Spatio-Temporal Selection of Reference Genes in the Two Congeneric Species of \u3ci\u3eGlycyrrhiza\u3c/i\u3e

Glycyrrhiza, a genus of perennial medicinal herbs, has been traditionally used to treat human diseases, including respiratory disorders. Functional analysis of genes involved in the synthesis, accumulation, and degradation of bioactive compounds in these medicinal plants requires accurate measurement of their expression profiles. Reverse transcription quantitative real-time PCR (RT-qPCR) is a primary tool, which requires stably expressed reference genes to serve as the internal references to normalize the target gene expression. In this study, the stability of 14 candidate reference genes from the two congeneric species G. uralensis and G. inflata, including ACT, CAC, CYP, DNAJ, DREB, EF1, RAN, TIF1, TUB, UBC2, ABCC2, COPS3, CS, R3HDM2, were evaluated across different tissues and throughout various developmental stages. More importantly, we investigated the impact of interactions between tissue and developmental stage on the performance of candidate reference genes. Four algorithms, including geNorm, NormFinder, BestKeeper, and Delta Ct, were used to analyze the expression stability and RefFinder, a comprehensive software, provided the final recommendation. Based on previous research and our preliminary data, we hypothesized that internal references for spatio-temporal gene expression are different from the reference genes suited for individual factors. In G. uralensis, the top three most stable reference genes across different tissues were R3HDM2, CAC and TUB, while CAC, CYP and ABCC2 were most suited for different developmental stages. CAC is the only candidate recommended for both biotic factors, which is reflected in the stability ranking for the spatio (tissue)-temporal (developmental stage) interactions (CAC, R3HDM2 and DNAJ). Similarly, in G. inflata, COPS3, R3HDM2 and DREB were selected for tissues, while RAN, COPS3 and CS were recommended for developmental stages. For the tissue-developmental stage interactions, COPS3, DREB and ABCC2 were the most suited reference genes. In both species, only one of the top three candidates was shared between the individual factors and their interactions, specifically, CAC in G. uralensis and COPS3 in G. inflata, which supports our overarching hypothesis. In summary, spatio-temporal selection of reference genes not only lays the foundation for functional genomics research in Glycyrrhiza, but also facilitates these traditional medicinal herbs to reach/maximize their pharmaceutical potential

Selection of Reference Genes for qRT-PCR Analysis in Medicinal Plant \u3cem\u3eGlycyrrhiza\u3c/em\u3e under Abiotic Stresses and Hormonal Treatments

Best known as licorice, Glycyrrhiza Linn., a genus of herbaceous perennial legume, has been used as a traditional herbal medicine in Asia and a flavoring agent for tobacco and food industry in Europe and America. Abiotic stresses and hormonal treatments can significantly impact the development and metabolism of secondary metabolites in Glycyrrhiza. To better understand the biosynthesis of the trace-amount bioactive compounds, we first screened for the suitable reference genes for quantitative real-time reverse transcription PCR (qRT-PCR) analysis in Glycyrrhiza. The expression profiles of 14 candidate reference genes, including Actin1 (ACT), Clathrin complex AP1 (CAC), Cyclophilin (CYP), Heat-shock protein 40 (DNAJ), Dehydration responsive element binding gene (DREB), Translation elongation factor1 (EF1), Ras related protein (RAN), Translation initiation factor (TIF1), β-Tubulin (TUB), Ubiquitin-conjugating enzyme E2 (UBC2), ATP binding-box transpoter 2 (ABCC2), COP9 signal compex subunit 3 (COPS3), Citrate synthase (CS), and R3H domain protein 2 (R3HDM2) from two congeneric species, Glycyrrhiza uralensis F. and Glycyrrhiza inflata B., were examined under abiotic stresses (osmotic and salinity) and hormonal treatments (Abscisic acid (ABA) and methyl jasmonic acid (MeJA)) using a panel of software, including geNorm, NormFinder, BestKeeper, and Delta CT. The overall stability, however, was provided by RefFinder, a comprehensive ranking system integrating inputs from all four algorithms. In G. uralensis, the most stable reference genes under osmotic stress, salt stress, ABA treatment, and MeJA treatment were TIF1, DNAJ, CS, and ABCC2 for leaves and DNAJ, DREB, CAC, and CAC for roots, respectively. In comparison, the top ranked genes were TUB, CAC, UBC2, and RAN for leaves and TIF1, ABCC2, CAC, and UBC2 for roots, respectively, under stress and hormonal treatments in G. inflata. ACT and TIF1, on the other hand, were the least stable genes under the most experimental conditions in the two congeneric species. Finally, our survey of the reference genes in legume shows that EF, ACT, UBC2, and TUB were the top choices for the abiotic stresses while EF, UBC2, CAC, and ABCC2 were recommended for the hormonal treatments in Leguminosae. Our combined results provide reliable normalizers for accurate gene quantifications in Glycyrrhiza species, which will allow us to exploit its medicinal potential in general and antiviral activities in particular

