Surface-enhanced Raman spectroscopy (SERS) has been widely applied for non-invasive, rapid measurement-based cancer detection; while its performance is highly dependent on sufficient training data for machine learning models. However, although large cancer cohorts are available in high-level research institutions, stringent privacy regulations and the sensitive nature of patient data limit cross-institutional sharing, constraining most SERS cancer detection studies to small sample sizes. This scarcity of training data results in model overfitting and insufficient generalization capability. To address these challenges, this study proposes CAM-Diffusion, a method that integrates Gradient-Weighted Class Activation Mapping (Grad-CAM) with conditional diffusion models to generate high-quality synthetic SERS spectra and provide interpretable localization of key features. The 1D-Grad-CAM module identifies discriminative Raman shift regions that contribute significantly to cancer classification through gradient-weighted mechanisms and constructs masks to guide the conditional diffusion process. CAM-Diffusion utilizes mask-guided cross-attention mechanisms to improve feature learning in key diagnostic regions through differential allocation of attention weights. Using a serum SERS dataset comprising four cancer types, two benign diseases, and healthy controls, five-fold cross-validation experiments under limited training data scenarios demonstrate that CAM-Diffusion achieved consistent performance improvements across eight classifiers, with accuracy gains ranging from 2.29% to 7.82%. In conclusion, CAM-Diffusion identifies key spectral regions, generates high-quality SERS spectral data, and substantially improves classifier performance through interpretability-guided synthesis.

Primary liver cancer is the fourth most common malignancy and the second leading cause of cancer-related death in China[1].Most patients are diagnosed at an advanced stage,which renders them ineligible for curative surgical resection.

OBJECTIVE:The impact of nucleos(t)ide analogues (NAs) therapy on the long-term outcomes in chronic HBV infection individuals outside 2025 European Association for the Study of the Liver (EASL) strongly recommended treatment indications remains uncertain. We aimed to assess the association between NAs therapy and the risks of cirrhosis and hepatocellular carcinoma (HCC) in Chinese chronic HBV infection individuals outside these criteria. DESIGN:We analysed data from 30 784 chronic HBV infection individuals across 12 centres in China. We categorised individuals outside 2025 EASL strongly recommended treatment criteria into four groups: (1) hepatitis B e antigen (HBeAg)-positive, high replicative (age <30, HBV DNA ≥8 log₁₀IU/mL, normal alanine aminotransferase (ALT), no/mild fibrosis, no family history of liver cancer); (2) HBeAg-positive, impending phase transition (typically age ≥30, HBV DNA ≥6 log₁₀IU/mL, normal ALT, no advanced fibrosis); (3) HBeAg-negative, low replicative (HBV DNA <3.3 log₁₀IU/mL, normal ALT, no/mild fibrosis); (4) HBeAg-negative, high replicative, low-risk (HBV DNA 3.3-4.3 log₁₀IU/mL, normal ALT, no/mild fibrosis). The primary endpoints were the incidence of cirrhosis and HCC. RESULTS:Up to 5 years of follow-up (117 814 person-years), we documented 635 incident cirrhosis and 164 HCC cases. In multivariable analyses, NA therapy significantly reduced risks of cirrhosis (HR 0.18, 95% CI 0.11 to 0.32) and HCC (HR 0.03, 95% CI 0.01 to 0.21) in Group 1, and cirrhosis (HR 0.50, 95% CI 0.41 to 0.61) and HCC (HR 0.25, 95% CI 0.16 to 0.41) in Group 2. No significant associations were observed in Groups 3 and 4. Findings were consistent in propensity score matching analyses. CONCLUSION:NAs therapy was associated with reduced risks of cirrhosis and HCC in HBeAg-positive individuals with high replicative or impending phase transition phenotypes, supporting the expansion of chronic HBV infection treatment criteria.

