ObjectiveTo establish an ultra-performance liquid fingerprint and multi-components determination method for Naomaili granules. To evaluate the quality of different batches by chemometrics， and the anti-neuroinflammatory effects of water extract and main components of Naomaili granules were tested in vitro. MethodsThe similarity and common peaks of 27 batches of Naomaili granules were evaluated by using Ultra performance liquid chromatography （UPLC） fingerprint detection. Ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） technology was used to determine the content of the index components in Naomaili granules and to evaluate the quality of different batches of Naomaili granules by chemometrics. LPS-induced BV-2 cell inflammation model was used to investigate the anti-neuroinflammatory effects of the water extract and main components of Naomaili granules. ResultsThe similarity of fingerprints of 27 batches of samples was > 0.90. A total of 32 common peaks were calibrated， and 23 of them were identified and assigned. In 27 batches of Naomaili granules， the mass fractions of 14 components that were stachydrine hydrochloride， leonurine hydrochloride， calycosin-7-O-glucoside， calycosin，tanshinoneⅠ， cryptotanshinone， tanshinoneⅡ_A， ginsenoside Rb₁， notoginsenoside R₁， ginsenoside Rg₁， paeoniflorin， albiflorin， lactiflorin， and salvianolic acid B were found to be 2.902-3.498， 0.233-0.343， 0.111-0.301， 0.07-0.152， 0.136-0.228， 0.195-0.390， 0.324-0.482， 1.056-1.435， 0.271-0.397， 1.318-1.649， 3.038-4.059， 2.263-3.455， 0.152-0.232， 2.931-3.991 mg∙g^-1， respectively. Multivariate statistical analysis showed that paeoniflorin， ginsenoside Rg₁， ginsenoside Rb₁ and staphylline hydrochloride were quality difference markers to control the stability of the preparation. The results of bioactive experiment showed that the water extract of Naomaili granules and the eight main components with high content in the prescription had a dose-dependent inhibitory effect on the release of NO in the cell supernatant. Among them， salvianolic acid B and ginsenoside Rb₁ had strong anti-inflammatory activity， with IC₅₀ values of （36.11±0.15） mg∙L^-1 and （27.24±0.54） mg∙L^-1， respectively. ConclusionThe quality evaluation method of Naomaili granules established in this study was accurate and reproducible. Four quality difference markers were screened out， and eight key pharmacodynamic substances of Naomaili granules against neuroinflammation were screened out by in vitro cell experiments.

Objective:To investigate the predictive value of microRNA-1(miR-1),brain natriuretic peptide(BNP)and ischemia-modified albumin(IMA)for unstable angina pectoris(UAP).Methods:We enrolled 237 UAP patients admitted to Xining Second People's Hospital between June 2018 and December 2020 as UAP group.Another 86 healthy subjects undergoing physical examination simultaneously were enrolled as control group.MiR-1 expression,BNP and IMA levels were measured.General data between UAP group and control group,serum miR-1 expression,BNP and IMA levels among different Braunwald class and prognosis were compared.Receiver operat-ing characteristic(ROC)curve was employed to analyze predictive value of miR-1,BNP,IMA and their combina-tion for prognosis in UAP patients.Results:Compared with participants in the control group,those in UAP group had significant higher serum miR-1 expression[(1.80±0.59)vs.(0.93±0.11)],BNP[(107.34±37.46)pg/ml vs.(52.31±10.64)pg/ml]and IMA[(79.76±19.29)g/L vs.(53.16±6.43)g/L](P＜0.001 all).As Braun-wald class increased(class Ⅰ～Ⅲ),serum miR-1 expression,BNP and IMA levels elevated(P＜0.001 all).Com-pared with patients in favorable outcome group,those in unfavorable outcome group had significant higher serum miR-1 expression[(2.31±0.54)vs.(1.53±0.41)],BNP[(147.03±29.63)pg/ml vs.(85.95±19.46)pg/ml]and IMA[(97.24±15.35)g/L vs.(70.35±13.88)g/L](P＜0.001 all).ROC curve indicated that AUC of com-bined detection for predicting unfavorable outcome in UAP patients was 0.925(95％CI 0.884～0.955),which was significantly higher than miR-1(AUC=0.880,95％CI 0.831～0.918),BNP(AUC=0.863,95％CI 0.813～0.904)and IMA(AUC=0.900,95％CI 0.854～0.935)alone(Z=2.884,3.130,2.090,P＜0.05 or＜0.01).Conclusion:MiR-1 expression,BNP and IMA levels significantly increase in UAP patients,and they are associated with the severity of disease.Combined detection has good predictive value for unfavorable outcome in UAP pa-tients.

