BACKGROUND:The histological differentiation grade of gastric cancer critically influences treatment and prognosis. While CT radiomics shows promise for noninvasive prediction, its relationship with gene expression remains unclear. PURPOSE:This study aimed to develop a clinical-radiomics model for predicting tumor differentiation in gastric cancer patients and to explore the underlying mechanisms. METHODS:We retrospectively analyzed clinical data and CT images from 162 gastric cancer patients, who were randomly assigned to training and validation cohorts. The least absolute shrinkage and selection operator (LASSO) method was used to select features and construct the Rad-score. Subsequently, clinical-radiomics models were built and evaluated for their predictive efficacy and clinical incremental value. Furthermore, hub genes were screened, and their associated pathways were investigated using machine learning, bioinformatics analysis, and experimental validation. RESULTS:A clinical-radiomics model based on N stage, M stage and Rad-score was developed. The receiver operating characteristic (ROC) curves indicated that the model had preliminary evidence of predictive potential within this single-center cohort (training AUC = 0.872, validation AUC = 0.935). The calibration curves indicated a reasonable concordance between the observed values and the predicted outcomes in this retrospective sample. The decision curve analysis demonstrated a net benefit that requires further confirmation in external cohorts. The clinical impact curve (CIC) demonstrated the model's potential clinical applicability, which warrants validation in prospective settings. Sequencing data further revealed that the key gene IGHG1 was significantly associated with the Rad-score, with potential mechanisms involving the TGF-beta signaling pathway. CONCLUSIONS:The clinical-radiomics model, incorporating N stage, M stage, and Rad-score, serves as a preliminary research tool for assessing tumor differentiation in gastric cancer patients within a single-center setting. External validation is required before any consideration of clinical generalization. Radiomics enables noninvasive evaluation of differentiation status while generating hypotheses regarding its underlying mechanisms.

Objectives To develop a CT-based radiomics model for predicting tumor differentiation in patients with gastric cancer. Exploring Rad-score correlation with gene expression and related mechanisms. Materials and Methods Clinical data and imaging of 162 gastric cancer patients were retrospectively analyzed. Patients were randomly allocated to training and validation cohorts. The least absolute shrinkage and selection operator (LASSO) methods were utilized to identify characteristics and develop the Rad-score. Clinical-radiomics models were developed and evaluated for predictive efficacy and clinical incremental value. Screening hub genes and exploring the pathways of hub genes through machine learning, bioinformatics analysis and experimental validation. Results Clinical-radiomics models based on N stage, M stage and Rad-score were developed. The receiver operating characteristic (ROC) curves indicated that the model had good predictive accuracy in the training (AUC = 0.872) and validation groups (AUC = 0.935). The calibration curves indicated a strong correlation between the observed values and the predicted outcomes. The decision curve analysis demonstrated a substantial net benefit associated with the clinical-radiomics model. The clinical impact curve (CIC) illustrated the effective clinical applicability of this model. Analysis of the sequencing data revealed that the key gene IGHG1 was significantly associated with Rad-score. The possible mechanisms are related to the TGF-β signaling, epithelial-mesenchymal transition and KRAS signaling pathway. Conclusions The predictive model based on N stage, M stage and Rad-score can effectively predict the differentiation in gastric cancer patients. Radiomics enables noninvasive prediction of tumor differentiation status while elucidating the expression levels of the IGHG1 and the underlying pathway.

