BACKGROUND:Hepatocellular carcinoma (HCC) develops in an immunosuppressive tumor microenvironment characterized by NLRP3 overexpression, in which myeloid-derived suppressor cells (MDSCs) are abundantly enriched and play a critical role in compromising T cell-mediated antitumor immunity. Arglabin (Arg), a natural sesquiterpene lactone derived from Artemisia glabella, shows potent anti-HCC potential, yet its direct molecular target responsible for the immunomodulatory effects in HCC remains poorly defined. PURPOSE:To identify its molecular target and delineate the pharmacological mechanisms underlying Arg-mediated anti-HCC immunity. METHODS:NLRP3 and CCR2 expression in myeloid-derived cells was investigated in HCC specimens by western blot, multiplex immunofluorescence and single-cell RNA sequencing data from the GEO database. Molecular interactions between Arg and NLRP3 were assessed through CETSA, SPR, LC-MS/MS, and pull-down assays. In vivo anti-tumor efficacy was evaluated in syngeneic murine models using H22 (BALB/c) and Hepa1-6 (C57BL/6) liver cancer cells. Functional studies included CCK8 and transwell migration assays. Tumor microenvironment composition was analyzed by flow cytometry, and tumor burden was monitored using in vivo bioluminescent imaging. Mechanistically, transcriptional regulation of CCR2 by NLRP3 was investigated through ChIP-qPCR. RESULTS:Here, we demonstrate that Arg suppresses HCC progression in syngeneic murine models by reducing MDSC infiltration and promoting CD8+ T-cell accumulation. MDSC co-implantation and depletion assays confirmed that the therapeutic efficacy of Arg functionally depends on MDSC modulation. Mechanistically, we identified NLRP3 as the direct molecular target of Arg with binding occurring specifically at the Cys280 residue; furthermore, experiments in Nlrp3-/- mice confirmed that NLRP3 is essential for the anti-HCC activity of Arg. Crucially, we reveal that Arg's antitumor effect is independent of the classical NLRP3 inflammasome/IL-1β pathway. Instead, NLRP3 functions through a non-canonical mechanism involving its nuclear translocation and recruitment to the Ccr2 promoter, thereby driving a transcriptional program that orchestrates MDSC trafficking into the tumor microenvironment. Furthermore, NLRP3/CCR2 blockade by Arg potentiates the therapeutic effects of immune checkpoint inhibitors (ICIs) to inhibit tumor growth. CONCLUSION:Our findings establish Arg as an NLRP3-targeting immunomodulator that reshapes the HCC microenvironment. Specifically, Arg binds to the Cys280 residue of NLRP3, thereby inhibiting its nuclear translocation and abrogating the transcriptional regulation of CCR2, which subsequently blocks MDSC trafficking. This effect works together with PD-1 blockade to overcome immune evasion in HCC, providing a strategy for reversing immunosuppression and potentiating immunotherapy.

The dysregulation of cellular epigenetic machinery has been established as a fundamental driver of oncogenesis. This recognition has propelled cancer epigenetics to the forefront of biomedical research, particularly regarding the mechanistic characterization of epigenetic switching events. These molecular switches represent critical regulatory nodes in the malignant transformation. The epigenetic switch is a complex structure formed through interactions between nucleic acid–protein complexes or protein–protein interaction complexes and specific DNA fragments. Triggered by a priming event, this molecular apparatus can reversibly activate or repress the transcription of multiple downstream genes. The inherent reversibility of these epigenetic switches presents novel therapeutic opportunities for targeted cancer intervention. Consequently, this review provides a systematic analysis of cancer-associated epigenetic switches identified in the past decade.

