Objective: To investigate the expression of anaplastic lymphoma kinase (ALK) in malignant melanoma and analyze its clinicopathological and molecular features. Methods: Eighty-seven malignant melanomas that were surgically resected at the First Affiliated Hospital, Zhejiang University School of Medicine between January 2015 and December 2019 were collected. Immunohistochemical staining was performed to evaluate ALK expression. Positive cases were further analyzed using clinical, pathological, and molecular testing data. Relevant literature was systematically reviewed, and follow-up was conducted. Results: Three cases showed ALK-positive expression. They were all in male patients, aged 52, 79, and 51 years, with tumors located in the left temporooccipital skin, maxillary gingiva, and left plantar skin, respectively. Cases 1 and 3 had a history of smoking and diabetes; case 2 had a history of alcohol use. Histologically, case 1 exhibited pagetoid spread with prominent lymphocytic infiltration; case 2 showed nodular growth with ulceration; while case 3, a recurrent lesion, was surrounded by abundant lymphocytes. All three cases displayed sheet-like proliferation of epithelioid or short spindle-shaped tumor cells, conspicuous nucleoli, and frequent mitoses. Marked pigmentation was observed in cases 1 and 3, but not in case 2. Fluorescence in situ hybridization (FISH) revealed ALK gene rearrangement in case 2, but not in cases 1 and 3. The nCounter analysis showed high expression of ALK exons 20-29 and intron 19 with low expression of exons 1-19 in cases 1 and 3, consistent with an alternative transcription initiation (ATI)-driven ALK expression pattern. In contrast, case 2 exhibited high expression of exons 20-29 but low expression of both exons 1-19 and intron 19, supporting an ALK fusion subtype. No mutations of KRAS, NRAS, BRAF, or PIK3CA were detected in any of the cases. Conclusions: Both ALK fusion and ATI-driven ALK expression patterns can present in malignant melanomas. Aberrant ALK activation suggests its potential role in molecular pathogenesis of malignant melanoma. Further investigation of ALK as a therapeutic target seems necessary.

Objective: To investigate the clinicopathological features and molecular genetics of large B-cell lymphoma with IRF4 gene rearrangements (LBL-IRF4) involving the head and neck region in adults. Methods: The clinicopathological and molecular genetics features of LBL-IRF4 in adults diagnosed at the First Affiliated Hospital of Zhengzhou University between January 2016 and December 2024 were analyzed using immunohistochemistry and fluorescence in situ hybridization (FISH). Clinical information was also collected and analyzed. Results: Seventeen cases were reviewed. There were 5 male and 12 female patients, aged 19 to 68 years, with media age of 42.0(21.0,61.5) years. All patients presented with head and neck lesions, especially Waldeyer's ring and cervical lymphadenopathy (15/17). No bone marrow involvement was detected in any of the patients while most of them were at the Ann Arbor stage Ⅰ-Ⅱ (16/17). Morphologically, 11 cases had a diffuse growth pattern, and 6 cases had a follicular/diffuse growth pattern, exhibited medium to large lymphoid cells. Two cases showed a "starry sky" appearance, and 4 cases exhibited coagulative necrosis. Tumor cells were positive for CD20 and CD79a. CD10 was positive in 13 cases (13/17) while bcl-2 was positive in 12 cases (12/17). Bcl-6 and MUM-1 were positive in all cases. CD5 was expressed in 5 cases (5/17). All cases were EBV-negative (0/16). IRF4 rearrangement was identified in all cases. No breaks in MYC or bcl-2 were detected in any of the cases. Bcl-6 rearrangements were found in 8 cases (8/17) by FISH. All 17 patients received chemotherapy after the diagnosis. Sixteen patients at follow up were alive (1 case was lost to follow-up). Conclusions: LBL-IRF4 involving the head and neck in adults is not uncommon. Adult cases mimic their pediatric counterparts, showing similar morphological and immunohistochemical features. Some cases are CD5 positive and harbor bcl-6-rearrangement. Overall, it carries a favorable prognosis. Routine testing of IRF4 rearrangement is recommended for cases with the following characteristics: in the head and neck region, morphologically high-grade follicular lymphoma or diffuse large B-cell lymphoma with strong MUM-1 expression irrespective of patient age.