Background: Stomach adenocarcinoma (STAD) is the fifth most common cancer and the third leading cause of cancer-related deaths worldwide. Cancer-testis antigens (CTAs) participate in the pathogenesis and development of multiple cancers and are aberrantly overexpressed in various types of cancer. This study aimed to develop a CTA-related gene signature (CTARSig) to predict prognosis in STAD patients and explore its underlying mechanisms. Methods: We performed differential and prognostic analyses of CTA-related genes and constructed a CTA-related signature (CTARSig) along with a novel nomogram to predict the prognosis of patients with STAD based on the Cox and The Least Absolute Shrinkage and Selection Operator. CTARSig was further validated in an external cohort (GSE84437). Additionally, univariate and multivariate Cox regression, as well as receiver operating characteristic (ROC) analyses, were performed to assess the CTARSig systematically. Single-sample gene set enrichment analysis and ESTIMATE were used to characterise the Tumor Immune Microenvironment (TIME) in patients with STAD. Furthermore, Gene Set Variation Analysis, Kyoto Encyclopedia of Genes and Genomes, and Gene Ontology analyses revealed the biological functions and signalling pathways associated with CTARSig. Finally, the human gastric cancer cell lines, HCG-27 and AGS, were used for in vitro and in vivo experiments, respectively, to further validate the role of ELOVL4. Results: Eleven CTA-related genes were identified to construct the CTARSig. Kaplan-Meier curves, independent prognostic analysis, and ROC curves revealed that CTARSig could better predict survival in patients with STAD. Moreover, in our study, we demonstrated that ELOVL4 is upregulated in gastric cancer tissues and that its high expression is associated with poor survival. Additionally, in vitro and in vivo experiments demonstrated that ELOVL4 promotes the metastatic and invasive potential of STAD cells, suggesting it may be a potential therapeutic target for STAD. Conclusion: In this study, a novel signature associated with CTAs was constructed for STAD, which may be a good predictor of patient prognosis. Thus, ELOVL4 may be a potential therapeutic target for gastric cancer. This study provides new insights into the potential roles of CTAs in gastric cancer.

Supplemental Figure 1. Th17 cells are not induced in the lung after the tumor cell challenge. Supplemental Figure 2. No significant difference in the production of IFN-gamma by NK cells in the Abt mice compared with the control.

Cancer-associated fibroblasts (CAFs) play a role in ovarian cancer (OV) evolution, immunosuppression and promotion of drug resistance. Exploring the value of CAFs-related biomarker in OV is of great importance. In the present work, we developed a CAFs-related index (CAFRI) based on an integrated analysis of single-cell and bulk RNA-sequencing and highlighted the value of CAFRI in predicting clinical outcomes in individuals with OV, tumour immune microenvironment (TIME) and response to immune checkpoint inhibitors (ICIs). The GSE151214 cohort was used for cell subpopulation localization and analysis, the TCGA-OV patients as a training set. Moreover, the ICGC-OV, GSE26193, GSE26712 and GSE19829 cohorts were used for the validation of CAFRI. The TIMER 2.0, CIBERSORT and ssGSEA algorithms were used for analysis of TIME characteristics based on the CAFRI. The GSVA, GSEA, GO, KEGG and tumour mutation burden (TMB) analyses were used for mechanistic exploration. Additionally, the IMvigor210 cohort was conducted to validate the predictive value of CAFRI on the efficacy of ICIs. Finally, CAFRI-based antitumour drug sensitivity was analysed. The findings demonstrate that the CAFRI can served as an excellent predictor of prognosis for individuals with OV, as well as identifying patients with different TIME characteristics, differentiating between immune 'hot' and 'cold' tumour populations, and providing new insights into the selection of ICIs and personalised treatment regimens. CAFRI provides new perspectives for the development of novel prognostic and immunotherapy efficacy predictive biomarkers for OV.

