Myricetin ameliorates airway inflammation and remodeling in asthma by activating Sirt1 to regulate the JNK/Smad3 pathway.

BACKGROUND: Myricetin has various biological activities and health benefits; however, its effects on airway remodeling in asthma have not been reported. PURPOSE: We aimed to investigate the possibility that myricetin improves airway remodeling by activating Sirt1 and has potential as a new treatment for asthma. METHODS: RAW 264.7 cells were stimulated with lipopolysaccharide and co-cultured with 3T6 cells in vitro to simulate the in vivo effects of inflammation on airway remodeling. Using an ovalbumin-induced chronic asthma mouse model, we compared changes in inflammatory factors and airway remodeling-related factors under treatment with myricetin and/or the Sirt1 inhibitor EX-527 using western blotting and quantitative PCR. Expression plasmids carrying Smad3 site mutations were transfected into 3T6 cells to identify the Sirt1 deacetylation site on Smad3 protein. RESULTS: Myricetin significantly reduced the infiltration of airway inflammatory cells and the production of interleukin (IL)-6 and IL-5, and inhibited mucus secretion by goblet cells, collagen fiber proliferation, and the increase in inflammatory cells in bronchoalveolar lavage fluid from asthmatic mice. Results of in vitro experiments were consistent with those conducted in vivo. Exploring the mechanism of action of myricetin, we found that myricetin downregulated the levels of phosphorylated (p)-JNK, p-Smad3, and acetylated Smad3 proteins by activating Sirt1 both in vivo and in vitro. K341 was identified as the main deacetylation site of Smad3 by myricetin-activated Sirt1. CONCLUSION: Myricetin ameliorates airway inflammation and remodeling in asthma by activating Sirt1 to regulate the JNK/Smad3 pathway.

Ferroptosis-related genes DUOX1 and HSD17B11 affect tumor microenvironment and predict overall survival of lung adenocarcinoma patients.

BACKGROUND: Recent studies have found that ferroptosis-related genes (FRGs) have broad applications in tumor therapy. However, the predictive potential of these genes in lung adenocarcinoma (LUAD) remains to be fully characterized. We aimed to investigate the FRGs that might be potential targets for LUAD. METHODS: We screened the RNA sequencing samples from LUAD patients from the GEO database and analyzed the ferroptosis-related differentially expressed genes (DEGs). A functional analysis of DEGs was performed. The risk model was constructed to evaluation and validation FRGs. We explored the immune landscape of LUAD and controls. The value of FRGs in diagnosing LUAD was tested in the GSE30219, GSE37745, GSE0081 datasets, and qPCR was used to verify their diagnostic value in LUAD patients in our hospital. RESULTS: A total of 1327 DEGs in quantitative proteomics were obtained, of which ferroptosis-related DEGs were 259. Enrichment analysis showed significant enrichment in the absorption and metabolism of fatty acids and arachidonic acid. The upregulated genes (GCLC, RRM2, AURKA, SLC7A5, and SLC2A1) and downregulated genes (ANGPTL7, ALOX15, ALOX15B, HSD17B11, IL33, TSC22D3, and DUOX1) were selected as core genes in tissue samples from 62 patients by qPCR. DUOX1 and HSD17B11 were obtained by bioinformatics analysis, both of which showed similar expression trends at the RNA and protein levels. The Kaplan-Meier method showed that DUOX1 and HSD17B11 were closely related to the overall survival (OS) of LUAD patients. CONCLUSION SUBSECTIONS: Ferroptosis-related genes DUOX1 and HSD17B11 are of considerable value in the diagnosis of LUAD patients. Their low expression suggests an increased recurrence rate and leads to a decrease in the patient quality of life.

Prognostic model for unresectable hepatocellular carcinoma treated with dual PD-1 and angiogenesis blockade therapy.

BACKGROUND AND AIMS: Dual programmed death 1 (PD-1) and angiogenesis blockade therapy is a frontline treatment for hepatocellular carcinoma (HCC). An accepted model for survival prediction and risk stratification in individual patients receiving this treatment is lacking. Aimed to develop a simple prognostic model specific to these patients. APPROACH AND RESULTS: Patients with unresectable HCC undergoing dual PD-1 and angiogenesis blockade therapy were included in training cohort (n=168) and validation cohort (n=72). We investigated the prognostic value of clinical variables on overall survival using a Cox model in the training set. A prognostic score model was then developed and validated. Predictive performance and discrimination were also evaluated. Largest tumor size and Alpha-fetoprotein concentration at baseline and Neutrophil count and Spleen volume change after 6 weeks of treatment were identified as independent predictors of overall survival in multivariable analysis and used to develop LANS score. Time-dependent receiver operating characteristic analysis, calibration curves, and C-index showed LANS score had favorable performance in survival prediction. Patients were divided into three risk categories based on LANS score. Median survival for patients with low, intermediate, and high LANS scores was 31.7, 23.5, and 11.5 months, respectively (p<0.0001). The disease control rates were 96.4%, 64.3%, and 32.1%, respectively (p<0.0001). The predictive performance and risk stratification ability of the LANS score were confirmed in validation and entire cohorts. CONCLUSION: The LANS score model can provide individualized survival prediction and risk stratification in patients with unresectable HCC undergoing dual PD-1 and angiogenesis blockade therapy.

