Methods: Sixty-two patients (2014–2019) were divided into the SC (n=32) and PC (n=30) groups. Clinical outcomes were evaluated using the Visual Analog Scale (VAS), modified Japanese Orthopedic Association (mJOA) score, and Neck Disability Index (NDI). Radiological parameters, including cervical lordosis (CL), fused segment angle (FSA), and disc height (DH), were assessed. Cage subsidence, adjacent segment degeneration (ASD), and other complications were also documented. The relationship between postoperative radiological changes and symptoms was also analyzed. Results: Both groups showed significant improvement in symptoms, with fusion achieved in all surgical segments. The SC group had a shorter operative time and reduced intraoperative blood loss. However, the SC group had a higher subsidence rate (16.7% vs. 5.6%, p=0.017). The PC group maintained better cervical alignment in terms of CL, FSA, and DH (p<0.05), with an increased incidence of dysphagia and ASD (p<0.05). Spearman correlation analyses showed that neither CL nor FSA loss at the 5-year follow-up was significantly associated with improvements in the mJOA, VAS, and NDI scores (p>0.05) in either group. Regression analyses indicated that subsidence or loss of lordosis did not correlate with the degree of postoperative symptoms (p<0.05). Conclusions Both SC and PC achieved sustained pain relief, neurological improvement, and solid fusion in three-level ACDF at the 5-year follow-up. SC demonstrated significant advantages in reducing dysphagia and ASD, while PC better maintained cervical alignment and segmental stability. Patient-specific factors should be used to guide implant selection. Further multicenter studies are required to validate these findings.

Objective To explore the role of chloride intracellular channel 5 (CLIC5) in the development of lung adenocarcinoma (LUAD). Methods The expression levels of CLIC5 in LUAD samples were determined using bioinformatics and qRT-PCR. The correlation between CLIC5 expression and LUAD progression was revealed through survival analysis and clinicopathological feature analysis. After the knockdown or overexpression of CLIC5 in the LUAD cells, cell proliferation was assessed through colony formation assay, cell apoptosis rate by flow cytometry, and cell migration and invasion by scratch and Transwell invasion assay. The protein levels of vimentin and Snail were detected by Western blot. Results Bioinformatics analysis results showed low CLIC5 expression levels in the LUAD samples. This finding was associated with the aggressive progression of LUAD and the poor survival of patients with LUAD. The qRT-PCR results confirmed that CLIC5 was mainly expressed in the cytoplasm of the LUAD cells. CLIC5 overexpression inhibited cell activity and proliferation and promoted the apoptosis of LUAD cells. The knockdown of CLIC5 exhibited the opposite effect. CLIC5 overexpression inhibited the migration and invasion of LUAD cells, downregulated Bcl-2, vimentin, and Snail and upregulated Bax, cleaved caspase-3 and E-cadherin in the LUAD cells. CLIC5 knockdown exhibited opposite effects. Conclusion CLIC5 overexpression promotes cell apoptosis and inhibits cell proliferation and invasion, thereby inhibiting the development of LUAD. Hence, CLIC5 is a novel biomarker of prognosis and therapeutic target for LUAD.

