ObjectiveTo investigate the heterogeneity of PD-L1 expression profiles between primary tumors and paired metastatic lymph nodes in advanced non-small cell lung cancer （NSCLC）， and to determine the predictive value of key immune marker expression for the efficacy of first-line targeted therapy and chemotherapy. MethodsA retrospective analysis was conducted on 128 patients with histologically confirmed NSCLC and corresponding metastatic lymph nodes， who received first-line tyrosine kinase inhibitors （TKIs） or platinum-based doublet chemotherapy at Guangdong Provincial People's Hospital between April 2017 and February 2020. Immunohistochemistry （IHC） was employed to evaluate PD-L1 expression in primary tumors and metastatic lymph nodes. The Wilcoxon signed-rank test was performed to compare PD-L1 expression characteristics between primary tumors and metastatic lymph nodes. Kaplan-Meier survival curves were constructed， and the Log-rank test was used to compare differences between groups. ResultsAmong 28 paired cases， PD-L1 expression levels were numerically higher in metastatic lymph nodes than in primary NSCLC tumors （median：32.5 vs. 10.0）， although the difference did not reach statistical significance （MD = 5.000， P = 0.083）. Survival analysis revealed that in patients with driver gene-positive NSCLC， high PD-L1 expression （TPS ≥ 50%） in lymph nodes was significantly associated with shorter progression-free survival （PFS） following first-line TKI therapy ［median PFS：4.0（TPS≥50%） vs. 8.9（1%≤TPS＜50%） vs. 18.0 （TPS＜1%） months， χ²=15.284， P＜0.001］. Conversely， in patients with driver gene-negative NSCLC， high PD-L1 expression in lymph nodes was associated with longer PFS ［median PFS：7.9 （TPS≥50%） vs. 3.0 （1%≤TPS＜50%）months， χ²=8.436， P=0.004］ and overall survival （OS） ［median OS：28.8 （TPS≥50%） vs. 14.2 （ 1%≤TPS＜50%） months， χ²=4.010， P=0.045］ after first-line chemotherapy. ConclusionPD-L1 expression in metastatic lymph nodes is largely consistent with that in primary tumors of patients with advanced NSCLC. High PD-L1 expression in lymph nodes is associated with improved survival outcomes in driver gene-negative patients undergoing chemotherapy， whereas it portends a poor prognosis with TKI treatment in driver gene-positive patients. These findings support the application of PD-L1 expression levels in metastatic lymph nodes to guide personalized precision medicine strategies for advanced NSCLC.

BACKGROUND:Previous studies have found that for postmenopausal osteoporosis patients,the Bushen Jianpi Formula can help regulate serum osteoclast-and osteoblast-related factors and intestinal flora levels,improve clinical symptoms of patients and further improve efficacy.OBJECTIVE:To investigate the effect of Bushen Jianpi Formula on bone metabolism and bone microarchitecture in ovariectomized rats.METHODS:Twenty-four female SD rats were randomly divided into four groups by random number table method:sham operation group(n=6)was subjected to periovarian fat removal,and bilateral ovariectomy was performed in model group(n=6),alendronate group(n=6),and Bushen Jianpi Formula group(n=6).Five days after modeling,the Bushen Jianpi Formula group was given Bushen Jianpi Formula by intragastric administration(once a day).The alendronate sodium group was given alendronate by intragastric administration(once a week).The sham operation group and model group were given an equal volume of normal saline by intragastric administration(once a day)for 12 consecutive weeks.After intragastric administration,serum levels of procollagen Ⅰ N-terminal peptide,β-isomerized C-terminal telopeptide of type Ⅰ collagen degradation product,and tumor necrosis factor α were detected.The distal femur microstructure was detected by Micro-CT.The femoral tissue morphology was observed by hematoxylin-eosin staining.The expression of type Ⅰ collagen and tumor necrosis factorα in the femur was detected by immunohistochemical staining.RESULTS AND CONCLUSION:(1)The serum ELISA test showed that compared with the sham operation group,the level of procollagen Ⅰ N-terminal peptide in the model group decreased(P＜0.05),and the levels of degradation products of β-isomerized C-terminal telopeptide of type Ⅰ collagen and tumor necrosis factor α increased(P＜0.05).Compared with the model group,the levels of the procollagen Ⅰ N-terminal peptide in the Bushen Jianpi Formula group and the alendronate sodium group increased(P＜0.05),and the levels of β-isomerized C-terminal telopeptide of type Ⅰ collagen degradation products and tumor necrosis factor α decreased(P＜0.05).