Production of Rhamnosyl Icariside II by snailase hydrolysis of Epimedium wushanense extracts.

Rhamnosyl Icariside II is a rare secondary flavonoid glycoside isolated from Epimedium L. plants. It has better stability and physiological activity than the primary flavonoid glycosides of Epimedium L., therefore, conversion of the primary flavonoid glycoside into Rhamnosyl Icariside II would be desirable. In this study, a method for the enzymatic production of Rhamnosyl Icariside II from the total flavonoids of Epimedium wushanense was established, and the conditions were optimized. Six commercial enzymes were screened, and the reaction conditions for the best enzyme were optimized. Snailase was the most effective hydrolase, and the highest yield was obtained under the optimized conditions. To facilitate industrial production of Rhamnosyl Icariside II, a scaled-up pilot test was performed. The reaction solution was extracted with n-butanol to obtain the Rhamnosyl Icariside II crude product, which was then subjected to silica gel column chromatography and preparative chromatography. Finally, a product of Rhamnosyl Icariside II with purity of 99.1 % was achieved, in a total yield of 46.8 %. Compared to direct extraction and acid hydrolysis, this method improves the product yield and purity, which is of great significance for the large-scale production of Rhamnosyl Icariside II. This study provides a basis for the physiological activity study of Rhamnosyl Icariside II, and offers possibilities for future applications in the healthcare sector.

Formation of key aroma-active and off-flavor components in concentrated peach puree.

Non-volatiles offer some insight into the formation of aroma-active components in peach puree (PP), but more depth investigation is still needed. Formation pathways of key aroma-active and off-flavor components in PP during thermal concentration (PP + C) and sterilization (PP + C + S) are unclear. Therefore, GC-O-MS combined with UPLC-MS/MS was used to identify the volatile and nonvolatile components and their formation pathways. Among the 36 aroma-active compounds, the contents of γ-decalactone, hexyl acetate, leaf acetate, hexanal, and 1-hexanol (odor activity value ≥ 1) decreased by 46 %, 100 %, 100 %, 92 %, and 100 % between PP and PP + C + S, causing the weakening of "green" and "fruity" attributes. Off-flavor components including 1-octen-3-one, isobutyric acid, isothiazole, and isovaleric acid were identified during thermal processing. 1-Octen-3-one content increased by 75 % from PP to PP + C + S through linolenic acid metabolism, which contributed to "cooked"; the formation of isobutyric and isovaleric acids, isothiazole, resulted in the enhancement of "sour/rancid" via serine and leucine metabolism.

Salivary Extracellular Vesicles: Biomarkers and Beyond in Human Diseases.

Extracellular vesicles, as bioactive molecules, have been extensively studied. There are abundant studies in the literature on their biogenesis, secretion, structure, and content, and their roles in pathophysiological processes. Extracellular vesicles have been reviewed as biomarkers for use in diagnostic tools. Saliva contains many extracellular vesicles, and compared with other body fluids, it is easier to obtain in a non-invasive way, making its acquisition more easily accepted by patients. In recent years, there have been numerous new studies investigating the role of salivary extracellular vesicles as biomarkers. These studies have significant implications for future clinical diagnosis. Therefore, in this paper, we summarize and review the potential applications of salivary extracellular vesicles as biomarkers, and we also describe their other functions (e.g., hemostasis, innate immune defense) in both oral and non-oral diseases.

Identification of CD147-positive extracellular vesicles as novel non-invasive biomarkers for the diagnosis and prognosis of colorectal cancer.

BACKGROUND: The mortality rate of colorectal cancer (CRC) can be decreased with effective screening and early diagnosis. Exosomes are released from cancer cells into the bloodstream, and circulating exosomes may serve as novel biomarkers. This study aimed to identify a sensitive and rapid method of exosome collection and measurement using specific antibodies. METHODS: ExoCounter, a high-sensitive exosome-counting system, allows the identification of exosomes without enrichment or purification, based on the identification of the transmembrane protein-CD147-on serum exosomes that are associated with CRC. RESULTS: Receiver operating characteristic curves between healthy donors and CRC patients were described and assessed by CD147-specific exosomes (exo-CD147), CEA, and CA19-9. And area under curves for exo-CD147, CEA, and CA19-9 were 0.827 (95%CI: 0.764-0.891), 0.630 (95%CI: 0.536-0.724), and 0.659 (95%CI: 0.559-0.759), respectively. Drawing a clinical decision curve of exo-CD147 for the diagnosis of CRC metastases showed that when the threshold probability of exo-CD147 was between 20% and 92%, the net clinical utilization rate was higher than for all patients with or without metastases. A nomogram was constructed using multivariate COX regression analysis to select significant variables such as the high CD147 group (>34 × 105 particles). Calibration curves for 1-, 3-, and 5-year survival rates of CRC patients showed that the actual 1-, 3-, and 5-year survival rates were in excellent agreement with the survival rates predicted by the nomogram. CONCLUSIONS: The increased CD147 expression in exosomes could serve as a diagnostic and prognostic biomarker for CRC.

