BACKGROUND:Renal fibrosis (RF) is a critical step in the progression of chronic kidney disease to end-stage renal failure, but its treatment remains highly challenging, highlighting the urgent clinical need for novel therapeutic strategies. Shikonin, a naturally derived compound, shows promise in attenuating the progression of RF. A thorough understanding of its specific molecular targets and mechanism of action is essential. PURPOSE:This study elucidates the molecular mechanism through which Shikonin modulates HSPA1A to suppress ferroptosis and regulate lipid metabolism in RF. METHODS:A unilateral ureteral obstruction (UUO) mouse model was used to assess the antifibrotic effects of Shikonin. Efficacy was evaluated by analyzing renal pathomorphological changes and fibrosis indicators, and by using ELISA to detect the inflammation levels and kidney function. We integrated results from network pharmacology, transcriptomics, and non-targeted metabolomics to explore Shikonin's mechanism in renal fibrosis and verified it through experiments including qRT-PCR, immunofluorescence, western blot, transmission electron microscopy, ROS staining, and Bodipy staining. In vitro, human renal tubule epithelial cells (HK-2) were stimulated with TGF-β1 and then treated with Shikonin. Lipid-MS, CETSA, SPR, and molecular dynamics simulations were jointly employed to identify Shikonin's target. After knocking down this target in vivo using AAV, the effect of Shikonin on renal fibrosis was observed. RESULTS:Shikonin notably ameliorated histopathological lesions, renal dysfunction, pathological damage and fibrosis in UUO model. Transcriptomic and non-targeted metabolomics analyses indicated that lipid metabolism specifically the arachidonic acid (AA) pathway, is a key pathway through which Shikonin exerted its effects. Mechanistically, Shikonin directly bind to HSPA1A and, through this interaction, downregulated the expression of key AA-metabolizing enzymes ACSL4 and ALOX15, thereby suppressing lipid peroxidation and ferroptosis. This mechanism was confirmed both in vivo and in vitro. Crucially, the anti-fibrosis efficacy of Shikonin was abolished upon HSPA1A knockdown or inhibition, and could be functionally reversed by exogenous AA supplementation. CONCLUSION:This study has revealed a novel mechanism by which Shikonin alleviates renal fibrosis: by targeting HSPA1A to reprogram AA metabolism and inhibit ferroptosis. These findings display the HSPA1A-AA metabolism-ferroptosis axis as a critical therapeutic pathway and provide a compelling mechanistic foundation for developing Shikonin as a promising candidate drug.

Epidermal growth factor receptor (EGFR) exon 19 deletions (19del) in non-small cell lung cancer (NSCLC) confer sensitivity to tyrosine kinase inhibitors (TKIs). However, the efficacy of the individual EGFR 19del variant remains unclear. This study aims to dissect the structural disparities of various EGFR 19del mutations and associate these variations with clinical outcomes. Here, a total of 399 patients from two cancer centers covering 35 distinct EGFR 19del variants were included in this study. Through molecular docking analysis, we identified a structure-based subgroup of EGFR 19del mutations, termed CLASS 3G (E746_A750>X, E746_S752>V, L747_A750>P, and L747_T751>P). In silico and in vitro analysis demonstrated that CLASS 3G mutations had reduced binding affinity to third-generation EGFR-TKIs. Clinically, patients with CLASS 3G showed inferior PFS to non-CLASS 3G after receiving third-generation EGFR-TKIs (first-line progression-free survival: 12.8 vs. 25.6 months, P=0.045; second-line progression-free survival: 7.7 vs. 15.2 months, P=0.039). This finding was further validated in an independent cohort, confirming consistent results with the primary analysis. Additionally, we found that CLASS 3G mutations derived significant benefits from third-generation EGFR-TKIs plus chemotherapy compared with third-generation EGFR-TKIs monotherapy (32.5 vs. 12.8 months, P=0.0038). Our study identified a subgroup of EGFR 19del that responded poorly to third-generation EGFR-TKIs based on structural features of the EGFR tyrosine kinase domain. These patients benefit substantially from the combination treatment of third-generation EGFR-TKIs plus chemotherapy.

