Objective To establish a genetic monitoring method for laboratory quails.Methods Quail microsatellite loci were searched in the literature,and microsatellite DNA loci suitable for quails were screened by an interspecific transfer method in closely related species,namely chickens and ducks.Quail liver DNA was extracted as a template,and the corresponding loci were screened by PCR amplification and agarose gel electrophoresis.On the basis of amplification of the selected microsatellite loci,the number of alleles,polymorphisms,and microsatellite loci combinations for quail genetic quality detection were selected and detection method were developed.Results We preliminary determined 23 microsatellite loci for genetic monitoring of closed-colony laboratory quails.Conclusions A genetic monitoring method for laboratory quails was preliminary developed.

Objective To identify the N6-methyladenosine (m6A)-related characteristic genes analyzed by gene clustering and immune cell infiltration in myocardial ischemia-reperfusion injury (MI/RI) after cardiopulmonary bypass through machine learning. Methods The differential genes associated with m6A methylation were screened by the dataset GSE132176 in GEO, the samples of the dataset were clustered based on the differential gene expression profile, and the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the differential genes of the m6A cluster after clustering were performed to determine the gene function of the m6A cluster. R software was used to determine the better models in machine learning of support vector machine (SVM) model and random forest (RF) model, which were used to screen m6A-related characteristic genes in MI/RI, and construct characteristic gene nomogram to predict the incidence of disease. R software was used to analyze the correlation between characteristic genes and immune cells, and the online website was used to build a characteristic gene regulatory network. Results In this dataset, a total of 5 m6A-related differential genes were screened, and the gene expression profiles were divided into two clusters for cluster analysis. The enrichment analysis of m6A clusters showed that these genes were mainly involved in regulating monocytes differentiation, response to lipopolysaccharides, response to bacteria-derived molecules, cellular response to decreased oxygen levels, DNA transcription factor binding, DNA-binding transcription activator activity, RNA polymerase Ⅱ specificity, NOD-like receptor signaling pathway, fluid shear stress and atherosclerosis, tumor necrosis factor signaling pathway, interleukin-17 signaling pathway. The RF model was determined by R software as the better model, which determined that METTL3, YTHDF1, RBM15B and METTL14 were characteristic genes of MI/RI, and mast cells, type 1 helper lymphocytes (Th1), type 17 helper lymphocytes (Th17), and macrophages were found to be associated with MI/RI after cardiopulmonary bypass in immune cell infiltration. Conclusion The four characteristic genes METTL3, YTHDF1, RBM15B and METTL14 are obtained by machine learning, while cluster analysis and immune cell infiltration analysis can better reveal the pathophysiological process of MI/RI.

Objective To explore the effects of isogenous micro-grafting(MG)technology and the combined use of botulinum toxin type A(BTX-A)on hair regeneration in mice.Methods Forty-five C57BL/6 mice were randomly divided into five groups:normal saline group(NS group),MG group(MB-0 group),MG+low-dose BTX-A group(MB-2 group),MG+medium dose BTX-A group(MB-10 group),MG+high-dose BTX-A group(MB-50 group).The growth of hair on the backs of the mice was observed after the administration of the drugs,and the hair follicles were evaluated by hematoxylin-eosin(HE)staining on the 7th,14th,and 21st days of the experiment,and the expression of vascular endothelial growth factor(VEGF)and β-catenin was evaluated by immunohisto-chemical staining on day 21.Results After treatment with micrograft and botulinum toxin type A,the skin darke-ning time was shortened(P＜0.05),and the coverage rate of new hair increased on the 14 th day(P＜0.05).Compared with the NS group,the number of hair follicles increased(P＜0.05)and the expression of VEGF andβ-catenin increased in each other group.Conclusion Isogenous micro-grafting technology combined with botuli-num toxin type A has a promotional effect on hair regeneration in mice,and the mechanism may be related to the promotion of vascular growth and activation of β-catenin signaling.