BACKGROUND Understanding the status and function of tumor-infiltrating immune cells is essential for improving immunotherapeutic effects and predicting the clinical response in human patients with carcinoma. However, little is known about tumor-infiltrating immune cells, and the corresponding research results in hepatocellular carcinoma (HCC) are limited. AIM To investigate potential biomarker genes that are important for the development of HCC and to understand how immune cell subsets react throughout this process. METHODS Using single-cell RNA sequencing and T-cell receptor sequencing, the heterogeneity and potential functions of immune cell subpopulations from HCC tissue and normal tissue adjacent to carcinoma, as well as their possible interactions, were analyzed. RESULTS Eight T-cell clusters from patients were analyzed and identified using bioinformatics, including six typical major T-cell clusters and two newly identified T-cell clusters, among which Fc epsilon receptor 1G+ T cells were characterized by the upregulation of Fc epsilon receptor 1G, tyrosine kinase binding protein, and T cell receptor delta constant, whereas metallothionein 1E+ T cells proliferated significantly in tumors. Differentially expressed genes, such as regulator of cell cycle, cysteine and serine rich nuclear protein 1, SMAD7 and metallothionein 1E, were identified as significantly upregulated in tumors and have potential as biomarkers. In association with T-cell receptor analysis, we inferred the clonal expansion characteristics of each T-cell cluster in HCC patients. CONCLUSION We identified lymphocyte subpopulations and potential biomarker genes critical for HCC development and revealed the clonal amplification of infiltrating T cells. These data provide valuable resources for understanding the response of immune cell subsets in HCC.

BACKGROUND:Gastric cancer is a common malignant tumor of the digestive system worldwide, and its early diagnosis is crucial to improve the survival rate of patients. Indocyanine green fluorescence imaging (ICG-FI), as a new imaging technology, has shown potential application prospects in oncology surgery. The meta-analysis to study the application value of ICG-FI in the diagnosis of gastric cancer sentinel lymph node biopsy is helpful to comprehensively evaluate the clinical effect of this technology and provide more reliable guidance for clinical practice. AIM:To assess the diagnostic efficacy of optical imaging in conjunction with indocyanine green (ICG)-guided sentinel lymph node (SLN) biopsy for gastric cancer. METHODS:Electronic databases such as PubMed, Embase, Medline, Web of Science, and the Cochrane Library were searched for prospective diagnostic tests of optical imaging combined with ICG-guided SLN biopsy. Stata 12.0 software was used for analysis by combining the "bivariable mixed effect model" with the "midas" command. The true positive value, false positive value, false negative value, true negative value, and other information from the included literature were extracted. A literature quality assessment map was drawn to describe the overall quality of the included literature. A forest plot was used for heterogeneity analysis, and P < 0.01 was considered to indicate statistical significance. A funnel plot was used to assess publication bias, and P < 0.1 was considered to indicate statistical significance. The summary receiver operating characteristic (SROC) curve was used to calculate the area under the curve (AUC) to determine the diagnostic accuracy. If there was interstudy heterogeneity (I 2 > 50%), meta-regression analysis and subgroup analysis were performed. RESULTS:Optical imaging involves two methods: Near-infrared (NIR) imaging and fluorescence imaging. A combination of optical imaging and ICG-guided SLN biopsy was useful for diagnosis. The positive likelihood ratio was 30.39 (95%CI: 0.92-1.00), the sensitivity was 0.95 (95%CI: 0.82-0.99), and the specificity was 1.00 (95%CI: 0.92-1.00). The negative likelihood ratio was 0.05 (95%CI: 0.01-0.20), the diagnostic odds ratio was 225.54 (95%CI: 88.81-572.77), and the SROC AUC was 1.00 (95%CI: The crucial values were sensitivity = 0.95 (95%CI: 0.82-0.99) and specificity = 1.00 (95%CI: 0.92-1.00). The Deeks method revealed that the "diagnostic odds ratio" funnel plot of SLN biopsy for gastric cancer was significantly asymmetrical (P = 0.01), suggesting significant publication bias. Further meta-subgroup analysis revealed that, compared with fluorescence imaging, NIR imaging had greater sensitivity (0.98 vs 0.73). Compared with optical imaging immediately after ICG injection, optical imaging after 20 minutes obtained greater sensitivity (0.98 vs 0.70). Compared with that of patients with an average SLN detection number < 4, the sensitivity of patients with a SLN detection number ≥ 4 was greater (0.96 vs 0.68). Compared with hematoxylin-eosin (HE) staining, immunohistochemical (+ HE) staining showed greater sensitivity (0.99 vs 0.84). Compared with subserous injection of ICG, submucosal injection achieved greater sensitivity (0.98 vs 0.40). Compared with 5 g/L ICG, 0.5 and 0.05 g/L ICG had greater sensitivity (0.98 vs 0.83), and cT1 stage had greater sensitivity (0.96 vs 0.72) than cT2 to cT3 clinical stage. Compared with that of patients ≤ 26, the sensitivity of patients > 26 was greater (0.96 vs 0.65). Compared with the literature published before 2010, the sensitivity of the literature published after 2010 was greater (0.97 vs 0.81), and the differences were statistically significant (all P < 0.05). CONCLUSION:For the diagnosis of stomach cancer, optical imaging in conjunction with ICG-guided SLN biopsy is a therapeutically viable approach, especially for early gastric cancer. The concentration of ICG used in the SLN biopsy of gastric cancer may be too high. Moreover, NIR imaging is better than fluorescence imaging and may obtain higher sensitivity.