(2)Micro-CT three-dimensional reconstruction showed that compared with the sham operation group,the number of bone trabeculae in the model group was significantly reduced,the continuity was worsened,and the gaps were increased.Both alendronate sodium and Bushen Jianpi Formula had a positive improvement effect on this phenomenon.(3)Hematoxylin-eosin staining showed that compared with the sham operation group,the number of bone trabeculae in the model group was reduced and the shape became thinner,and the network structure was destroyed.Compared with the model group,the number of bone trabeculae in the Bushen Jianpi Formula group and the alendronate sodium group increased,the shape became thicker,and the reticular structure was restored.(4)Immunohistochemical staining showed that compared with the sham operation group,the expression of type Ⅰ collagen was reduced and tumor necrosis factor α expression was increased in the model group.Compared with the model group,the expression of type Ⅰ collagen increased and the expression of tumor necrosis factor α decreased in the Bushen Jianpi Formula group and the alendronate sodium group.The results show that the Bushen Jianpi Formula can effectively improve bone metabolism and bone microstructure in ovariectomized rats.

BACKGROUND: Double-stranded RNA-specific adenosine deaminase 1 (ADAR1) binds to double-stranded RNA and catalyzes the deamination of adenosine (A) to inosine (I). The functional mechanism of ADAR1 in non-small cell lung cancer (NSCLC) remains incompletely understood. This study aimed to investigate the prognostic significance of ADAR1 in NSCLC and to elucidate its potential role in regulating tumor cell proliferation and migration. METHODS: Data from The Cancer Genome Atlas (TCGA) and cBioPortal were analyzed to assess the correlation between high ADAR1 expression and clinicopathological features as well as prognosis in lung cancer. We performed Western blot (WB), cell proliferation assays, Transwell invasion/migration assays, and nude mouse xenograft modeling to examine the phenotypic changes and molecular mechanisms induced by ADAR1 knockdown. Furthermore, the ADAR1 p150 overexpression model was utilized to validate the proposed mechanism. RESULTS: ADAR1 expression was significantly elevated in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) tissues compared with adjacent non-tumor tissues (LUAD: P=3.70×10-15, LUSC: P=0.016). High ADAR1 expression was associated with poor prognosis (LUAD: P=2.03×10-2, LUSC: P=2.81×10-2) and distant metastasis (P=0.003). Gene Set Enrichment Analysis (GSEA) indicated that elevated ADAR1 was associated with mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway activation, matrix metalloproteinase-9 (MMP-9) expression, and cell adhesion. ADAR1 and MMP-9 levels showed a strongly positive correlation (P=6.45×10-34) in 10 lung cancer cell lines, highest in H1581. Knockdown of ADAR1 in H1581 cells induced a rounded cellular morphology with reduced pseudopodia. Concomitantly, it suppressed cell proliferation, invasion, migration, and in vivo tumorigenesis. It also suppressed ERK phosphorylation and downregulated cellular Finkel-Biskis-Jinkins murine osteosarcoma viral oncogene homolog (c-FOS), MMP-9, N-cadherin, and Vimentin. Conversely, ADAR1 p150 overexpression in PC9 cells enhanced ERK phosphorylation and increased c-FOS and MMP-9 expression. CONCLUSIONS High ADAR1 expression is closely associated with poor prognosis and distant metastasis in NSCLC patients. Mechanistically, ADAR1 may promote proliferation, invasion, migration, and tumorigenesis in lung cancer cells via the ERK/c-FOS/MMP-9 axis.