The multifaceted role of placental growth factor in the pathogenesis and progression of bronchial asthma and pulmonary fibrosis: Therapeutic implications.

Placental growth factor (PlGF) is a glycosylated dimeric protein that is homologous to vascular endothelial growth factor (VEGF). PlGF expression is upregulated in patients with bronchial asthma, suggesting that it plays a role in the pathogenesis of asthma. Bronchial asthma is characterized by chronic airway inflammation and airway hyperresponsiveness (AHR). After recurrent asthma attacks, pulmonary fibrosis develops and leads to airway remodeling and a further decline in lung function. In this review, we focused on the pivotal role of PlGF in chronic airway inflammation, AHR, and airway remodeling during bronchial asthma. Furthermore, we summarized data showing that PlGF may be a potential therapeutic target in bronchial asthma.

Pharmacokinetic and Pharmacodynamic Interactions Between Henagliflozin, a Novel Selective SGLT-2 Inhibitor, and Warfarin in Healthy Chinese Subjects.

PURPOSE: While controlling blood glucose, patients with diabetes and abnormal coagulation should be treated with positive anticoagulation because the hypercoagulable state of their blood is the primary cause of macroangiopathy. The goal of this study was to evaluate the pharmacokinetic and pharmacodynamic (PK/PD) interactions between henagliflozin, a novel selective sodium-glucose cotransporter 2 inhibitor, and warfarin in healthy subjects. METHODS: This single-center, open-label, single-arm clinical study was conducted in 16 healthy male Chinese subjects. According to the study protocol, the PK properties of henagliflozin 10 mg/d and warfarin 5 mg/d were collected and tabulated in accordance with sampling time. All study drugs were given with once-daily administration. Subjects were monitored for adverse reactions and their severity, outcomes, and relationship to study drug. This influences of warfarin on the PK properties of henagliflozin (Cmax,ss and AUCτ,ss), the effects of henagliflozin on the PK properties of warfarin (Cmax, AUC0-t, and AUC0-∞), and the influences of henagliflozin on the PD properties of warfarin (PTmax, PTAUC, INRmax, and INRAUC) were evaluated. FINDINGS: The geometric mean ratios (GMRs; 90% CIs) of henagliflozin Cmax,ss and AUCτ,ss were 101.75% (96.11%-107.72%) and 102.21% (100.04%-104.42%), respectively. The GMRs (90% CIs) of S- and R-warfarin Cmax, AUC0-t, and AUC0-∞ were as follows: Cmax, 114.31% (106.30%-122.91%) and 115.09% (109.46%-121.01%), respectively; AUC0-t, 120.15% (116.71%-123.69%) and 119.01% (116.32%-121.76%); and AUC0-∞, 120.81% (117.17%-124.58%) and 121.94% (118.90%-125.05%). The GMRs (90% CIs) of warfarin PTmax and PTAUC were 92.73% (91.25%-94.22%) and 97.42% (96.61%-98.24%). The GMRs (90% CIs) of warfarin INRmax and INRAUC were 92.66% (91.17%-94.17%) and 97.36% (96.52%-98.21%). A total of 32 cases of mild adverse events were reported, and were recovered/resolved. There were no serious adverse events reported. IMPLICATIONS: No significant clinically relevant effects on the PK/PD properties of henagliflozin or warfarin were found with coadministration of the two drugs in these healthy male Chinese subjects. Based on these findings, it is expected that henagliflozin and warfarin can be used in combination without dose adjustment. Chinadrugtrials.org.cn identifier: CTR20190240.

A comprehensive investigation on source apportionment and multi-directional regional transport of volatile organic compounds and ozone in urban Zhengzhou.