Objective To evaluate the prognostic value of radiation-induced lymphocytopenia in the survival of patients with primary hepatocellular carcinoma receiving radiation therapy.Methods The clinical data of 98 patients with unresectable primary hepatocellular carcinoma who received radiotherapy were retros-pectively analyzed.The minimum absolute lymphocyte count(min ALC)was graded in accordance with CTCAE V4.0.The optimal threshold of min ALC for prognosis was calculated by using the receiver operating characteristic curve,and the correlation of min ALC with clinical characteristics and dosimetry parameters was analyzed.The Kaplan-Meier method was employed to analyze the survival of patients with different levels of min ALC.Univariate and multivariate Cox proportional regression models were applied to analyze prognostic factors.Results The baseline and min ALC of 98 patients during radiotherapy were 1.52×109/L and 0.45×109/L,respecti vely(P＜().()0 1).The optimal cut-off value of min ALC for the prediction of the one-year survival rate was 0.38×109/L.GTV,the mean dose of the liver and spleen,the V5 and V10 of the liver and spleen,and the V15 of the spleen were correlated with min ALC,and the V5 of the liver was an independent predictor of min ALC.The overall survival of patients with high min ALC was higher than that of patients with low min ALC.Independent prognostic factors were min ALC≤0.38×109/L(HR=0.515,P=0.024),min ALC ≥ grade 3(HR=0.576,P=0.032),tumor thrombus in the portal/vena cava,Child-Pugh grade A,increase of ≥2 points in the Child-Pugh score after radiotherapy,and received more than two other therapies.Conclusion Min ALC≤0.38×109/L and min ALC≥grade 3 have independent prognostic value in patients with unresectable hepatocellular carcinoma receiving radiotherapy.

Supplementary Figure S1: Optimization of PEGylated cholesterol amount for cell labeling. Supplementary Figure S2: Morphology and characterization of isolated bone marrow dendritic cells. Supplementary Figure S3: Characterization of AS1411-ENVs prepared by direct incubation of ENVs with AS1411-PEG2000-Chol. Supplementary Figure S4: HE staining of major organs and tumor in three negative control groups.

Objective To evaluate the short-term efficacy and quality of life of primary hepatocellular carcinoma patients after radiotherapy and pregabalin treatment for neuropathic pain with bone metastasis. Methods 32 patients with primary hepatocellular carcinoma bone metastases were treated with radiotherapy combined with pregabalin treatment.Then, we prospectively studied the analgesic efficacy for neuropathic pain and quality of life, used the brief pain inventory and douleur neuropathique 4 questionnaire (DN4) to evaluate pain at baseline, one and two months after radiotherapy, assessed pain response using the international consensus endpoint definition of bone metastasis, and used European Organization for Research and Treatment of Cancer Research and Treatment Quality of Life Questionnaire (EORTC QLQ-C30) and bone metastasis module (QLQ-BM22) for quality of life assessment. Results One and two months after radiotherapy, the average DN4 score of neuropathic pain decreased, and the objective pain relief rates were 62.8% and 68.6%, respectively.The physical, emotional, social, and role functional scores of EORTC QLQ-C30 functional scale significantly increased in the first month after radiotherapy.Symptom scale of pain (P=0.015), insomnia (P=0.035), and loss of appetite (P=0.022) improved, and fatigue was aggravated (P < 0.05).Two months after radiotherapy, the mean overall health score and all functional scale scores significantly increased than those at baseline.The scores of all symptom scales decreased, except fatigue, constipation, and financial difficulties (P < 0.05).In addition, pain responders showed significant improvement in emotional function (P=0.025) and physical function (P=0.029) in the functional scale and in pain (P=0.014) and fatigue (P=0.035) in the symptom scale.The QLQ-BM22 score showed that the painful sites (P=0.021) and pain characteristics (P=0.04) of the responders significantly improved compared with those of nonresponders two months after radiotherapy. Conclusion Radiotherapy combined with pregabalin can relieve neuropathic pain caused by bone metastasis from primary hepatocellular carcinoma and greatly improve the quality of life, particularly in pain responders.