ObjectiveTo systematically clarify the expression profile of splicing factor 3B subunit 6 （SF3B6） in pancreatic cancer， its clinical prognostic value， and its regulatory effect on the malignant phenotype of cancer cells through bioinformatics analysis， clinical sample validation， and in vitro functional experiments， and to assess its potential as a novel tumor biomarker. MethodsTranscriptomic and clinical data were downloaded from The Cancer Genome Atlas and Xena public databases at The University of California Santa Cruz to investigate the difference in the expression of SF3B6 between pancreatic cancer tissue and normal pancreatic tissue， and the association of SF3B6 with the prognosis and clinical indices of patients with pancreatic cancer was analyzed. Three pancreatic ductal adenocarcinoma cell lines and one normal pancreatic ductal epithelial cell line were cultured， and RT-qPCR was used to measure the expression level of SF3B6 in pancreatic cancer. A total of 43 patients with pancreatic cancer who underwent surgery in Binzhou Medical University Hospital from January 2024 to December 2025 were enrolled as subjects， and Western blot and RT-qPCR were used to measure the expression of SF3B6 in pancreatic cancer tissue. Based on the median expression level of SF3B6 measured by RT-qPCR， the patients were divided into high and low expression groups. The association of SF3B6 expression with clinical features and prognosis was assessed. A pancreatic cancer cell line with stable low SF3B6 expression was constructed， and CCK-8 assay and Transwell migration and invasion assays were used to observe the impact of SF3B6 on the biological behaviors of the pancreatic cancer cell line， including proliferation， migration， and invasion. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and a one-way analysis of variance was used for comparison between multiple groups， while the least significant difference t-test or the Games-Howell test was used for further comparison between two groups； the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups. The chi-square test was used for comparison of categorical data between two groups. A Logistic regression analysis was used to investigate the influencing factors for the expression of SF3B6， and the univariate and multivariate Cox proportional-hazards regression model analyses were used to investigate the risk factors for prognosis. ResultsThe bioinformatics analysis showed that SF3B6 was highly expressed in pancreatic cancer tissue （P<2.2×10^-16） and significantly affected the overall survival （P=0.011） and progression-free survival （P=0.002） of patients with pancreatic cancer. Sex， age， clinical stage， and TNM stage were not influencing factors for SF3B6 expression （all P>0.05）， and SF3B6 could be used as an independent risk factor for the survival outcome of patients with pancreatic cancer （overall survival： hazard ratio=2.26， 95% confidence interval： 1.13 — 4.50， P=0.021； progression-free survival： hazard ratio=2.37，95% confidence interval： 1.22 — 4.58， P=0.010）. Cellular and tissue experiments confirmed the high expression level of SF3B6 in pancreatic cancer cell lines （P<0.05）， and the expression level of SF3B6 increased with the increase in the malignancy of pancreatic cancer， showing the highest relative expression level in PANC-1 cells with the highest degree of malignancy； the expression level of SF3B6 in the solid tissue of pancreatic cancer was significantly higher than that in paracancerous tissue （P<0.01）. The expression level of SF3B6 might affect the overall survival of patients with pancreatic cancer （P<0.05）， while it showed no significant association with age， sex， pathological grade， maximum tumor diameter， or clinical stage （all P>0.05）. Knockdown of SF3B6 significantly inhibited the proliferation， migration， and invasion of pancreatic cancer cells （all P<0.01）. ConclusionSF3B6 is highly expressed in pancreatic cancer， and the high expression of SF3B6 can promote the proliferation， migration， and invasion of pancreatic cancer cells. Therefore， SF3B6 can be used as a potential biomarker for prognostic evaluation.

This study examined the effect of islet macrophages on β-cell function changes during type 2 diabetes mellitus (T2DM) progression based on the traditional Chinese medicine theory that " moderate fire generating qi, hyperactive fire consuming qi" . T2DM is closely associated with chronic low-grade inflammation, with islet macrophages playing a central role in this process. Under physiological conditions, islet macrophages secrete anti-inflammatory and growth factors to regulate the immune response, promote cell proliferation, and support islet β-cell survival and function, reflecting the concept of " moderate fire generating qi" . However, during the pathological process of T2DM, islet macrophages become over-activated and dysfunctional, secreting large amounts of pro-inflammatory factors that trigger severe inflammatory responses and oxidative stress. This process damages islet β-cells, disrupts the islet microenvironment and blood supply, exacerbates local inflammation and structural damage, and worsens the survival environment of β-cells. Ultimately, this leads to fewer β-cells and function loss, aligning with the " hyperactive fire consuming qi" theory, where excessive fire depletes qi and blood. This study enhances the understanding and application of traditional Chinese medicine theories in modern medicine, offering a new perspective on T2DM prevention and treatment. Regulating islet macrophage function and reducing their pro-inflammatory responses may become key strategies for preserving β-cell function and slowing T2DM progression.