To understand the characteristics, source apportionment, and regional transport of volatile organic compounds (VOCs) and ozone (O3) in a typical city with severe air pollution in central China, we observed and analyzed 115 VOC species at an urban site in Zhengzhou from 29 July to 26 September 2021. During this period, observation- and emission-based approaches revealed that Zhengzhou was in a VOC-limited regime. The average concentration of total VOCs (TVOCs) was 162.25 ± 71.42 µg/m3, dominated by oxygenated VOCs (OVOCs, 34.49%), alkanes (24.29%), and aromatics (19.49%). Six VOC sources were identified using positive matrix factorization (PMF) model, including paint solvent usage (25.32%), secondary production (24.11%), industrial production (19.22%), vehicle exhaust (16.18%), biogenic emission (8.87%), and combustion (6.30%). To assess the regional contribution and source apportionment of VOCs and O3, Comprehensive Air Quality Model with Extensions (CAMx) with the Ozone Source Apportionment Technology (OSAT) was used for simulation. Results showed that the VOCs were significantly affected by local emissions (about 70%), while O3 was mainly attributed to regional and super-regional transport. Regarding multi-directional regional transport of VOCs and O3, dominant contributions were from the northeast and east-northeast directions, and O3 contributions were also predominantly from the east and east-southeast directions. In terms of source apportionment, the transportation and industrial sectors (including solvent usage) were the major contributors to O3 and VOCs. To alleviate VOCs and O3 pollution, transportation and industrial emission reduction should be strengthened, and regional coordination, especially from the northeast to east-southeast directions, should be emphasized in addition to local management.

Quantification of cefaclor in human plasma using SIL-IS LC-ESI-MS/MS for pharmacokinetics study in healthy Chinese volunteers.

A steady, high-efficiency, and precise liquid chromatography-electrospray ionization-tandem mass spectrometry method was established and validated using cefaclor-d5 as the stable isotope-labeled internal standard for quantification of cefaclor in human plasma. One-step protein precipitation was applied to extract human plasma samples using methanol as precipitant. An Ultimate XB C18 column (2.1 × 50.0 mm, 5.0 µm) was used to achieve chromatographic separation. Mobile phases of gradient elution were an aqueous solution containing 0.1% formic acid (mobile phase A) and an acetonitrile solution containing 0.1% formic acid (mobile phase B). Electrospray ionization in positive-ion mode was applied to detect under multiple reaction monitoring mode. Target fragment ion pairs of cefaclor and stable isotope-labeled internal standard, respectively, were m/z 368.2 → 191.1 and m/z 373.2 → 196.1. Linear range of this method was between 20.0 and 10,000.0 ng/ml, with coefficient of determination (R2 ) >0.9900. Seven concentrations of quality control samples were used: 20.0 ng/ml (lower limit of quantitation), 60.0 ng/ml (low quality control), 650 ng/ml (middle quality control), 5000 ng/ml (arithmetic average middle quality control [AMQC]), 7500 ng/ml (high quality control), 10,000 ng/ml (upper limit of quantification), and 40,000 ng/ml (dilution quality control [DQC]). The method was validated for selectivity, lower limit of quantitation, linearity, accuracy, precision, recovery, matrix effect, dilution reliability, stability, carryover, and incurred sample reanalysis. This stable isotope-labeled internal standard liquid chromatography-electrospray ionization-tandem mass spectrometry approach has been successfully applied to study the pharmacokinetics of cefaclor dry suspension among healthy Chinese volunteers.

Adult Triploid Rainbow Trout Can Adapt to Various Dietary Lipid Levels by Coordinating Metabolism in Different Tissues.