DNA hypomethylating agents (HMAs) are widely used to treat acute myeloid leukemia (AML)/myelodysplastic syndrome (MDS), but most treated patients relapse and lack standard treatment options. Using high-throughput screening, the approved all-trans retinoic acid (ATRA) is identified that exhibit high selectivity in killing HMA-resistant AML cells compared to parental cells. Mechanistically, HMA-resistant cells are overloaded with DNA hypomethylation-associated endogenous viral double-stranded RNA (dsRNA) which, however, fails to trigger an anticancer interferon (IFN) immune response due to downregulation of dsRNA sensor retinoic acid-inducible gene I (RIG-I). ATRA compensates for RIG-I expression, thereby re-triggering IFN response and potently inhibiting HMA-resistant AML cell lines, xenograft mice, and patient-derived primary cells. A library of potential RIG-I-inducing compounds is rationally constructed and screened, in which the approved M3 AML treatment drug tamibarotene (TAM) exhibits strikingly 28036-fold selectivity and 779 pm IC50 in killing HMA-resistant AML cells. ATRA and TAM do not selectively inhibit p53-mutant cancer cells. Together, this study uncovers a common resistance mechanism in HMA-treated AML patients and, in addition, provides highly potent and selective agents that can overcome resistance through re-triggering IFN anticancer immune response.

BackgroundThe utility of pre- and post-operative alpha-fetoprotein (AFP) and des-gamma (γ)-carboxy prothrombin (DCP) expression patterns and their dynamic changes as predictors of the outcome of hepatic resection for hepatocellular carcinoma (HCC) has yet to be well elucidated.MethodsFrom a multicenter database, AFP and DCP data during the week prior to surgery and the first post-discharge outpatient visit (within 1-2 months after surgery) were collected from patients with HCC who underwent hepatectomy. AFP-DCP expression patterns were categorized according to the number of positive tumor markers (AFP ≥ 20ng/mL, DCP ≥ 40mAU/mL), including double-negative, single-positive, and double-positive. Changes in the AFP-DCP expression patterns were delineated based on variations in the number of positive tumor markers when comparing pre- and post-operative patterns.ResultsPreoperatively, 53 patients (8.3%), 337 patients (52.8%), and 248 patients (38.9%) exhibited double-negative, single-positive, and double-positive AFP-DCP expression patterns, respectively. Postoperatively, 463 patients (72.6%), 130 patients (20.4%), and 45 patients (7.0%) showed double-negative, single-positive, and double-positive AFP-DCP expression patterns, respectively. Survival analysis showed a progressive decrease in recurrence-free (RFS) and overall survival (OS) as the number of postoperative positive tumor markers increased (both P < 0.001). Multivariate analysis showed that postoperative AFP-DCP expression pattern, but not preoperative AFP-DCP expression pattern, was an independent risk factor for RFS and OS. Further analysis showed that for patients with positive preoperative markers, prognosis gradually improves as positive markers decrease postoperatively. In particular, when all postoperative markers turned negative, the prognosis was consistent with that of preoperative double-negative patients, regardless of the initial number of positive markers.ConclusionsAFP-DCP expression patterns, particularly postoperative patterns, serve as vital sources of information for prognostic evaluation following hepatectomy for HCC. Moreover, changes in AFP-DCP expression patterns from pre- to post-operation enable dynamic prognostic risk stratification postoperatively, aiding the development of individualized follow-up strategies.

目的 探讨腹腔镜胆囊切除+胆总管切开取石+胆管一期缝合术(LC+LCBDE+PDC)在胆总管结石合并胆囊结石患者中的应用价值.方法 选取2020年6月—2022年6月分别接受腹腔镜胆囊切除+胆总管切开取石+胆管一期缝合术(LC+LCBDE+PDC)治疗的51例胆总管结石合并胆囊结石患者作为LCBDE组,同期选取接受内镜逆行性胰胆管造影(ERCP)+LC治疗的29例胆总管结石合并胆囊结石患者作为ERCP组;对两组患者临床资料进行回顾性分析.结果 LCBDE组手术时间、住院时间、结石清除率均显著优于ERCP组(P<0.05),两组术后肛门排气时间比较差异不显著(P>0.05);治疗前两组焦虑自评量表(SAS)、抑郁自评量表(SDS)、SF-36评分比较差异不显著(P>0.05),治疗后两组SAS、SDS、SF-36评分均得到显著改善(P<0.05),且术后LCBDE组SAS、SDS、SF-36评分均显著优于ERCP组(P<0.05);LCBDE组并发症发生率显著低于ERCP组(P<0.05).结论 与ERCP+LC相比,LC+LCBDE+PDC用于治疗胆总管结石合并胆囊结石有效性和安全性均更高,且能更好地改善患者负面情况和生活质量,值得临床借鉴推广.