Objective To investigate the effects and mechanisms of oridonin(ORI)on human keloid-derived fi-broblasts(HKF).Methods The effects of ORI on the proliferation activity of HKF were assessed using the CCK-8 assay.The experiment was divided into control and experimental groups.Cell scratch and Transwell assays were conducted to evaluate the migration and invasion capabilities of HKF.Real-time quantitative PCR(RT-qPCR)and Western blot were used to examine the impact of ORI on the expression of extracellular matrix-related mRNA and fi-bronectin 1(FN1),α-smooth muscle actin(α-SMA),type Ⅰ collagen(COL Ⅰ),and COL Ⅲ in HKF.The ex-periment was also divided into control,model,and transforming growth factor(TGF)-β1+ORI groups.RT-qPCR and Western blot were utilized to determine the effects of ORI on the expression of TGF-β1-induced mRNA and nu-cleotide-binding oligomerization domain-like receptor protein 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC),Smad2,Smad3,phosphorylated Smad2(p-Smad2),and p-Smad3 in HKF.Results CCK-8 assay demonstrated that the cell inhibition rate of HKF progressively increased with increasing concentra-tions of ORI.Compared with the control group,the experimental group exhibited a significant reduction in the mi-gration area at 24 hours and a decrease in the number of invasive cells.Furthermore,there was a significant down-regulation in the expression levels of FN1,α-SMA,COL Ⅰ,and COL Ⅲ(P＜0.05).In comparison with the con-trol group,the model group showed a significant upregulation in the expression of NLRP3,ASC,Smad2,Smad3,p-Smad2,and p-Smad3(P＜0.05).Relative to the model group,the TGF-β1+ORI group demonstrated a signifi-cant downregulation in the expression of NLRP3,ASC,Smad2,Smad3,p-Smad2,and p-Smad3(P＜0.05).Conclusion ORI the proliferation,migration,and invasiveness of HKF,as well as the formation and deposition of the extracellular matrix,through the blockade of the TGF-β1/Smad signaling pathway and the NLRP3-mediated in-flammatory response.

Objective To investigate the effect of cornus officinalis glycosides on apoptosis and oxidative damage of human renal tubular epithelial cells line induced by high glucose by regulating E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)pathway.Methods Human renal tubule epithelial cell line HK-2 was divided into control group(control,5.5 mmol/L glucose),model group(HG,30 mmol/L glucose),and total cornus offici-nalis glycosides groups with low,medium and high dose(L-COG,M-COG,H-COG).After Nrf2 pathway inhibitors were added,HK-2 cells were divided into H-COG group and ML385 group.CCK8 method was used to detect cell activity.Apoptosis was detected by flow cytometry.The expressions of Bcl-2 associated X protein(Bax),B-cell lymphoma/leukemia 2(Bcl-2),activated caspase3(cleaved caspase3),Nrf2/HO-1 pathway protein were detected by Western blot.Reactive oxygen species(ROS)content was detected by DCFH-DA,glutathione peroxi-dase(GSH-Px)activity,superoxide dismutase(SOD)activity,interleukin-6(IL-6)content and tumor necrosis factor α(TNF-α)content were detected by ELISA.Results Compared with control group,the cell activity,Bcl-2 protein expression,GSH-Px and SOD activity in model group were significantly decreased,and the apoptosis rate,Bax,Nrf2,HO-1 and cleaved caspase3 protein expression,ROS,IL-6 and TNF-α content were significantly in-creased(P＜0.05).Compared with model group,the cell activity,Bcl-2 protein expression,GSH-Px and SOD ac-tivity of L-COG,M-COG,and H-COG were significantly increased.Apoptosis rate,Bax,Nrf2,HO-1,cleave-caspase3 protein expression,ROS,IL-6,TNF-α content were significantly decreased(P＜0.05).Compared with H-COG,the cell activity,Bcl-2 protein expression,GSH-Px and SOD activity in ML385 group were significantly increased.The apoptosis rate,Bax and cleaved caspase3 protein expression,ROS,IL-6,TNF-α content were sig-nificantly decreased(P＜0.05).Conclusions The total cornus officinis glucoside may reduce the oxidative stress and inflammatory response of renal tubular epithelial cells induced by high glucose through inhibiting Nrf2/HO-1 pathway and then reduce cell apoptosis.