Objective:To explore the correlation of apparent diffusion coefficient (ADC) of magnetic resonance diffusion weighted imaging (DWI) examination before radiotherapy in patients with advanced cervical squamous cell carcinoma with clinicopathological characteristics and radiotherapy efficacy.Methods:The clinical data of 80 patients with advanced cervical cancer who were admitted to the Second Hospital of Nanjing from September 2019 to March 2022 were retrospectively analyzed. All patients underwent magnetic resonance imaging (MRI) DWI examination. The differences in ADC values among cervical squamous cell carcinoma patients with different clinicopathological characteristics were analyzed. The patients were divided into the effective group (complete remission+partial remission) and the ineffective group (stable disease+progressive disease) based on the radiotherapy effect, and the differences in ADC values between the two groups were compared. The logistic regression model was used to analyze the factors affecting the radiotherapy efficacy of patients with advanced cervical squamous cell carcinoma.Results:Among 80 patients with advanced cervical squamous cell carcinoma, 21 achieved complete remission, 31 achieved partial remission, 25 achieved stable disease, and 3 achieved progressive disease after radiotherapy; there were 52 cases in the effective group and 28 cases in the ineffective group. The ADC value of the effective group before radiotherapy was higher than that of the ineffective group [(0.99±0.14)×10 -3mm 2/s vs. (0.76±0.20)×10 -3mm 2/s], and the difference was statistically significant ( t = 6.01, P < 0.001); after radiotherapy, the ADC value of the effective group was also higher than that of the ineffective group [(1.43±0.25)×10 -3mm 2/s vs. (1.11±0.23)×10 -3mm 2/s), and the difference was statistically significant ( t = 5.61, P < 0.001); the ADC values of both the effective and ineffective groups increased after radiotherapy compared to before radiotherapy (both P < 0.05). The ADC values of patients with different International Federation of Obstetrics and Gynecology (FIGO) stage, degree of pathological differentiation, depth of lesion infiltration, Ki-67 expression, lymph node metastasis, and distant metastasis were statistically significant (all P < 0.05). The results of multivariate logistic regression analysis showed that ≥FIGO stage Ⅲ, low differentiation, lymph node metastasis, lymphatic vessel infiltration, distant metastasis, and low ADC value before radiotherapy were independent risk factors for efficacy of radiotherapy in patients with advanced cervical squamous cell carcinoma (all P < 0.05). Conclusions:The ADC value before radiotherapy is a factor that affects the radiotherapy effect of patients with advanced cervical squamous cell carcinoma. The lower the ADC value before radiotherapy is, the worse the radiotherapy effect of patients will be.

Objective To explore the clinical features and risk factors of preoperative anemia in patients with gastric cancer.Methods The clinical data of 218gastric cancer patients who received gastric cancer surgery in the Second Affiliated Hospital of Nanjing Medical University from January 2015 to December 2018 were retrospectively analyzed.Patients were divided into the anemia group(n = 84)and non-anemia group(n =134)based on hemoglobin levels.The gender,age,hypertension,diabetes,body mass index(BMI),carcinoembryonic antigen(CEA),cancer antigen 199(CA199),cancer antigen 724(CA724),tumor location,tumor size,clinical stage and lymph node metastasis were collected.The Logistic regression analysis was performed to analyze the risk factors of preoperative anemia in patients with gastric cancer.Results Among 218 patients with gastric cancer,the incidence of preoperative anemia was 38.53%,including 51 cases(60.7%)of mild anemia and 50 cases(59.5%)of normocytic anemia.There were no statistically signifi-cant differences in BMI,hypertension,diabetes,CEA,CA199,CA724,tumor location,and lymph node metastasis between the two groups(P>0.05),while there were statistically significant differences in the terms of gender,age,tumor size,and clinical stage be-tween the two groups(P<0.05).Multivariate Logistic regression analysis showed that gender(P = 0.002),age(P = 0.023),tumor size(P =0.001),and clinical stage(P =0.003)were independent risk factors for preoperative anemia in patients with gastric cancer.Conclusion Gender,age,tumor size and clinical stage are independent risk factors for preoperative anemia in patients with gastric canc-er,which is helpful for clinicians to take some intervention strategies for patients.