Objective To develop a novel protective ventilator circuit component and to verify its performance by water seal and anti-splash experiments.Methods A novel protective ventilator circuit component had a design scheme with the multifunctional joint,and consisted of a tee connection tube,an isolation sleeve and a stop sleeve,of which,the tee connection tube was made of polyethylene polymer material and the others were made of silicone material.The tee connection tube had a T-shaped structure with two standard connection ports,which was composed of an adapter,a sealing cap,a plug and a sealing ring;the isolation sleeve was in the shape of a cylinder with a raised bottom,which was inserted into the adapter;the stop sleeve was located in the isolation sleeve,with an inverted frustum of a cone at the bottom and a rounded hole in the middle of the inverted frustum.An open ventilator circuit tube was involved in the performance verification of the circuit component developed.In the water seal experiment,sputum aspiration was simulated and the heights of the liquid level drop in the L-shaped tubes were compared after sputum aspiration.In the anti-splash experiment,the infection rates on the surfaces of the sterile hole towels and gloves were calculated.Results Water seal experiment showed after sputum aspiration the open ventilator circuit tube had the liquid level at the L-shaped tube higher significantly than that of the circuit component;the anti-splash experiment indicated sputum aspiration resulted in the occurance of the splashing out of the secretion and 77.5%infection rate by the open ventilator circuit tube,while no splashing out and 0%infection rate by the circuit component developed.Conclusion The novel protective ventilator circuit component behaves well in sealing and anti-splashing,and thus is worthy of clinical application for sputum aspiration.[Chinese Medical Equipment Journal,2025,46(4):113-117]

