目的 探讨老年胃癌患者血清长基因间非蛋白编码RNA(LINC)00511和LINC01133的表达及与临床特征的关系,分析二者单项及与糖类抗原(CA)72-4联合检测对老年人胃癌的临床诊断价值.方法 选取老年胃癌患者100例作为实验组,再选取同期老年健康体检者76例作为对照组,通过定量逆转录聚合酶链反应检测血清LINC00511和LINC01133的表达,分析二者血清表达水平的相关性及与肿瘤临床特征的关系;采用受试者工作特征(ROC)曲线评估其对胃癌的临床诊断效能;通过计算ROC曲线下面积(AUC)、敏感度、特异度、阳性预测值及阴性预测值,评估单项及联合检测的临床应用价值.结果 实验组LINC00511和LINC01133表达量显著高于对照组(均P<0.01),实验组二者表达水平呈显著正相关(r=0.304,P<0.01).LINC01133血清水平与性别、TNM临床分期和淋巴结转移均无统计学差异(P>0.05),但与肿瘤合并远处转移呈正相关(P<0.01).LINC00511、LINC01133及CA72-4诊断胃癌的AUC分别为0.659、0.807和0.764,其中LINC01133诊断的敏感度(81.0％)最高,CA72-4诊断的特异度(84.2％)最高.LINC00511与LINC01133具有显著诊断互补性(P<0.05).而LINC00511和CA72-4、LINC01133和CA72-4之间均无显著互补性(P>0.05).LINC00511和LINC01133联合可将诊断的敏感度提高至92.0％,二者与CA72-4联合可将敏感度和阴性预测值分别提高至94.0％和85.7％.结论 LINC00511和LINC01133在老年胃癌患者血清中高表达,与临床进展显著相关,可能是老年胃癌辅助诊断的潜在血清学标志物.

Objective: To investigate the clinical diagnostic value of multi-target stool fecal immunochemical test-DNA (FIT-DNA) test in colorectal cancer (CRC) and advanced adenoma (AA). Methods: A total of 235 patients who were undergoing colonoscopy or colorectal cancer surgery in the Cancer Hospital, Chinese Academy of Medical Sciences from April 2021 to January 2022 were prospectively enrolled. There were 141 males and 94 females, with an average age of (55±13) years (22-86). The patients were divided into two groups, including 215 patients who were first diagnosed but not treated (86 cases of CRC, 12 cases of AA, 25 cases of non-advanced adenoma, 8 cases of hyperplastic or other polyps and 84 apparently healthy cases) and 20 patients in the intervention group (2 cases with a history of CRC surgery, 6 cases with a history of endoscopic surgery, 4 non-CRC patients with special diseases and 8 cases with a history of neoadjuvant chemoradiotherapy). Fresh stool samples were collected before intestinal preparation or surgery for FIT-DNA test using the matching kit for sample processing and nucleic acid purification. KRAS mutation and methylation of BMP3 and NDRG4 genes were detected by fluorescence probe method, and FIT method was employed to detect fecal occult blood. Colonoscopy or pathological biopsy results were used as the gold standard. And the screening and diagnostic efficacy of FIT-DNA test for colorectal cancer and advanced adenoma were evaluated by receiver operating curve (ROC). Results: The sensitivity of FIT-DNA test for early colorectal cancer and advanced adenoma was 7/7 and 8/12, respectively. And the negative predictive value was 98.1% (104/106) and 93.7% (104/111), respectively. The overall screening sensitivity for both early colorectal cancer and advanced adenoma was 15/19, and the negative predictive value was 96.3% (104/108). Besides, the area under the curves (AUCs) were 0.982 (95%CI: 0.960-1.000, P<0.05), 0.758 (95%CI: 0.592-0.924, P<0.05) and 0.841 (95%CI: 0.724-0.957, P<0.05), respectively. Moreover, the diagnostic sensitivity of FIT-DNA test was 98.8% (85/86) for colorectal cancer, 8/12 for advanced adenoma, and 94.9% (93/98) for both colorectal cancer and advanced adenoma, with a specificity of 88.9% (104/117). The AUCs were 0.968 (95%CI: 0.937-0.997, P<0.05), 0.758 (95%CI: 0.592-0.924, P<0.05) and 0.942 (95%CI: 0.905-0.979, P<0.05), respectively. After the inclusion of intervention group, the overall diagnostic sensitivity and specificity of FIT-DNA test was 91.6% (98/107) and 89.1% (114/128), respectively. Conclusion: FIT-DNA test has a high early screening and diagnostic efficacy for colorectal cancer.