Objective: This study aimed to develop a methodology for detecting FMS-like tyrosine kinase 3 (FLT3) gene internal tandem duplication (ITD) mutation burden using polymerase chain reaction-next-generation sequencing (PCR-NGS). The prognostic value of FLT3-ITD mutation burden detected with PCR-NGS in patients with acute leukemia (AL) before allogeneic hematopoietic cell transplantation (allo-HSCT) is investigated. Methods: This retrospective study developed a methodology for detecting FLT3-ITD mutational burden in bone marrow samples isolated from 65 patients with AL with FLT3-ITD mutations who received allo-HSCT from January 2021 to June 2024 at Ruijin Hospital. PCR-NGS was used for data analysis, and results were compared with conventional quantitative PCR (qPCR) . Results: The PCR-NGS assay demonstrated robust performance, with a sensitivity of 10(-6). Pretransplant complete remission with FLT3-ITD negativity via qPCR was achieved in 58 patients, comprising 25 with FLT3-ITD positivity in PCR-NGS [median VAF:0.629% (0.004% -26.350% ) ]. The 2-year probability of relapses and event-free survival (EFS) were 11.2% and 86.2% for patients with FLT3-ITD VAF of <0.1% and 30.6% and 64.5% for those having FLT3-ITD VAF of ≥0.1%, respectively (relapse: HR=3.159, 95% CI: 0.950-10.510, P=0.048; EFS: HR=2.846, 95% CI: 0.953-8.500, P=0.050). Two-year probability of relapses and EFS were 12.5% and 84.4% for patients with both negative PCR-NGS and qPCR, 26.2% and 73.8% for those with positive PCR-NGS and negative qPCR, and 28.6% and 57.1% for patients with both positive PCR-NGS and qPCR (relapse: HR=2.892, 95% CI: 1.122-7.451, P=0.0321; EFS: HR=1.784, 95% CI: 0.880-3.615, P=0.248), respectively, before allo-HSCT. Maintenance therapy with FLT3 inhibitors is a protective factor for reduced relapse rate and better overall survival rate after allo-HSCT. Conclusion: A highly sensitive PCR-NGS assay for FLT3-ITD mutational burden has been developed. Compared with qPCR, PCR-NGS demonstrates superior sensitivity and enables more accurate prediction of posttransplant outcomes in patients with AL undergoing allo-HSCT.