Supplemental materials and methods

OBJECTIVES:HORMAD1 is a cancer/testis antigen (CTAs) that regulates DNA homologous recombination, mismatch repair, and other tumor characteristics. However, its role and regulatory mechanisms in gastric cancer remain unclear.METHODS:We performed transcriptomic profiling on seven gastric cancers and paired tissues; HORMAD1 was significantly upregulated in gastric cancer samples and was related to poor prognosis survival. Furthermore, cancer pathway microarray, bioinformatic analysis, western blot, and immunochemistry assay demonstrated that HORMAD1 affected the NF-κB signaling pathway.RESULTS:In vitro and vivo studies confirmed that HORMAD1 knockdown inhibited cell growth and invasion, whereas overexpression reversed these effects. Mechanistically, HORMAD1 regulates the epithelial-mesenchymal transition process (EMT) via the NF-κB pathway by increasing the phosphorylation levels of NF-κB (p-65) and Iκκ-β. Downstream target genes of the NF-κB signaling pathway, such as c-Myc, CyclinD1, may be involved in HORMAD1-induced tumorigenesis in gastric cancer (GC).CONCLUSIONS:HORMAD1 plays an important role in gastric cancer progression and could be a promising prognostic biomarker and therapeutic target.

Objective:To evaluate the clinical efficacy and safety of fecal microbiota transplantation(FMT)for the treatment of chronic functional constipation in the elderly.Methods:A total of 33 elderly patients with chronic functional constipation were included and given three sessions of FMT.Changes in fecal characteristics, constipation, mood and quality of life in these patients were evaluated using the Bristol stool form scale(BSFS), the constipation assessment scale(CAS), patient assessment of constipation symptoms(PAC-SYM), the Zung self-rating anxiety scale(SAS), the Zung self-rating depression scale(SDS), and the patient assessment of constipation quality of life(PAC-QOL)before and 12 weeks after treatment.The clinical efficacy was based on comparison between pre-and post-treatment results for each patient.Results:Clear improvement was achieved in 33 patients 12 weeks after treatment, compared with before transplantation.Post-treatment scores of the constipation assessment scale and symptom self-assessment questionnaire for patients with constipation were(8.9±1.2)scores and(26.5±2.4)scores, respectively, significantly lower than pre-transplantation scores of(12.2±1.1)scores and(32.4±2.4)scores( t=15.034, 13.904, both P<0.001). Similarly, post-treatment scores were also lower than pre-transplantation levels for the self-rating anxiety scale[(50.4±8.4)scores vs.(57.5±9.0)scores, t=10.333, P<0.001], the self-rating depression scale[(50.6±8.4)% vs.(55.0±10.5)%, t=5.301, P<0.001], and self-assessment questionnaire for quality of life[(88.2±7.3)scores vs.(103.7±7.3)scores, t=23.300, P<0.001]. Conclusions:FMT can improve fecal characteristics and constipation symptoms, relieve anxiety and depression, improve the quality of life, and provide a new option for the treatment for elderly patients with chronic functional constipation.

目的 探讨N6-甲基腺嘌呤(m6 A)去甲基酶肥胖相关蛋白(FTO)在乳腺癌细胞对曲妥珠单抗耐药中的作用及新型抗人表皮生长因子受体2(HER2)人源化A21抗体(HuA21)对耐药性乳腺癌细胞增殖的影响.方法 制备曲妥珠单抗耐药细胞株(BT474/TR),光学显微镜观察BT474/TR与BT474细胞形态,四甲基偶氮唑盐(MTT)法检测细胞增殖,免疫荧光法检测增殖相关抗原(Ki67)表达,流式细胞术检测细胞周期,qPCR和Western blot法检测FTO与甲基转移酶样蛋白3抗体(METTL3)表达.结果 BT474/TR出现细胞核固缩、伪足消失,细胞增殖速度下降.曲妥珠单抗处理BT474不同时间至形成耐药细胞,Ki67表达逐渐降低(P<0.05),S期细胞比例逐渐减小(P<0.05),FTO表达逐渐增加(P<0.05),但是METTL3表达没有变化.FTO抑制剂甲氯芬那酸乙酯(MA2)促进BT474/TR细胞增殖,且提高其对曲妥珠单抗的敏感性(P<0.05).HuA21抗体对BT474/TR细胞增殖具有抑制作用(P<0.05),且联合MA2和曲妥珠单抗对BT474/TR细胞抑制效果更好(P<0.05).结论 HER2阳性乳腺癌细胞FTO基因表达与其对曲妥珠单抗耐药之间存在相关性,抑制FTO活性可以增强乳腺癌耐药细胞对曲妥珠单抗治疗的敏感性;HuA21抗体对曲妥珠单抗耐药性乳腺癌细胞有抑制作用.