Objective:To establish a method of dual nueleic acid test strips for rapid detection of Bacillus cereus cytK and nhe toxin genes based on polymerase chain reaction(PCR)and colloidal gold technique,and to evaluate its specificity,sensitivity,repeatability and stability.Methods:Bacillus cereus DNA was extracted by boiling method.Specific primers were designed with Bacillus cereus cytK and nhe as the target genes.Clonal transformation was used to identify the PCR products.The optimal labeling amounts of colloidal gold-labeled streptavidin,quality control line(C line),cytK detection line(T1)and nhe detection line(T2)were determined.The nucleic acid test strips were assembled and its specificity,sensitivity,reproducibility and stability were evaluated.Results:The DNA concentration of Bacillus cereus was 248 mg·L-1,and the purities were 1.8-2.0.After cloning and plasmid sequencing,the similarities between the PCR products and the sequences of cytK and nhe registered in the GenBank database were 100％.Under the condition of pH 7.0,the optimal amount of streptavidin labeling per 200 μL of colloidal gold solution was 6.0 μL;the optimal marking amount was 2.00 g·L-1 for the quality control line(C line),0.550 g·L-1 for cytK gene detection line(T1)and 0.2 g·L-1 for nhe gene detection line(T2).In the specificity test,positive result on the test strips was seen only for Bacillus cereus,and no cross-reactivity was observed for Staphylococcus aureus,Escherichia coli,Pseudomonas aeruginosa and Bacillus subtilis,which were consistent with the electrophoresis results.Sensitivity assay showed that even when DNA concentration was reduced to 10-2 mg·L-1,three bands(C line,T1 line and T2 line)could be observed,and the detection limit of the test strip was one-tenth of agarose gel electrophoresis(10-1 mg·L-1).The nucleic acid test strips were verified by different operators in different laboratories,and the results were consistent.The stability of the test strips was verified at the 6th,9th and 12th months,and the results showed good stability.Conclusion:The dual nucleic acid test strip method established in this study can simutaneously detect the cytK and nhe toxin genes of Bacillus cereus with high sensitivity and specificity,achieving short-term visual detection.

Objective To investigate the changes of 22 bile acid sub components and 17 traditional bio-chemical indicators in serum of patients with non-alcoholic fatty liver disease(NAFLD),and the diagnostic value of detecting the above indicators alone or in combination for NAFLD.Methods A total of 168 NAFLD patients(NAFLD group)and 216 non-NAFLD apparently healthy individuals(non-NAFLD group)were se-lected,bile acid sub components were determined by liquid chromatography tandem mass spectrometry,and traditional biochemical indicators were detected by automatic biochemical analyzer.Results There were sta-tistically significant differences in the levels of 12 bile acid sub components and 12 traditional biochemical indi-cators between NAFLD group and non-NAFLD group(P<0.05).Compared to traditional biochemical indica-tors,bile acid sub components were less affected by body mass index(BMI).The area under the curve for di-agnosing NAFLD by combining three bile acid sub components[taurocholic acid(TCA),sodium taurodeoxy-cholate(TDCA),and tauroursodeoxycholic acid(UDCA)]with three traditional biochemical indicators[ala-nine aminotransferase(ALT),5'Nucleotidase(5'-NT),and small and dense low-density lipoprotein cholester-ol(sd-LDL-C)]was the largest,which was 0.810.Conclusion Twelve kinds of bile acid sub components in the blood of NAFLD patients have changed,and the combined detection of bile acid sub components and tradi-tional biochemical indicators could improve the diagnostic efficacy of NAFLD to a certain extent.

In recent years, the role of cerebral venous outflow (VO) in the assessment of outcome in patients with acute ischemic stroke (AIS) has been receiving increasing attention. As a key component of the neurovascular compensation mechanism, good VO is of great significance for the functional outcome in patients with AIS. This article systematically reviews the methods and clinical applications of using CT angiography to evaluate VO, with a focus on exploring the association of VO characteristics with clinical outcome, symptomatic intracranial hemorrhage and cerebral edema progression, aiming to provide imaging evidence for optimizing individualized treatment and outcome assessment in patients with large vessel occlusive stroke.