Epigenetic variants carried by circulating tumor DNA can be used as biomarkers for early detection of hepatocellular carcinoma (HCC) by noninvasive liquid biopsy. However, traditional methylation analysis method, bisulfite sequencing, with disadvantages of severe DNA damage, is limited in application of low-amount cfDNA analysis. Through mild enzyme-mediated conversion, enzymatic methyl sequencing (EM-seq) is ideal for precise determination of cell-free DNA methylation and provides an opportunity for HCC early detection. EM-seq of methylation control DNA showed that enzymatic conversion of unmethylated C to U was more efficient than bisulfite conversion. Moreover, a relatively large proportion of incomplete converted EM-seq reads contains more than 3 unconverted CH site (CH = CC, CT or CA), which can be removed by filtering to improve accuracy of methylation detection by EM-seq. A cohort of 241 HCC, 76 liver disease, and 279 normal plasma samples were analyzed for methylation value on 1595 CpGs using EM-seq and targeted capture. Model training identified 283 CpGs with significant differences in methylation levels between HCC and non-HCC samples. A HCC screening model based on these markers can efficiently distinguish HCC sample from non-HCC samples, with area under the curve of 0.957 (sensitivity = 90%, specificity = 97%) in the test set, performing well in different stages as well as in serum α-fetoprotein/protein induced by vitamin K absence-II negative samples. Filtering of reads with ≥ 3 CHs derived from incomplete conversion can significantly reduce the noise of EM-seq detection. Based on targeted EM-seq analysis of plasma cell-free DNA, our HCC screening model can efficiently distinguish HCC patients from non-HCC individuals with high sensitivity and specificity.

Background Routine clinical staging for hepatocellular carcinoma (HCC) incorporates liver function, general health, and tumor morphology. Further refinement of prognostic assessments and treatment decisions may benefit from the inclusion of tumor biological marker alpha-fetoprotein (AFP) and systemic inflammation indicator C-reactive protein (CRP). Methods Data from a multicenter cohort of 2770 HCC patients undergoing hepatectomy were analyzed. We developed the PACE risk score (Prognostic implications of AFP and CRP Elevation) after initially assessing preoperative AFP and CRP’s prognostic value. Subgroup analyzes were performed in BCLC cohorts A and B using multivariable Cox analysis to evaluate the prognostic stratification ability of the PACE risk score and its complementary utility for BCLC staging. Results Preoperative AFP ≥ 400ng/mL and CRP ≥ 10 mg/L emerged as independent predictors of poorer prognosis in HCC patients who underwent hepatectomy, leading to the creation of the PACE risk score. PACE risk score stratified patients into low, intermediate, and high-risk groups with cumulative 5-year overall (OS) and recurrence-free survival (RFS) rates of 59.6%/44.9%, 43.9%/38.4%, and 20.6%/18.0% respectively (all P < 0.001 ). Increased PACE risk scores correlated significantly with early recurrence and extrahepatic metastases frequency (all P < 0.001 ). The multivariable analysis identified intermediate and high-risk PACE scores as independently correlating with poor postoperative OS and RFS. Furthermore, the PACE risk score proficiently stratified the prognosis of BCLC stages A and B patients, with multivariable analyses demonstrating it as an independent prognostic determinant for both stages. Conclusion The PACE risk score serves as an effective tool for postoperative risk stratification, potentially supplementing the BCLC staging system.