Objective: To summarize the clinical and genetic characteristics of Chinese patients with Birt-Hogg-Dubé syndrome (BHD). Methods: We retrospectively analyzed the clinical data of patients diagnosed with BHD between January 1, 2015, and April 30, 2025, at Peking Union Medical College Hospital, Chinese Academy of Medical Science (Beijing center); the First Affiliated Hospital of USTC, University of Science and Technology of China (Hefei center); and the First Affiliated Hospital of Guangzhou Medical University (Guangzhou center). Epidemiological characteristics, clinical manifestations, genetic variants were summarized, and the correlation between genetic mutations and phenotypic presentations were explored. Statistical analyses were performed using SAS 9.4 software. Results: A total of 168 BHD families comprising 256 patients (76 from Beijing, 141 from Hefei, and 39 from Guangzhou) were enrolled, including 201 (78.5%) probands and 55 (21.5%) family screening participants. The cohort included 188 females, with a male-to-female ratio of 1∶2.8. The mean age at diagnosis was 18-81 (44.1±12.6) years, with a median misdiagnosis duration of 12 (0, 72) months. Among 256 patients, family history was present in 195 patients (76.2%), including 71 (36.4%) pulmonary cysts with pneumothorax, 60 (30.8%) pulmonary cysts alone, 53 (27.2%) pneumothorax alone, and 11 (5.6%) renal tumors. Pulmonary cysts were present in 254 patients (99.2%), most commonly presenting as extensive cysts (≥10 cysts, 68.5%, 174/254) followed by moderate cysts (5-9 cysts, 24%, 61/254). Cysts were bilateral in 248 patients (97.6%), while only 6 (2.4%) had unilateral involvement. Lower lobe predominance was noted in 237 patients (93.3%), and paramediastinal distribution in 158 (62.2%). Pneumothorax occurred in 170 patients (66.4%), with 113 (66.5%) experiencing unilateral events and 57 (33.5%) bilateral events. The mean age at first pneumothorax was (37.4±11.6) years, with an average of 2.4 episodes per patient. Cutaneous manifestations were noted in 55.9% (143/256), but only 15.4% (22/143) underwent biopsy, confirming fibrofolliculomas (9 cases) and trichodiscomas (2 cases). Significant regional differences in skin lesion prevalence were observed (Beijing 48.7% (37/76) vs. Hefei 72.3% (102/141) vs. Guangzhou 10.3% (4/39), χ2=50.01, P<0.001). Renal imaging was performed in 232 patients, revealing renal tumors in 10 patients (4.3%), including 6 cases of renal cell carcinoma and 4 cases of angiomyolipoma. Ninety-two distinct genetic variants were identified, including 38 novel mutations. The most common variants in Chinese patients were exon 11 c.1285dupC (13.5%, 27/200), exon 11 c.1285delC (7.5%, 15/200), and exon 14 c.1579_1580insA (12/200, 6.0%). Variants associated with higher pneumothorax rates included exon 9 c.929_930insTT (3/3), large deletions in exons 1-3 (8/9) and exon 6 c.469_471delTTC (4/5). Cutaneous manifestations were more common with exon 11 flanking sequence c.1177-5_1177-3delCTC (5/6), exon 6 c.469_471del TTC (4/5), and exon 9 c.1015C>T (11/14). Conclusions: Chinese patients with BHD predominantly present with pulmonary cysts and pneumothorax, with a relatively low incidence of renal tumors. Although the detection rate of skin lesions is higher than previously reported, significant regional variations persist. Multidisciplinary collaboration is recommended in medical centers to improve diagnostic accuracy. Furthermore, Chinese patients exhibit a distinct genetic variant profile, with five variants potentially associated with pneumothorax and skin lesion phenotypes; however, further studies are required to validate these findings.

To explore the clinical features and genetic etiology of two Chinese patients with Cowden syndrome (CS).

To assess the clinical value of non-invasive prenatal testing (NIPT) for identifying maternal malignant tumors.

Objective: To explore the pathogenesis, clinicopathological and molecular genetic features of H3/IDH wildtype, paediatric-type high-grade glioma (RTK1). Methods: A total of five cases diagnosed by the clinical features,imaging,histopathology,molecular genetics and prognosis from the Department of Pathology, the First Medical Center of PLA General Hospital were collected(2022-2025). Results: Among the five cases, three were female and two were male, aged 5-38 years,the median age is 8 years old.Tumors were located in the left/right frontal lobe, cerebellum brainstem, and right temporal lobe, respectively. The poor limb movement, unstable walking, headache accompanied by nausea, vomiting in five cases. Histopathology shows features of high-grade gliomas histological changes characterized by densely arranged cells with cell atypia, vascular proliferation,necrosis and mitotic activity. Molecular showed of PDGFRA amplification or mutation in five cases, accompanied by MGMT methylation, TERT, TP53 mutation. The total course of disease from onset to death in one case is about 10 years, indicating that the progression of the disease is slower than that of adult high-grade gliomas. Conclusions: Pediatric-type gliomas occur predominantly in children but can also be observed in adults. Their disease progression and prognosis are generally more favorable compared to adult-type high-grade gliomas. Molecular testing plays a crucial role in diagnosis and differential diagnosis, holding significant importance for treatment and prognosis evaluation.