Although fertility-preserving treatment strategies have demonstrated significant clinical efficacy in patients with early-stage endometrial cancer(EC)and atypical endometrial hyperplasia(AEH),some patients who experience recurrence still express a strong desire for fertility and request conservative retreatment.This poses new challenges for clinical management.This article reviews the latest research advances in retreatment for recurrent patients and synthesizes findings from previous studies to draw the following conclusions.First,before formulating a retreatment plan,it is crucial to comprehensively evaluate key factors such as tumor characteristics,fertility intentions,and overall health status of the patient.These factors collectively determine the feasibility and appropri-ateness of an individualized retreatment strategy.Second,studies have shown that retreatment options for AEH/EC are diverse,with high-dose oral progestin remaining the primary approach.Combining progestin with metformin,GnRH-a,and hysteroscopic resection can enhance treatment outcomes.Individualized treatment plans should be tailored to the specific conditions of each patient.Based on literature analysis,the complete remission(CR)rate of retreatment ranges from 81.1%to 88.6%,with a pregnancy rate of 26.5%-50.0%and a live birth rate of 14.3%-29.0%.However,the recurrence rate remains high at 24.5%-45.5%,significantly higher than that of initial treatment.This indicates that while retreatment has achieved some success in disease control and fertility preservation,the risk of recurrence still requires significant attention.Therefore,a strict monitoring and follow-up system must be established during retreatment.Additionally,studies have identified factors associated with a higher risk of recurrence,including age over 35,overweight or obesity,polycystic ovarian syndrome,metabolic syndrome,high pathological grade,and advanced clinical stage.On the other hand,pregnancy following CR,weight loss,and maintenance therapy serve as protective factors against disease recurrence.Patients are encouraged to actively engage in weight management during treatment and consider initiating assisted reproductive technology promptly after achieving CR to optimize pregnancy outcomes while minimizing the risk of recurrence.Future research should prioritize investigating precision treatment strategies informed by molecular classification and identifying predictive biomarkers,thereby enabling the development of more personalized and precise treatment plans tailored to indi-vidual patients.

Glioblastoma (GBM) is a highly aggressive primary brain tumor characterized by poor prognosis. Conventional chemo-radiotherapy demonstrates limited therapeutic efficacy and is often accompanied by significant side effects, largely due to factors such as drug resistance, radiation resistance, the presence of the blood-brain barrier (BBB), and the activation of DNA damage repair mechanisms. There is a pressing need to enhance treatment efficacy, with BRD4 identified as a promising target for increasing GBM sensitivity to therapy. Lacking small molecule inhibitors, BRD4 can be degraded using PROteolysis Targeting Chimera (PROTAC), thereby inhibiting DNA damage repair. To deliver PROTAC, SIAIS171142 (SIS) effectively, we designed a responsive nanocapsule, MPL_(SS)P@SIS, featuring GBM-targeting and GSH-responsive drug release. Modified with 1-methyl-l-tryptophan (MLT), nanocapsules facilitate targeted delivery of SIS, downregulating BRD4 and sensitizing GBM cells to radiotherapy and chemotherapy. After intravenous administration, MPL_(SS)P@SIS selectively accumulates in tumor tissue, enhancing the effects of radiotherapy and temozolomide (TMZ) by increasing DNA damage and oxidative stress. GSH activates the nanocapsules, triggering BRD4 degradation and hindering DNA repair. In mouse models, the nanosensitizer, combined with TMZ and X-ray irradiation, efficiently inhibited the growth of GBM. These findings demonstrate a novel PROTAC-based sensitization strategy targeting BRD4, offering a promising approach for effective GBM therapy.