Objective:To retrospectively analyze the clinical efficacy and safety of biological agents in the treatment of psoriasis in the elderly.Methods:A retrospective study was conducted. A total of 124 patients aged ≥ 65 years with moderate-to-severe psoriasis (psoriasis area and severity index [PASI] score ≥ 3 points or body surface area [BSA] ≥ 3%), who were treated with biological agents at the Department of Dermatology and Venereology, People's Hospital of Xinjiang Uygur Autonomous Region, from June 2020 to December 2023, were collected. PASI, BSA, and dermatology life quality index (DLQI) scores were recorded at weeks 0 (pre-treatment), 4, 12, and 24 after the beginning of treatment to evaluate the efficacy. The proportions of patients achieving ≥ 75% and 90% improvements in PASI scores (PASI75 and PASI90, respectively) were calculated, and adverse reactions were recorded. Data were processed using SPSS 29.0 and GraphPad Prism 10.0 software. Normally distributed continuous variables were presented as mean ± standard deviation, while non-normally distributed continuous variables were presented as median with upper and lower quartiles ( Q1, Q3). Wilcoxon signed-rank test was used for comparisons of non-normally distributed data between groups, and chi-square test and Fisher's exact test were used for comparing categorical data. Results:Among the 124 patients, there were 72 males (58.1%) and 52 females (41.9%), and their ages ranged from 65 to 87 years. Treatment regimens included secukinumab for 86 patients (69.4%), ustekinumab for 15 (12.1%), ixekizumab for 14 (11.3%), guselkumab for 5 (4.0%), and adalimumab for 4 patients (3.2%). The pre-treatment PASI, BSA, and DLQI scores [ M ( Q1, Q3) ] were 12.2 (7.8, 19.6) points, 16.0% (10.2%, 25.0%), and 16 (11, 20) points respectively, which significantly decreased to 0.8 (0, 1.2) points, 1.0% (0, 2.0%), and 0 (0, 3) points respectively at week 24 after the start of treatment ( Z = 9.66, 9.66, 9.63, respectively, all P < 0.01). The proportions of patients achieving PASI75 at weeks 4, 12, and 24 were 42.7% (53/124), 80.6% (100/124), and 93.5% (116/124) respectively, and those achieving PASI90 were 13.7% (17/124), 48.4% (60/124), and 85.5% (106/124) respectively. There were no significant differences in the proportions of patients achieving PASI75 and PASI90 between the subgroups with and without comorbidities, as well as between the early-onset psoriasis subgroup and late-onset psoriasis subgroup at weeks 4, 12, and 24 (all P > 0.05). Among the 124 patients, 42 were followed up for over 2 years, and the proportions of patients achieving PASI75 and PASI90 after 2 years of treatment were 71.4% (30 cases) and 54.8% (23 cases), respectively. Adverse reactions occurred in 13 patients, including upper respiratory infections in 4 patients (3.2%), elevated aminotransferase levels in 3 patients (2.4%), eczematoid dermatitis in 2 patients (1.6%), pruritus in 2 patients (1.6%), nodular prurigo in 1 patient (0.1%), and vitiligo in 1 patient (0.1%) . Conclusion:Biological agents showed rapid and marked efficacy in the treatment of moderate-to-severe psoriasis in the elderly, with few and mild adverse reactions, suggesting high safety and overall marked efficacy.

BACKGROUND:Previously,a SENP1 gene-silenced human alveolar epithelial cell line was successfully constructed in vitro,and the role of SENP1 in hyperoxic lung injury was investigated at the cellular level.OBJECTIVE:To construct a mouse model of alveolar type II epithelial cell-specific knockout of SENP1 gene based on the Cre-loxP recombinase system.METHODS:SENP1flox/-mice were self-crossed to obtain SENP1flox/flox and SENP1flox/-mice;Sftpc-Cre+/+mice were crossed with wild-type mice to obtain more Sftpc-Cre+/-mice.Sftpc-Cre+/+or offspring Sftpc-Cre+/-mice were crossed with SENP1flox/-or offspring SENP1flox/flox mice to obtain SENP1flox/-Sftpc-Cre+/-double heterozygous mice.SENP1flox/-Sftpc-Cre+/-mice were then crossed with SENP1flox/flox mice to obtain SENP1flox/floxSftpc-Cre+/-mice.The genomic DNA was extracted by tail clipping and amplified by PCR.The amplified product was subjected to agarose gel electrophoresis to determine the mouse genotypes.Lung tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were subjected to immunofluorescence double-labelling and western blot assay to verify the knockdown effect of SENP1 gene.Heart,liver,lung and kidney tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were stained with hematoxylin-eosin to observe the histomorphology of each organ in the two groups of mice.RESULTS AND CONCLUSION:SENP1flox/floxSftpc-Cre+/-mice were correctly screened by agarose gel electrophoresis.Immunofluorescence double-labeling experiments showed that the mean fluorescence intensity of SENP1 was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with that of SENP1flox/flox mice(P<0.01)and no significant co-localization of SENP1 and Sftpc was observed(P<0.01).Western blot results showed that SENP1 protein expression was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with SENP1flox/flox mice(P<0.001).Hematoxylin-eosin staining showed no significant alterations in the histomorphology of heart,liver,lung and kidney tissues in SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice.This study successfully constructed alveolar type II epithelial cell-specific knockout SENP1 gene mice using the Cre-loxP recombinase system,which provides a good tool for the subsequent study of the role of SENP1 gene in lung diseases such as bronchopulmonary dysplasia and idiopathic pulmonary fibrosis,in which alveolar type II epithelial cells are the main damage cells.