Triploid rainbow trout can adapt to various dietary lipid levels; however, the mechanisms of systematic adaptation are not well understood. To investigate how adult triploid rainbow trout maintains lipid hemostasis under different exogenous lipid intake, a 77-day feeding trial was conducted. Diets with lipid contents of 20%, 25%, and 30% were formulated and fed to triploid rainbow trout with an initial weight of 3 ± 0.02 kg, and they were named L20, L25, and L30 group, respectively. Results showed that the condition factor, hepatosomatic index, liver color, and plasma triglyceride were comparable among three groups (p > 0.05), whereas the value of specific growth rate, viscerosomatic index, and liver glycogen content gradually increased with increasing dietary lipid level (p < 0.05). A significantly highest value of plasma glucose and nonesterified fatty acids were found in the L30 group (p < 0.05), whereas the significantly higher content of plasma total cholesterol, high-density lipoprotein-cholesterol, and low-density lipoprotein-cholesterol was found in the L25 group compared with those in L20 group (p < 0.05). As for lipid deposition, abdominal adipose tissue, and muscle were the main lipid storage place for triploid rainbow trout when tissues' weight is taken into consideration. Overall quantitative PCR showed that the lipid transport and glycolysis were upregulated, and fatty acids oxidative was downregulated in liver when fish were fed low lipid diets. It meant that the liver was the primary lipid metabolizing organ to low lipid diet feeding, which could switch energy supply between glycolysis and fatty acids oxidation. Fish fed with a moderate dietary lipid level diet could increase lipid uptake and promote lipogenesis in muscle. Abdominal adipose tissue could efficiently uptake excess exogenous free fatty acid through upregulating fatty acid uptake and synthesis de novo and then storing it in the form of triglyceride. Excess lipid uptake is preferentially stored in abdominal adipose tissue through coordinated fatty acid uptake and fatty acid synthesis de novo as dietary lipid levels increased. In summary, triploid rainbow trout can adapt to various dietary lipid levels by coordinating metabolism in different tissues.

Three-Period Bioequivalence Study of Sodium Levofolinate Injection With Calcium Levofolinate for Injection and Sodium Folinate for Injection in Healthy Chinese Subjects.

The aim of this study was to compare the bioequivalence and safety of test preparation sodium levofolinate injection with reference preparations of calcium levofolinate for injection and sodium folinate for injection in China. A single-center, randomized, open-label, 3-period, crossover test was conducted on 24 healthy subjects. Plasma concentration of levofolinate, dextrofolinate, and their metabolites l-5-methyltetrahydrofolate and d-5-methyltetrahydrofolate were quantified by a validated chiral-liquid chromatography-tandem mass spectrometry method. All adverse events (AEs) were documented to evaluate safety as they occurred and evaluated descriptively. Pharmacokinetic parameters (maximum plasma concentration, time to maximum concentration, area under the plasma concentration-time curve over the dosing interval, area under the plasma concentration-time curve from time 0 to infinity, terminal elimination half-life, and terminal rate constant) of 3 preparations were calculated. A total of 8 subjects (10 cases) of AEs occurred in this trial. No serious AEs or unexpected serious adverse reactions were observed. Sodium levofolinate was bioequivalent to calcium levofolinate and sodium folinate in Chinese subjects, and the 3 preparations were all well tolerated.

Novel insight into the evolution of volatile compounds during dynamic freeze-drying of Ziziphus jujuba cv. Huizao based on GC-MS combined with multivariate data analysis.

To understand the evolution of aroma in jujubes during dynamic freeze drying (FD), the relationship between aroma compounds, precursors, and related enzyme activities were analyzed. Fifty-three volatiles were identified during FD processing. After FD, the total aroma contents were increased from 11,004 to 14,603 µg/kg, ketones content was significantly decreased by 54.11 %, resulted in the loss of creamy note in freeze-dried jujube (FDJ). Through the network analysis, serine, glycine, proline, valine, cysteine, arginine, glutamic acid, lysine and leucine had the significant correlation with pyrazines, dominated the roasty note of FDJ. Linoleic acid, α-linolenic acid and oleic acid with lipoxygenase had important effects on the increase of esters (from 412 to 9,486 µg/kg), contributed fruity and sweet notes of FDJ. Besides, through the Mantel test, the influence degree of factors on the formation of FDJ aroma was ranked as temperature > enzyme activity > fatty acids > amino acids.

Clinical and economic burden of anxiety/depression among older adult COPD patients: evidence from the COPD-AD China Registry study.