目的 探讨中老年人群营养不良与骨质疏松症的相关性.方法 选取中国科学技术大学附属第一医院(安徽省立医院)老年医学科收治的78例中老年骨质疏松患者,运用双能X线法测定所选取患者的骨密度(BMD),根据体质指数将78例患者分为营养不良组和营养正常组.对于两组患者的性别、不同年龄骨密度、骨盐含量以及骨密度的T值等相关指标进行比较.结果 在中老年患者中,营养不良组左上臂、右侧股骨、正位脊柱的骨密度均显著低于营养正常组(P均<0.05);两组上述各部位T值比较,差异也均有统计学意义(P均<0.01).各部位骨盐含量,营养不良组较营养正常组明显更低(P<0.01).在中老年人群中,营养不良组左上臂、正位脊柱部位的骨密度、骨盐含量都显著低于营养正常组(P均<0.05);而右侧股骨的骨密度两组间比较,差异不显著(P均>0.05).但在老年男性中,右侧股骨营养不良较营养正常组T值显著降低(P<0.05).在80岁以下中老年人群中,营养不良组的左上臂、右侧股骨、正位脊柱骨密度、骨盐含量、T值均显著低于营养状态正常组(P均<0.05).在80岁及以上老年患者中,营养不良组的左上臂及正位脊柱中的骨密度与营养正常组对比显著降低,差异有统计学意义(P均<0.05),T值变化不明显,差异不显著(P均>0.05);正位脊柱中骨盐含量营养不良较正常组显著降低,差异有统计学意义(P均<0.05).结论 营养不良是高龄患者骨质疏松的重要危险因素之一.

Background Drug resistance prevents the effective treatment of cancers. DNA methylation has been found to participate in the development of cancer drug resistance. Methods We performed the wound-healing and invasion assays to test the effect of the paraoxonase gene PON3 on esophageal cancer (EC) cells. In addition, in vivo EC-derived tumor xenografts in nude mice were generated to test the effect of PON3 on the chemoresistance of EC cells. Results We found that PON3 is hypermethylated in drug-resistant EC cell line K150, which in-return down-regulates its expression. The following experiments by the forced changes of PON3 level in vitro and in vivo demonstrated that the PON3 expression negatively correlates with drug resistance in EC cells. Further wound-healing and invasion assays showed that PON3 suppresses the migration and invasion of EC cells. Conclusion Our data established that PON3 is associated with the EC drug resistance, which may serve as a biomarker for the potential therapeutic treatment of EC.

Background: MicroRNAs (miRNAs) was reported to be involved in cancer radio-resistance, which remains a major obstacle for effective cancer therapy.Methods: The differently expressed miRNAs were detected by RNA-seq experiment in nasopharyngeal cancer (NPC) cells. MiR-20a-5p was selected as our target, which was subject to finding its target gene Rab27B via bioinformatics analysis. The qRT-PCR, western blot and the luciferase reporter assays were performed to confirm Rab27B as the target of miR-20a-5p. In addition, the roles of miR-20a-5p in NPC radio-resistance were detected by transfection of either miR-20a-5p-mimic or miR-20a-5p-antagomiR. The involvement of Rab27B with NPC radio-resistance was also detected by the experiments with siRNA-mediated repression of Rab27B or over-expression of GFP-Rab27B. Wound healing and invasion assays were performed to detect the roles of both miR-20a-5p and Rab27B.Results: MiR-20a-5p promotes NPC radio-resistance. We identified that its target gene Rab27B negatively correlates with miR-20a-5p-mediated NPC radio-resistance by systematic studies of a radio-sensitive (CNE-2) and resistant (CNE-1) NPC cell lines. Repression of Rab27B by siRNA suppresses cell apoptosis and passivates CNE-2 cells, whereas overexpression of Rab27B triggered cell apoptosis and sensitizes CNE-1 cells.Conclusions: MiR-20a-5p and its target gene Rab27B might be involved in the NPC radio-resistance. Thus the key players and regulators involved in this pathway might be the potential targets for developing effective therapeutic strategies against NPC.