Objective: To investigate the expression of microfibril-associated protein 5 (MFAP5) in ovarian cancer and its influence on malignant behavior of ovarian cancer cells. Methods: GEPIA, CSIOVDB and Kaplan-Meier Plotter online databases were used to analyze the expression of MFAP5 in various tumor tissues, especially in ovarian cancer. Immunohistochemistry was used to detect the expression level of MFAP5 in ovarian cancer tissue chip (The tissue microarray was commissioned to Zhongke Guanghua [Xi'an] Intelligent Biotechnology Co., Ltd. for processing. The specimens were sourced from Henan Provincial People's Hospital from January 2018 to March 2023), and the relationship between MFAP5 expression and the clinical characteristics of patients with ovarian cancer was analyzed using SPSS software. Kaplan-Meier Plotter online database was used to analyze the relationship between MFAP5 expression and survival prognosis of ovarian cancer patients. The ovarian cancer data sets GSE9891 and TCGA594 were downloaded from GEO and TCGA respectively, and ssGSEA was used to analyze the scores of gene sets related to epithelial mesenchymal transition, migration and invasion in ovarian cancer data sets. Next, the relationship between MFAP5 expression and the above scores was analyzed using GraphPad Prism. The expression of MFAP5 in normal fibroblasts (NFs) and cancer associated fibroblasts (CAFs) was verified by western blot. MFAP5 expression in CAFs was reduced by MFAP5-si RNA, and influence of CAFs with high or low expression of MFAP5 on malignant behavior of ovarian cancer cell SKOV3 was verified by transwell test. The influence of CAFs with high or low expression of MFAP5 on epithelial mesenchymal transition markers of ovarian cancer cell SKOV3 was verified by western blot. The CCK8 assay and 3D co-culture model were used to verify the effects of CAFs with high and low MFAP5 expression on the chemoresistance of ovarian cancer cells SKOV3. Transwell assay, western blot, and 3D co-culture model were used to explore and verify the related pathways through which MFAP5 affects the malignant behavior and drug resistance of ovarian cancer cells SKOV3. Results: Online data analysis showed that the expression of MFAP5 in ovarian cancer tissue was significantly higher than that in normal ovarian tissue (P＜0.001). Immunohistochemistry showed that the expression of MFAP5 was mainly concentrated in ovarian cancer stroma, and the later stage and the higher grade ovarian cancer tissues showed higher MFAP5 expression. Survival analysis showed that the high expression of MFAP5 was related to the poor prognosis of patients, and it was only significant in later stages and higher grades. ssGSEA analysis showed that the expression of MFAP5 was positively correlated with the scores of gene sets related to epithelial-mesenchymal transition, migration and invasion of ovarian cancer. In vitro experiments showed that reducing the expression of MFAP5 in CAFs could not only partially reduce its supportive effect on the malignant behaviors such as migration and invasion of ovarian cancer epithelial cells SKOV3 (The number of migrating cells in the control group, si-NC group, and si-MFAP5-group was 73.44±7.80, 199.74±18.26, and 91.21±6.70, respectively. The number of invading cells in the control group, si-NC group, and si-MFAP5-group was 61.62±8.76, 174.81±15.23, and 67.17±9.83, respectively), but also increase the sensitivity of ovarian cancer epithelial cells SKOV3 to chemotherapy and maintenance therapy. Further research showed that MFAP5 could activate the AKT signaling pathway in ovarian cancer cells SKOV3, and inhibiting the AKT pathway could block the supportive effect of MFAP5 on the malignant behavior and drug resistance of ovarian cancer cells. Conclusions: MFAP5 is highly expressed in ovarian cancer, especially in the stromal tissue of ovarian cancer, where the expression level is the highest. High expression of MFAP5 often indicates a poor prognosis for ovarian cancer patients. By reducing the expression of MFAP5 in CAFs, the supportive effect on the malignant biological behavior and drug resistance of ovarian cancer epithelial cells can be weakened. The underlying mechanism of this phenomenon may be related to the blockade of the AKT signaling pathway in ovarian cancer cells.