Background: Anxiety and depression are common in patients with chronic obstructive pulmonary disease (COPD), especially older adult patients. This can complicate the disease progression and lead to increased clinical and economic burden. We sought to investigate the clinical and economic burdens associated with the presence of anxious and/or depressive symptoms among older adult COPD patients. Methods: We screened 579 patients aged over 60 years and diagnosed with COPD via a lung function test following the 2017 Global Initiative Chronic Obstructive Lung Disease (GOLD) guidelines. Anxiety and depression were measured using the Hospital Anxiety and Depression Scale (HADS) through face-to-face interviews at admission. Follow-up was conducted by telephone calls at 6, 12, 18, 24, and 36 months after discharge to assess clinical and economic burden. COPD-anxiety and/or depression patients were matched to patients without anxiety and depression (COPD-only) using propensity scores. Multivariate regression models were used to compare clinical and economic burden between COPD-anxiety and/or depression and COPD-only groups. Results: Compared with COPD-only patients, COPD patients complicated with anxiety and/or depression had increased clinical burden, including higher COPD-related outpatient visits, COPD-related hospitalizations, and length of COPD-related hospitalizations (p < 0.001). Moreover, they also had an increased economic burden, including higher annual total healthcare costs, medical costs, and pharmacy costs (p < 0.001). Conclusion: Older adult COPD patients with anxiety or depression had significantly higher clinical and economic burdens than patients without these comorbidities. These findings deserve further exploration and may be useful for the formulation of relevant healthcare policies.

The potential therapeutic value and application prospect of engineered exosomes in human diseases.

Exosomes are tiny vesicles produced by a wide range of cells that contain complex RNA and protein. In the diagnosis, treatment, and prevention of illness, they offer great potential. In vitro engineering technique modifies exosomes to produce designed exosomes that include nucleic acids, proteins, and medicines, and are targeted to particular tissues or cells. Their applications range from tumor imaging and gene therapy to vaccine production and regenerative medicine to targeted medication delivery. Many disciplines have promising futures for using this technology. In this review, we'll look at the potential therapeutic usefulness and use of engineered exosomes in a variety of human illnesses with various systemic manifestations.

PTBPs: An immunomodulatory-related prognostic biomarker in pan-cancer.

Background: The polypyrimidine tract-binding protein (PTBP) nuclear ribonucleoprotein family of proteins, including PTBP1, PTBP2 and PTBP3, regulate the process of cell proliferation, differentiation, apoptosis and carcinogenesis. PTBPs exhibit oncogenic effects in certain tumors. However, the role of PTBPs in pan-cancer remains unclear. Our study examined the clinical significance and mechanism of PTBPs in pan-cancer. Methods: We compared the expression of PTBPs in paired and unpaired tissue samples from the Cancer Genome Atlas (TCGA) database. Univariate and multivariate Cox regression, Kaplan-Meier curves, and time-dependent receiver operating characteristic (ROC) curves were used to assess the prognostic significance of PTBPs in pan-cancer. The cBioPortal database also identified genomic abnormalities in PTBPs. TISIDB, TCGA, and Cellminer were used to investigate the relationship between PTBP expression and immune subtypes, immune checkpoint (ICP) genes, tumor mutational burden (TMB), microsatellite instability (MSI), tumor-infiltrating immune cells, and chemosensitivity. cBioPortal was used to search for PTBP co-expressing genes in pan-cancer, and GO and KEGG enrichment analyses were performed to search for PTBP-related signaling pathways. Results: PTBPs were shown to be widely upregulated in human tumor tissues. PTBP1 showed good prognostic value in ACC, KIRP, and LGG; PTBP2 in ACC and KICH; and PTBP3 in ACC, LGG, and PAAD, with AUC >0.7. PTBPs were differentially expressed in tumor immune subtypes and had a strong correlation with tumor-infiltrating lymphocytes (TILs) in the tumor microenvironment (TME). In addition, PTBP expressions were related to ICP, TMB, and MSI, suggesting that these three PTBPs may be potential tumor immunotherapeutic targets and predict the efficacy of immunotherapy. Enrichment analysis of co-expressed genes of PTBPs showed that they may be involved in alternative splicing, cell cycle, cellular senescence, and protein modification. Conclusion: PTBPs are involved in the malignant progression of tumors. PTBP1, PTBP2 and PTBP3 may be potential biomarkers for prognosis and immunotherapy in pan-cancer and may be novel immunotherapeutic targets.

Evaluation of Pharmacokinetic Interactions Between the New SGLT2 Inhibitor SHR3824 and Valsartan in Healthy Chinese Volunteers.