Placental expression of AChE, α7nAChR and NF-κB in patients with preeclampsia

Objectives: This study aimed to investigate placental expression of AChE, α7nAChR and NF-κB in patients with preeclampsia and discuss about its clinical significance. Material and methods: mRNA expression levels of acetylcholine (AChE), alpha-7 nicotinic acetylcholine receptor (α7nAChR) and nuclear factor-kB (NF-κB) in placenta were detected by qRT-PCR, and protein levels were determined by immunohis­tological analysis and Western Blot in 35 women with preeclampsia (including 20 cases of mild preeclampsia and 15 cases of severe preeclampsia) and 30 cases in control group, respectively. Results: The expression of AChE mRNA and protein in placenta increased significantly in patients with preeclampsia compared with the control group (p < 0.01). It was lower in patients with severe preeclampsia than in patients with mild preeclampsia (p < 0.05). The expression of α7nAChR mRNA and protein in placenta decreased significantly in patients with preeclampsia compared with the control group (p < 0.01). However, the expression of α7nAChR mRNA and protein in patients with severe preeclampsia was higher than that in patients with mild preeclampsia, without significant difference(p > 0.05). The expression of NF-κB protein in placenta decreased significantly in patients with preeclampsia compared with the control group(p < 0.01). It was higher in patients with severe preeclampsia than in patients with mild preeclampsia (p < 0.05), but there was no significant difference between preeclampsia group and control group in the expression of NF-κB mRNA in placenta (p > 0.05). The results of Western blotting assay were consistent with those of immunohistochemistry. Conclusions: Abnormal expression of AChE, α7nAChR and NF-κB in placenta may be associated with preeclampsia. Cho­linergic anti-inflammatory pathway may play an important role in the pathogenesis of preeclampsia

Analysis and Design of Wind Turbine Monitoring System Based on Edge Computing

INTRODUCTION: A wind turbine data analysis method based on the combination of Hadoop and edge computing is proposed. OBJECTIVES: Solve the wind turbine health status monitoring system large data, time extension, energy consumption and other problems. METHODS: By analysing the technical requirements and business processes of the system, the overall framework of the system was designed and a deep reinforcement learning algorithm based on big data was proposed. RESULTS: It solves the problem of insufficient computing resources as well as energy consumption and latency problems occurring in the data analysis layer, solves the problems in WTG task offloading, and improves the computational offloading efficiency of the edge nodes to complete the collection, storage, and analysis of WTG data. CONCLUSION: The data analysis and experimental simulation platform is built through Python, and the results show that the application of Hadoop and the edge computing offloading strategy based on the DDPG algorithm to the system improves the system's quality of service and computational performance, and the method is applicable to the distributed storage and analysis of the device in the massive monitoring data

KIAA1199 Correlates With Tumor Microenvironment and Immune Infiltration in Lung Adenocarcinoma as a Potential Prognostic Biomarker

Background: KIAA1199 has been considered a key regulator of carcinogenesis. However, the relationship between KIAA1199 and immune infiltrates, as well as its prognostic value in lung adenocarcinoma (LUAD) remains unclear.Methods: The expression of KIAA1199 and its influence on tumor prognosis were analyzed using a series of databases, comprising TIMER, GEPIA, UALCAN, LCE, Prognoscan and Kaplan-Meier Plotter. Further, immunohistochemistry (IHC), western blot (WB) and receiver operating characteristic (ROC) curve analyses were performed to verify our findings. The cBioPortal was used to investigate the genomic alterations of KIAA1199. Prediction of candidate microRNA (miRNAs) and transcription factor (TF) targeting KIAA1199, as well as GO and KEGG analyses, were performed based on LinkedOmics. TIMER and TISIDB databases were used to explore the relationship between KIAA1199 and tumor immune infiltration.Results: High expression of KIAA1199 was identified in LUAD and Lung squamous cell carcinoma (LUSC) patients. High expression of KIAA1199 indicated a worse prognosis in LUAD patients. The results of IHC and WB analyses showed that the expression level of KIAA1199 in tumor tissues was higher than that in adjacent tissues. GO and KEGG analyses indicated KIAA1199 was mainly involved in extracellular matrix (ECM)-receptor interaction and extracellular matrix structure constituent. KIAA1199 was positively correlated with infiltrating levels of CD4+ T cells, macrophages, neutrophil cells, dendritic cells, and showed positive relationship with immune marker subsets expression of a variety of immunosuppressive cells.Conclusion: High expression of KIAA1199 predicts a poor prognosis of LUAD patients. KIAA1199 might exert its carcinogenic role in the tumor microenvironment via participating in the extracellular matrix formation and regulating the infiltration of immune cells in LUAD. The results indicate that KIAA1199 might be a novel biomarker for evaluating prognosis and immune cell infiltration in LUAD