Emetine dihydrochloride alleviated radiation-induced lung injury through inhibiting EMT.

Radiation-induced lung injury (RILI), divided into early radiation pneumonia (RP) and late radiation-induced pulmonary fibrosis (RIPF), is a common serious disease after clinical chest radiotherapy or nuclear accident, which seriously threatens the life safety of patients. There has been no effective prevention or treatment strategy till now. Epithelial-mesenchymal transition (EMT) is a key step in the occurrence and development of RILI. In this study, we demonstrated that emetine dihydrochloride (EDD) alleviated RILI through inhibiting EMT. We found that EDD significantly attenuated EMT-related markers, reduced Smad3 phosphorylation expression after radiation. Then, for the first time, we observed EDD alleviated lung hyperaemia and reduced collagen deposit induced by irradiation, providing protection against RILI. Finally, it was found that EDD inhibited radiation-induced EMT in lung tissues. Our study suggested that EDD alleviated RILI through inhibiting EMT by blocking Smad3 signalling pathways. In summary, our results indicated that EDD is a novel potential radioprotector for RILI.

Evaluation of optic nerve head vessels density changes after phacoemulsification cataract surgery using optical coherence tomography angiography.

AIM: To evaluate optic nerve head (ONH) vessel density (VD) changes after cataract surgery using optical coherence tomography angiography (OCTA). METHODS: This was a prospective observational study. Thirty-four eyes with mild/moderate cataracts were included. ONH scans were obtained before and 3mo after cataract surgery using OCTA. Radial peripapillary capillary (RPC) density, all VD, large VD and retinal nerve fiber layer thickness (RNFLT) in total disc, inside disc, and different peripapillary sectors were assessed and analyzed. Image quality score (QS), fundus photography grading and best-corrected visual acuity (BCVA) were also collected, and correlation analyses were performed between VD change and these parameters. RESULTS: Compared with baseline, both RPC and all VD increased in inside disc area 3mo postoperatively (from 47.5%±5.3% to 50.2%±3.7%, and from 57.87%±4.30% to 60.47%±3.10%, all P<0.001), but no differences were observed in peripapillary area. However, large VD increased from 5.63%±0.77% to 6.47%±0.72% in peripapillary ONH region (P<0.001). RPC decreased in inferior and superior peripapillary ONH parts (P=0.019, <0.001 respectively). There were obvious negative correlations between RPC change and large VD change in inside disc, superior-hemi, and inferior-hemi (r=-0.419, -0.370, and -0.439, P=0.017, 0.044, and 0.015, respectively). No correlations were found between VD change and other parameters including QS change, fundus photography grading, postoperative BCVA, and postoperative peripapillary RNFLT. CONCLUSION: RPC density and all VD in the inside disc ONH region increase 3mo after surgery in patients with mild to moderate cataract. No obvious VD changes are found in peripapillary area postoperatively.

Accuracy of newer generation intraocular lens power calculation formulas in pediatric cataract patients.

PURPOSE: To evaluate the accuracy of newer generation intraocular lens (IOL) power calculation formulas (EVO 2.0 and Kane) with established formulas (Barrett Universal II, Haigis and SRK/T) in pediatric cataract patients. METHODS: Retrospective study. We enrolled 110 eyes (110 patients) in Eye Hospital of Wenzhou Medical University. All patients underwent uneventful cataract surgery and implanted with posterior chamber IOL in the bag. We calculate the mean prediction errors (PE) and percentage within 1 diopter (D) at 1 month to assess the accuracy, and percentage > 2D was defined as prediction accident. Then, we performed subgroup analysis according to age and axial length (AL). RESULTS: The mean age and AL were 37.45 ± 23.28 months and 21.16 ± 1.29 mm. The mean PE for all patients was as follows: Barrett (- 0.30), EVO (0.18), Haigis (- 0.74), Kane (- 0.36), and SRK/T (0.58), p < 0.001. In addition, EVO and SRK/T formulas were relatively accurate in patients younger than 24 months and with AL ≤ 21 mm, while EVO got lower prediction accident rate than SRK/T (3/41 vs 8/41, 4/52 vs 5/52). Moreover, Barrett, EVO, and Kane formulas achieved better accuracy and lower prediction accident rate in patients older than 24 months and with AL > 21 mm (both > 51/69 and 43/58, and < 3/69 and 3/58). CONCLUSIONS: In patients older than 24 months and with AL > 21 mm, Barrett, EVO, and Kane formulas were relatively accurate, while in patients younger than 24 months and with AL ≤ 21 mm, EVO was more accurate, followed by SRK/T formula.

