity to HSV did not reduce the therapeutic efficacy of DISC/mGM-CSF, and, when administered in combination with syngeneic dendritic cells, further decreased tumor growth and increased the incidence of complete tumor regression. Direct intratumor injection of DISC/mGM-CSF also inhibited the growth of CT26 tumor cells implanted on the contralateral flank or seeded into the lungs following i.v. injection of tumor cells (experimental lung metastasis). Proliferation of splenocytes in response to Con A was impaired in progressor and tumor-bearer, but not regressor, mice. A potent tumor-specific CTL response was generated from splenocytes of all mice with regressing, but not progressing tumors following in vitro peptide stimulation; this response was specific for the gp70 AH-1 peptide SPSYVYHQF and correlated with IFN-gamma, but not IL-4 cytokine production. Depletion of CD8(+) T cells from regressor splenocytes before in vitro stimulation with the relevant peptide abolished their cytolytic activity, while depletion of CD4(+) T cells only partially inhibited CTL generation. Tumor regression induced by DISC/mGM-CSF virus immunotherapy provides a unique model for evaluating the immune mechanism(s) involved in tumor rejection, upon which tumor immunotherapy regimes may be based.     

Effects of Salt-Alkaline Stress on Carbohydrate Metabolism in Rice Seedlings

The aim of this study was to investigate carbohydrate metabolism in rice seedlings subjected to salt-alkaline stress. Two relatively salt-alkaline tolerant (Changbai 9) and sensitive (Jinongda 138) rice cultivars, grown hydroponically, were subjected to salt-alkaline stress via 50 mM of salt-alkaline solution. The carbohydrate content and the activities of metabolism-related enzymes in the leaves and roots were investigated. The results showed that the contents of sucrose, fructose, and glucose in the leaves and roots increased under salt-alkaline stress. Starch content increased in the leaves but decreased in the roots under salt-alkaline stress. The activities of sucrose-phosphate synthase, sucrose synthase, amylase, and ADP-glucose pyrophosphorylase increased whereas the activities of neutral invertase and acid invertase decreased in the leaves under salt-alkaline stress. The activities of sucrose-phosphate synthase, sucrose synthase, amylase, neutral invertase, and acid invertase increased in the roots under salt-alkaline stress. In conclusion, salt-alkaline stress caused the accumulation of photosynthetic assimilates in the leaves and decreased assimilation export to the roots.     

Emerging Landscape of SARS-CoV-2 Variants and Detection Technologies

Molecular Diagnosis & Therapy - In 2019, a new coronavirus was identified that has caused significant morbidity and mortality worldwide. Like all RNA viruses, severe acute respiratory...     

广西宁明第三纪类黄杞翅果化石

广西宁明西南约4km处新发现的一个植物群中,含有一些与胡桃科黄杞属现生种的翅果在形态上近似的化石。它们的果翅三裂,裂片具有羽状脉序。这些化石的一部分特征与黄杞属的裸果黄杞组的相似,另一部分特征与该属的黄杞组的相似,因而难以将它们归于其中的任何一组,作者将其归于类黄杞属。后者是胡桃科黄杞族的一个器官属,与目前分布于我国长江以南及东南亚地区的黄杞属以及分布于美洲新大陆的美黄杞属有密切亲缘关系。在地史时期,类黄杞属的代表较广泛地分布于北半球。然而,当前化石却是这类化石在我国南方首次被发现,这为探讨黄杞族的演化提供了重要线索。从当前化石的研究和有关文献中得知,黄杞族祖先的翅果形态与裸果黄杞组的现生种可能有某种程度的近似。     

Downregulation of cPLA2γ expression inhibits EGF-induced chemotaxis of human breast cancer cells through Akt pathway

Phospholipids play an important role in mediating cell migration. In the present study, we investigated the role of cPLA₂γ in chemotaxis of human breast cancer cells. Inhibition of cPLA₂γ expression by small interference RNA severely inhibits EGF-induced chemotaxis in a dose-dependent manner in MDA-MB-231, MCF-7, T47D and ZR-75-30 cells. Furthermore, silencing cPLA₂γ expression also impaired directional migration, adhesion and invasion in MDA-MB-231 cells. In addition, we investigated the molecular mechanism by which cPLA₂γ regulated migration. Knockdown of cPLA₂γ suppressed the phosphorylation of Akt at both Thr308 and Ser473. Phosphorylation of PKCζ, downstream of Akt, was also dampened. Knockdown of cPLA₂γ also impaired the phosphorylation of integrin β1 and cofilin, key regulators of cell adhesion and actin polymerization, respectively. Taken together, our results suggest that cPLA₂γ plays an important role in cancer cell chemotaxis.