NaoXinTong Inhibits the Development of Diabetic Retinopathy in d

Buchang NaoXinTong capsule (NXT) is a Chinese Materia Medica standardized product extracted from 16 Chinese traditional medical herbs and widely used for treatment of patients with cerebrovascular and cardiovascular diseases in China. Formation of microaneurysms plays an important role in the development of diabetic retinopathy. In this study, we investigated if  NXT can protect diabetic mice against the development of diabetic retinopathy. The db/db mice (~6 weeks old), a diabetic animal model, were divided into two groups and fed normal chow or plus NXT for 14 weeks. During the treatment, fasting blood glucose levels were monthly determined. After treatment, retinas were collected to determine retinal thickness, accumulation of carbohydrate macromolecules, and caspase-3 (CAS-3) expression. Our results demonstrate that administration of NXT decreased fasting blood glucose levels. Associated with the decreased glucose levels, NXT blocked the diabetes-induced shrink of multiple layers, such as photoreceptor layer and outer nuclear/plexiform layers, in the retina. NXT also inhibited the diabetes-induced expression of CAS-3 protein and mRNA, MMP-2/9 and TNFα mRNA, accumulation of carbohydrate macromolecules, and formation of acellular capillaries in the retina. Taken together, our study shows that NXT can inhibit the development of diabetic retinopathy and suggests a new potential application of NXT in clinic

Polycomb group proteins EZH2 and EED directly regulate androgen receptor in advanced prostate cancer

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/149265/1/ijc32118.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/149265/2/ijc32118_am.pd

Attenuating Immune Response of Macrophage by Enhancing Hydrophilicity of Ti Surface

Immune responses can determine the in vivo fate of implanted materials. The strategy for developing implants has shifted towards using materials with immunomodulatory activity. However, the immunoregulatory effect of hydrophilicity of titanium surface on the macrophage behavior and its underlying mechanism remain poorly understood. Here, the Ti surface hydrophilicity-dependent behavior of murine RAW264.7 macrophages was investigated in vitro. Two laboratory models with significantly different surface hydrophilicity and similar roughness were established with Ti-polished and Ti-H2O2 surfaces. The results of cell morphology observation showed that the Ti-H2O2 surface yielded enhanced cell adhesion and less multinucleated cell formation. CCK-8 assay indicated that the growth rate of macrophage on Ti-H2O2 surface is higher than that of Ti-polished. ELISA assay result revealed lower level of proinflammatory factor TNF-α and higher level of anti-inflammatory factor IL-10 on the Ti-H2O2 surface compared to Ti-polished. Subsequently, immunofluorescence and western blotting analysis showed that activation of the NF-κB-TNF-α pathway might be involved in the modulation of the immune response by surface hydrophilicity. Together, these results suggested that relative high hydrophilic Ti surface might attenuate the immune response of macrophage by activating NF-κB signaling. These findings could provide new insights into designing implant devices for orthopedic applications

Room-temperature conversion of ethane and the mechanism understanding over single iron atoms confined in graphene

Abstract(#br)The catalytic conversion of ethane to high value-added chemicals is significantly important for utilization of hydrocarbon resources. However, it is a great challenge due to the typically required high temperature (> 400 °C) conditions. Herein, a highly active catalytic conversion process of ethane at room temperature (25 °C) is reported on single iron atoms confined in graphene via the porphyrin-like N 4 -coordination structures. Combining with the operando time of flight mass spectrometer and density functional theory calculations, the reaction is identified as a radical mechanism, in which the C–H bonds of the same C atom are preferentially and sequentially activated, generating the value-added C 2 chemicals, simultaneously avoiding the over-oxidation of the products to CO 2 . The in-situ formed O–FeN 4 –O structure at the single iron atom serves as the active center for the reaction and facilitates the formation of ethyl radicals. This work deepens the understanding of alkane C–H activation on the FeN 4 center and provides the reference in development of efficient catalyst for selective oxidation of light alkane