Humans ; Lung Neoplasms/physiopathology* ; Adenosine Deaminase/genetics* ; Matrix Metalloproteinase 9/genetics* ; Cell Proliferation ; Carcinoma, Non-Small-Cell Lung/physiopathology* ; Cell Movement ; Animals ; Mice ; RNA-Binding Proteins/genetics* ; Female ; Male ; Cell Line, Tumor ; Proto-Oncogene Proteins c-fos/genetics* ; Middle Aged ; MAP Kinase Signaling System ; Gene Expression Regulation, Neoplastic ; Mice, Nude ; Extracellular Signal-Regulated MAP Kinases/genetics*

Diabetic kidney disease （DKD） stands as one of the most prevalent microvascular complications of diabetes，noted for its concealed onset and tendency to evolve into end-stage renal disease，profoundly impacting patients' life expectancy and quality of life. Epithelial-to-mesenchymal transition （EMT） is a central pathological process in the initiation and progression of DKD，facilitating disease advancement and renal fibrosis，thus representing a crucial focus of research into the pathological mechanisms of DKD. EMT is driven by the abnormal activation of signaling pathways，including transforming growth factor-β （TGF-β）/Smad，secreted glycoprotein/β-catenin，Notch，tumor necrosis factor-α （TNF-α）/nuclear factor-κB （NF-κB），and phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR），leading to renal cellular injury and subsequently accelerating renal fibrosis and the progression of DKD. Traditional Chinese medicine （TCM），characterized by its multi-target and multi-pathway therapeutic approach，demonstrates unique advantages in addressing DKD and EMT. Recent research has shown that active ingredients in TCM，including glycosides，flavonoids，and polyphenols，as well as TCM formulas，can precisely target these relevant signaling pathways，effectively inhibiting cellular injury in DKD and intervening in the EMT process. These findings not only underscore the potential of TCM monomers and formulas in treating DKD and EMT but also pave new directions for research in this field within TCM. This paper systematically reviewed the signaling pathways associated with EMT and provided an in-depth analysis of the research achievements and underlying mechanisms of TCM monomers and formulas in treating DKD and intervening in EMT，aiming to offer new insights and directions for TCM in the treatment of DKD and research on EMT，thereby further promoting the modernization and development of TCM.

BACKGROUND:Previous studies have found that for postmenopausal osteoporosis patients,the Bushen Jianpi Formula can help regulate serum osteoclast-and osteoblast-related factors and intestinal flora levels,improve clinical symptoms of patients and further improve efficacy.OBJECTIVE:To investigate the effect of Bushen Jianpi Formula on bone metabolism and bone microarchitecture in ovariectomized rats.METHODS:Twenty-four female SD rats were randomly divided into four groups by random number table method:sham operation group(n=6)was subjected to periovarian fat removal,and bilateral ovariectomy was performed in model group(n=6),alendronate group(n=6),and Bushen Jianpi Formula group(n=6).Five days after modeling,the Bushen Jianpi Formula group was given Bushen Jianpi Formula by intragastric administration(once a day).The alendronate sodium group was given alendronate by intragastric administration(once a week).The sham operation group and model group were given an equal volume of normal saline by intragastric administration(once a day)for 12 consecutive weeks.After intragastric administration,serum levels of procollagen Ⅰ N-terminal peptide,β-isomerized C-terminal telopeptide of type Ⅰ collagen degradation product,and tumor necrosis factor α were detected.The distal femur microstructure was detected by Micro-CT.The femoral tissue morphology was observed by hematoxylin-eosin staining.The expression of type Ⅰ collagen and tumor necrosis factorα in the femur was detected by immunohistochemical staining.RESULTS AND CONCLUSION:(1)The serum ELISA test showed that compared with the sham operation group,the level of procollagen Ⅰ N-terminal peptide in the model group decreased(P＜0.05),and the levels of degradation products of β-isomerized C-terminal telopeptide of type Ⅰ collagen and tumor necrosis factor α increased(P＜0.05).Compared with the model group,the levels of the procollagen Ⅰ N-terminal peptide in the Bushen Jianpi Formula group and the alendronate sodium group increased(P＜0.05),and the levels of β-isomerized C-terminal telopeptide of type Ⅰ collagen degradation products and tumor necrosis factor α decreased(P＜0.05).