BACKGROUND: Molecular subtyping is an essential complementarity after pathological analyses for targeted therapy. This study aimed to investigate the consistency of next-generation sequencing (NGS) results between circulating tumor DNA (ctDNA)-based and tissue-based in non-small cell lung cancer (NSCLC) and identify the patient characteristics that favor ctDNA testing. METHODS: Patients who diagnosed with NSCLC and received both ctDNA- and cancer tissue-based NGS before surgery or systemic treatment in Lung Cancer Center, Sichuan University West China Hospital between December 2017 and August 2022 were enrolled. A 425-cancer panel with a HiSeq 4000 NGS platform was used for NGS. The unweighted Cohen's kappa coefficient was employed to discriminate the high-concordance group from the low-concordance group with a cutoff value of 0.6. Six machine learning models were used to identify patient characteristics that relate to high concordance between ctDNA-based and tissue-based NGS. RESULTS: A total of 85 patients were enrolled, of which 22.4% (19/85) had stage III disease and 56.5% (48/85) had stage IV disease. Forty-four patients (51.8%) showed consistent gene mutation types between ctDNA-based and tissue-based NGS, while one patient (1.2%) tested negative in both approaches. Patients with advanced diseases and metastases to other organs would be suitable for the ctDNA-based NGS, and the generalized linear model showed that T stage, M stage, and tumor mutation burden were the critical discriminators to predict the consistency of results between ctDNA-based and tissue-based NGS. CONCLUSION ctDNA-based NGS showed comparable detection performance in the targeted gene mutations compared with tissue-based NGS, and it could be considered in advanced or metastatic NSCLC.
Humans ; Carcinoma, Non-Small-Cell Lung/pathology* ; Circulating Tumor DNA/blood* ; High-Throughput Nucleotide Sequencing/methods* ; Female ; Male ; Lung Neoplasms/pathology* ; Middle Aged ; Mutation/genetics* ; Aged ; Adult ; Aged, 80 and over

OBJECTIVES: To assess the efficacy and safety of Tiaozhou Ziyin (TZZY) recipe for treatment of premature ovarian insufficiency (POI) and explore the possible mechanisms. METHODS: We used bioinformatics analyses and network pharmacology to identify the main active ingredients in TZZY recipe and their core targets, which were verified by Western blotting. We tested the efficacy and safety of the recipe in 60 POI patients, who were randomized into control group (n=30) with Femoston treatment and TZZY group (n=30) with additional TZZY recipe treatment for 3 menstrual cycles. RESULTS: The core active ingredients of TZZY recipe included kaempferol, β-sitosterol, luteolin, and quercetin. The core targets included SRC, TP53, STAT3, PIK3CA, and MAPK3, which were involved in positive regulation of cell movement and protein phosphorylation, the cancer pathways and the PI3K-Akt signaling pathway. Molecular docking showed that the core active ingredients had good binding ability with the core targets. In female rat models of POI, TZZY recipe treatment significantly up-regulated ovarian expressions of p-PI3K and p-Akt proteins. In the clinical trial, treatment with Femoston and Femoston plus TZZY recipe both significantly increased E₂ levels and reduced FSH and LH levels and Kupperman scores of the patients, and the combined treatment produced significantly stronger effects. Both treatments increased the number of antral follicles of the patients, but the combined treatment also significantly increased the levels of AMH. CONCLUSIONS The therapeutic mechanism of TZZY recipe for POI involves multiple active ingredients, multiple therapeutic targets and multiple pathways, and activating the PI3K /Akt pathway is one of its main mechanisms of action, to improve ovarian reserve function, alleviate clinical symptoms, and enhance clinical efficacy in POI patients.
Female ; Primary Ovarian Insufficiency/drug therapy* ; Humans ; Drugs, Chinese Herbal/therapeutic use* ; Animals ; Rats ; Molecular Docking Simulation ; Signal Transduction ; Sitosterols/therapeutic use* ; Kaempferols/therapeutic use*