Objective:To investigate the predictive value of microRNA-1(miR-1),brain natriuretic peptide(BNP)and ischemia-modified albumin(IMA)for unstable angina pectoris(UAP).Methods:We enrolled 237 UAP patients admitted to Xining Second People's Hospital between June 2018 and December 2020 as UAP group.Another 86 healthy subjects undergoing physical examination simultaneously were enrolled as control group.MiR-1 expression,BNP and IMA levels were measured.General data between UAP group and control group,serum miR-1 expression,BNP and IMA levels among different Braunwald class and prognosis were compared.Receiver operat-ing characteristic(ROC)curve was employed to analyze predictive value of miR-1,BNP,IMA and their combina-tion for prognosis in UAP patients.Results:Compared with participants in the control group,those in UAP group had significant higher serum miR-1 expression[(1.80±0.59)vs.(0.93±0.11)],BNP[(107.34±37.46)pg/ml vs.(52.31±10.64)pg/ml]and IMA[(79.76±19.29)g/L vs.(53.16±6.43)g/L](P＜0.001 all).As Braun-wald class increased(class Ⅰ～Ⅲ),serum miR-1 expression,BNP and IMA levels elevated(P＜0.001 all).Com-pared with patients in favorable outcome group,those in unfavorable outcome group had significant higher serum miR-1 expression[(2.31±0.54)vs.(1.53±0.41)],BNP[(147.03±29.63)pg/ml vs.(85.95±19.46)pg/ml]and IMA[(97.24±15.35)g/L vs.(70.35±13.88)g/L](P＜0.001 all).ROC curve indicated that AUC of com-bined detection for predicting unfavorable outcome in UAP patients was 0.925(95％CI 0.884～0.955),which was significantly higher than miR-1(AUC=0.880,95％CI 0.831～0.918),BNP(AUC=0.863,95％CI 0.813～0.904)and IMA(AUC=0.900,95％CI 0.854～0.935)alone(Z=2.884,3.130,2.090,P＜0.05 or＜0.01).Conclusion:MiR-1 expression,BNP and IMA levels significantly increase in UAP patients,and they are associated with the severity of disease.Combined detection has good predictive value for unfavorable outcome in UAP pa-tients.

BACKGROUND:Previously,a SENP1 gene-silenced human alveolar epithelial cell line was successfully constructed in vitro,and the role of SENP1 in hyperoxic lung injury was investigated at the cellular level.OBJECTIVE:To construct a mouse model of alveolar type II epithelial cell-specific knockout of SENP1 gene based on the Cre-loxP recombinase system.METHODS:SENP1flox/-mice were self-crossed to obtain SENP1flox/flox and SENP1flox/-mice;Sftpc-Cre+/+mice were crossed with wild-type mice to obtain more Sftpc-Cre+/-mice.Sftpc-Cre+/+or offspring Sftpc-Cre+/-mice were crossed with SENP1flox/-or offspring SENP1flox/flox mice to obtain SENP1flox/-Sftpc-Cre+/-double heterozygous mice.SENP1flox/-Sftpc-Cre+/-mice were then crossed with SENP1flox/flox mice to obtain SENP1flox/floxSftpc-Cre+/-mice.The genomic DNA was extracted by tail clipping and amplified by PCR.The amplified product was subjected to agarose gel electrophoresis to determine the mouse genotypes.Lung tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were subjected to immunofluorescence double-labelling and western blot assay to verify the knockdown effect of SENP1 gene.Heart,liver,lung and kidney tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were stained with hematoxylin-eosin to observe the histomorphology of each organ in the two groups of mice.RESULTS AND CONCLUSION:SENP1flox/floxSftpc-Cre+/-mice were correctly screened by agarose gel electrophoresis.Immunofluorescence double-labeling experiments showed that the mean fluorescence intensity of SENP1 was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with that of SENP1flox/flox mice(P<0.01)and no significant co-localization of SENP1 and Sftpc was observed(P<0.01).Western blot results showed that SENP1 protein expression was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with SENP1flox/flox mice(P<0.001).Hematoxylin-eosin staining showed no significant alterations in the histomorphology of heart,liver,lung and kidney tissues in SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice.This study successfully constructed alveolar type II epithelial cell-specific knockout SENP1 gene mice using the Cre-loxP recombinase system,which provides a good tool for the subsequent study of the role of SENP1 gene in lung diseases such as bronchopulmonary dysplasia and idiopathic pulmonary fibrosis,in which alveolar type II epithelial cells are the main damage cells.