(2)Micro-CT three-dimensional reconstruction showed that compared with the sham operation group,the number of bone trabeculae in the model group was significantly reduced,the continuity was worsened,and the gaps were increased.Both alendronate sodium and Bushen Jianpi Formula had a positive improvement effect on this phenomenon.(3)Hematoxylin-eosin staining showed that compared with the sham operation group,the number of bone trabeculae in the model group was reduced and the shape became thinner,and the network structure was destroyed.Compared with the model group,the number of bone trabeculae in the Bushen Jianpi Formula group and the alendronate sodium group increased,the shape became thicker,and the reticular structure was restored.(4)Immunohistochemical staining showed that compared with the sham operation group,the expression of type Ⅰ collagen was reduced and tumor necrosis factor α expression was increased in the model group.Compared with the model group,the expression of type Ⅰ collagen increased and the expression of tumor necrosis factor α decreased in the Bushen Jianpi Formula group and the alendronate sodium group.The results show that the Bushen Jianpi Formula can effectively improve bone metabolism and bone microstructure in ovariectomized rats.

The incidence of osteoporotic thoracolumbar vertebral fracture (OTLVF) in the elderly is gradually increasing. The kyphotic deformity caused by various factors has become an important characteristic of OTLVF and has received increasing attention. Its clinical manifestations include pain, delayed nerve damage, sagittal imbalance, etc. Currently, the definition and diagnosis of OTLVF with kyphotic deformity in the elderly are still unclear. Although there are many treatment options, they are controversial. Existing guidelines or consensuses pay little attention to this type of fracture with kyphotic deformity. To this end, the Lumbar Education Working Group of the Spine Branch of the Chinese Medicine Education Association and Editorial Committee of Chinese Journal of Trauma organized the experts in the relevant fields to jointly develop Clinical guidelines for the diagnosis and treatment of osteoporotic thoracolumbar vertebral fractures with kyphotic deformity in the elderly ( version 2024), based on evidence-based medical advancements and the principles of scientificity, practicality, and advanced nature, which provided 18 recommendations to standardize the clinical diagnosis and treatment.

Objective To prepare decellularized scaffolds from rabbit cartilage at various concentrations and assess their physicochemical properties and compatibility with stem cells to provide an experimental basis for cartilage repair.Methods Bone marrow mesenchymal stem cells(BMSCs)were cultured using the Percoll density gradient separation method,and this was followed by flow cytometric analysis and testing of their osteogenic and chondrogenic differentiation capabilities.Cartilage pieces were excised from rabbit knees and hip joints and subjected to physical crushing,repeated freeze-thaw cycles,and mixed enzymatic digestion for decellularization.To compare and observe the physicochemical properties of the decellularized scaffolds at different concentrations,three groups of scaffolds(labelwd A,B,and C)were designed with concentrations of 100％,50％and 30％,with three replicates each.Third-generation PKH26-labeled BMSCs were seeded onto optimally concentrated scaffolds and cultured for 1 week to observe cell growth.Results Flow cytometry detected BMSC surface antigens with positive expression of CD44 and CD90 and negative expression of CD45.Osteogenic induction stained with alizarin red showed red calcific nodules,and chondrogenic induction stained with alcian blue showed blue cartilaginous nodules.No apparent cell morphology was observed in the three groups of scaffolds stained with hematoxylin-eosin,and toluidine blue.There was a significant difference in DNA concentration between decellularized samples and non-decellularized scaffolds(P＜0.05).The content of glycosaminoglycans was slightly lower than the normal values.Significant differences were observed between the three groups of scaffolds in terms of pore size,water absorption,porosity,tensile strength,and Young's modulus(P＜0.05).After co-cultivation of stem cells with the scaffolds,cell adhesion was found to be good.Conclusions Percoll density gradient separation can obtain high-purity rabbit BMSCs,and the mixed decellularization method is superior.Group B scaffolds were the most suitable for tissue-engineered cartilage repair.BMSCs cultured in vitro grew well on Group B scaffolds.

