Screening and functional analysis of StMYB transcription factors in pigmented potato under low-temperature treatment.

MYB transcription factors play an extremely important regulatory role in plant responses to stress and anthocyanin synthesis. Cloning of potato StMYB-related genes can provide a theoretical basis for the genetic improvement of pigmented potatoes. In this study, two MYB transcription factors, StMYB113 and StMYB308, possibly related to anthocyanin synthesis, were screened under low-temperature conditions based on the low-temperature-responsive potato StMYB genes family analysis obtained by transcriptome sequencing. By analyzed the protein properties and promoters of StMYB113 and StMYB308 and their relative expression levels at different low-temperature treatment periods, it is speculated that StMYB113 and StMYB308 can be expressed in response to low temperature and can promote anthocyanin synthesis. The overexpression vectors of StMYB113 and StMYB308 were constructed for transient transformation tobacco. Color changes were observed, and the expression levels of the structural genes of tobacco anthocyanin synthesis were determined. The results showed that StMYB113 lacking the complete MYB domain could not promote the accumulation of tobacco anthocyanins, while StMYB308 could significantly promote the accumulation involved in tobacco anthocyanins. This study provides a theoretical reference for further study of the mechanism of StMYB113 and StMYB308 transcription factors in potato anthocyanin synthesis.

Fecal microbiota transplantation plus tislelizumab and fruquintinib in refractory microsatellite stable metastatic colorectal cancer: an open-label, single-arm, phase II trial (RENMIN-215).

Background: Immunotherapy has revolutionized the treatment of cancer. However, microsatellite stable (MSS) metastatic colorectal cancer (mCRC) shows a low response to PD-1 inhibitors. Antiangiogenic therapy can enhance anti-PD-1 efficacy, but it still cannot meet clinical needs. Increasing evidence supported a close relationship between gut microbiome and anti-PD-1 efficacy. This study aimed to explore the efficacy and safety of the combination of fecal microbiota transplantation (FMT) and tislelizumab and fruquintinib in refractory MSS mCRC. Methods: In the phase II trial, MSS mCRC patients were administered FMT plus tislelizumab and fruquintinib as a third-line or above treatment. The primary endpoint was progression-free survival (PFS). Secondary endpoints were overall survival (OS), objective response rate (ORR), disease control rate (DCR), duration of response (DoR), clinical benefit rate (CBR), safety and quality of life. Feces and peripheral blood were collected for exploratory biomarker analysis. This study is registered with Chictr.org.cn, identifier ChiCTR2100046768. Findings: From May 10, 2021 to January 17, 2022, 20 patients were enrolled. Median follow-up was 13.7 months. Median PFS was 9.6 months (95% CI 4.1-15.1). Median OS was 13.7 months (95% CI 9.3-17.7). Median DoR was 8.1 months (95% CI 1.7-10.6). ORR was 20% (95% CI 5.7-43.7). DCR was 95% (95% CI 75.1-99.9). CBR was 60% (95% CI 36.1-80.9). Nineteen patients (95%) experienced at least one treatment-related adverse event (TRAE). Six patients (30%) had grade 3-4 TRAEs, with the most common being albuminuria (10%), urine occult blood (10%), fecal occult blood (10%), hypertension (5%), hyperglycemia (5%), liver dysfunction (5%), hand-foot skin reaction (5%), and hypothyroidism (5%). No treatment-related deaths occurred. Responders had a high-abundance of Proteobacteria and Lachnospiraceae family and a low-abundance of Actinobacteriota and Bifidobacterium. The treatment did not change the structure of peripheral blood TCR repertoire. However, the expanded TCRs exhibited the characteristics of antigen-driven responses in responders. Interpretation: FMT plus tislelizumab and fruquintinib as third-line or above treatment showed improved survival and manageable safety in refractory MSS mCRC, suggesting a valuable new treatment option for this patient population. Funding: This study was supported by the National Natural Science Foundation of China (82102954 to Wensi Zhao) and the Special Project of Central Government for Local Science and Technology Development of Hubei Province (ZYYD2020000169 to Yongshun Chen).

Different doses of UV-B radiation affect pigmented potatoes' growth and quality during the whole growth period.