Objective To assess the efficacy and safety of Tiaozhou Ziyin(TZZY)recipe for treatment of premature ovarian insufficiency(POI)and explore the possible mechanisms.Methods We used bioinformatics analyses and network pharmacology to identify the main active ingredients in TZZY recipe and their core targets,which were verified by Western blotting.We tested the efficacy and safety of the recipe in 60 POI patients,who were randomized into control group(n=30)with Femoston treatment and TZZY group(n=30)with additional TZZY recipe treatment for 3 menstrual cycles.Results The core active ingredients of TZZY recipe included kaempferol,β-sitosterol,luteolin,and quercetin.The core targets included SRC,TP53,STAT3,PIK3CA,and MAPK3,which were involved in positive regulation of cell movement and protein phosphorylation,the cancer pathways and the PI3K-Akt signaling pathway.Molecular docking showed that the core active ingredients had good binding ability with the core targets.In female rat models of POI,TZZY recipe treatment significantly up-regulated ovarian expressions of p-PI3K and p-Akt proteins.In the clinical trial,treatment with Femoston and Femoston plus TZZY recipe both significantly increased E2 levels and reduced FSH and LH levels and Kupperman scores of the patients,and the combined treatment produced significantly stronger effects.Both treatments increased the number of antral follicles of the patients,but the combined treatment also significantly increased the levels of AMH.Conclusion The therapeutic mechanism of TZZY recipe for POI involves multiple active ingredients,multiple therapeutic targets and multiple pathways,and activating the PI3K/Akt pathway is one of its main mechanisms of action,to improve ovarian reserve function,alleviate clinical symptoms,and enhance clinical efficacy in POI patients.

Objective:To analyze the relationship between 24-hour urinary calcium (24 h UCa) level and the risk of end-stage kidney disease (ESKD), cardiovascular disease (CVD), and all-cause mortality.Methods:In the Chinese Cohort Study of Chronic Kidney Disease, we examined 3 375 patients aged 18-74 years with CKD stages 1-4. Kaplan-Meier survival and Cox proportional hazard regression models were used to test a time-to-event association between levels of 24 h UCa and incidence of ESKD, CVD, and all-cause mortality.Results:During a follow-up of 4.17 (3.37, 5.20) years, 179, 145, 104 and 38 ESKD events occurred in <0.60, 0.60-, 1.20-, ≥2.32 mmol 24 h UCa groups. Higher levels of 24 h UCa (1.20-,≥2.32 mmol) were independently associated with a lower incidence of ESKD events in patients with CKD, with HR (95% CI) of 0.71 (0.54-0.93) and 0.43 (0.29-0.64), respectively. No significant associations with CVD and all-cause mortality endpoints were detected. Conclusion:Among patients with CKD, levels of 24 h UCa displayed an association with the risk of ESKD among patients with CKD stages 1-4.

Objective:To observe changes in peripapillary blood flow before and after combined treatment with anti-vascular endothelial growth factor (VEGF) drugs and Dexamethasone intravitreal implant (DEX) in patients with central retinal vein occlusion (CRVO).Methods:A prospective clinical study. Thirty-three eyes of 33 patients with newly diagnosed non-ischemic CRVO and macular edema (ME) were enrolled from Shanxi Eye Hospital between April 2023 and April 2024. All patients underwent best-corrected visual acuity (BCVA) and swept-source optical coherence tomography angiography (SS-OCTA) examinations. The treatment regimen consisted of three intravitreal injections of ranibizumab and one DEX implant. SS-OCTA was used to scan a 3 mm×3 mm area centered on the optic disc to measure peripapillary retinal nerve fiber layer (RNFL) thickness and blood flow density in the superficial vascular complex (SVC), deep vascular complex (DVC), and radial peripapillary capillaries (RPC). Changes in SVC, DVC, and RPC blood flow density and RNFL thickness were evaluated at 3 and 6 months post-treatment. Shapiro-Wilk test was used to assess normality, and Spearman's rank correlation coefficient was applied for correlation analysis.Results:Compared with before treatment, the blood flow density changes of SVC and RPC showed a downward trend at 3 and 6 months after treatment. Among them, the difference was statistically significant at 6 months after treatment ( Z=?2.592, ?2.070, P=0.012, 0.042), while there was no statistically significant difference at 3 months after treatment ( P>0.05). The blood flow density of DVC showed an upward trend at 3 and 6 months after treatment, but the differences were not statistically significant ( P>0.05). The results of the correlation analysis showed that the thickness of RNFL was negatively correlated with the blood flow density of DVC ( r=?0.768, P<0.001). It was positively correlated with the blood flow densities of SVC and RPC ( r=0.288, 0.398; P=0.040, 0.004). Conclusion:Anti-VEGF drugs combined with DEX treatment can significantly improve the perioptic disc blood flow distribution in eyes with CRVO, manifested as a decrease in blood flow density of SVC and RPC, while a compensatory increase in blood flow of DVC. The thickness variation of RNFL is closely related to the blood flow density of different vascular layers.