Abstract Commensal bacteria are crucial to maintain immune homeostasis in mucosal tissues and disturbances in their ecology can affect disease susceptibility. Here, we report evidence that commensal bacteria shape the efficiency of immune surveillance in mucosal tissues. Antibiotic-treated (Abt) mice were more susceptible to development of engrafted B16/F10 melanoma and Lewis lung carcinoma, exhibiting a shortened mean survival time with more numerous and larger tumor foci in the lungs. The defective antitumor response of Abt mice was independent of dehydration caused by antibiotics. Host defenses relied upon intact commensal bacteria with no class specificity. Mechanistic investigations revealed a defective induction of the γδT17 cell response in lungs of Abt mice; here, more aggressive tumor development was observed, possibly related to a reduction in IL6 and IL23 expression there. Adding normal γδT cells or supplementing IL17 restored the impaired immune surveillance phenotype in Abt mice. Overall, our results demonstrated the importance of commensal bacteria in supporting the host immune response against cancer, defined an important role for γδT17 responses in the mechanism, and suggested deleterious effects of antibiotic treatment on cancer susceptibility and progression. Cancer Res; 74(15); 4030–41. ©2014 AACR.

Objective To establish the methods of cytokine-induced killer (CIK)cells and evaluate the characteristics,anti-tumor activity in vitro and in vivo and the safety.Methods Anti-CD3 mAb,recombinant human interleukin 2 (rhIL-2 )and recombinant human interferon-γ(rhIFN-γ)were used to induce the peripheral blood mononuclear cells (PBMCs)proliferation abundantly in vitro.Flow cytometry assay (FACS),in vitro cytotoxicity as-say and tumor mouse model xenografted with Ho-8910 (human ovarian cancer cell line)were used to detect the char-acteristics and the anti-tumor activityof CIK cells.Additionally,the contamination of exogenous agents and the acute toxicity effect of CIK cells were also determined.Results After 21 days in vitro culture,the total number of CIK cells were more than 1010,among which there were mainly CD3 +CD8 +T cells and CD3 +CD56 +NKT cells.CIK cells dis-played high cytotoxicity against Ho8910,A549,K562 and HepG2 cells in vitro,and they also showed therapeutic effects in tumor xenografted mice.Additionally,no acute toxicity was observed in CIK cells-treated mice,and there was also no contamination of bacteria,fungus,mycoplasma and virus in CIK cells.Conclusion CIK cells have high anti-tumor activity without any toxic side effects.

Whether cisplatin plus vinorelbine (VC) or cisplatin plus docetaxel (DC) are equally effective in the treatment of advanced non-small-cell lung cancer (NSCLC) remains controversial. The aim of this study was to compare the VC and DC regimens in the first-line treatment of advanced NSCLC. A search was conducted through PubMed, the Cochrane Central Register of Controlled Trials (CENTRAL), EMBASE and the Chinese Biomedical Literature database (CBM). The language of the publication was not considered to be a limitation. The recruited trials were evaluated for eligibility and quality and the data were extracted and analyzed. The endpoints were overall response, survival rate and toxicity. We analyzed 9 randomized controlled trials (RCTs), including a total of 1,886 patients. Patients receiving DC therapy exhibited a significantly higher response rate [relative risk (RR)=0.83, 95% CI: 0.73-0.95 and P=0.007] and 2-year survival rate (RR=0.65, 95% CI: 0.50-0.84 and P=0.001). However, the 1-year survival rate for the two cisplatin-based regimens were comparable (RR=0.90, 95% CI: 0.81-1.01 and P=0.07). Patients receiving the VC regimen more frequently developed grade 3/4 leucopenia, anemia and vomiting, whereas those receiving DC chemotherapy were more prone to grade 3/4 diarrhea. The incidence of grade 3/4 neutropenia, thrombocytopenia and nausea were similar between the two arms. In conclusion, our study indicated that DC is superior to the VC regimen in terms of tumor response rate, 2-year survival rate and safety for the first-line treatment of advanced NSCLC.