PURPOSE: Hypertension is often observed in patients with diabetes, and the progression of diabetic nephropathy is closely related to blood pressure elevation. Thus, the effects of hypoglycemic drugs on kidney function and pharmacokinetic interactions in combination with antihypertensive and hypoglycemic drugs are of great clinical value. The purpose of this study was to evaluate the pharmacokinetic interactions between henagliflozin (SHR3824), a new sodium-dependent glucose transporter 2 (SGLT2) inhibitor class drug, and valsartan, an angiotensin II receptor blocker. METHODS: A single-center, single-arm, open-label, self-controlled study was conducted in healthy Chinese volunteers. The pharmacokinetic parameters were calculated with Phoenix WinNonlin version 7.0, and the statistical analysis was performed with SAS version 9.4. Data on pharmacokinetic parameters (single and/or steady-state) were collected and tabulated for different analytes (valsartan and SHR3824) according to the sampling time specified in the protocol. Continuous attention was paid to the safety of all subjects. The aim of the study was to evaluate the effect of a single dose of valsartan on the pharmacokinetic behavior of SHR3824 after multiple doses of SHR3824 (Cmax,ss and AUCτ,ss) and the effect of multiple doses of SHR3824 on the pharmacokinetic behavior of valsartan (Cmax, AUC0-24h, and AUC0-∞). A mixed effect model was used to estimate the point estimation and 90% CI of the geometric mean ratio of the corresponding pharmacokinetic indices at the combined-medication stage (SHR3824 + valsartan) and the single-medication stage (SHR3824 or valsartan). FINDINGS: Twelve volunteers were screened into this experiment and underwent blood sampling. The pharmacokinetic properties of SHR3824 were evaluated after its administration alone or in combination with valsartan. Point estimates and 90% CIs of the geometric mean ratio of SHR3824 Cmax,ss and AUCτ,ss were within the conventional bioequivalence range of 80% to 125%. The pharmacokinetic properties of valsartan were evaluated after its administration alone or in combination with SHR3824. The geometric mean ratios and 90% CIs of the valsartan Cmax, AUC0-24h, and AUC0-∞ were also within the range of 80% to 125%. Thirty-four mild adverse events were reported, with no serious adverse events or suspected unexpected serious adverse reactions. IMPLICATIONS: This study provides basis for the clinical co-administration of SHR3824 with angiotensin II receptor blockers represented by valsartan. Based on these findings, co-administration of SHR3824 and valsartan seemed to have no effect on the pharmacokinetic properties of either drug. Chinadrugtrials.org.cn Identifier: CTR20180002.

Tumor Microenvironment: Lactic Acid Promotes Tumor Development.

Lactic acid is a "metabolic waste" product of glycolysis that is produced in the body. However, the role of lactic acid in the development of human malignancies has gained increasing interest lately as a multifunctional small molecule chemical. There is evidence that tumor cells may create a large amount of lactic acid through glycolysis even when they have abundant oxygen. Tumor tissues have a higher quantity of lactic acid than normal tissues. Lactic acid is required for tumor development. Lactate is an immunomodulatory chemical that affects both innate and adaptive immune cells' effector functions. In immune cells, the lactate signaling pathway may potentially serve as a link between metabolism and immunity. Lactate homeostasis is significantly disrupted in the TME. Lactate accumulation results in acidosis, angiogenesis, immunosuppression, and tumor cell proliferation and survival, all of which are deleterious to health. Thus, augmenting anticancer immune responses by lactate metabolism inhibition may modify lactate levels in the tumor microenvironment. This review will evaluate the role of lactic acid in tumor formation, metastasis, prognosis, treatment, and histone modification. Our findings will be of considerable interest to readers, particularly those engaged in the therapeutic treatment of cancer patients. Treatments targeting the inhibition of lactate synthesis and blocking the source of lactate have emerged as a potential new therapeutic option for oncology patients. Additionally, lactic acid levels in the plasma may serve as biomarkers for disease stage and may be beneficial for evaluating therapy effectiveness in individuals with tumors.

NCOA4: An Immunomodulation-Related Prognostic Biomarker in Colon Adenocarcinoma and Pan-Cancer.

Treatment of cancer in humans requires a thorough understanding of the multiple pathways by which it develops. Recent studies suggest that nuclear receptor coactivator 4 (NCOA4) may be a predictive biomarker for renal cancer. In the present work, TCGA, GEPIA, and several bioinformatics approaches were used to analyze the NCOA4 expression patterns, prognostic relevance, and association between NCOA4 and clinicopathological features and immune cell infiltration. We investigated NCOA4 expression in malignancies. Low NCOA4 expression was associated with poor overall survival in individuals with malignancies such as cholangiocarcinoma, colon adenocarcinoma, and clear cell renal carcinoma. We also analyzed NCOA4 DNA methylation in normal and primary tumor tissues and investigated possible functional pathways underlying NCOA4-mediated oncogenesis. In conclusion, downregulation of NCOA4 is associated with poor prognosis, and NCOA4 may be a predictive biomarker for COAD.