Amivantamab is the first fully humanized bispecific antibody approved for the treatment of non-small cell lung cancer (NSCLC) in the world. Amivantamab can block epidermal growth factor receptor (EGFR) pathway and mesenchymal-epithelial transformation factor (MET) pathway simultaneously, trigger the internalization and degradation of EGFR and MET, and activate the antitumor immune response. Amivantamab has been approved by the National Medical Products Administration for the treatment of adult patients with locally advanced or metastatic NSCLC with EGFR exon 20 insertion mutation, EGFR exon 19 deletion or exon 21 L858R substitution mutation, and is expected to be widely used in clinical practice soon. How to reasonably manage the adverse reactions related to amivantamab and maximize its efficacy is an urgent problem to be solved. Based on the existing medical evidence, combined with clinical experience, the expert group of this consensus finally formulated this "Expert consensus on amivantamab clinical application and adverse reaction management (2025 edition)" after multiple discussions. The contents of the consensus include the clinical use and management of adverse reactions of amivantamab. The recommendations focus on the prevention of infusion-related reactions, skin adverse reactions, venous thromboembolism, peripheral edema, oral mucositis, ocular toxicity and interstitial lung disease, amivantamab dose adjustment and treatment when adverse events occur, in order to provide guidance for clinicians to correctly use amivantamab and manage related adverse reactions.

Retroperitoneal soft tissue sarcomas (RPS) represent a clinically challenging group of heterogeneous mesenchymal malignancies, predominantly comprising liposarcoma and leiomyosarcoma subtypes. These tumors are characterized by aggressive biological behavior, high rates of local recurrence, and unfavorable clinical outcomes. The nuclear receptor peroxisome proliferator-activated receptor γ (PPARγ) serves as a critical regulator of diverse physiological processes, including adipocyte differentiation, glucose/lipid metabolism, inflammatory responses, and immune homeostasis. Emerging evidence demonstrates that dysregulation of PPARγ signaling is closely associated with RPS pathogenesis, particularly in retroperitoneal liposarcoma (RPLS), where PPARγ functional inactivation or aberrant expression correlates significantly with tumor grade and clinical progression. While preclinical studies have demonstrated the therapeutic potential of PPARγ agonists in suppressing tumor proliferation and inducing apoptosis, clinical translation has been limited by intertumoral heterogeneity in drug responsiveness and dose-limiting adverse effects. This review systematically examines the molecular biology of PPARγ and its emerging role in RPS pathobiology, with the aim of informing precision diagnostic and therapeutic strategies for this complex disease entity.

The circadian rhythm system coordinates physiological functions such as gene expression, hormone release, energy consumption, cell growth and metabolism through a multi-level molecular network to maintain homeostasis. Circadian core genes regulate cell cycle checkpoints, metabolic homeostasis, and key signaling pathways through a transcription-translation negative feedback loop, and their dysfunction can drive malignant progression of tumors. At the same time, circadian rhythm affects the immune surveillance and immune escape process in the tumor immune microenvironment by regulating the development, differentiation and effector function of immune cells, and further promotes the occurrence and development of tumors. At present, chronotherapy based on circadian rhythm has shown significant advantages in clinical practice. Optimizing the dosage regimen and implementing the chronotherapy strategy can significantly improve the anti-tumor efficacy and reduce the side effects of drugs. Small molecule modulators targeting the biological clock or novel sequential treatment regimens combined with immune checkpoint inhibitors are being evaluated in a number of clinical trials. In view of the great potential value of circadian rhythm regulation in precision medicine, this paper systematically explores the interaction network of circadian rhythm, tumor and immune response, and provides a new perspective for tumor treatment strategy based on biological clock regulation.