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by persistent synovial inflammation and progressive joint destruction. Although methotrexate (MTX) remains a first-line therapy for RA, its clinical use may be limited by inadequate response, secondary loss of efficacy, and adverse effects, highlighting the need for safer and more effective therapeutic strategies. Corylifol A (CA), a bioactive monomer derived from traditional Chinese medicine, has been reported to exert immunomodulatory and anti-inflammatory effects; however, its therapeutic role and underlying mechanisms in RA remain insufficiently understood. In this study, THP-1-Lucia™ NF-κB reporter cells were used to screen traditional Chinese medicine-derived monomers with anti-inflammatory potential, and CA was selected for further evaluation in RA fibroblast-like synoviocytes (RA FLSs) and experimental arthritis models. Integrated transcriptomic and network pharmacology analyses were performed to identify the potential molecular targets and pathways involved in the therapeutic effects of CA in RA. CA significantly inhibited the proliferation, migration, invasion, and inflammatory cytokine production of RA FLSs, while alleviating inflammation and bone destruction in arthritic mice. Mechanistically, CA inactivated heat shock protein 90 alpha (HSP90α) and consequently suppressed downstream Wnt signaling. Molecular docking and Asp93 mutation-based rescue experiments further suggested that Asp93 may be involved in the interaction between CA and HSP90α. Moreover, CA enhanced the therapeutic effects of MTX, resulting in more obvious suppression of RA FLSs inflammatory activation and arthritis progression in vivo. These findings suggest that CA may represent a potential therapeutic candidate for RA, and the enhanced therapeutic effects observed with CA plus MTX further support its therapeutic relevance.

Background: P97 regulates diverse cellular processes, including proteostasis, metabolism and pluripotency. Pharmacological inhibitors of P97 protein have shown antitumor activity in various cancers. However, the downstream pathways and effectors remain poorly characterized, limiting clinical translation. Methods: We performed a quantitative proteomic analysis using liquid chromatography–mass spectrometry (LC–MS) to identify differentially expressed proteins (DEPs) (fold change > 1.5, p < 0.05) and pathway alterations in gastric cancer AGS cells following treatment with two distinct P97 inhibitors, Eeyarestatin I (EerI) and NMS873. Results: Proteomic analysis revealed that both inhibitors significantly perturbed multiple signaling pathways. The endoplasmic reticulum (ER) proteostasis network was markedly disrupted, evidenced by activation of the unfolded protein response (UPR) and upregulation of ER-associated degradation (ERAD) components. Pathway enrichment further indicated that cell death processes, particularly ferroptosis, were prominently affected (p < 0.05). Key ferroptosis regulators showed altered expression, which was validated by Western blotting. Functional assays demonstrated that P97 inhibition suppressed cell proliferation and increased the accumulation of ferroptosis-associated lipid hydroperoxides in vitro, suggesting execution of ferroptotic cell death. In vivo, using a xenograft mouse model established by subcutaneous injection of AGS cells into nude mice, P97 inhibition significantly reduced tumor growth, confirming its antitumor efficacy. Conclusions: This integrated proteomic and functional study delineates that pharmacological P97 inhibitors disrupt ER proteostasis and simultaneously trigger ferroptosis in gastric cancer cells. These findings identify ferroptosis as a key downstream mechanism of P97 inhibition and provide molecular mechanisms for developing P97-targeted therapies in gastric cancer.