Transcriptome profiling of mRNAs in muscle tissue of Pinan cattle and Nanyang cattle.

Mammalian muscle development is regulated by complex gene networks at the molecular level. The revelation of gene regulatory mechanisms is an important basis for the study of muscle development and molecular breeding. To analyze the excellent meat performance of Pinan cattle at the molecular level, we performed high-throughput RNA sequencing to analyze the key regulatory genes that determine the muscle quality traits in Pinan cattle (n = 3) and Nanyang cattle (n = 3). We identified 57 differentially expressed genes in muscle tissue of Pinan cattle compared to that of Nanyang cattle, including 32 upregulated and 25 downregulated genes. GO enrichment analysis showed that these genes were significantly enriched in 'molecular function', including voltage-gated ion channel activity, calcium channel activity and calcium ion binding, and KEGG pathway analysis results revealed that adrenergic signaling in cardio myocytes, cell adhesion molecules and inositol phosphate metabolism pathway were significantly enriched. We identified the reliability of RNA-Seq data through RT-qPCR. Meanwhile, we found that GSTA3, PLCB1 and ISYNA1 genes are highly expressed in muscle tissue of Pinan cattle, and these genes play important roles in PI3K/Akt, MEK1/2-ERK and p53-ISYNA1 signaling pathway. In summary, our results suggested that these differentially expressed genes may play important roles in muscle development in Pinan cattle. However, the functions and mechanism of these significantly differential expressed genes should be investigated in future studies.

Knockdown of a ß-Adrenergic-Like Octopamine Receptor Affects Locomotion and Reproduction of Tribolium castaneum.

The neurohormone octopamine regulates many crucial physiological processes in insects and exerts its activity via typical G-protein coupled receptors. The roles of octopamine receptors in regulating behavior and physiology in Coleoptera (beetles) need better understanding. We used the red flour beetle, Tribolium castaneum, as a model species to study the contribution of the octopamine receptor to behavior and physiology. We cloned the cDNA of a ß-adrenergic-like octopamine receptor (TcOctß2R). This was heterologously expressed in human embryonic kidney (HEK) 293 cells and was demonstrated to be functional using an in vitro cyclic AMP assay. In an RNAi assay, injection of dsRNA demonstrated that TcOctß2R modulates beetle locomotion, mating duration, and fertility. These data present some roles of the octopaminergic signaling system in T. castaneum. Our findings will also help to elucidate the potential functions of individual octopamine receptors in other insects.

A circular RNA from NFIX facilitates oxidative stress-induced H9c2 cells apoptosis.

Myocardial infarction is the leading cause of death worldwide, and cardiomyocyte apoptosis during myocardial infarction and reperfusion is a significant factor of poor prognosis. As important regulatory molecules, biofunctions of circRNAs in the pathogenesis of myocardial infarction remain elusive. To confirm the expression level and biological function of circNFIX in cardiomyocytes upon oxidative stress. Divergent polymerase chain reaction and Sanger sequencing were performed to verify the circular structure. The stability of circNFIX was confirmed by RNase R treatment and actinomycin D assay. In order to simulate oxidative stress during myocardial infarction, H9c2 cells were subjected to hydrogen peroxide and hypoxia stimulation. In vivo, mouse models of myocardial ischemia were established. The biological function of circNFIX in cardiomyocytes was investigated through loss- and gain-of-function assays, and cardiomyocyte apoptosis level was detected by the terminal deoxyribonucleotidyl transferase-mediated TdT-mediated dUTP nick end labeling assay and Western blot. CircNFIX is abundant, conserved, and stable in H9c2 cells. The expression of circNFIX was significantly downregulated in cardiomyocytes subjected to oxidative stress. Enforced CircNFIX promotes H9c2 cells apoptosis induced by hydrogen peroxide, in sharp contrast to circNFIX knockdown. In this study, we found that circNFIX served as a pro-apoptosis factor in cardiomyocyte apoptosis. CircNFIX possesses potential to be the biomarker and therapeutic target in myocardial infarction.