ObjectiveTo improve the therapeutic effect of Buyang Huanwutang（BYHW） on diabetic kidney disease （DKD） and explore new methods for developing new Chinese medicine decoctions，we utilized 5-hydroxymethylcytosine （5hmC）-Seal sequencing technology and network pharmacology to modify BYHW. MethodsWe selected 14 diabetes mellitus （DM） patients and 15 DKD patients hospitalized in the Department of Endocrinology of Peking University Third Hospital in 2021. Circulating free DNA （cfDNA） in the patients’ plasma was sequenced. After data processing and screening， we performed temporal clustering analysis to select a DKD 5hmC gene set， which was then cross-validated with a DKD database gene set to obtain the DKD gene set. We retrieved target genes of the seven herbal components of BYHW from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and the Encyclopedia of Traditional Chinese Medicine （ETCM）， and performed cross-analysis with the DKD gene set to identify common genes shared by the disease and the Chinese medicines. A protein-protein interaction （PPI） network was constructed for the common genes to screen out the key genes. Chinese medicines targeting these key genes were searched against ETCM to identify removable Chinese medicines. Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis was performed on non-common DKD genes， and key genes in DKD-related pathways were selected based on machine learning. The GSE30529 dataset was used to verify the expression trends of 5hmC-modified genes and the feasibility of target genes as drug targets. TCMBank was used to search for target genes and obtain compounds targeting these genes and the corresponding Chinese medicines to construct a "key target-compound-Chinese medicine" network. Molecular docking was employed to verify the binding affinity of compounds with key targets. TCMSP and ETCM were used to search and count the candidate Chinese medicines targeting DKD-related genes， and a new decoction was formed by adding the selected Chinese medicines. A mouse model of DKD was established to examine the efficacy of the new decoction based on the mouse body mass， random blood glucose， urinary microalbumin （mALB）， serum creatinine （Scr）， and blood urea nitrogen （BUN） and by hematoxylin-eosin staining， periodic acid-Schiff staining， Masson staining， immunofluorescence assay， and Real-time PCR. ResultsThe cross-analysis results showed that the DKD gene set included 507 genes， of which 30 were target genes of BYHW. The PPI analysis indicated that the top 15% target genes regarding the degree were interleukin-6 （IL-6）， Toll-like receptor 4 （TLR4）， lactotransferrin （LTF）， lipoprotein lipase （LPL）， and sterol regulatory element-binding transcription factor 1 （SREBF1）. Persicae Semen and Pheretima in BYHW were unrelated to key genes and removed. Machine learning identified 10 potential target genes， among which TBC1 domain family member 5 （TBC1D5）， RAD51 paralog B （RAD51B）， and proteasome 20S subunit alpha 6 （PSMA6） had expression trends consistent with the GSE30529 dataset and could serve as drug targets. The "key target-compound-Chinese medicine" network and molecular docking results indicated that the compounds with good binding affinity to target proteins were arginine， glycine， myristicin， serine， and tyrosine， corresponding to 121 Chinese medicines. The top 10 Chinese medicines targeting DKD-related genes were Lycii Fructus， Ginseng Radix et Rhizoma， Dioscoreae Rhizoma， Rehmanniae Radix Praeparata， Isatidis Radix， Glehniae Radix， Ophiopogonis Radix， Allii Sativi Bulbus， Isatidis Folium， and Bolbostemmatis Rhizoma. Based on traditional Chinese medicine theory， the new decoction was obtained after removal of Persicae Semen and Pheretima and addition of Rehmanniae Radix Praeparata and Dioscoreae Rhizoma. Animal experiment results indicated that the modified BYHW improved the kidney function and inhibited renal fibrosis in DKD mice， with better effects than the original decoction. ConclusionThe BYHW modified based on 5hmC-Seal sequencing demonstrates better performance in inhibiting fibrosis and ameliorating DKD than the original decoction. This elucidates the biomedical theory behind the epigenetic modification of traditional Chinese medicine prescriptions， potentially offering new perspectives for the exploration of these prescriptions