Monomeric Far-red and Near-infrared Fluorescent Biliproteins of Ultrahigh Brightness.

Far-red and near-infrared fluorescent proteins have regions of maximum transmission in most tissues and can be widely used as fluorescent biomarkers. We report that fluorescent phycobiliproteins originating from the phycobilisome core subunit ApcF2 can covalently bind biliverdin, named BDFPs. To further improve BDFPs, we conducted a series of studies. Firstly, we mutated K53Q and T144A of BDFPs to increase their effective brightness up to 190 % in vivo. Secondly, by homochromatic tandem fusion of high-brightness BDFPs to achieve monomerization, which increases the effective brightness by up to 180 % in vivo, and can effectively improve the labeling effect. By combining the above two approaches, the brightness of the tandem BDFPs was much improved compared with that of the previously reported fluorescent proteins in a similar spectral range. The tandem BDFPs were expressed stably while maintaining fluorescence in mammalian cells and Caenorhabditis elegans. They were also photostable and resistant to high temperature, low pH, and chemical denaturation. The tandem BDFPs advantages were proved in applications as biomarkers for imaging in super-resolution microscopy.

Identification of Vernonia patula Merr. and Its Similar Varieties Based on a Combination of HPLC Fingerprinting and Chemical Pattern Recognition.

Vernonia patula Merr. (VP) is a traditional medicine used by the Zhuang and Yao people, known for its therapeutic properties in treating anemopyretic cold and other diseases. Distinguishing VP from similar varieties such as Praxelis clematidea (PC), Ageratum conyzoides L. (AC) and Ageratum houstonianum Mill (AH) was challenging due to their similar traits and plant morphology. The HPLC fingerprints of 40 batches of VP and three similar varieties were established. SPSS 20.0 and SIMCA-P 13.0 were used to statistically analyze the chromatographic peak areas of 37 components. The results showed that the similarity of the HPLC fingerprints for each of the four varieties was >0.9, while the similarity between the control chromatogram of VP and its similar varieties was <0.678. Cluster analysis and partial least squares discriminant analysis provided consistent results, indicating that all four varieties could be individually clustered together. Through further analysis, we found isochlorogenic acid A and isochlorogenic acid C were present only in the original VP, while preconene II was present in the three similar varieties of VP. These three components are expected to be identification points for accurately distinguishing VP from PC, AC and AH.

Expression and Correlation of Hypoxia-Inducible Factor-1α (HIF-1α) with Pulmonary Artery Remodeling and Right Ventricular Hypertrophy in Experimental Pulmonary Embolism.

BACKGROUND We investigated the expression of HIF-1α (hypoxia-inducible factor-1α) and its correlations with pulmonary artery remodeling and right ventricular hypertrophy in PE (pulmonary embolism) rats. MATERIAL AND METHODS After the PE rat model was established, the dynamic changes of mPAP (mean pulmonary artery pressure), PAMT (relative medial thickness of small pulmonary arteries), WA/TA (vessel wall area/total area), and RVHI (right ventricular hypertrophy index) were detected. Then, histomorphology of pulmonary vascular was observed, followed by HIF-1α mRNA and protein for pulmonary artery and right ventricle were checked via in situ hybridization and immunohistochemistry, respectively. The correlations between HIF-1α and mPAP, PAMT, WA/TA, and RVHI were analyzed. RESULTS The mPAP level increased from the initial time to 12 weeks of PE and reached the peak at 12 weeks. After 4 weeks of PE, PAMT increased compared with the initial control group (p<0.05), and increased further after 8 weeks and 12 weeks. Changes of the vessel WA/TA were the same as PAMT. Compared with the initial control group, RVHI increased at 8 weeks (p<0.05) and 12 weeks of PE (p<0.01). HIF-1α mRNA and protein were positively correlated with mPAP, PAMT, and WA/TA in pulmonary arteries. HIF-1α mRNA and protein were positively correlated with mPAP, PAMT, WA/TA, and RVHI in right ventricles (p<0.01). CONCLUSIONS HIF-1α mRNA and protein are expressed in the pulmonary artery and right ventricular of PE rats, and HIF-1α may correlate with pulmonary artery remodeling and right ventricular hypertrophy in rats with PE.