目的 研究肝细胞肝癌(HCC)组织中 APOBEC1 互补因子(A1CF)和上移码蛋白 1(UPF1)的表达及临床意义.方法 选取 2017 年 1 月至 2020 年 1 月在该院诊治的 118 例HCC患者作为研究对象.应用荧光定量PCR检测组织中A1CF、UPF1 mRNA 表达.应用免疫组织化学检测组织中 A1CF、UPF1 蛋白表达.A1CF、UPF1 mRNA表达的相关性采用 Pearson 相关分析.A1CF、UPF1 蛋白表达的相关性采用 Spearman秩相关分析.分析 HCC 癌组织中 A1CF、UPF1 蛋白表达与临床病理特征的关系.Kaplan-Meier 生存分析A1CF、UPF1 蛋白表达对 HCC患者生存预后的影响.单因素及多因素 COX 回归分析影响 HCC 患者临床生存预后的因素.结果 癌组织中 A1CF mRNA 相对表达量显著高于癌旁组织,差异有统计学意义(P＜0.001).癌组织中UPF1 mRNA相对表达量显著低于癌旁组织,差异有统计学意义P＜0.001).HCC癌组织中A1CF蛋白表达阳性率明显高于癌旁组织(P＜0.001),HCC癌组织中UPF1 蛋白表达阳性率明显低于癌旁组织(P＜0.001).HCC 癌组织中 A1CF 与 UPF1 mRNA 表达呈显著负相关(r=-0.713,P＜0.001).A1CF与UPF1 蛋白表达亦呈显著负相关(rs=-0.782,P＜0.001).不同肿瘤分期、组织学分级 HCC癌组织中A1CF、UPF1 蛋白表达阳性率比较,差异有统计学意义(P＜0.05).A1CF 阳性组患者累积生存明显低于A1CF阴性组,差异有统计学意义(P＜0.05);UPF1 阴性组患者累积生存低于UPF1 阳性组差异有统计学意义(P＜0.05).肿瘤分期Ⅲ期、组织学分级Ⅲ级、A1CF蛋白阳性、UPF1 蛋白阴性是影响患者不良生存预后的独立危险因素.结论 HCC中A1CF表达升高,UPF1 表达降低,二者表达与肿瘤 TNM分期、病理分级有关,是影响 HCC患者不良预后的独立因素.

Additional file 2: Table S1. C sites prone to incomplete conversion in EM-seq. Table S2. Quality control metrics for Targeted EM-seq using HCC cell line input from 5 to 20 ng. Table S3. Quality control metrics for Targeted EM-seq from 596 clinical samples. Table S4. Comparison of methylation levels detected from two hybridization batches. Table S5. Summary of patient characteristics. Table S6. Summary of 283 CpG markers. Table S7. Methylation levels of 283 CpGs from clinical samples.

Background and Aims:Hepatectomy is an effective treatment for selected patients with large hepatocellular carcinoma (HCC). This study aimed to develop a nomogram incorporating non-tumoral liver volume (non-TLV) and liver function markers to predict the patients' overall survival (OS) and disease-free survival (DFS).Methods:Data of 198 consecutive large HCC patients who underwent hepatectomy at the Zhongshan Hospital Xiamen University were collected. Another 68 patients from the Mengchao Hepatobiliary Surgery Hospital served as an external validation cohort. The nomograms were developed based on the independent prognostic factors screened by multivariate Cox regression analyses. Concordance index (C-index), calibration curves, and time-dependent receiver operating characteristic (ROC) curves were used to measure the discrimination and predictive accuracy of the models.Results:High HBV DNA level, low non-TLV/ICG, vascular invasion, and a poorly differentiated tumor were confirmed as independent risk factors for both OS and DFS. The model established in this study predicted 5-year post-operative survival and DFS in good agreement with the actual observation confirmed by the calibration curves. The C-indexes of the nomograms in predicting OS and DFS were 0.812 and 0.823 in the training cohort, 0.821 and 0.846 in the internal validation cohort, and 0.724 and 0.755 in the external validation cohort. The areas under the ROC curves (AUCs) of nomograms for predicted OS and DFS at 1, 3, and 5 year were 0.85, 0.86, 0.83 and 0.76, 0.76, 0.63, respectively.Conclusions:Nomograms with non-TLV/ICG predicted the prognosis of single large HCC patients accurately and effectively.

Tumor suppressor p53 is inactivated by thousands of heterogeneous mutations in cancer, but their individual druggability remains largely elusive. Here, we evaluated 800 common p53 mutants for their rescue potencies by the representative generic rescue compound arsenic trioxide (ATO) in terms of transactivation activity, cell growth inhibition, and mouse tumor–suppressive activities. The rescue potencies were mainly determined by the solvent accessibility of the mutated residue, a key factor determining whether a mutation is a structural one, and the temperature sensitivity, the ability to reassemble the wild-type DNA binding surface at a low temperature, of the mutant protein. A total of 390 p53 mutants were rescued to varying degrees and thus were termed as type 1, type 2a, and type 2b mutations, depending on the degree to which they were rescued. The 33 type 1 mutations were rescued to amounts comparable to the wild type. In PDX mouse trials, ATO preferentially inhibited growth of tumors harboring type 1 and type 2a mutants. In an ATO clinical trial, we report the first-in-human mutant p53 reactivation in a patient harboring the type 1 V272M mutant. In 47 cell lines derived from 10 cancer types, ATO preferentially and effectively rescued type 1 and type 2a mutants, supporting the broad applicability of ATO in rescuing mutant p53. Our study provides the scientific and clinical communities with a resource of the druggabilities of numerous p53 mutations ( www.rescuep53.net ) and proposes a conceptual p53-targeting strategy based on individual mutant alleles rather than mutation type.

