EGFR immunohistochemical assay of the tumor sections from different groups.

<p>A: 0 mg/kg As₂O₃ (control). The strongest red-brownish membrane-bound immunostaining on the A431 tumor tissue slice reflected the abundant over expression of EGFR (+++). B: 0.5 mg/kg As₂O₃ treatment group. Strong brownish membrane staining indicated plentiful of EGFR expression (++∼+++). C: 2.5 mg/kg As₂O₃ treatment group demonstrated moderate to low EGFR expression (+∼++). D: 5.0 mg/kg As₂O₃ treatment group showed weak EGFR expression (+). (Scale bar = 100 µm).</p

Data_Sheet_1_Randomized trial of transcutaneous auricular vagus nerve stimulation on patients with disorders of consciousness: A study protocol.doc

BackgroundTranscutaneous auricular vagus nerve stimulation (taVNS) has recently been explored for the treatment of Disorders of consciousness (DoC) caused by traumatic brain injury. The evidence of taVNS during the consciousness recovery has been recently reported. However, the mechanism of taVNS in the recovery of consciousness is not clear. This study attempts to investigate the effectiveness of taVNS in DoC by means of Coma Recovery Scale-Revised (CRS-R), Magnetic resonance imaging (MRI), Electrophysiology (EEG), and Single-molecular array (Simoa).Methods/designNighty patients with DoC acquired brain injury are randomized into one of three groups receiving sham taVNS or active taVNS (just left and left or right), respectively. Each of the three groups will experience a 40 days cycle (every 10 days for a small period, baseline 2 weeks, intervention 2 weeks, 40 min per day, 5 days per week, then no intervention for 2 weeks, intervention 2 weeks, 40 min per day, and 5 days per week). Primary outcomes (CRS-R) will be recorded five times during every period. Secondary outcomes will be recorded at the first and at the last period [MRI, EEG, Phosphorylated tau (P-tau), and Neurofilament light chain (NFL)]. We will take notes the adverse events and untoward effects during all cycles.DiscussionTranscutaneous auricular vagus nerve stimulation as a painless, non-invasive, easily applied, and effective therapy was applied for treatment of patients with depression and epilepsy several decades ago. Recent progress showed that taVNS has behavioral effects in the consciousness recovery. However, there is no clinical evidence to support the effects of taVNS on brain activity. Therefore, we will design a randomized controlled trial to evaluate the effectiveness and safety of taVNS therapy for DoC, and explore neural anatomy correlated to taVNS during the consciousness recovery. Finally, this protocol also tests some biomarkers along with the recovery of consciousness.Clinical Trial RegistrationChinese Clinical Trial Registry, ChiCTR2100045161. Registered on 9 April 2021.</p

Table_1_Relationship between self-care compliance, trust, and satisfaction among hypertensive patients in China.pdf

IntroductionHypertension is a growing public health concern worldwide. It is a leading risk factor for all-cause mortality and may lead to complications such as cardiovascular disease, stroke, and kidney failure. Poor compliance of hypertensive patients is one of the major barriers to controlling high blood pressure. Compliance is not ideal among Chinese patients, and increasing patient self-care compliance with hypertension is necessary.MethodsThis article analyzes the status of self-care compliance, trust, and satisfaction among Chinese hypertensive patients using cross-sectional data from Zhejiang Province. We use a multi-group structural equation model (MGSEM) to compare the interrelationships across genders.ResultsThe study's findings show that the average trust, satisfaction, and compliance scores are 3.92 ± 0.55, 3.98 ± 0.61, and 3.33 ± 0.41, respectively. Female patients exhibit higher average total scores for trust and compliance than male patients. The SEM results indicate that trust has a direct positive association with compliance [β = 0.242, 95% CI: (0.068, 0.402)] and satisfaction [β = 0.260, 95% CI: (0.145, 0.367)], while their satisfaction is not directly associated with compliance. The results of MGSEM show that trust has an indirect effect on compliance in the male group through satisfaction [β = 0.051, P DiscussionThis study reveals the mechanisms of self-care compliance, trust, and satisfaction among Chinese hypertensive patients. Its findings may serve as a reference for guiding primary healthcare providers to improve hypertension patients' compliance and implement gender-targeted health interventions.</p

The cellular EGFR expression percentage of different treatment groups <i>in vitro</i> assessed by flow cytometry.