Objective Through integrated omics analysis of melanoma clinical samples, this study aims to investigate the clinical significance of solute carrier family 1 member 5 (SLC1A5) expression and its correlation with the tumor immune microenvironment. Methods Transcriptomic data from melanoma tissues in The Cancer Genome Atlas (TCGA) and other databases were analyzed. Cox regression and log-rank tests were used to evaluate the correlation between SLC1A5 expression and patient survival. Gene set enrichment analysis (GSEA) was performed to explore potential functional mechanisms. The correlation between SLC1A5 expression and clinicopathological characteristics, immune infiltration and immunomodulatory molecule expression were systematically analyzed. Results Metastatic melanoma exhibited significantly higher SLC1A5 expression than primary melanoma. Elevated SLC1A5 expression correlated with worse overall survival (OS), progression-free interval (PFI) and disease-free interval (DFI) in melanoma patients. GSEA revealed significant enrichment of metastasis-related pathway, anti-apoptotic process, vascular endothelial growth factor (VEGF) signaling and immunomodulatory pathway in SLC1A5-high tumors. Furthermore, SLC1A5 expression correlated with immune infiltration (T cells, monocytes and M2-type macrophages), and positively correlated with immune checkpoint molecules [cytotoxic T-lymphocyte-associated antigen 4 (CTLA4), poliovirus receptor-like 2 (PVRL2)], chemokines [C-C motif chemokine ligand 18 (CCL18), CCL20] and transforming growth factor-β1 (TGF-β1), while negatively correlating with T cell chemokines [C-X-C motif chemokine ligand 9 (CXCL9)/CXCL10/CXCL11]. Conclusion SLC1A5 serves as an independent prognostic biomarker in melanoma. Its overexpression may shape an immunesuppressive microenvironment by dysregulating immune molecules expression and cellular infiltration, ultimately facilitating immune escape and malignant progression.

To investigate the causal relationship between gut microbiota, T-cell function, and the risk of colorectal cancer.

Objective: Profiling tumor cell heterogeneity before and after chemotherapy in breast cancer patients to delineate the cellular evolutionary trajectory at single-cell resolution, thereby identifying potential targets for intervention. Methods: Using a case-control study design, a female patient with breast cancer admitted to the Department of Breast Surgery, The First Affiliated Hospital of Hunan University of Chinese Medicine in September 2020 was enrolled as the subject. Fresh tumor tissue samples, collected both before and after chemotherapy, were subjected to single-cell RNA sequencing to assess transcriptomic profiles and observe the impact of chemotherapy on the intratumoral microenvironment. Specifically, a pre-chemotherapy biopsy sample was obtained in June 2020, and a post-chemotherapy surgical resection sample was obtained in September 2020. Pathological diagnosis confirmed Grade Ⅲ invasive ductal carcinoma for both samples, with a molecular subtype of Luminal B. Results: A significance threshold of |log₂FC|>2 and a P-value <0.05 were set to define statistically significant differences for subsequent bioinformatic analysis. Sequencing data revealed that a total of 8 599 cells were profiled in this study, with 4 180 (48.6%) and 4 419 (51.4%) cells derived from pre- and post-chemotherapy tumor tissues, respectively. It characterized the cellular composition of the tumor microenvironment and identified 13 distinct cell clusters. These included basal cells, pericytes, plasma cells, T cells, B cells, fibroblasts, endothelial cells, NK cells, mast cells, epithelial cells, macrophages, cycling cells, and plasmacytoid dendritic cells. Signaling pathways and transcription factors associated with these cell clusters were subsequently analyzed and subjected to enrichment analysis. Furthermore, this study delineated the precise cellular architecture and developmental trajectories of breast cancer before and after chemotherapy. It also predicted that the APOD, ELN, and F2R genes may play pivotal roles in disease progression. Conclusion: This study utilized single-cell RNA sequencing to analyze intra-tumoral cellular heterogeneity in a breast cancer patient before and after chemotherapy. The findings may provide a clinically informative direction for identifying novel potential therapeutic targets during chemotherapy, prior to primary tumor resection.