Plasma Inflammatory Cytokine IL-4, IL-8, IL-10, and TNF-α Levels Correlate with Pulmonary Function in Patients with Asthma-Chronic Obstructive Pulmonary Disease (COPD) Overlap Syndrome.

BACKGROUND The aim of this study was to investigate the plasma inflammatory cytokine levels and their correlations with pulmonary function in patients with asthma-chronic obstructive pulmonary disease overlap syndrome (ACOS). MATERIAL AND METHODS Between January 2013 and December 2014, a total of 96 patients with asthma, acute exacerbation of chronic obstructive pulmonary disease (AECOPD), or ACOS were enrolled, and 35 healthy people were included as a control group. Fasting plasma interleukin (IL)-4, IL-8, IL-10, and tumor necrosis factor alpha (TNF-α) levels were detected using enzyme-linked immunosorbent assay (ELISA). Correlations between the plasma inflammatory cytokine levels and forced expiratory volume in 1 second (FEV1), FEV1/predicted value ratio (FEV1%pred), and FEV1/forced vital capacity (FVC) were analyzed. RESULTS IL-4 and IL-8 levels showed statistically significant differences among the 3 groups of patients (both P<0.001); IL-4 level was significantly lower, while IL-8 level was significantly higher in the AECOPD group and ACOS group than those in the asthma group (all P<0.05). IL-10 level and TNF-α level were significantly different among the 3 patient groups (both P<0.001). IL-10 level was significantly different between each of the 2 groups (all P<0.001). TNF-α level in the asthma group was higher than in the AECOPD group and ACOS group (both P<0.001). IL-4 and IL-10 were positively and IL-8 and TNF-α were negatively related with FEV1, FEV1%pred, and FEV1/FVC. CONCLUSIONS Plasma levels of inflammatory cytokines IL-4, IL-8, IL-10, and TNF-α are related with severity of airway diseases and could be potential markers for the evaluation of asthma, COPD, and ACOS.

LRIG1 regulates cadherin-dependent contact inhibition directing epithelial homeostasis and pre-invasive squamous cell carcinoma development.

Epidermal growth factor receptor (EGFR) pathway activation is a frequent event in human carcinomas. Mutations in EGFR itself are, however, rare, and the mechanisms regulating EGFR activation remain elusive. Leucine-rich immunoglobulin repeats-1 (LRIG1), an inhibitor of EGFR activity, is one of four genes identified that predict patient survival across solid tumour types including breast, lung, melanoma, glioma, and bladder. We show that deletion of Lrig1 is sufficient to promote murine airway hyperplasia through loss of contact inhibition and that re-expression of LRIG1 in human lung cancer cells inhibits tumourigenesis. LRIG1 regulation of contact inhibition occurs via ternary complex formation with EGFR and E-cadherin with downstream modulation of EGFR activity. We find that LRIG1 LOH is frequent across cancers and its loss is an early event in the development of human squamous carcinomas. Our findings imply that the early stages of squamous carcinoma development are driven by a change in amplitude of EGFR signalling governed by the loss of contact inhibition.

Circadian clock gene expression regulates cancer cell growth through glutaminase.