Introduction: UltraViolet- Biological (UV-B) plays an important role in plant growth and the formation of nutrients, especially secondary metabolites. Methods: To investigate the phenotypic changes, physiological responses, and internal genes expression of potatoes under enhanced UV-B radiation, two Yunnan native pigmented potatoes varieties named "Huaxinyangyu" and "Jianchuanhong" were exposed to different UV-B doses during whole growth duration. Results: Pearson correlation analysis and principal component analysis showed that the agronomic characters (i.e. plant height, pitch, stem diameter, and root shoot ratio) of plants treated with low dose ultraviolet (T1) did not change significantly compared with the absence of ultraviolet radiation (CK), even unit yield increased slightly; Similarly, under low UV-B radiation, photosynthetic and physiological parameters (photosynthetic rate, stomatal conductance, respiration rate, and transpiration rate) of leaves were significantly increased. In addition, low-dose UV-B treatment promoted the synthesis of tuber nutrients (e.g. phenols, chlorogenic acids, flavonoids, vitamin C, anthocyanins) and increased the expression of structural genes for anthocyanin synthesis. The number of nutrients and gene expression in tubers raised by the "Huaxinyangyu" was the highest at 84 days, and "Jianchuanhong" was the highest at 72 days. However, the higher dose of UV-B radiation (T2) will cause greater damage to the pigmented potatoes plants, making the plants reduce the yield, and significantly reduce the tuber nutrients. Discussion: This study showed that proper ultraviolet radiation will not harm pigmented potatoes, but also improve their oxidative stress tolerance, increase the structure genes expression of anthocyanins and continuously synthesize beneficial substances to improve the yield and quality of potato tubers.

Paeonin extracted from potatoes protects gastric epithelial cells from H2O2-induced oxidative damage in vitro by PI3K/Akt-mediated Nrf2 signaling pathway.

In this study, it is aimed to investigate the antioxidant mechanism of new extracts from potatoes. Four pigments, namely, Petunin, Paeonin, Malvidin and Pelargonidin, were extracted from potatoes by high performance liquid chromatography (HPLC). Our results showed that the cellular morphology and cell viability were significantly altered in gastric mucosal epithelial cells (GES-1) treated with different hydrogen peroxide (H2O2) concentrations over time (P < 0.05). Paeonin presented the strongest anti-oxidative effects on H2O2-treated cells, in both a dose- and time-dependent manner, determined by ARE-luciferase activity and HO-1 mRNA expression. After pre-treatment with Paeonin in H2O2-exposed cells, Keap1, Nrf2, HO-1 and NQO1 protein expressions were remarkably up-regulated. Furthermore, immunostaining of Nrf2 expression was obviously elevated in the H2O2 + Paeonin group over time. The GSH content in the H2O2 + Paeonin group was notably lower than that in the H2O2 + Paeonin + GSK690693 group. Paeonin promoted cell cycle with augmented Cyclin D1 and p27 protein expressions. Moreover, Paeonin suppressed apoptosis with increased Bcl2, total Caspase3 and total Caspase8 protein expressions and decreased Bax, p-Caspase3 and p-Caspase8 protein expression in H2O2-treated cells. These results suggested that Paeonin might exert an anti-oxidative role by activating Nrf2 signaling pathway with the changes of cell cycle and apoptosis.

A novel tumor-promoting role for nuclear factor IA in glioblastomas is mediated through negative regulation of p53, p21, and PAI1.

Background Nuclear factor IA (NFIA), a transcription factor and essential regulator in embryonic glial development, is highly expressed in human glioblastoma (GBM) compared with normal brain, but its contribution to GBM and cancer pathogenesis is unknown. Here we demonstrate a novel role for NFIA in promoting growth and migration of GBM and establish the molecular mechanisms mediating these functions. Methods To determine the role of NFIA in glioma, we examined the effects of NFIA in growth, proliferation, apoptosis, and migration. We used gain-of-function (overexpression) and loss-of-function (shRNA knockdown) of NFIA in primary patient-derived GBM cells and established glioma cell lines in culture and in intracranial xenografts in mouse brains. Results Knockdown of native NFIA blocked tumor growth and induced cell death and apoptosis. Complementing this, NFIA overexpression accelerated growth, proliferation, and migration of GBM in cell culture and in mouse brains. These NFIA tumor-promoting effects were mediated via transcriptional repression of p53, p21, and plasminogen activator inhibitor 1 (PAI1) through specific NFIA-recognition sequences in their promoters. Importantly, the effects of NFIA on proliferation and apoptosis were independent of TP53 mutation status, a finding especially relevant for GBM, in which TP53 is frequently mutated. Conclusion NFIA is a modulator of GBM growth and migration, and functions by distinct regulation of critical oncogenic pathways that govern the malignant behavior of GBM.