p53 is mutated in half of cancer cases. However, no p53-targeting drugs have been approved. Here, we reposition decitabine for triple-negative breast cancer (TNBC), a subtype with frequent p53 mutations and extremely poor prognosis. In a retrospective study on tissue microarrays with 132 TNBC cases, DNMT1 overexpression was associated with p53 mutations ( P = 0.037) and poor overall survival (OS) ( P = 0.010). In a prospective DEciTabinE and Carboplatin in TNBC (DETECT) trial (NCT03295552), decitabine with carboplatin produced an objective response rate (ORR) of 42% in 12 patients with stage IV TNBC. Among the 9 trialed patients with available TP53 sequencing results, the 6 patients with p53 mutations had higher ORR (3/6 vs. 0/3) and better OS (16.0 vs. 4.0 months) than the patients with wild-type p53. In a mechanistic study, isogenic TNBC cell lines harboring DETECT-derived p53 mutations exhibited higher DNMT1 expression and decitabine sensitivity than the cell line with wild-type p53. In the DETECT trial, decitabine induced strong immune responses featuring the striking upregulation of the innate immune player IRF7 in the p53-mutated TNBC cell line (upregulation by 16-fold) and the most responsive patient with TNBC. Our integrative studies reveal the potential of repurposing decitabine for the treatment of p53-mutated TNBC and suggest IRF7 as a potential biomarker for decitabine-based treatments.

Developmental arsenic exposure has been associated with cognitive deficits in epidemiological studies, but the underlying mechanisms remain poorly understood. Here, we establish a mouse model of developmental arsenic exposure exhibiting deficits of recognition and spatial memory in the offspring. These deficits are associated with genome-wide DNA hypomethylation and abnormal expression of cognition-related genes in the hippocampus. Arsenic atoms directly bind to the cysteine-rich ADD domain of DNA methyltransferase 3A (DNMT3A), triggering ubiquitin- and proteasome-mediated degradation of DNMT3A in different cellular contexts. DNMT3A degradation leads to genome-wide DNA hypomethylation in mouse embryonic fibroblasts but not in non-embryonic cell lines. Treatment with metformin, a first-line antidiabetic agent reported to increase DNA methylation, ameliorates the behavioral deficits and normalizes the aberrant expression of cognition-related genes and DNA methylation in the hippocampus of arsenic-exposed offspring. Our study establishes a DNA hypomethylation effect of developmental arsenic exposure and proposes a potential treatment against cognitive deficits in the offspring of pregnant women in arsenic-contaminated areas.

The tumor suppressor p53 is inactivated by over hundreds of heterogenous mutations in cancer. Here, we purposefully selected phenotypically reversible temperature-sensitive (TS) p53 mutations for pharmacological rescue with thermostability as the compound-screening readout. This rational screening identified antiparasitic drug potassium antimony tartrate (PAT) as an agent that can thermostabilize the representative TS mutant p53-V272M via noncovalent binding. PAT met the three basic criteria for a targeted drug: availability of a co-crystal structure, compatible structure-activity relationship, and intracellular target specificity, consequently exhibiting antitumor activity in a xenograft mouse model. At the antimony dose in clinical antiparasitic therapy, PAT effectively and specifically rescued p53-V272M in patient-derived primary leukemia cells in single-cell RNA sequencing. Further scanning of 815 frequent p53-missense mutations identified 65 potential PAT-treatable mutations, most of which were temperature sensitive. These results lay the groundwork for repurposing noncovalent antiparasitic antimonials for precisely treating cancers with the 65 p53 mutations.