Background: ASTRUM-002 met the primary endpoint of progression-free survival (PFS) with the combination of serplulimab plus HLX04 (bevacizumab biosimilar) and chemotherapy at interim analysis. Here, we report the results of the final survival analysis. Methods: A total of 636 patients with treatment-naïve, locally advanced or metastatic nonsquamous non-small cell lung cancer (nsq-NSCLC) without epidermal growth factor receptor (EGFR) or anaplastic lymphoma kinase (ALK)/ROS proto-oncogene 1 receptor tyrosine kinase (ROS1) genetic alterations were randomized 1:1:1 to receive serplulimab plus HLX04 and chemotherapy (group A), serplulimab plus HLX04 placebo and chemotherapy (group B), or double placebo plus chemotherapy (group C). Patients and the investigators were blinded to the group assignments. The primary endpoint was blinded independent central review-assessed PFS. Overall survival (OS) was the key secondary endpoint. Results: At the final analysis, median OS was 23.7 (95% confidence interval [CI] 20.5 to 27.5) months, 26.8 (95% CI 21.2 to 30.9) months, and 20.3 (95% CI 16.2 to 24.6) months in groups A (n = 212), B (n = 214), and C (n = 210), respectively. A significant reduction in risk of death for group B compared to group C was observed (hazard ratio [HR] = 0.66, 95% CI 0.52 to 0.83; P < 0.001). A total of 79 (37.6%) patients in group C had crossed over to serplulimab plus HLX04 treatment. Median OS in group C adjusted by the 2-stage model was 14.2 months (95% CI 11.9 to 17.0), corresponding to an adjusted HR of 0.53 (95% CI 0.42 to 0.68; P < 0.001) for group B versus group C. Using the rank-preserving structural failure time model, the adjusted median OS in group C was 17.9 months (95% CI 14.2 to 20.3), with a corresponding adjusted HR of 0.65 (95% CI 0.51 to 0.83; P < 0.001). No statistical difference in median OS for group A compared to group B (HR = 1.12, 95% CI 0.88 to 1.42; P = 0.363) was found. Conclusions: Serplulimab plus chemotherapy significantly prolonged OS and maintained PFS benefit compared to chemotherapy; however, the addition of bevacizumab biosimilar HLX04 did not yield further improvement for the first-line treatment of nsq-NSCLC without EGFR or ALK/ROS1 genetic alterations. Trial registration: This trial was registered at ClinicalTrials.gov (NCT03952403, date of registration: 2019 May 14).

This study aimed to analyze and compare the proportion of patients with different types of inflammatory arthritis and investigate the clinical characteristics, including symptoms and signs, medication choices, and disease activity, in the daily clinical practice of China. Patients with inflammatory arthritis were recruited from 16 Grade-A tertiary hospitals between August 2021 and April 2022. The medical profiles, encompassing sociodemographic characteristics, clinical and laboratory date, were collected. This study included 2,693 patients with arthritis, with rheumatoid arthritis (RA) accounting for the highest proportion (50.50

Background:Diabetic retinopathy (DR) is a leading cause of blindness in diabetic patients, driven by inflammation, oxidative stress, and hypoxia. The hemoglobin-to-red cell distribution width ratio (HRR) is a novel inflammatory marker reflecting these pathological mechanisms. This study aimed to investigated the association of HRR with DR risk and severity. Methods:Data from the 2005-2008 National Health and Nutrition Examination Survey were analyzed using weighted logistic regression, subgroup analysis, restricted cubic splines, mediation analysis, and other methods. Results:Among 1,260 diabetic patients, HRR was inversely associated with DR development (OR = 0.85, P = 0.008), remaining significant post-propensity score matching. A non-linear relationship was identified, with an inflection point at HRR = 10.81 (P for non-linearity < 0.001), above which the protective effect strengthened with increasing HRR. Mediation analyses revealed diastolic blood pressure (15.9% mediation) and HbA1c (60.5% competitive mediation) as partial mediators of the HRR-DR association. HRR was also inversely associated with DR severity, particularly proliferative DR (vs. mild non-proliferative DR: OR = 0.67, P = 0.030; vs. severe non-proliferative DR: OR = 0.04, P = 0.002). Conclusions:HRR is negatively correlated with DR onset and progression, highlighting its potential as a cost-effective biomarker for DR risk stratification.

Abstract Alternative splicing derived neoantigens can be served as promising immunotherapeutic targets for tumor. However, little is known about the AS milieu in Pediatric glioma, including diffuse midline glioma (DMG), pediatric high grade glioma (pHGG) and medulloblastoma (MB). We performed integrated analysis of alternative splicing using bulk RNA-seq and single cell RNA-seq datasets of pediatric brain tumor specimens, including 9 DMG, 25 pHGG and 19 MB cases. We also re-analyzed RNA-sequencing data from non-malignant human brain tissues. We reconstructed the landscape of AS in Pediatric glioma and contrasted that to observed isoforms in non-malignant brain. We found that AS score can be used to stratify the MB patients samples, and high AS score was significantly associated with worse survival in MB. We identified novel splicing events in cell-surface proteins that are suitable targets for engineered T-cell therapies in DMG, pHGG and MB. We also identified novel splicing events derived neoantigens in DMG, pHGG and MB respectively, which are potential immunotherapy targets. We found MB specific isoforms of transforming growth factor beta (TGFβ) signaling pathway genes which are expressed exclusively by MB tumor cells. Our studies shed light on the effect of immunotherapy on AS and identify novel targets for emerging immunotherapies in pediatric brain tumor.