A genome-wide microarray highlights the antiinflammatory genes targeted by oolong tea theasinensin A in macrophages.

Theasinensin A is one of the oolong tea theasinensins, which differ from green tea catechins and black tea theaflavins. In a previous study, we found that theasinesin A had a potential effect on antiinflammation since theasinensin A suppressed LPS-induced COX2 and PGE(2) production. To clarify the molecular mechanisms, we investigated the gene expression profiling in macrophage-like cells treated with theasinensin A through a genome-wide DNA microarray in the present study. Among 22,050 oligonucleotides, the expression levels of 406 genes were increased by ≥3-fold in LPS-activated RAW264 cells, 259 gene signals of which were attenuated by theasinensin A treatment (≥2-fold). Expression levels of 717 genes were decreased by ≥3-fold in LPS-activated cells, of which 471 gene signals were restored by theasinensin A treatment (≥2-fold). These genes were further categorized as "defense, inflammatory response, cytokines activities, and receptor activities," and some of them were confirmed by real-time polymerase chain reaction. Furthermore, pathways analysis revealed that theasinensin A regulated the relevant expression networks of chemokines, interleukins, and interferons to exert its antiinflammatory effects.

CCM3 signaling through sterile 20-like kinases plays an essential role during zebrafish cardiovascular development and cerebral cavernous malformations.

Cerebral cavernous malformation is a common human vascular disease that arises due to loss-of-function mutations in genes encoding three intracellular adaptor proteins, cerebral cavernous malformations 1 protein (CCM1), CCM2, and CCM3. CCM1, CCM2, and CCM3 interact biochemically in a pathway required in endothelial cells during cardiovascular development in mice and zebrafish. The downstream effectors by which this signaling pathway regulates endothelial function have not yet been identified. Here we have shown in zebrafish that expression of mutant ccm3 proteins (ccm3Delta) known to cause cerebral cavernous malformation in humans confers cardiovascular phenotypes identical to those associated with loss of ccm1 and ccm2. CCM3Delta proteins interacted with CCM1 and CCM2, but not with other proteins known to bind wild-type CCM3, serine/threonine protein kinase MST4 (MST4), sterile 20-like serine/threonine kinase 24 (STK24), and STK25, all of which have poorly defined biological functions. Cardiovascular phenotypes characteristic of CCM deficiency arose due to stk deficiency and combined low-level deficiency of stks and ccm3 in zebrafish embryos. In cultured human endothelial cells, CCM3 and STK25 regulated barrier function in a manner similar to CCM2, and STKs negatively regulated Rho by directly activating moesin. These studies identify STKs as essential downstream effectors of CCM signaling in development and disease that may regulate both endothelial and epithelial cell junctions.

[Relation between angiogenesis, fibrinolysis and invasion/metastasis in breast cancer].

OBJECTIVE: To investigate the relation between angiogensis, fibrinolysis and invasion/metastasis in breast cancer. METHODS: The expression of urokinase-type plasminogen activator (uPA) and microvascular density (MVD) was immunohistochemically studied in 110 patients with primary breast cancer. RESULTS: High uPA expression was found in 59 patients (53.6%), and weak expression in 51 patients (46.4%). Strong MVD expression was found in 53 patients (48.2%), and weak expression in 57 patients (51.8%). The correlation between uPA expression and tumor size, lymph node status, TNM stage was statistically significant. Expression of MVD was also significantly associated with tumor size and TNM stage. Neither age related to GDDP, menopausal status nor PR ER status was significantly with uPA and MVD expression. Patients with strong expression of uPA and/or MVD had a significantly shorter relapse-free survival than those with weak expression of uPA and/or MVD. Especially, patients with strong expression of both uPA and MVD were likely to develop recurrence and metastasis. Multivariate analysis showed that uPA and MVD were two independent prognostic factors affecting the relapse-free survival. CONCLUSION: Angiogensis and fibrinolysis were closely associated with invasion and metastasis of breast cancer. uPA and MVD may be two strong and independent biologic markers in predicting postoperative relapse and metastasis of breast cancer.