Dear Editor, p53 is the most frequently mutated protein in cancers. We recently reported that arsenic trioxide (ATO) can efficiently restore tumour-suppressor function to structural mutant p53,1,2 spawning a series of p53-targeted ATO clinical trials registered on ClinicalTrials.gov. However, ATO, together with other p53-targeted agents, still lacks reports for statistically significant clinical efficacy at present. Targeting the molecular alterations caused by p53 mutations (for example, become addicted to a signaling), rather than directly targeting mutant p53 itself, is an alternative strategy to treat p53-mutated patients. We and others previously reported a high complete response rate of the decitabine (DAC) in treating p53mutated acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS).3–5 This observation is particularly encouraging considering the high hazard ratio of TP53 mutation for overall survival in myeloid malignancies (Figure S1A-D). However, all responding p53-mutated patients relapsed in a few months.4,5 Thus, understandingmechanisms underlying the high response rate and the inevitable relapse is critical. DAC and analogues were reported to trigger interferon (IFN) signaling in mouse embryonic fibroblasts (MEFs) and cancer cells.6–8 We confirmed the preferentially inhibition of cell growth and upregulation of IFN signaling genes byDAC inTrp53−/−MEFs (Figure S2A,B).Using isogenic myeloid malignant THP-1 cells (originally null p53) infected with wild-type (WT) p53, mutant p53 (R282W, a hotspot p53 mutation frequently detected in AML/MDS5) or vector (Figure S2C), we found that DAC also preferentially inhibited p53-deficient THP-1 lines (Figure S2D). In RNA-seq, 5 μMDAC significantly upregulated 216 and 332 genes in p53-R282W cells and p53 WT cells, respectively. The encoded proteins were highly enriched in an interaction network centering type I IFN signaling in p53-R282W cells, but not in p53 WT cells (Figure 1A,B). Consistently, IFN signaling was themost enriched gene ontology biolog-

Current tumor imaging agents are often limited by their liability to dissipate from tumor tissues. As cell sugar sorting enables exogenous sugars to be delivered into predetermined subcellular locations, we synthesized sialic acid (Sia) derivatives with rhodamine-X conjugated at C-9 (ROXSia), which hitchhikes cell sialic acid sorting to target tumor cell lysosomes, exhibiting pH-independent long-term probe retention in lysosomes. ROXSia gives selective, bright, and endured fluorescence signals in subcutaneous tumors and orthotopic tumors in mice models. These results indicate the potential of ROXSia as a lysosome-targeted optical agent for fluorescence-guided tumor resection.

A newly identified lncRNA designated as RP11-284P20.2 has been identified to be up-regulated in hepatocellular carcinoma (HCC), but its role in HCC remain poorly understood. Quantitative PCR and immunocytochemical analysis were performed using the HCC tissues to identify the potential interaction partners of RP11-284P20.2. Moreover, RP11-284P20.2 was knocked down in HCC cell lines, HepG2 and SMMC7721, to investigate the influence of this lncRNA on cell growth properties. Additionally, RNA fluorescence in situ hybridization and immunofluorescence, RNA immunoprecipitation, and RNA pull-down assays were performed to determine the interaction of RP11-284P20.2 with c-met mRNA and eukaryotic translation initiation factor 3b (EIF3b). Silencing RP11-284P20.2 inhibited cell viability, migration, invasion, and colony formation, and increased apoptosis. Overexpression of c-met abolished these effects of RP11-284P20.2 in HCC cells. Histopathological examination showed that HCC tissues with high RP11-284P20.2 expression had higher c-met protein level than that in HCC tissues with low RP11-284P20.2 expression. However, there was no positive correlation between the expression levels of RP11-284P20.2 and c-met mRNA. RP11-284P20.2 knockdown led to a decease in c-met protein expression level, but did not affect the c-met mRNA expression level. These data suggest that RP11-284P20.2 regulates c-met protein expression level, which is independent of c-Met mRNA expression level. It was also confirmed that RP11-284P20.2 has high affinity toward both c-met mRNA and EIF3b protein, and hence RP11-284P20.2 probably recruits EIF3b protein to c-met mRNA and further facilitates its translation. RP11-284P20.2 promotes cell proliferation and invasion in hepatocellular carcinoma by recruiting EIF3b to induce c-met protein synthesis.