目的 探讨胃癌淋巴结转移风险预测模型的构建及评价.方法 回顾分析244例胃癌患者临床资料,根据是否发生淋巴结转移将患者分为转移组(n=171)和无转移组(n=73).采用Logistic回归分析胃癌淋巴结转移的危险因素,构建胃癌淋巴结转移的预测模型,采用受试者工作特征(ROC)曲线评价预测模型的价值.结果 Logistic回归分析发现,浸润深度、临床分期是胃癌淋巴结转移的独立危险因素(P<0.05).建立回归模型Logit(P)=-1.833+3.121×浸润深度(肌层)+2.799×浸润深度(穿透浆膜层)+5.468×临床分期,预测模型的ROC曲线下面积为0.958(95％CI:0.925～0.980,P<0.001),约登指数:0.815,cut-off值为0.715,敏感度和特异性分别为84.2％和97.3％.结论 浸润深度、临床分期是胃癌患者淋巴结转移的独立危险因素,该模型有助于临床预测胃癌患者是否发生淋巴结转移.

目的:探讨影响非小细胞肺癌骨转移的危险因素.方法:选取2015年至2019年在南京医科大学第二附属医院住院治疗的非小细胞肺癌患者207例[骨转移组(n=113)、无骨转移组(n=94)].检测血清CA125、CEA、NSE、CYFRA21-1、LDH和ALP的水平,记录年龄、性别、吸烟史、高血压、糖尿病、临床分期、肿瘤位置、组织学类型和淋巴结转移等因素.采用单因素和Logistic多因素回归分析筛选影响非小细胞肺癌患者骨转移的危险因素.结果:单因素分析结果发现两组临床分期、淋巴结转移、CA125、CEA、NSE、CYFRA21-1、LDH比较具有统计学差异(P<0.05);NSE、CYFRA21-1在腺癌和鳞状细胞癌血清中水平无统计学差异(P>0.05);Logistic多因素回归分析结果显示,临床分期、淋巴结转移和CA125是影响非小细胞肺癌患者骨转移的主要因素(P<0.01).结论:临床分期、淋巴结转移和CA125是影响非小细胞肺癌患者骨转移的独立危险因素,为临床早期治疗提供重要的指导意义.

目的:评估姑息放疗(三维适形/适形调强放疗)治疗晚期肝脏恶性肿瘤(包括原发性肝癌、转移性肝癌)的疗效、不良反应以及生活质量变化.方法:选择2015年6月至2018年12月采用三维适形/适形调强放疗治疗的南京中医药大学附属南京医院126例晚期肝脏恶性肿瘤患者,前瞻性研究放疗近期疗效及不良反应,并于放疗前、放疗结束、放疗后3个月、放疗后6个月,运用EORTC QLQ-C30量表动态评估患者生活质量,并计算半年、1年生存率.结果:放疗后3个月,完全缓解4例(3.2％),部分缓解57例(45.2％),疾病稳定29例(23.0％),疾病进展36例(28.6％),客观缓解率为48.4％,疾病控制率为71.4％;6个月、1年生存率分别为66.7％、17.5％,中位生存时间为6.8个月,不良反应以疲乏、食欲下降、肝功能异常、腹水为主;与放疗前相比,在放疗结束时,整体健康状况评分有所降低(P =0.007),放疗后3个月、6个月,QLQ-C30整体健康状态评分,相对于基线分别改善32.5％、37.5％,恶化22.5％、27.5％,保持稳定45％、35％.多因素Logistic回归分析显示近期疗效、ECOG评分、肝内病灶数目以及放疗生物等效剂量对肝脏恶性肿瘤放疗后生活质量影响具有统计学意义.结论:姑息放疗有利于降低晚期肝脏恶性肿瘤负荷,放疗不良反应可接受,与放疗前相比,放疗后3个月患者生活质量显著提高.