Glutamine is an essential amino acid for malignant tumor cells. Glutaminase that metabolizes glutamine reaches a maximum expression in tumors immediately before the maximum proliferation rate. Tumor cells grow at different rates during the day. We postulated that the activity of glutaminase in tumor cells is subject to the regulation of circadian clock gene. We measured glutaminase by western blot analysis and circadian clock gene expression by real-time polymerase chain reaction in the liver and tumor cells at six equispaced time points of the day in individual mice of a 12/12 h light/dark schedule. The results showed that the tumor-bearing mice, under normal diurnal conditions, are circadianly entrained, as reflected by the normal host locomotor activity rhythms and rhythmic liver clock gene expression. The tumors within these mice are also circadianly organized, as reflected by circadian clock gene (Bmal1) expression. What is most remarkable is that kidney-type glutaminase also showed circadian rhythms in the same pattern with tumor circadian clock gene expression in liver cancer xenograft model, indicating that conditionally inhibiting glutaminase activity may provide a new target for cancer therapy.

[Study on the value of systemic immune inflammation index and nomogram in predicting the prognosis of patients with mucoepidermoid carcinoma].

PURPOSE: To investigate the relationship between preoperative systemic immune-inflammation index (SII) and relapse-free survival (RFS) after surgical resection of mucoepidermoid carcinoma(MEC). METHODS: The data of 135 patients with MEC who underwent surgical resection in the First Affiliated Hospital of Zhengzhou University from January 2016 to July 2019 were collected, and the receiver operating characteristic(ROC) curve was performed on the SII of patients. The optimal cut-off value was obtained by ROC analysis. Therefore, the patients' SII index was divided into high and low group, and survival analysis was performed by Kaplan-Meier method. Cox proportional regression model and least absolute shrinkage and selection operator (LASSO) were used to analyze the factors influencing prognosis, and a nomogram model was built to predict patients' relapse-free survival(RFS). Area under curve (AUC) and correction curve were used to evaluate the model and verify the consistency. RESULTS: Survival analysis showed that the RFS rate in low SII group was significantly higher than that in high SII group. Cox proportional hazard regression model showed high SII(HR=2.179, 95%CI: 1.072-4.426, P=0.031) and low tumor differentiation(HR=6.894, 95%CI: 2.770-17.158, P=0.000) and cervical lymph node metastasis (HR=2.091, 95%CI: 1.034-4.230, P=0.040) were significant predictors of poor RFS. CONCLUSIONS: The lower the preoperative SII, the better the prognosis of patients. The nomogram prognosis of MEC based on SII is effective.

Comprehensive safety evaluation of a novel multitargeting compound XYY-CP1106: A candidate for Alzheimer's disease.

Multitargeting has become a promising strategy for the development of anti-Alzheimer's disease (AD) drugs, considering the complexity of molecular mechanisms in AD pathology. In most pre-clinical studies, the effectiveness of these multi-targeted anti-AD drugs has been demonstrated but comprehensive safety assessments are lacking. Here, the safety evaluation of a novel multi-targeted candidate in AD (XYY-CP1106), characterized by its dual-property of iron chelation and monoamine oxidase B inhibition, was conducted by multifaceted analysis. Acute toxicity in mice was conducted to investigate the safety of oral administration and the maximum tolerated dose of the agent. In vitro Ames analysis, CHL chromosomal aberration analysis, and bone marrow micronucleus analysis were executed to evaluate the genotoxicity. A teratogenesis investigation in pregnant mice were meticulously performed to evaluate the teratogenesis of XYY-CP1106. Furthermore, a 90-day long-term toxicity analysis in rats was investigated to evaluate the cumulative toxicity after long-term administration. Strikingly, no toxic phenomena were found in all investigations, demonstrating relatively high safety profile of the candidate compound. The securing of safety heightened the translational significance of XYY-CP1106 as a novel multi-targeted anti-AD candidate, supporting the rationality of multitargeting strategy in the designs of smart anti-AD drugs.

ß-Catenin determines upper airway progenitor cell fate and preinvasive squamous lung cancer progression by modulating epithelial-mesenchymal transition.