PBK/TOPK promotes chemoresistance to oxaliplatin in hepatocellular carcinoma cells by regulating PTEN.

Oxaliplatin (OXA) resistance limits the efficiency of treatment for hepatocellular carcinoma (HCC). Studies have shown that the PDZ-binding kinase (PBK) plays important roles in tumors. However, the role of PBK in HCC is still a problem. In this study, we explored whether PBK is involved in the chemoresistance to OXA in HCC. Expressions of PBK in six HCC cell lines and one human hepatocytes line were determined by real-time quantitative PCR and western blot analysis. SNU-182 and HepG2 cells were chosen to induce OXA resistance. PBK was silenced or overexpressed in OXA-resistant and sensitive cell lines. Then, cell proliferation, migration, and invasion were measured by cholecystokinin-8 assay and Transwell assay, respectively. The Cancer Genome Atlas dataset showed that PBK is highly expressed in HCC and signifies poor prognosis to patient with HCC. Results showed that expression of PBK in HCC cells was significantly higher than that in THLE2 cells, and it was further increased in OXA-resistant HCC cells. Silencing of PBK promoted the sensitivity of drug-resistant HCC cells to OXA. Overexpression of PBK relieved the apoptosis induced by OXA and promoted the migration and invasion of OXA-sensitive HCC cells. Thus, this study revealed that high PBK expression is correlated with OXA resistance in HCC cells, which may provide a promising therapeutic target for treating HCC.

The variation in the major constituents of the dried rhizome of Ligusticum chuanxiong (Chuanxiong) after herbal processing.

BACKGROUND: Rhizoma Chuanxiong (RC; Chuanxiong), which is the dried rhizome of Ligusticum chuanxiong (Umbelliferae), is commonly used in Chinese medicine (CM) for improving blood circulation and dispersing blood stasis. RC is usually processed before use in clinical practice to enhance its therapeutic efficacy. This study aimed to investigate the temporal variations of the major constituents of RC by HPLC-DAD-MS during herbal processing to investigate the effects of an adjuvant (e.g., wine), steaming vs stir-frying and the optimal processing time. METHODS: An HPLC-DAD-MS method was developed to determine the major constituents of the RC processed by one of the four processing methods, i.e., stir-frying, steaming, stir-frying with rice wine and steaming with rice wine. Processing was conducted over 60 min. Six major compounds, namely ferulic acid, senkyunolide I, senkyunolide H, senkyunolide A, Z-ligustilide and levistolide A, were selected as markers to analyze the effects on the markers' levels of the different processing methods and optimize the processing time. RESULTS: The results indicated that (a) processing with wine had no discernible impact on the amounts of the six chemical markers in RC; (b) the amounts of the major constituents of RC subjected to steam processing were higher than those of the RC subjected to stir-fry processing. CONCLUSION: Among the four different methods evaluated for RC processing, steaming was better and the optimal time for steaming RC was 40 min.

Fingerprint analysis of processed Rhizoma Chuanxiong by high-performance liquid chromatography coupled with diode array detection.

BACKGROUND: Rhizoma Chuanxiong (RC) is the dried rhizome of Ligusticum chuanxiong Hort., and various types of processed Rhizoma Chuanxiong (PRC) are widely used in China. However, quality assurance and quality control of these processed medicines remain challenging. This study aims to investigate the chemical compositions of various PRC preparations by a high-performance liquid chromatography (HPLC) coupled with diode array detection (DAD) method. METHODS: A HPLC-DAD method with validation was developed for PRC samples. Seven batches of plant samples from two processing methods, stir-frying and steaming, were analyzed by the HPLC-DAD method. Common peaks in PRC chromatograms were chosen to calculate their relative retention time (RRT) and relative peak area (RPA), and similarity analyses of the chromatographic fingerprints were conducted by Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine software (Version 2004 A). RESULTS: In the 24-h stability test, the relative standard deviation for the RRT and RPA was less than 0.07% and 2.57%, respectively. The precision was less than 0.08% for the RRT and 2.48% for the RPA. The repeatability for the RRT and RPA was less than 0.03% and 2.64%, respectively. The similarities between the seven PRC batches were range from 0.956 to 0.990. After stir-frying or steaming, the amount of ferulic acid in PRC was much higher than that in the raw material. CONCLUSIONS: The fingerprint analysis of PRC by different processing methods was feasible by HPLC-DAD.