目的:探究在胸腔内注射不同剂量的顺铂用于治疗肺癌的恶性胸水的临床疗效.方法:选取我院2019年4月至2020年4月的94例肺癌恶性胸水的患者,随机的方式分组,其中47例为对照组,给予低剂量的胸腔内注射顺铂治疗,另47例为研究组,给予高剂量的顺铂治疗,对比治疗效果.结果:与对照组比,研究组的治疗总有效率更高,不良反应的发生率更低,差异有统计学意义(P<0.05);与对照组比较,研究组的生存质量评分、胸水完全消失时间、药物治疗总时间均更好,差异有统计学意义(P<0.05).结论:肺癌恶性胸水的治疗中采用高剂量的胸腔内顺铂注射治疗,疗效确切,安全可靠,值得在临床上进行推广应用.

目的:探究单次大分割放疗在体外对树突状细胞(DC)激活T细胞的影响.方法:收集肝癌患者外周血单核细胞(PBMC),在细胞因子刺激下体外培养获得DC;分别在15Gy/1f和5Gy/3f条件下对HepG2细胞进行照射,通过离心获得肿瘤相关抗原;采用流式细胞术及ELISA法,检测负载了这两种抗原的DC细胞表型HLA-DR、CD40、CD80、CD83、CD86以及细胞因子IL-12 p70、IL-1β、IL-6、TNF-α分泌变化;通过胞内染色法检测负载这两种抗原的DC对特异性T细胞的激活情况;利用MTT法检测上述被活化的T细胞对HepG2细胞的杀伤活性.结果:15 Gy/1 f组负载肿瘤相关抗原的DC表达细胞表型HLA-DR、CD40、CD80、CD83及CD86均高于5 Gy/3 f组(P<0.05);15 Gy/1 f组负载肿瘤相关抗原的DC分泌细胞因子IL-12 p70和IL-6高于5Gy/3f组(P<0.05);胞内染色结果显示,15Gy/1f组负载肿瘤相关抗原的DC相比5Gy/3f组更能刺激CD4+T细胞[(0.392±0.187)％vs(0.315±0.118)％]和CD8+T细胞[(0.362±0.159)％vs(0.119±0.090)％]分泌IFN-γ(P<0.05);MTT实验结果显示,15 Gy/1 f组激活的T细胞对HepG2细胞的杀伤活性更强(P<0.05).结论:单次大分割放疗更能刺激DC对特异性T细胞的活化作用,这为今后肝癌的治疗提供了理论依据.

目的 探讨肿瘤标志物联合ALP在非小细胞肺癌骨转移中的诊断价值.方法 选取非小细胞肺癌患者159例[骨转移组(n= 82)、无骨转移组(n =77)]和83例肺炎患者(肺炎组).检测血清CEA、CYFRA21-1和ALP的水平,记录年龄、性别等信息.采用Pearson相关分析评价各指标间的相关性.非小细胞肺癌患者骨转移的影响因素采用Logistic回归分析.采用受试者工作特征(ROC)曲线评价血清CEA、CYFRA21-1和ALP诊断非小细胞肺癌患者骨转移的价值.结果 本研究共纳入159例非小细胞肺癌患者,未发生骨转移77例,发生骨转移82例,骨转移发生率为51.6％,腺癌与鳞状细胞癌骨转移发生率比较,差异无统计学意义(P＞0.05).骨转移组CEA、CYFRA21-1和ALP的水平均高于非骨转移组和肺炎组(P＜0.05),非骨转移组血清CEA、CYFRA21-1和ALP的水平均高于肺炎组,差异有统计学意义(P＜0.05).ALP与CYFRA21-1、CEA呈正相关(r= 0.278,P＜0.01;r= 0.343,P＜0.01).Logistic 回归分析结果显示,CYFRA21-1、CEA 和 ALP 是 NSCLC 患者发生骨转移的危险因素.ROC曲线结果显示,CEA、CYFRA21-1和ALP联合诊断非小细胞肺癌患者骨转移的曲线下面积(AUC)为0.829,95％CI:0.761～0.884,约登指数为0.512,敏感度和特异性分别为72.0％和79.2％.结论 肿瘤标志物联合ALP在非小细胞肺癌患者骨转移的诊断中具有重要价值.