[This corrects the article DOI: 10.3389/fphar.2021.692806.].

Participating in scientific research is essential for undergraduate students that major in biotechnology, public health, biomedical science, and medical laboratory technology to develop their research skill. Course-based undergraduate research experiences (CUREs) represent a transformative pedagogical approach that integrates authentic research projects into conventional curricula. However, a systematic experimental training system is lacking. Here, we present the Antimicrobial Resistance-focused Pharmaceutical Developing Research (APDR) program, which broadly covers multiple courses and key topics, such as microorganism isolation and identification, drug sensitivity detection, protein expression and homology modeling, and molecular docking and new drug screening, with the potential for publishable research outcomes. Students enrolled in this program mastered various research skills and demonstrated a 2.4-fold increase in published articles per capita compared with those not enrolled in this program. Significant improvements in follow-up course scores in genetic engineering (18.5%), microbiology (6.9%), and molecular biology (10.7%) were observed among participants. Furthermore, 44.1% of participants demonstrated scholarly productivity as exemplified by authored or co-authored publications. Therefore, a systematic APDR program offers students a positive learning experience to enhance students' research skills. and provides a pedagogical tool that can be adapted to improve students' ability to engage in biomedical research.

Background and objectivesProtein disulfide isomerases (PDIs) are essential enzymes that facilitate the proper folding of proteins and maintain protein quality within the endoplasmic reticulum. Dysregulation of PDIs has been correlated with numerous disorders, including cancer and rheumatoid arthritis (RA). E64FC26 (EFC), a small molecule that inhibits a wide range of PDI family members, has shown promise as a therapeutic agent in oncology. However, its effects on RA have not yet been studied. This research investigates the efficacy of EFC as a potential treatment for RA.

BACKGROUND:Real-world data on afatinib for patients with non-small cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR) G719X + S768I co-mutations remains limited. This study aims to evaluate the efficacy and safety of afatinib in a relatively large cohort of NSCLC patients with these uncommon mutations in Xuanwei and Fuyuan counties, China. METHODS:A retrospective study was conducted on 96 NSCLC patients from Xuanwei and Fuyuan, China, between August 2019 and March 2024. Patients with advanced-stage NSCLC harboring EGFR G719X + S768I co-mutations or single EGFR G719X/S768I mutations who received first-line afatinib treatment were included in the study. The parameters analyzed included the progression-free survival (PFS), and overall survival (OS). While treatment-related adverse events (AEs) were assessed for the safety of afatinib. RESULTS:All of the 96 patients, 59 were female and 37 were male, with a median age was 59.2±9.0 years. The mutations were classified as single EGFR G719X/S768I mutations (n=34; 35.4%) and G719X + S768I co-mutations (n=62; 64.6%). The median progression-free survival (mPFS) for all patients was 12 months (95% CI: 9.5-14.5), and median overall survival (mOS) was 29 months (95% CI: 18.0-40.0). Patients with the G719X + S768I co-mutations exhibited significantly longer mPFS (17 vs. 8 months, P<0.001) and mOS (33 vs. 15 months, P=0.04) compared to those with and single EGFR G719X/S768I mutation. Common treatment-related AEs included diarrhea (n=80; 83.3%), skin rash/acne (n=77; 80.2%), paronychia (n=53; 55.2%), and stomatitis (n=33; 34.4%), all of which were manageable. There was no significant difference in AEs between the two mutation groups. CONCLUSIONS:This study represents the largest cohort of NSCLC patients with EGFR G719X + S768I co-mutations treated with first-line afatinib. Our findings confirmed the effectiveness and safety of afatinib in this patient population. Furthermore, the presence of the G719X + S768I co-mutations serves as an independent predictor of favorable PFS for NSCLC patients. This study will provide new clinical evidence supporting afatinib therapy for patients with EGFR G719X + S768I co-mutations, both in China and globally. This study fills an important gap in the existing literature by providing robust, large-scale clinical data, offering new insights for the treatment of NSCLC patients with these uncommon mutations.