Pyroptosis is an identified programmed cell death that has been highly linked to endoplasmic reticulum (ER) dynamics. However, the crucial proteins for modulating dynamic ER membrane curvature change that trigger pyroptosis are currently not well understood. In this study, a biotin-labeled chemical probe of potent pyroptosis inducer α-mangostin (α-MG) was synthesized. Through protein microarray analysis, reticulon-4 (RTN4/Nogo), a crucial regulator of ER membrane curvature, was identified as a target of α-MG. We observed that chemically induced proteasome degradation of RTN4 by α-MG through recruiting E3 ligase UBR5 significantly enhances the pyroptosis phenotype in cancer cells. Interestingly, the downregulation of RTN4 expression significantly facilitated a dynamic remodeling of ER membrane curvature through a transition from tubules to sheets, consequently leading to rapid fusion of the ER with the cell plasma membrane. In particular, the ER-to-plasma membrane fusion process is supported by the observed translocation of several crucial ER markers to the "bubble" structures of pyroptotic cells. Furthermore, α-MG-induced RTN4 knockdown leads to pyruvate kinase M2 (PKM2)-dependent conventional caspase-3/gasdermin E (GSDME) cleavages for pyroptosis progression. In vivo, we observed that chemical or genetic RTN4 knockdown significantly inhibited cancer cells growth, which further exhibited an antitumor immune response with anti-programmed death-1 (anti-PD-1). In translational research, RTN4 high expression was closely correlated with the tumor metastasis and death of patients. Taken together, RTN4 plays a fundamental role in inducing pyroptosis through the modulation of ER membrane curvature remodeling, thus representing a prospective druggable target for anticancer immunotherapy.
Pyroptosis/immunology* ; Humans ; Endoplasmic Reticulum/immunology* ; Animals ; Nogo Proteins/antagonists & inhibitors* ; Mice ; Cell Line, Tumor ; Xanthones/pharmacology* ; Neoplasms/pathology* ; Mice, Nude

Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*

Vertebral refracture following percutaneous vertebral augmentation (PVA) is commonly seen in elderly patients with osteoporotic thoracolumbar compression fractures (OTLCF). It can lead to recurrent pain, loss of vertebral height, progression of kyphosis, and even neurological dysfunction, significantly impairing patients′ quality of life. Current diagnosis and treatment face multiple challenges, including high misdiagnosis rate, difficulty in choosing between surgical and non-surgical treatment options, lack of standardized surgical protocols, interference from intralesional bone cement during procedures, inadequate stability of internal fixation in osteoporotic bone, and suboptimal compliance of anti-osteoporotic therapy. Establishing a standardized diagnostic and therapeutic framework is urgently needed. To standardize the management process and improve outcomes for vertebral refractures after PVA in elderly OTLCF patients, Spinal Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized experts in the field to develop Guideline for the diagnosis and treatment of vertebral refracture after percutaneous vertebral augmentation in elderly patients with osteoporotic thoracolumbar compression fractures ( version 2025), based on current literature and clinical experience, and adhering to principles of scientific rigor and clinical applicability. A total of 11 recommendations were proposed, encompassing diagnosis, treatment, and rehabilitation of vertebral refracture after PVA in elderly patients with OTLCF, aiming to provide a foundation for a standardized management.