Purpose: Stereotactic body radiation therapy (SBRT) is emerging as a new noninvasive treatment in patients with primary liver carcinoma or liver-confined metastatic cancer. However, the radiobiological targets remain a subject of debate. Here, we investigated the potential biological effects of the radiation on the human hepatocellular carcinoma HepG2 cells. Materials and methods: Firstly, HepG2 cells were divided into three groups: control group, 3.5 Gy*8f group (L group), and 15 Gy*1f group (H group). After treatment, cell proliferation was examined using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and plate colony formation assays. Cell cycle and apoptosis were assessed using propidium iodide and Hoechst 33258 staining, respectively. Furthermore, the mechanisms underlying irradiation-induced cell cycle arrest and cell apoptosis were investigated by Western blot assay. Results: Irradiation could effectively inhibit the proliferation and colony formation of HepG2 cells, and the single high dose irradiation showed stronger inhibitory effects. Irradiation-induced cell cycle arrest at G2/M phase in HepG2 cell, during which the expression levels of cyclin B1, CDK1, and p-CDK1 proteins were downregulated, whereas expression of p21 was upregulated in the irradiated HepG2 cells. After irradiation, typical morphological changes of apoptosis in HepG2 cells were observed; the number of cell apoptosis and the expression of apoptosis associated proteins were significantly increased in HepG2 cells by high dose irradiation compared with low dose irradiation. Additionally, compared with low dose irradiation, high dose irradiation significantly downregulated the phosphorylated proteins in the Ras/Raf/MEK/ERK signaling pathway. Conclusions: Our results suggest that irradiation applied in SBRT, particularly single high dose irradiation, mediates its anti-tumor effects by inducing cell cycle arrest and apoptosis via modulation of the Ras/Raf/MEK/ERK signaling pathway.

Extracellular vesicles (EVs) are lipid-bilayer-enclosed vesicles of submicron size that are secreted by various cells. As mediators of intercellular communication, EVs can alter the physiological state of recipient cells by delivering encapsulated proteins and nucleic acids. Incontestably, growing evidence has shown important biological roles and the clinical relevance of EVs. The use of stem cell-derived EVs as a cell-free therapeutic modality for skin treatment has emerged as a promising application in dermatology. However, the moderate isolation efficiency of prevalent ultracentrifugation and low secretion rate make the massive low-cost production of EVs difficult. Here, we report development of engineered EVs (eEV) derived from human umbilical cord mesenchymal stem cells (hucMSCs) for skin treatment. Ultrasonication was used to shear intact hucMSCs for only 1 min, followed by regular centrifugation and filtration for producing nanoscale eEVs. This approach has ∼20-fold higher yield and ∼100-fold faster production than that of naturally secreted EVs (nsEV), while the production cost decreased to less than 10%. The eEVs have similar morphology, size distribution, and typical protein markers compared to nsEVs. Moreover, in vitro, both nsEVs and eEVs promote the proliferation and migration of dermal fibroblasts and increase in the expression of collagen, elastin, and fibronectin, whereas the matrix metalloproteinases-1 (MMP-1) and MMP-3 production can be significantly reduced. The wound-healing study in mice showed that both nsEVs and eEVs promote wound recovery in comparison with the controls. In sum, our results indicate that hucMSC-derived eEVs prepared by ultrasonication potentially can be used to increase skin extracellular matrix and enhance skin rejuvenation.