Objective:To analyze the relationship between 24-hour urinary calcium (24 h UCa) level and the risk of end-stage kidney disease (ESKD), cardiovascular disease (CVD), and all-cause mortality.Methods:In the Chinese Cohort Study of Chronic Kidney Disease, we examined 3 375 patients aged 18-74 years with CKD stages 1-4. Kaplan-Meier survival and Cox proportional hazard regression models were used to test a time-to-event association between levels of 24 h UCa and incidence of ESKD, CVD, and all-cause mortality.Results:During a follow-up of 4.17 (3.37, 5.20) years, 179, 145, 104 and 38 ESKD events occurred in <0.60, 0.60-, 1.20-, ≥2.32 mmol 24 h UCa groups. Higher levels of 24 h UCa (1.20-,≥2.32 mmol) were independently associated with a lower incidence of ESKD events in patients with CKD, with HR (95% CI) of 0.71 (0.54-0.93) and 0.43 (0.29-0.64), respectively. No significant associations with CVD and all-cause mortality endpoints were detected. Conclusion:Among patients with CKD, levels of 24 h UCa displayed an association with the risk of ESKD among patients with CKD stages 1-4.

Objective:To observe changes in peripapillary blood flow before and after combined treatment with anti-vascular endothelial growth factor (VEGF) drugs and Dexamethasone intravitreal implant (DEX) in patients with central retinal vein occlusion (CRVO).Methods:A prospective clinical study. Thirty-three eyes of 33 patients with newly diagnosed non-ischemic CRVO and macular edema (ME) were enrolled from Shanxi Eye Hospital between April 2023 and April 2024. All patients underwent best-corrected visual acuity (BCVA) and swept-source optical coherence tomography angiography (SS-OCTA) examinations. The treatment regimen consisted of three intravitreal injections of ranibizumab and one DEX implant. SS-OCTA was used to scan a 3 mm×3 mm area centered on the optic disc to measure peripapillary retinal nerve fiber layer (RNFL) thickness and blood flow density in the superficial vascular complex (SVC), deep vascular complex (DVC), and radial peripapillary capillaries (RPC). Changes in SVC, DVC, and RPC blood flow density and RNFL thickness were evaluated at 3 and 6 months post-treatment. Shapiro-Wilk test was used to assess normality, and Spearman's rank correlation coefficient was applied for correlation analysis.Results:Compared with before treatment, the blood flow density changes of SVC and RPC showed a downward trend at 3 and 6 months after treatment. Among them, the difference was statistically significant at 6 months after treatment ( Z=?2.592, ?2.070, P=0.012, 0.042), while there was no statistically significant difference at 3 months after treatment ( P>0.05). The blood flow density of DVC showed an upward trend at 3 and 6 months after treatment, but the differences were not statistically significant ( P>0.05). The results of the correlation analysis showed that the thickness of RNFL was negatively correlated with the blood flow density of DVC ( r=?0.768, P<0.001). It was positively correlated with the blood flow densities of SVC and RPC ( r=0.288, 0.398; P=0.040, 0.004). Conclusion:Anti-VEGF drugs combined with DEX treatment can significantly improve the perioptic disc blood flow distribution in eyes with CRVO, manifested as a decrease in blood flow density of SVC and RPC, while a compensatory increase in blood flow of DVC. The thickness variation of RNFL is closely related to the blood flow density of different vascular layers.