Human lung cancers, including squamous cell carcinoma (SCC) are a leading cause of death and, whilst evidence suggests that basal stem cells drive SCC initiation and progression, the mechanisms regulating these processes remain unknown. In this study we show that ß-catenin signalling regulates basal progenitor cell fate and subsequent SCC progression. In a cohort of preinvasive SCCs we established that elevated basal cell ß-catenin signalling is positively associated with increased disease severity, epithelial proliferation and reduced intercellular adhesiveness. We demonstrate that transgene-mediated ß-catenin inhibition within keratin 14-expressing basal cells delayed normal airway repair while basal cell-specific ß-catenin activation increased cell proliferation, directed differentiation and promoted elements of early epithelial-mesenchymal transition (EMT), including increased Snail transcription and reduced E-cadherin expression. These observations are recapitulated in normal human bronchial epithelial cells in vitro following both pharmacological ß-catenin activation and E-cadherin inhibition, and mirrored our findings in preinvasive SCCs. Overall, the data show that airway basal cell ß-catenin determines cell fate and its mis-expression is associated with the development of human lung cancer.

Myd88 deficiency influences murine tracheal epithelial metaplasia and submucosal gland abundance.

Tracheal epithelial remodelling, excess mucus production, and submucosal gland hyperplasia are features of numerous lung diseases, yet their origins remain poorly understood. Previous studies have suggested that NF-κB signalling may regulate airway epithelial homeostasis. The purpose of this study was to determine whether deletion of the NF-κB signalling pathway protein myeloid differentiation factor 88 (Myd88) influenced tracheal epithelial cell phenotype. We compared wild-type and Myd88-deficient or pharmacologically inhibited adult mouse tracheas and determined that in vivo Myd88 deletion resulted in increased submucosal gland number, secretory cell metaplasia, and excess mucus cell abundance. We also found that Myd88 was required for normal resolution after acute tracheal epithelial injury. Microarray analysis revealed that uninjured Myd88-deficient tracheas contained 103 transcripts that were differentially expressed relative to wild-type and all injured whole tracheal samples. These clustered into several ontologies and networks that are known to functionally influence epithelial cell phenotype. Comparing these transcripts to those expressed in airway progenitor cells revealed only five common genes, suggesting that Myd88 influences tracheal epithelial homeostasis through an extrinsic mechanism. Overall, this study represents the first identification of Myd88 as a regulator of adult tracheal epithelial cell phenotype.

Xylariaopyrones E-I, five new α-pyrone derivatives from the endophytic fungus Xylariales sp. (HM-1).

Five new α-pyrones, xylariaopyrones E-I (1-5), along with three known analogues (6-8) were isolated from the cultivation broth of the endophytic fungus Xylariales sp. (HM-1). The structures of the new compounds including their absolute configurations were elucidated by comprehensive spectroscopic methods and quantum ECD calculations. Xylariaopyrone E (1) is the first example of α-pyrone derivative with a novel [3, 2, 0] bridge ring system via a ketal function group in the side chain. In bioactivity assays, xylariaopyrones E-G (1-3) showed moderate inhibiting activities against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa with MIC values from 25.4 to 64.5 µg/mL, whereras xylariaopyrone G (3) exhibited significant inhibition of monoamine oxidase B with an IC50 value of 15.6 µmol/L. Xylariaopyrone H (4) and the known compound 7 showed moderate toxicity against brine shrimp larvae with inhibition rates of 42.8% and 44.5%, respectively.

Comparison of the Clinical Outcomes Between Nebulized and Systemic Corticosteroids in the Treatment of Acute Exacerbation of COPD in China (CONTAIN Study): A Post Hoc Analysis.