8024 Background: Ensartinib has been approved as ALK-positive locally advanced/metastatic NSCLC, but its role in adjuvant therapy remains unknown. This real-world study aimed to evaluate the efficacy and safety of ensartinib as adjuvant therapy in ALK-positive NSCLC. Methods: Data were retrospectively collected from “Ensacove Patient Assistance Program" held by Betta pharmaceutical company. The patients who were ≥18 years-old and had completely resected, histologically confirmed stage I-III NSCLC as classified according to the eighth edition of AJCC/UICC, were documented ALK-positive, and had at least one post-baseline CT scan. The primary endpoint is 2-year disease-free survival (DFS) rate. Secondary endpoints include DFS, safety and OS. Results: From November 19, 2020, to May 31, 2024, a total of 296 patients were screened. 222 patients were enrolled. The median age was 55 years-old. 128 females. 98 (44.1%) were stage I, 42 (18.9%) were stage II, 82 (36.9%) were stage III. 98.2% were lung adenocarcinoma. Thirty-two patients (14.4%) received postoperative chemotherapy. The median duration of ensartinib treatment was 25.3 (95% confidence intervals [CI], 3.1-47.2) months. At the data-cutoff date of December 29, 2024, median follow-up time was 23.5 months (95% CI, 21.7-26.2). The median DFS was immature. The 2-year DFS rate was 92.1% (95% CI, 86.6%-95.5%) for all patients, and 89.3% (95% CI, 77.9%-95.0%), 100.0% (95% CI, NR-NR) and 91.0% (95% CI, 80.6%-95.9%) for patients with stage I, stage II, and stage III, respectively. 16 patients had disease recurrence or death, including 7 patients with local recurrence and 9 patients with distant metastasis. The OS data was immature with only 2 deaths occurred. The treatment-related adverse events (TRAEs) of any grade occurred in 169 patients (76.1%), and 11 patients (5.0%) experienced ≥grade 3 TRAEs. The most common TRAEs were rash (60.8%). Serious TRAEs were 8 (3.6%) patients. TRAEs led to dose reductions, dose interruptions and permanent discontinuation were 21.2%, 9.0% and 3.2% of the patients, respectively. No deaths due to TRAEs were reported. Conclusions: To our knowledge, this real-world study has the largest sample size on postoperative adjuvant therapy with ALK inhibitors. In this real-world setting, ensartinib demonstrated encouraging efficacy and well-tolerated safety profile among stage IA1-IIIB ALK-positive NSCLC, providing a potential adjuvant therapy option in this patient population.

Background:Metastasis is a hallmark of cancer and the leading cause of cancer-related mortality. However, the mechanism underlying liver metastasis in colorectal cancer (CRC) remains incompletely understood. This study explores the role of long non-coding RNA (lncRNA) SLERT in promoting CRC liver metastasis by downregulating HUNK expression. Methods:SLERT expression levels in CRC tissues were analyzed and correlated with patient survival outcomes. Functional assays, including migration and invasion assays, were performed to assess the impact of SLERT knockdown and overexpression on metastatic behavior. Mechanistic studies examined SLERT's interaction with the RNA-binding protein RBM15 and its effect on HUNK mRNA stability. The subcellular localization of SLERT was also determined. Results:SLERT was significantly upregulated in CRC tissues and associated with poor survival outcomes. Silencing SLERT inhibited CRC cell migration and invasion, whereas its overexpression enhanced these metastatic properties. Mechanistically, SLERT interacted with RBM15, impairing its ability to stabilize HUNK mRNA, leading to decreased HUNK expression and increased metastatic potential. SLERT was primarily localized in the cytoplasm, indicating its active role in gene regulation within the tumor microenvironment. Conclusion:LERT promotes liver metastasis in CRC by downregulating HUNK expression through RBM15-mediated mRNA destabilization. These findings suggest that SLERT could serve as a diagnostic biomarker and therapeutic target. Targeting SLERT or restoring HUNK expression may provide novel strategies to combat CRC liver metastasis and improve patient prognosis.