<p>The dynamic EGFR cellular expression percentage after treatment of different concentrations of As₂O₃ (0 µM, 0.5 µM, 2.5 µM, 5.0 µM) varied with time (*<i>p</i><0.05, **<i>p</i><0.01). All experiments were carried out in triplicate; each point represents the mean ± standard error values.</p

Effects of As₂O₃ on A431 cell EGFR expression.

<p>Cells were exposed to different concentrations of As₂O₃. At 48 h post-treatment, cells were assessed by fluorescence microscopy for visualization of the intake of EGF-Cy5.5 and cell immunohistochemistry for assay of EGFR expression. A–D, representative fluorescence images of different groups (Scale bar = 100 µm), Cy5.5 was pseudo-colored red, DAPI was pseudo-colored blue; E-H, representative images of cellular EGFR Immunohistochemistry assay (Scale bar = 100 µm), diaminobenzidine (DAB) showed as brown color represented EGFR expression and hematoxylin showed as blue color indicated the cellular nuclear. Interestingly, cell numbers decreased as the arsenic trioxide concentration increased from 0 µM to 5 µM in both fluorescent and immunostained images, indicating that As₂O₃ induced a dose-dependent inhibition on tumor cell proliferation as previously reported <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046255#pone.0046255-Kumar1" target="_blank">[31]</a>.</p

Monitoring tumor growth and body weight of tumor-bearing mice during treatment.

<p>Tumor growth (A) and body weight (B) of tumor-bearing mice treated with saline (as untreated control group), arsenic trioxide (ATO) at 0.5 mg/kg, 2.5 mg/kg or 5.0 mg/kg daily for 12 days. Six mice were used in each group. The tumor volume and body weight of all four groups were also measured every two days. Tumor volume was calculated according to the formula V = (a×b²)/2 where a and b represent the length and width of the tumor. Measurements were continued to 12th day. <i>p</i><0.05 is a significant difference between control and treatment groups.</p

<i>In vivo</i> dynamic near-infrared fluorescent imaging of A-431 tumor models.

<p>A: The representative fluorescence images of the tumor regions in mice were acquired at 4 h post injection of EGF-Cy5.5. Fluorescence signal from Cy5.5 was pseudo-colored red. B: The dynamic measurement comparison of fluorescence intensity of tumor in different groups. It was demonstrated that the fluorescence intensity in the tumor regions were changed with time (<i>p</i><0.05). On day 0 (before As₂O₃ treatment), there was no significant difference of signal intensity of tumors between treatment and control groups (<i>p</i>>0.05). On day 4, 8, 12 (after As₂O₃ treatment), the signal intensity of EGF-Cy5.5 uptake by control group (0 mg/kg/day As₂O₃) gradually increased, while the intensities in other three groups with different concentrations (0.5 mg/kg/day, 2.5 mg/kg/day, 5.0 mg/kg/day) of As₂O₃ treatment gradually decreased (<i>p</i><0.05). C: The <i>in vivo</i> fluorescence intensity was compared between post-treatment (on day 12) in four different groups compared with respective pre-treatment (on day 0). All plots are representative of results from groups of mice treated under the same experimental conditions. Each point represents the mean values (n = 6/group, *<i>p</i><0.05, **<i>p</i><0.01).</p

Image_1_Genome, transcriptome, and metabolome analyses provide new insights into the resource development in an edible fungus Dictyophora indusiata.JPEG