Objective:To evaluate the impact of bladder volume on dosimetric parameters of clinical target volume (CTV) and organs at risk (OAR) of intensity modulated proton therapy (IMPT) for localized prostate cancer during the treatment planning and daily treatment.Methods:Clinical data of 25 patients with localized prostate cancer admitted to Ruijin Hospital affiliated to Shanghai Jiao Tong University School of Medicine from November 2021 to June 2022 and enrolled in the "Proton Therapy System" (SAPT-PS-01) registered clinical trial were retrospectively analyzed. All patients were male and the median age was 72 years old. A total of 30 sets of IMPT plans were obtained. Based on the planning CT (30 sets) and weekly verification CT during treatment (172 sets), bladder volume, CTV and OAR dose parameters were collected. Spearman correlation analysis was used to evaluate the correlation between bladder volume in CT and the dosimetric parameters of CTV and OAR during IMPT plans, and Wilcoxon-Mann-Whitney test was adopted to compare the dosimetric parameters of CTV and OAR among different bladder volume change groups.Results:The V 95% of CTV1 and CTV2 were both 100.0%±0.0% in IMPT plans. Bladder volume was significantly negatively correlated with D mean, V 70 Gy(RBE), V 60 Gy(RBE), V 50 Gy(RBE), V 40 Gy(RBE) of the bladder ( P<0.001, 0.003, <0.001, <0.001,<0.001), and D mean, V 50 Gy(RBE) of the small intestine (both P<0.001). During treatment, bladder D mean, V 70 Gy(RBE), V 60 Gy(RBE), V 50 Gy(RBE), V 40 Gy(RBE)( P<0.001, 0.001, <0.001, <0.001, <0.001), rectal D mean, V 50 Gy(RBE), V 40 Gy(RBE) (all P<0.001), small intestine D mean, V 50 Gy (RBE) (both P<0.001) of patients with bladder volume increase >20% compared to baseline were significantly decreased compared to those in IMPT plans. But CTV1 V 100%, and CTV2 V 95% were significantly decreased too( P=0.029, 0.020). In the bladder volume decreased>20% patients, the D mean, V 70 Gy(RBE), V 60 Gy(RBE), V 50 Gy(RBE), V 40 Gy(RBE) of the bladder were significantly increased compared to those in IMPT plans (all P<0.001). However, a bladder volume reduction of ≤20% and increase of ≤20% from baseline had no significant impact on CTV and OAR dosimetric parameters during treatment. Conclusions:For patients with localized prostate cancer undergoing proton therapy, a certain bladder volume should be ensured during planning CT scans. During the daily treatment, the bladder volume should be maintained between 80%-120% of the baseline level to ensure CTV coverage and good dose sparing to OAR.

Objective:To review the risk factors for early and medium-term complications of fenestration-branch endovascular thoracic aortic repair (F/B-TEVAR) in patients with complex aortic arch disease.Methods:The clinical and follow-up data of 202 patients undergoing F/B-TEVAR treatment from Feb 2019 to Sep 2023 in these centers were retrospectively analyzed .Results:There were 46 cases suffering from postoperative complications (22.8%). The risk factors with statistical significance included aortic atherosclerotic plaque [ OR=2.843; 95% CI (1.4-5.6); P<0.01], aortic intramural thrombosis [ OR=2.358; 95% CI (1.2-4.6), P=0.011], the aortic dilatation [ OR=4.219; 95% CI (1.6-11.3), P<0.01], the history of stroke [ OR=2.088; 95% CI (1.1-4.1), P=0.032], smoking history [ OR=2.680; 95% CI: (1.3-5.5); P<0.01], duration of surgery [ OR=1.9; 95% CI: (1.2-2.9); P=0.042].While the application of 3D printing assistive technology [ OR=0.392; 95% CI: (0.2-0.9); P=0.048] was in a negative correlation with postoperative complication. Conclusions:The independent risk factors for complications after F/B-TVAR included aortic atherosclerotic plaque, aortic intramural thrombosis, the aortic dilatation, the history of stroke, smoking history,duration of surgery.The application of 3D printing technology can effectively reduce the complication rate.

Objective To investigate the effect and mechanism of miR-27a-3p on nerve cell apoptosis induced by oxygen glucose deprivation/reoxygenation(OGD/R)through regulation of Rho family GTPase 3(Rnd3)expression.Methods PC12 neurons were cultured in vitro and reoxygenated for 3,6,9 h and 12 h after 2 h oxygen glucose deprivation.Cell viability,miR-27a-3p expression and Rnd3 mRNA expression were assessed at each time point and the optimal reoxygenation time point was screened.After transfection of miR-27a-3p Mimic,miR-27a-3p Inhibitor and their negative control,transfection of shRnd3 and its negative control,or co-transfection of shRnd3 and miR-27a-3p Inhibitor through lentivirus,CCK-8 assay was used to detect cell activity.The apoptosis rate of the cells was detected using flow cytometry.Expression of miR-27a-3p and Rnd3 mRNA was detected by RT-qPCR.Expression of apoptosis-related protein and Rnd3 protein was detected by Western blot.The dual luciferase reporter assay confirmed the targeting relationship between miR-27a-3p and Rnd3.Results Upregulation of miR-27a-3p increased cell viability,decreased total cell apoptosis rate,suppressed pro-apoptotic proteins Cleaved Caspase-3(C-caspase-3)and Bax,and promoted expression of anti-apoptotic protein Bcl-2(P<0.05);The opposite result was found when down-regulating miR-27a-3p.The double luciferase reporter gene assay showed that Rnd3 was the target gene of miR-27a-3p.Down-regulation of Rnd3 increased cell viability,decreased the total rate of apoptosis,suppressed the pro-apoptotic protein C-caspase-3,Bax,and promoted expression of the anti-apoptotic protein Bcl-2(P<0.05).However,miR-27a-3p Inhibitor reversed the protective effect of shRnd3.Conclusion miR-27a-3p alleviates OGD/R-induced damage to PC12 neurons by targeting Rnd3 to inhibit cell apoptosis.