BACKGROUND:Whether the addition of bevacizumab to a programmed cell death protein-1 (PD-1) inhibitor plus chemotherapy can provide further survival benefits as first-line treatment in non-squamous non-small-cell lung cancer (NSCLC) without EGFR sensitising mutations, or ALK/ROS1 rearrangements, is unknown. We evaluated serplulimab (an anti-PD-1 antibody) plus HLX04 (a bevacizumab biosimilar) and chemotherapy in non-squamous NSCLC. METHODS:ASTRUM-002 is a randomised, double-blind, multicentre, phase 3 trial with a three-arm design. Patients (aged ≥18 years and ≤75 years) with locally advanced or metastatic non-squamous NSCLC without EGFR sensitising mutations or ALK/ROS1 rearrangements and previous systemic therapy were randomly assigned (1:1:1; stratified by PD-L1 expression, smoking history, and brain metastasis) to receive serplulimab (4·5 mg/kg intravenously) plus HLX04 (15 mg/kg intravenously) and chemotherapy (pemetrexed and carboplatin; group A), serplulimab plus chemotherapy plus HLX04 placebo (group B), or chemotherapy plus serplulimab placebo plus HLX04 placebo (group C). The primary endpoint was the blinded independent central review assessed progression-free survival per the Response Evaluation Criteria in Solid Tumors version 1.1, assessed in the intention-to-treat population. This study is registered with ClinicalTrials.gov, NCT03952403, and is complete. FINDINGS:Between Nov 25, 2019, and June 15, 2022, 642 patients were enrolled across 72 hospitals in China; six in the safety run-in phase and the remaining 636 patients were randomised to group A (212 patients), B (214), or C (210). 465 (73%) were male, 171 (27%) were female, and 599 (94%) were of Han ethnicity. By the data cutoff date of June 15, 2023, the median follow-up duration was 23·4 months (95% CI 21·6-24·9) in group A, 23·1 (21·4-25·6) in group B, and 23·0 (20·7-25·6) in group C. Median progression-free survival was 12·6 months (95% CI 8·7-14·0; 123 events) in group A, 11·0 months (95% CI 8·4-12·7; 130 events) in group B, and 5·6 months (95% CI 4·8-6·8; 156 events) in group C. A significant reduction was seen in risk of progressive disease or death for patients in group B compared with group C (hazard ratio [HR] 0·55, 95% CI 0·43-0·69; p<0·0001). No significant improvement in progression-free survival was seen for group A compared with group B (HR 0·86, 0·67-1·11; p=0·25). Treatment-related serious adverse events occurred in 82 (39%) patients in group A, 79 (37%) in group B, and 51 (24%) in group C. Grade 3 or worse treatment-related adverse events occurred in 149 (71%) patients in group A, 142 (66%) in group B, and 119 (57%) in group C. Treatment-related adverse events leading to death occurred in ten (5%) patients in group A, five (2%) in group B, and seven (3%) in group C. INTERPRETATION:The addition of serplulimab to chemotherapy led to significantly longer progression-free survival in patients with locally advanced or metastatic non-squamous NSCLC compared with chemotherapy alone and represents an alternative first-line treatment option for this patient population. HLX04 plus serplulimab and chemotherapy did not confer further statistical benefit compared with serplulimab plus chemotherapy. FUNDING:Shanghai Henlius Biotech.

Tumor-associated macrophages (TAMs) account for up to 50% of tumor mass and serve as key innate immune cells in the tumor microenvironment (Kzhyshkowska et al., 2024). TAMs are primarily M2-like phenotypes that contribute to tumor immunosuppression and promote immune escape by secreting anti-inflammatory fac tors, thereby supporting tumor growth, metastasis, and angiogenesis. TAMs can be reprogrammed into M1-like phenotypes by immunostimulatory signals, promoting pro-inflammatory cytokines such as TNF-α and IL-12 that suppress tumor growth. Growing evidence highlights TAM modulation as an effective anticancer approach by boosting immunotherapeutic responses, primarily through facilitating M2-to-M1 polarization (Mantovani et al., 2022). However, fully harnessing the therapeu tic potential of TAMs for cancer therapy requires both precise targeting and functional reprogramming, which remains a significant challenge.