Dictyophora indusiata (Vent. Ex Pers.) Fisch. (DI) is an edible and medicinal fungus widely used in East Asian countries. However, during DI cultivation, the formation of fruiting bodies cannot be regulated, which leads to yield and quality losses. The present study performed a combined genome, transcriptome, and metabolome analysis of DI. Using Nanopore and Illumina sequencing approaches, we created the DI reference genome, which was 67.32 Mb long with 323 contigs. We identified 19,909 coding genes on this genome, of which 46 gene clusters were related to terpenoid synthesis. Subsequent transcriptome sequencing using five DI tissues (cap, indusia, mycelia, stipe, and volva) showed high expression levels of genes in the cap, indicating the tissue’s importance in regulating the fruiting body formation. Meanwhile, the metabolome analysis identified 728 metabolites from the five tissues. Mycelium was rich in choline, while volva was rich in dendronobilin; stipe had monosaccharides as the primary component, and the cap was the main source of indole acetic acid (IAA) synthesis. We confirmed the importance of tryptophan metabolism for DI fruiting body differentiation based on KEGG pathway analysis. Finally, the combined multiomics analysis identified three new genes related to IAA synthesis of the tryptophan metabolic pathway in the cap, which may regulate DI fruiting body synthesis and improve DI quality. Thus, the study’s findings expand our understanding of resource development and the molecular mechanisms underlying DI development and differentiation. However, the current genome is still a rough draft that needs to be strengthened.</p

Identification of Potential Biomarkers for Ovarian Cancer by Urinary Metabolomic Profiling

To evaluate the application of urinary metabolomics on discovering potential biomarkers for epithelial ovarian cancer (EOC), urine samples from 40 preoperative EOC patients, 62 benign ovarian tumor (BOT) patients, and 54 healthy controls were collected and analyzed with ultraperformance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS). Good separations were obtained for EOC vs BOT, EOC vs healthy controls analyzed by partial least-squares discriminant analysis, or principal component analysis. Twenty-two ascertained metabolomic biomarkers were found to be disturbed in several metabolic pathways among EOC patients, including nucleotide metabolism (pseudouridine, N4-acetylcytidine), histidine metabolism (l-histidine, imidazol-5-yl-pyruvate), tryptophan metabolism (3-indolelactic acid), and mucin metabolism (3′-sialyllactose and 3-sialyl-N-acetyllactosamine). In addition, the concentrations of some urinary metabolites of 18 postoperative EOC patients among the 40 EOC patients changed significantly compared with those of their preoperative condition, and four of them suggested recovery tendency toward normal level after surgical operation, including N4-acetylcytidine, pseudouridine, urate-3-ribonucleoside, and succinic acid. These metabolites would be highly postulated to be associated with EOC. In conclusion, our study demonstrated that urinary metabolomics analysis by UPLC-QTOF/MS, performed in a minimally noninvasive and convenient manner, possessed great potential in biomarker discovery for EOC

Table_1_Genome, transcriptome, and metabolome analyses provide new insights into the resource development in an edible fungus Dictyophora indusiata.XLSX

Dictyophora indusiata (Vent. Ex Pers.) Fisch. (DI) is an edible and medicinal fungus widely used in East Asian countries. However, during DI cultivation, the formation of fruiting bodies cannot be regulated, which leads to yield and quality losses. The present study performed a combined genome, transcriptome, and metabolome analysis of DI. Using Nanopore and Illumina sequencing approaches, we created the DI reference genome, which was 67.32 Mb long with 323 contigs. We identified 19,909 coding genes on this genome, of which 46 gene clusters were related to terpenoid synthesis. Subsequent transcriptome sequencing using five DI tissues (cap, indusia, mycelia, stipe, and volva) showed high expression levels of genes in the cap, indicating the tissue’s importance in regulating the fruiting body formation. Meanwhile, the metabolome analysis identified 728 metabolites from the five tissues. Mycelium was rich in choline, while volva was rich in dendronobilin; stipe had monosaccharides as the primary component, and the cap was the main source of indole acetic acid (IAA) synthesis. We confirmed the importance of tryptophan metabolism for DI fruiting body differentiation based on KEGG pathway analysis. Finally, the combined multiomics analysis identified three new genes related to IAA synthesis of the tryptophan metabolic pathway in the cap, which may regulate DI fruiting body synthesis and improve DI quality. Thus, the study’s findings expand our understanding of resource development and the molecular mechanisms underlying DI development and differentiation. However, the current genome is still a rough draft that needs to be strengthened.</p