Background and Objective: Although corticosteroids have been widely used in the treatment of acute exacerbation of chronic obstructive pulmonary disease (AECOPD), few studies have evaluated the effectiveness of nebulized corticosteroids (NCS), systemic corticosteroids (SCS), and NCS plus SCS in the management of AECOPD in China. This study aimed to evaluate the effectiveness of NCS, SCS, and NCS plus SCS in Chinese patients with AECOPD. Patients and Methods: This was a real-world study of AECOPD patients at 43 sites from January to September 2014. During hospitalization, patients treated with nebulized budesonide (NCS group, n=1091), SCS (SCS group, n=709), or both (NCS+SCS group, n=1846) were included. Propensity score matching (PSM) and subgroup analyses were performed. The primary outcomes were the length of hospital stay, mortality, and change in arterial blood gases from baseline. Results: Multivariable analysis showed that the three treatments at the same severity of AECOPD were not significantly different regarding intubation rates, rates of pneumonia improvement at discharge, rates of new-onset pneumonia in hospital, and mortality. Following PSM, NCS+SCS was associated with greater length of hospital stay than both NCS and SCS (in patients without respiratory failure [RF, P<0.001] and with type I RF [P=0.022]), and more hospitalization costs than the other two treatments (in patients without RF [P<0.001]). Conclusion: NCS is effective for patients with AECOPD, which may be an alternative treatment option. Further clinical trials are urgently needed to better understand the efficacy of NCS, SCS, and NCS+SCS in AECOPD management in China.

The Distribution and Health Risk Assessment of Metals in Soils in the Vicinity of Industrial Sites in Dongguan, China.

Exponential industrialization and rapid urbanization have resulted in contamination of soil by metals from anthropogenic sources in Dongguan, China. The aims of this research were to determine the concentration and distribution of various metals (arsenic (As), cadmium (Cd), chromium (Cr), copper (Cu), mercury (Hg), nickel (Ni), lead (Pb) and zinc (Zn)) in soils and identify their potential health risks for local residents. A total of 106 soil samples were collected from the vicinity of industrial sites in Dongguan. Two types of samples were collected from each site: topsoil (0-20 cm, TS) and shallow soil (20-50 cm, SS). Results showed that the soils were contaminated by metals and pollution was mainly focused on TS. The geoaccumulation index (Igeo) and pollution indexes (PI) implied that there was a slight increase in the concentrations of Cd, Cu, Hg, Ni, and Pb, but the metal pollution caused by industrial activities was less severe, and elements of As and Cr exhibited non-pollution level. The risk assessment results suggested that there was a potential health risk associated with As and Cr exposure for residents because the carcinogenic risks of As and Cr via corresponding exposure pathways exceeded the safety limit of 10(-6) (the acceptable level of carcinogenic risk for humans). Furthermore, oral ingestion and inhalation of soil particles are the main exposure pathways for As and Cr to enter the human body. This study may provide basic information of metal pollution control and human health protection in the vicinity of industrial regions.

Quantitative assessment of the variation in IGF2BP2 gene and type 2 diabetes risk.

Insulin-like growth factor 2 mRNA-binding protein 2 (IFG2BP2) belongs to an mRNA-binding protein family involved in the development and stimulation of insulin action, which has attracted considerable attention as a candidate gene for type 2 diabetes (T2D) since it was first identified through genome-wide association approach. The relationship between IFG2BP2 and T2D has been reported in various ethnic groups; however, these studies have yielded contradictory results. To investigate this inconsistency, we performed a meta-analysis of 35 studies involving a total of 175,965 subjects for two wildly studied polymorphisms (rs4402960 and rs1470579) of the IFG2BP2 to evaluate the effect of IFG2BP2 on genetic susceptibility for T2D. An overall random-effects per-allele OR of 1.13 (95% CI: 1.12-1.15; P < 10(-5)) and 1.09 (95% CI: 1.07-1.12; P < 10(-5)) was found for the two variants, respectively. Significant results were also observed using dominant or recessive genetic model. No significant results between study heterogeneity were found in most of the comparison. In the subgroup analysis by ethnicity, sample size, diagnostic criterion and mean age and BMI of cases, significantly increased risks were found for these polymorphisms in almost all genetic models. This meta-analysis demonstrated that these two common polymorphisms is a risk factor for developing T2D, but these associations vary in different ethnic populations.