OBJECTIVE: To observe the correlation between the four limbs perfusion index (PI) and blood lactic acid in patients with neurosis, and evaluate the predictive value of PI on microcirculation perfusion metabolic disorder in patients with neurosis. METHODS: A prospective observational study was conducted. Adult patients admitted to the department of neurological intensive care unit (NICU) of the First Affiliated Hospital of Xinjiang Medical University from July 1 to August 20 in 2020 were enrolled. Under the condition of indoor temperature controlled at 25 centigrade, all patients were placed in the supine position, and the blood pressure, heart rate, PI of both fingers and thumb toes and arterial blood lactic acid were measured within 24 hours and 24-48 hours after NICU. The difference of four limbs PI at different time periods and its correlation with lactic acid were compared. Receiver operator characteristic curve (ROC curve) was used to evaluate the predictive value of four limbs PI on patients with microcirculatory perfusion metabolic disorder. RESULTS: A total of 44 patients with neurosis were enrolled, including 28 males and 16 females; average age (61.2±16.5) years old. There were no significant differences in PI of the left index finger and the right index finger [2.57 (1.44, 4.79) vs. 2.70 (1.25, 5.33)], PI of the left toe and the right toe [2.09 (0.85, 4.76) vs. 1.88 (0.74, 4.32)] within 24 hours after entering the NICU, and the PI of the left index finger and the right index finger [3.17 (1.49, 5.07) vs. 3.14 (1.33, 5.36)], PI of the left toe and the right toe [2.07 (0.75, 5.20) vs. 2.07 (0.68, 4.67)] at 24-48 hours after NICU admission (all P > 0.05). However, compared to the PI of the upper and lower limbs on the same side, except for the 24-48 hours after ICU of the PI difference between the left index finger and the left toe (P > 0.05), the PI of the toe was lower than that of the index finger at the other time periods (all P < 0.05). The correlation analysis showed that the PI value of four limbs of patients in both time periods were significantly negatively correlated with arterial blood lactic acid (the r values of the left index finger, the right index finger, the left toe and the right toe were -0.549, -0.482, -0.392 and -0.343 respectively within 24 hours after entering the NICU; the r values of the left index finger, the right index finger, the left toe and the right toe were -0.331, -0.292, -0.402 and -0.442 respectively after entering the NICU 24-48 hours, all P < 0.05). Taking lactic acid ≥ 2 mmol/L as the diagnostic standard for metabolic disorder of microcirculation perfusion (total 27 times, accounting for 30.7%). The efficacy of four limbs PI in predicting microcirculation perfusion metabolic disorder were compared. ROC curve analysis showed that the area under the curve (AUC) and 95% confidence interval (95%CI) of left index finger, right index finger, left toe and right toe predicting microcirculation perfusion metabolic disorder were 0.729 (0.609-0.850), 0.767 (0.662-0.871), 0.722 (0.609-0.835), 0.718 (0.593-0.842), respectively. There was no significant difference in AUC compare with each other (all P > 0.05). The cut-off value of PI of right index finger for predicting microcirculation perfusion metabolic disorder was 2.46, the sensitivity was 70.4%, the specificity was 75.4%, the positive likelihood ratio was 2.86, and the negative likelihood ratio was 0.30. CONCLUSIONS There are no significant differences in PI of bilateral index fingers, bilateral toes in patients with neurosis. However, unilateral upper and lower limbs showed lower PI in the toe than in the index finger. There is a significantly negatively correlation between PI and arterial blood lactic acid in all four limbs. PI can predict the metabolic disorder of microcirculation perfusion, and its cut-off value is 2.46.