A retrospective analysis of clinicopathological features was conducted on 10 cases of thyroid tumors with PTEN mutations from Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Fujian Provincial Hospital, and the Affiliated Hospital of Qingdao University from 2020 to 2024. Histomorphological features were quantified using the Thyroid Histomorphological PTEN Hamartoma Syndrome (PHTS) scoring system(THiPS), immunophenotypic profiling and genetic features were examined via immunohistochemical staining and next-generation sequencing respectively. The cohort included 3 males and 7 females, with the age [M(Q1,Q3)] of 68.0(53.0, 74.0) years. Those 10 cases included papillary thyroid carcinomas (3 cases), oncocytic carcinoma of the thyroid (2 cases), follicular thyroid carcinoma (1 case), differentiated high-grade thyroid carcinoma (1 case), and thyroid low risk neoplasm (3 cases). The cases in this study had low THiPS scores [(1.0±1.15) scores], and all lacked the characteristic morphological features of PHTS. Immunohistochemical staining showed that loss of PTEN expression was confirmed in 7 cases, positive expression was found in 3 cases with non-sense PTEN mutation. Molecular analysis revealed co-mutations with PTEN in 80% of cases (8/10), including BRAFV600E(3 cases), TERT promoter (4 cases), and TP53 mutations (2 cases), while 2 cases harbored PTEN-only mutations. Follow-up data were obtained for 6 cases, with a follow-up period of 15.0 (9.0, 20.0) months. Among them, 1 case was initially diagnosed with OCT pulmonary metastasis, while the remaining 5 cases showed no recurrence or metastatic lesions during the follow-up period. The THiPS score has limited diagnostic value for thyroid tumors with PTEN alteration. Negative PTEN immunohistochemical staining may suggest a PTEN mutation, but positive staining does not exclude a genetic alteration. PTEN gene mutations can be combined with BRAF, TERT, and TP53 alteration.

Multiple myeloma (MM) is a highly het1076erogeneous hematologic malignancy driven by complex genetic and molecular abnormalities. Driver gene mutations, particularly in the RAS/MAPK, DNA damage repair, and NF-κB pathways, are central to MM pathogenesis, progression, and prognosis. Existing risk stratification systems based on cytogenetics and clinical features remain limited in predictive accuracy. Emerging genomic prognostic models and targeted therapies offer new precision treatment strategies. Integrating gene mutation analysis into prognostic frameworks may improve outcome prediction and guide therapy. This review summarizes current advances on gene mutations in MM, their prognostic implications, and potential therapeutic targets.

Here we report the case of a CBF-AML patient with KIT p.D816V mutation who failed avapritinib induction therapy and subsequently underwent bridging allogeneic hematopoietic stem cell transplantation (allo-HSCT), along with a literature review. The patient was a 64-year-old male who did not achieve remission after induction therapy with the DA regimen (Daunorubicin + Cytarabine). After reinduction with avapritinib combined with the DCHG regimen (Decitabine + Homoharringtonine + Cytarabine + Granulocyte colony-stimulating factor), he attained complete remission (CR) and flow cytometry minimal residual disease (MRD) negativity, but the RUNX1::RUNX1T1 fusion gene remained positive. During consolidation therapy, flow MRD reappeared, and the fusion gene level progressively increased. The patient then underwent a 9/10 HLA-matched unrelated donor allo-HSCT. Post-transplant, the fusion gene persisted, prompting treatment with the "ITI" regimen (with dose adjustments, including sequential addition of interferon and interleukin-2, pomalidomide incorporation, and standard vs. escalated dosing of "ITI" regimen). Currently, MRD negativity has been maintained for over 5 months, with good treatment tolerance. This finding suggest that the "ITI" regimen may serve as a novel and well-tolerated therapeutic option for CBF-AML patients with persistent RUNX1::RUNX1T1 fusion gene positivity after allo-HSCT and KIT p.D816V mutation, particularly in cases of avapritinib treatment failure.