The aim of this study was to evaluate tumor markers of molecular abnormalities that display tissue specificity, as to detect circulating tumor cells (CTCs) in breast cancer patients. Quantitative real-time RT-PCR was used to determine h-MAM, BCSG1, CK19, and c-erbB2 mRNA levels in peripheral blood (PB) of breast cancer patients. Results were compared with other epithelial cancers (lung or esophagus cancer), benign breast disease, and healthy individuals. We found that h-MAM mRNA was only detectable in the PB of patients with breast cancer (49 of 65, 75.4%), but not in patients with other epithelial cancers, benign breast disease, or healthy individuals. No significant differences in the expression level and positive detection rate of BCSG1, CK19, and c-erbB2 mRNA were observed between breast cancer and other epithelial cancers. Furthermore, the expression level and positive detection rate of h-MAM mRNA in PB were significantly correlated to the breast cancer pathologic stage (p=0.012 and p=0.015, respectively). Chemotherapy, radiotherapy, or total tumor resection (after 7 days of treatment) resulted in a significant decrease in the expression level of h-MAM mRNA in PB compared to the levels prior to the treatment (p<0.001). Importantly, an increase in h-MAM mRNA expression was detected in patients immediately after surgery, as well as 3 days post-surgery. These results indicate that the quantitative analysis of h-MAM mRNA is a useful tool for detecting CTCs in breast cancer patients, and can have a potential diagnostic utility in early micrometastasis, clinical evaluation of cancer treatment efficacy, and post-treatment monitoring of breast cancer patients.

目的 探讨5-氮杂-2'-脱氧胞苷(5-Aza)联合临床化疗药物紫杉醇与氟尿嘧啶对于胃癌裸鼠移植瘤生长的抑制作用和抑癌基因RUNX3 mRNA以及肿瘤转移相关蛋白E-cadherin与Vimentin表达的影响.方法 建立胃癌细胞株MKN-45的裸鼠移植瘤模型,分为0.9%氯化钠注射溶液对照、5-Aza(2 mg/kg,每天1次)、化疗药物(1 mg/kg紫杉醇与5 mg/kg氟尿嘧啶,分别于治疗周期第1、8天用)以及5-Aza与紫杉醇及氟尿嘧啶联合用药,采用腹腔注射方法给药3周.观察各组裸鼠移植瘤的生长速度变化,并用RT-PCR方法检测肿瘤RUNX3基因的mRNA表达水平,免疫组织化学方法检测E-cadherin与Vimentin表达情况.结果 胃癌细胞MKN-45裸鼠移植瘤模型经药物治疗3周后,与空白对照组相比,5-Aza组或T+F组的移植瘤生长速度明显减慢(P<0.05),而且5-Aza与T+F联合治疗组比单独处理组的肿瘤生长速度下降更为显著(P<0.05).通过作用机制分析发现,与对照组相比,化疗药物T+F组RUNX3 mRNA及E-cadherin与Vimentin蛋白表达水平无明显变化;5-Aza处理组RUNX3 mRNA与E-cadherin蛋白表达水平有明显升高,Vimentin表达水平有明显下降;药物联合组RUNX3 mRNA与E-cadherin蛋白表达水平升高明显,而Vimentin表达水平下降更为显著.结论 5-Aza具有抑制胃癌裸鼠移植瘤生长与转移的能力,联合化疗药物紫杉醇与氟尿嘧啶处理效果更显著.5-Aza发挥肿瘤抑制功能可能是通过提高抑癌基因RUNX3基因表达实现的.