OBJECTIVE To investigate the effect of Guanxinning tablets(GXN) on early heart failure model rats, and to explore the protective mechanism of GXN on heart failure rats from the perspective of intestinal flora. METHODS Six rats who underwent sham operation were set as sham operation group. Took 80 SD rats to undergo aortic arch stenosis and established a heart failure rat model. The surviving rats were divided into 4 groups, namely the model control group, the positive control group(captopril tablets 12.5 mg·kg–1), high-dose and low-dose of GXN group(600, 1 200 mg·kg–1). The 4 groups were administered continuously for 8 weeks. Cardiac ultrasonography was performed every 4 week. Serum NT-proBNP, hs-CRP, IL-6, TNF-α, SOD and MDA levels were measured. The effects of GXN on the structure and function of intestinal flora were observed based on the high-throughput sequencing technology and bioinformatics analysis of 16S gut microbiome. RESULTS Compared to the model control group, after giving different doses of GXN, the survival rate of rats increased, and the thickness of the ventricular wall decreased to varying degrees. The weight of the heart and coefficient of the heart were all reduced. GXN could also reduce the level of inflammatory factors, inhibit the level increase of NT-proBNP in rats, and increase the activity of serum SOD. In addition, GXN intervention could significantly improve the intestinal flora diversity of rats with heart failure, the possible target genera of GXN were Akkermansia genera, Phascolarctobacterium genera and Oxalobacter genera. The effect of GXN on intestinal function in rats with heart failure might be concentrated in non-homologous end-joining, influenza A, carotenoid synthesis, indole alkaloids biosynthesis, betalain biosynthesis, renin-angiotensin system and other biological pathways. CONCLUSION The protective effect of GXN on early heart failure rats may be related to the regulation of intestinal flora pathway.

Objective To investigate the expression and clinical significance of circular RNA(circRNA)051778 in lung adenocarcinoma-malignant pleural effusion(LA-MPE)and tuberculous pleural effusion(TPE).Methods This is a cross-sectional study.A total of 212 patients were recruited from the Jiangxi Chest Hospital between October 2018 and September 2019,and their pleural effusion samples and/or plasma samples were collected.The exosomal circRNA profile was sketched by circRNA microarray.Differentially expressed circRNAs(DECs)were verified by droplet digital PCR.In addition,a putative circRNA-miRNA-mRNA network was constructed,and Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses were performed to predict the functions of the DECs.The diagnostic value of circRNA_051778 was evaluated by binary logistic regression and receiver operating characteristic curve.Results The expression level of circRNA_051778 in the LA-MPE samples was(3.92±0.48)copies/100 ng cDNA,while that in the TPE samples was(21.53±2.22)copies/100 ng cDNA.Compared to that in the TPE samples,circRNA_051778 was significantly downregulated in the LA-MPE samples(P＜0.001).The potential targets of circRNA_051778 were enriched in positive regulation of GTPase activity,cytoplasm,protein binding,and cancer-related pathways.The area under the curve(AUC)for the combined assessment of circRNA_051778 with liquid-based thin-layer cytology(TCT),erythrocyte sedimentation rate(ESR),and tuberculosis antibody(TBA)was 0.98(95％confidence interval:0.97-1.00),with the sensitivity being 88.0％and the specificity being 100.0％.Conclusion Exosomal circRNA_051778 is downregulated in LA-MPE.According to the findings from the GO and KEGG analyses,exosomal circRNA_051778 may play a role in cancer development and has the potential to serve as a marker for differential diagnostic of LA-MPE and TPE when it is used in combination with TCT,ESR,and TBA.

Background: Androgenetic alopecia (AGA) leads to thinning of scalp hair and affects 60%~70% of the adult population worldwide. Developing more effective treatments and studying its mechanism are of great significance. Previous clinical studies have revealed that hair growth is stimulated by 650-nm red light. Objective: This study aimed to explore the effect and mechanism of 650-nm red light on the treatment of AGA by using ex vivo hair follicle culture. Methods: Human hair follicles were obtained from hair transplant patients with AGA. Hair follicles were cultured in Williams E medium and treated with or without 650-nm red light.Real-time RT-PCR and immunofluorescence staining were used to detect the expression level of genes and proteins in hair follicles, respectively. RNA-sequencing analysis was carried out to reveal the distinct gene signatures upon 650 nm treatment. Results: Low-level 650 nm red light promoted the proliferation of human hair follicles in the experimental cultured-tissue model. Consistently, 650 nm red light significantly delayed the transition of hair cycle from anagen to catagen in vitro. RNA-seq analysis and gene clustering for the differentially expressed genes suggests that leukocyte transendothelial migration, metabolism, adherens junction and other biological process maybe involved in stimulation of hair follicles by 650-nm red light treatment. Conclusion The effect of 650-nm red light on ex vivo hair follicles and the transcriptome set which implicates the role of red light in promoting hair growth and reversing of miniaturization process of AGA were identified.