Objective: To investigate the role and molecular mechanisms of transcription factor EB (TFEB) and its target genes in the treatment of multiple myeloma (MM) with bortezomib. Methods: TFEB target genes were predicted using the GTRD database (http://gtrd.biouml.org/), identifying Ptch1 gene for further study. Expression changes of Ptch1 in RPMI8226 and U266 MM cell lines after bortezomib treatment were assessed by real time fluorogenic quantitative PCR (RT-qPCR) and Western blot. RPMI8226 and U266 cell lines were transfected with siRNA-TFEB, and mRNA and protein levels of key factors (Ptch1, Gli1) in the Ptch1/Hedgehog signaling pathway were measured by RT-qPCR and Western blot. Furthermore, Ptch1 was overexpressed in MM cell lines via lentiviral transduction. Autophagy was evaluated by acridine orange staining, and protein levels of LC3B, Beclin-1, and Lamp-1 were measured by Western blot. Lysosomal quantity changes were assessed by lysosomal fluorescent probes. Results: Bortezomib (6.0×10(-6) mmol/L, 24 h) significantly reduced Ptch1 mRNA and protein levels in both cell lines compared with blank control group (all P<0.05). siRNA-TFEB transfection reversed bortezomib's inhibition of Hedgehog pathway key factors Ptch1 and Gli. Ptch1 overexpression in bortezomib-treated RPMI8226 and U266 cells significantly reduced the relative expression of autophagy-related proteins LC3B, Beclin-1, and Lamp-1 (all P=0.001). Acridine orange staining showed fewer acidic vesicular organelles within two cell lines (all P=0.001). The relative fluorescence expressions of lysosomal probes reflecting the number of lysosomes were also decreased (P values of RPMI8226 and U266 cell lines were 0.001 and 0.007, respectively) . Conclusion: The knockdown of TFEB can specifically promote the expression of the Ptch1/Hedgehog signaling pathway, thereby reducing bortezomib-induced autophagy in MM cells and reversing the inhibitory effect of bortezomib on the proliferation of MM cell lines.

Objective: To investigate the clinical, laboratory, and prognostic features of myelodysplastic neoplasm (MDS) patients harboring acute myeloid leukemia (AML) -like mutations. Methods: We retrospectively analyzed clinical, molecular, and outcome data from 1 464 adults with primary MDS diagnosed at the Institute of Hematology and Blood Diseases Hospital from August 2016 to June 2024. Results: AML-like mutations were detected in 64 patients (4.4% ). Compared with patients without AML-like mutations, those with AML-like mutations were younger [median 50 (IQR 39-60) vs 56 (45, 65) years; P=0.001], more often female (51.6% vs 35.4% ; P=0.009), had higher bone marrow blast percentage [6.5% (3.0%, 10.5% ) vs 2.5% (1.0%, 7.0% ) ; P<0.001], a higher rate of normal karyotype (75.0% vs 48.1% ; P<0.001), and lower hemoglobin levels [73 (67, 82) g/L vs 80 (66, 98) g/L; P=0.006]. The AML-like group had a higher number of gene mutations than the non-AML-like group [3 (IQR 2-4) vs 2 (1, 3) ; P<0.001). It was enriched for mutations in NPM1, DNMT3A, WT1, PTPN11, NRAS, BCOR, FLT3, CEBPA, and MYC (all P<0.05) and had lower rates of U2AF1, ASXL1, and TP53 mutations (all P<0.05). Overall survival (OS) did not differ between groups (P=0.730) ; however, the AML-like group had significantly shorter leukemia-free survival (LFS) [19 months (95% CI: 13-25) vs 46 months (95% CI: 38-54) ; P=0.012] and a higher 2-year cumulative incidence of AML transformation [ (41.7±9.1) % vs (10.4±1.1) % ; P<0.001]. Within the AML-like group, OS, LFS, and cumulative incidence of AML transformation did not differ between patients with low blasts and those with excess blasts (IB). Multivariable Cox regression identified age ≥60 years and PTPN11 mutations as independent adverse prognostic factors for OS, while DNMT3A, PTPN11, and FLT3 mutations independently predicted leukemic transformation. Conclusions: MDS patients harboring AML-like mutations exhibit distinct clinical and molecular features and a higher risk of progression to AML.