Hypoxia, a state of low oxygen tension in solid tumors, is not only closely correlated with resistance to both radiotherapy and chemotherapy, but also associated with poor prognosis of tumors and regional lymph node status. Herein, based on the analysis of cell samples from tumor patients, low-density lipoprotein receptor (LDLR) was found to be overexpressed on the surface of hypoxic tumor cell membranes, and confirmed to be an effective hypoxia marker through specific binding with anti-LDLR antibody in solid tumors. In addition, using the special therapeutic microenvironment of hypoxia, tirapazamine (TPZ, which can be used as both a hypoxia-activated chemotherapy prodrug and radiotherapy sensitizer) was integrated with PEGylated photosensitizer chlorin e6 (Ce6-PEG) by self-assembly, and anti-LDLR was then modified on the surface to form tumor hypoxia-targeting multifunctional nanoparticles (CPTA). CPTA possesses a multimodal antitumor effect via a simultaneous photothermal therapy (PTT)/photodynamic therapy (PDT) effect generated by Ce6, and chemotherapy/radiotherapy actions sensitized by TPZ. It is noteworthy that tumor oxygen was consumed in the process of PDT and the hypoxia was subsequently exacerbated, which can greatly increase the TPZ-sensitized chemotherapy and lead to a synergistic antitumor effect. Both in vitro and in vivo experiments demonstrated that CPTA possesses an excellent therapeutic effect through PTT, PDT, and TPZ sensitized radiotherapy and chemotherapy. This hypoxic tumor targeting synergetic therapeutic strategy has great potential for future clinical transformation.

Objective: The aims of this study were to compare the clinical outcomes between ultrasound-guided percutaneous microwave ablation (US-PMWA) and surgical resection (SR) in patients with recurrent intrahepatic cholangiocarcinoma (ICC) and to identify the prognostic factors associated with the two treatment methods. Methods: This retrospective study was institutional review board approved. A total of 121 patients (102 men and 19 women) with 136 ICCs after hepatectomy from April 2011 to January 2017 were reviewed. Fifty-six patients underwent US-PMWA and 65 patients underwent SR. Survival, recurrence and liver function were compared between the two groups. Effect of changes in key parameters [i.e., overall survival (OS) and recurrence-free survival (RFS)] was statistically analyzed with the log-rank test. Univariate and multivariate analysis were performed on clinicopathological variables to identify factors affecting long-term outcome. Results: The OS and RFS after MWA were comparable to that of SR (p = .405, and p = .589, respectively). Estimated 5-year OS rates were 23.7% after MWA and 21.8% after SR; for RFS, estimated 3-year RFS rates were 33.1% after MWA and 30.6% after SR. Major complication rates in SR group were higher than that in MWA (p < .001) (SR, 13.8% vs. MWA, 5.3%). Multivariate analysis showed tumor number (p = .012), ALBI grade (p = .007), and metastasis (p = .016), may become OS rate predictors. Conclusions: US-PMWA had comparable oncologic outcomes with SR and could be a safe and effective treatment for recurrent ICC after hepatectomy.

目的:探讨三维适形/适形调强放疗对不可手术的原发性肝癌的治疗效果、不良反应以及影响疗效和预后的相关因素.方法:回顾性分析2013年7月至2016年5月我院45例行放疗的不可手术的原发性肝癌患者(其中12例合并静脉癌栓,33例曾行肝动脉化疗栓塞术)的临床资料.患者放疗结束后1～3个月观察近期疗效,随访1、2、3年,对影响疗效和预后的相关因素进行分析.结果:放疗后肝癌原发灶完全缓解(CR)4例(8.9％),部分缓解(PR)16例(35.5％),疾病稳定(SD)17例(37.8％),疾病进展(PD)8例(17.8％);静脉癌栓者CR 1例(8.33％),PR 3例(25.0％),SD 6例(50.0％)和PD 2例(16.67％).肝癌原发灶及静脉癌栓放疗客观缓解率分别为44.4％和33.3％.放疗期间出现Ⅲ度以上的不良反应有恶心呕吐、骨髓抑制、食欲下降和疲乏等.影响放疗效果的因素有肿瘤直径、ΔCT值、肿瘤处方剂量以及是否曾接受介入治疗.45例患者1、2、3年生存率分别为59.8％、40.9％和2.22％.多因素分析结果显示,肿瘤体积、肿瘤分期、静脉癌栓是影响预后的重要因素.结论:三维适形/适形调强放疗对不可手术原发性肝癌疗效较好,不良反应患者可耐受.肿瘤的直径、血供、肿瘤处方剂量是影响肝癌放疗效果的关键因素,肿瘤体积、分期及是否合并静脉癌栓是影响预后的独立因素.