Rheumatoid arthritis (RA) is a common chronic, inflammatory autoimmune disease, and early clinical diagnosis is crucial for its treatment. CXCR4 expression was characterized in arthritic mouse models and joints of RA patients, and [18 F]AIF-NOTA-QHA-04 specificity was assessed in non-malignant cells with elevated CXCR4 expression. This study explored the application of CXCR4-targeted PET probe [18 F]AIF-NOTA-QHA-04 in monitoring disease activity and therapeutic efficacy in RA. To this aim, the metabolic characteristics of [18 F]AIF-NOTA-QHA-04 and correlation of [18 F]AIF-NOTA-QHA-04 uptake with arthritis severity were evaluated by PET imaging in arthritic mice. [18 F]AIF-NOTA-QHA-04 potential in evaluating therapeutic efficacy was further investigated in arthritic mice following methotrexate (MTX) and etanercept (ETC) treatment. CXCR4 expression was significantly increased in the inflamed joints of collagen-induced arthritis (CIA) and collagen-antibody induced arthritic (CAIA) mice, and in synovial tissues of RA patients. [18 F]AIF-NOTA-QHA-04 showed high specificity for CXCR4, with increased probe uptake in arthritic joints that was strongly correlated with arthritis severity scores. PET imaging revealed that increased uptake of [18 F]AIF-NOTA-QHA-04 in arthritic joints paralleled disease activity, with uptake decreasing upon remission. Furthermore, [18 F]AIF-NOTA-QHA-04 PET imaging provided earlier and more sensitive assessments of the efficacy of MTX and ETC compared to traditional methods. The CXCR4-targeted PET probe [18 F]AIF-NOTA-QHA-04 is a promising tool for RA diagnosis and monitoring, with high specificity and sensitivity. The potential of this probe as a biomarker for disease activity and therapeutic response underscores its value in personalized medication strategies for the management of RA.

Background: Real-world data on afatinib for patients with non-small cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR) G719X + S768I co-mutations remains limited. This study aims to evaluate the efficacy and safety of afatinib in a relatively large cohort of NSCLC patients with these uncommon mutations in Xuanwei and Fuyuan counties, China. Methods: A retrospective study was conducted on 96 NSCLC patients from Xuanwei and Fuyuan, China, between August 2019 and March 2024. Patients with advanced-stage NSCLC harboring EGFR G719X + S768I co-mutations or single EGFR G719X/S768I mutations who received first-line afatinib treatment were included in the study. The parameters analyzed included the progression-free survival (PFS), and overall survival (OS). While treatment-related adverse events (AEs) were assessed for the safety of afatinib. Results: All of the 96 patients, 59 were female and 37 were male, with a median age was 59.2 +/- 9.0 years. The mutations were classified as single EGFR G719X/S768I mutations (n=34; 35.4%) and G719X + S768I co-mutations (n=62; 64.6%). The median progression-free survival (mPFS) for all patients was 12 months (95% CI: 9.5-14.5), and median overall survival (mOS) was 29 months (95% CI: 18.0-40.0). Patients with the G719X + S768I co-mutations exhibited significantly longer mPFS (17 vs. 8 months, P<0.001) and mOS (33 vs. 15 months, P=0.04) compared to those with and single EGFR G719X/S768I mutation. Common treatment-related AEs included diarrhea (n=80; 83.3%), skin rash/acne (n=77; 80.2%), paronychia (n=53; 55.2%), and stomatitis (n=33; 34.4%), all of which were manageable. There was no significant difference in AEs between the two mutation groups. Conclusions: This study represents the largest cohort of NSCLC patients with EGFR G719X + S768I co-mutations treated with first-line afatinib. Our findings confirmed the effectiveness and safety of afatinib in this patient population. Furthermore, the presence of the G719X + S768I co-mutations serves as an independent predictor of favorable PFS for NSCLC patients. This study will provide new clinical evidence supporting afatinib therapy for patients with EGFR G719X + S768I co-mutations, both in China and globally. This study fills an important gap in the existing literature by providing robust, large-scale clinical data, offering new insights for the treatment of NSCLC patients with these uncommon mutations.