Adult ; Female ; Male ; Humans ; Middle Aged ; Aged ; Lactic Acid ; Microcirculation ; Perfusion Index ; Lower Extremity ; Area Under Curve ; Nervous System Diseases

BACKGROUND: The switch/sucrose nonfermentable chromatin-remodeling (SWI/SNF) complex is a pivotal chromatin remodeling complex, and the genomic alterations (GAs) of the SWI/SNF complex are observed in several cancer types, correlating with multiple biological features of tumor cells. However, their role in liver metastasis of non-small cell lung cancer (NSCLC) remains unclear. Our study aims to investigate the role and potential mechanisms underlying NSCLC liver metastasis induced by the GAs of SWI/SNF complex. METHODS: The GAs of SWI/SNF complex in NSCLC cell lines (H1299, H23 and H460) were identified by whole-exome sequencing (WES). ARID1A knockout H1299 cell was constructed with the CRISPR/Cas9 technology. The mouse model of liver metastasis from NSCLC was established to simulate lung cancer liver metastasis and observe the metastasis rate under different gene mutation conditions. RNA sequencing and Western blot were conducted for differential gene expression analysis. Immunohistochemistry (IHC) analysis was used to assess protein expression levels of SWI/SNF-regulated target molecules in mouse liver metastases. RESULTS: WES analysis revealed intracellular gene mutations. The animal experiments demonstrated a correlation between the GAs of SWI/SNF complex and a higher liver metastasis rate in immunodeficient mice. Transcriptome sequencing and Western blot analysis showed upregulated expression of ALDH1A1 and APOBEC3B in SWI/SNF-mut cells, particularly in ARID1A-deficient H460 and H1299 sgARID1A cells. IHC staining of mouse liver metastases further demonstrated elevated expression of ALDH1A1 in the H460 and H1299 sgARID1A group. CONCLUSIONS This study underscores the critical role of the GAs of SWI/SNF complex, such as ARID1A and SMARCA4, in promoting liver metastasis of lung cancer cells. The GAs of SWI/SNF complex may promote liver-specific metastasis by upregulating ALDH1A1 and APOBEC3B expression, providing novel insights into the molecular mechanisms underlying lung cancer liver metastasis.
Animals ; Mice ; Carcinoma, Non-Small-Cell Lung/genetics* ; Lung Neoplasms/genetics* ; Mutation ; Liver Neoplasms/genetics*

Objective:To prepare a phase-change lipid nanoparticle modified by tumor homing membrane-penetrating peptide (tLyP-1) and carrying paclitaxel (PTX) engineered by metal polyphenol network (TA-Fe 3+ ), and evaluate the therapeutic effects of tumor targeting, ultrasound/photoacoustic imaging and photothermal combined chemotherapy in vitro. Methods:Phase-change lipid nanoparticles (t-P@TFP) with TA-Fe 3+ engineered PTX mediated by tLyP-1 were prepared by solvent replacement method, thin film hydration method and double emulsification method. Its detection and characterization, in vitro targeting ability, photothermal conversion ability, in vitro photoacoustic and ultrasonic imaging ability, CCK-8 method, cell live and death double staining method and flow cytometry method were used to detect the safety of nanoparticles and the killing effects of different nanoparticles on 4T1 cells. Results:t-P@TFP nanoparticles were successfully prepared. Transmission electron microscopy showed that the nanoparticles were spherical with uniform shape and size, with a particle size of (209.8±1.56)nm and a potential of (-25.9±1.36)mV. Laser confocal scanning microscopy showed that t-P@TFP nanoparticles could gather around 4T1 cells in a targeted manner. It had an efficient photothermal conversion effect, and nanoparticles could quickly become microbubbles after being irradiated by near-infrared laser, which enhanced the in vitro ultrasonic imaging effect; The photoacoustic signal of nanoparticles increased with the increase of concentration. CCK-8 method, double staining of living and dead cells and flow cytometry showed that t-P@TFP combined photothermal chemotherapy had the best anti-tumor effect. Conclusions:t-P@TFP nanoparticles are successfully prepared. The nanoparticles have good targeting ability for photoacoustic and ultrasonic imaging and have good photothermal effect, killing breast cancer cells, which is expected to realize the integration of diagnosis and treatment.