Objective:To evaluate the real-world short-term effectiveness of ixekizumab in the treatment of psoriasis, and to investigate factors influencing the effectiveness.Methods:Baseline data and short-term effectiveness evaluation results were retrospectively collected from patients with psoriasis, who received ixekizumab treatment in Department of Dermatology, Xiangya Hospital from November 2019 to September 2021. A descriptive analysis was performed on the baseline characteristics of patients, continuous data were described as median (lower quartile, upper quartile), and categorical data were described as percentages. Comparisons of disease severity scores before and after the treatment with ixekizumab were performed using Wilcoxon signed-rank test or paired McNemar test. Multivariable logistic regression analysis was conducted to explore factors influencing the effectiveness of 4-week ixekizumab treatment.Results:A total of 118 patients with psoriasis were included, including 94 males and 24 females, and their age [ M ( Q1, Q3) ] was 43.4 (32.5, 53.0) years; plaque psoriasis (99 cases, 83.9%) and severe psoriasis (72 cases, 68.6%) predominated among the 118 patients, and skin lesions were mainly located on the scalp (59/116, 50.9%). Among the 49 patients who had received 2-week ixekizumab treatment, 27 (55.1%) achieved a 50% improvement in the psoriasis area and severity index (PASI) score (PASI50) ; after 4-week treatment, 44 (89.8%), 30 (61.2%), 13 (26.5%) and 10 (20.4%) patients achieved PASI50/75/90/100 respectively, and their PASI scores (2.1 [1.1, 7.1]), involved body surface area (3.9% [0.5%, 14.5%]), dermatology life quality index scores (1.0 [0.0, 2.0]) and physician global assessment (PGA) scores (1.0 [1.0, 3.0]) were significantly lower than the corresponding scores at baseline (12.4 [8.8, 23.2], 22.0% [11.3%, 43.4%], 6.0 [3.0, 11.0], 4.0 [3.0, 5.0], respectively; all P < 0.001]. Multivariable logistic regression analysis showed that the baseline body mass index was significantly associated with the PASI75 response rate ( OR = 0.814, 95% CI: 0.659 - 0.958, P = 0.029) and the proportion of patients with PGA0/1 ( OR = 0.743, 95% CI: 0.562 - 0.917, P = 0.017) after 4-week ixekizumab treatment, and the baseline BSA score was significantly associated with the proportion of patients with PGA0/1 after 4-week ixekizumab treatment ( OR = 0.924, 95% CI: 0.870 - 0.968, P = 0.003) . Conclusion:The 4-week ixekizumab treatment significantly decreased the severity of psoriasis, and may be more effective in patients with lower disease severity and lower body mass index at baseline.

Molecular knowledge of human gastric corpus epithelium remains incomplete. Here, by integrated analyses using single-cell RNA sequencing (scRNA-seq), spatial transcriptomics, and single-cell assay for transposase accessible chromatin sequencing (scATAC-seq) techniques, we uncovered the spatially resolved expression landscape and gene-regulatory network of human gastric corpus epithelium. Specifically, we identified a stem/progenitor cell population in the isthmus of human gastric corpus, where EGF and WNT signaling pathways were activated. Meanwhile, LGR4, but not LGR5, was responsible for the activation of WNT signaling pathway. Importantly, FABP5 and NME1 were identified and validated as crucial for both normal gastric stem/progenitor cells and gastric cancer cells. Finally, we explored the epigenetic regulation of critical genes for gastric corpus epithelium at chromatin state level, and identified several important cell-type-specific transcription factors. In summary, our work provides novel insights to systematically understand the cellular diversity and homeostasis of human gastric corpus epithelium in vivo.
Humans ; Epigenesis, Genetic ; Gastric Mucosa/metabolism* ; Chromatin/metabolism* ; Stem Cells ; Epithelium/metabolism* ; Fatty Acid-Binding Proteins/metabolism*