目的 检测微小RNA-4491(miR-4491)在胃癌患者血清中的表达水平,分析其与胃癌临床病理特征的相关性,探讨其单独检测及其与CA72-4联合检测在胃癌诊断中的临床价值.方法 实时荧光定量聚合酶链反应检测胃癌患者和健康受试者血清中miR-4491的表达水平,通过受试者工作特征(ROC)曲线、曲线下面积(AUC)、危险分数分析法评估miR-4491对胃癌的诊断效能,计算灵敏度、特异度、阳性预测值(PPV)和阴性预测值(NPV),探讨miR-4491独立及与CA72-4联合检测在胃癌诊断中的应用价值.结果 胃癌组和健康对照组血清miR-4491的相对表达水平分别为3.65±4.04和1.29±1.21,二者比较,差异有统计学意义(P<0.01),miR-4491的表达量与胃癌临床分期、淋巴结及远处转移均不相关(P>0.05),但与其病理分化程度呈负相关(P<0.05).miR-4491独立检测诊断胃癌的AUC为0.78,敏感度、特异度、PPV、NPV分别为84.5％,59.4％,74.0％和72.9％,而CA72-4诊断胃癌的AUC为0.71,敏感度、特异度、PPV和NPV分别为20.9％,78.3％,51.9％和47.0％.MiR-4491与CA72-4联合诊断的AUC为0.83,PPV和NPV分别提高至87.1％和89.7％.结论 miR-4491在胃癌血清中高表达,是胃癌临床筛查的潜在血清学标志物.

Abstract Background Identifying breast cancer patients with DNA repair pathway-related germline pathogenic variants (GPVs) is important for effectively employing systemic treatment strategies and risk-reducing interventions. However, current criteria and risk prediction models for prioritizing genetic testing among breast cancer patients do not meet the demands of clinical practice due to insufficient accuracy. Methods The study population comprised 3041 breast cancer patients enrolled from seven hospitals between October 2017 and 11 August 2019, who underwent germline genetic testing of 50 cancer predisposition genes (CPGs). Associations among GPVs in different CPGs and endophenotypes were evaluated using a case-control analysis. A phenotype-based GPV risk prediction model named DNA-repair Associated Breast Cancer (DrABC) was developed based on hierarchical neural network architecture and validated in an independent multicenter cohort. The predictive performance of DrABC was compared with currently used models including BRCAPRO, BOADICEA, Myriad, PENN II, and the NCCN criteria. Results In total, 332 (11.3%) patients harbored GPVs in CPGs, including 134 (4.6%) in BRCA2, 131 (4.5%) in BRCA1, 33 (1.1%) in PALB2, and 37 (1.3%) in other CPGs. GPVs in CPGs were associated with distinct endophenotypes including the age at diagnosis, cancer history, family cancer history, and pathological characteristics. We developed a DrABC model to predict the risk of GPV carrier status in BRCA1/2 and other important CPGs. In predicting GPVs in BRCA1/2, the performance of DrABC (AUC = 0.79 [95% CI, 0.74–0.85], sensitivity = 82.1%, specificity = 63.1% in the independent validation cohort) was better than that of previous models (AUC range = 0.57–0.70). In predicting GPVs in any CPG, DrABC (AUC = 0.74 [95% CI, 0.69–0.79], sensitivity = 83.8%, specificity = 51.3% in the independent validation cohort) was also superior to previous models in their current versions (AUC range = 0.55–0.65). After training these previous models with the Chinese-specific dataset, DrABC still outperformed all other methods except for BOADICEA, which was the only previous model with the inclusion of pathological features. The DrABC model also showed higher sensitivity and specificity than the NCCN criteria in the multi-center validation cohort (83.8% and 51.3% vs. 78.8% and 31.2%, respectively, in predicting GPVs in any CPG). The DrABC model implementation is available online at http://gifts.bio-data.cn/ . Conclusions By considering the distinct endophenotypes associated with different CPGs in breast cancer patients, a phenotype-driven prediction model based on hierarchical neural network architecture was created for identification of hereditary breast cancer. The model achieved superior performance in identifying GPV carriers among Chinese breast cancer patients.

Objective To investigate the effect of NAIF1 in gastric cancer cell lines MKN45. Methods We constructed pLVX-Tight-Flag-NAIF1-puro plasmid with Tet-on system. DOX was added to induce NAIF1 expression in MKN45 cells. The cells were collected at 0, 6, 12 and 24 hours after DOX addition for gene expression microarray detection and biological analysis of differentially expressed genes. qRT-PCR and Western blot were used to verify the changes in mRNA and protein levels of the selected target differential genes. Results The biological analysis of gene microarray hybridization results showed that IFIT1, IFIT2 and IFIT3 expression significantly increased at 24h, qRT-PCR also showed this change, and Western blot further verified the change in protein level. However, IFIT5 showed no significant change in mRNA and gene expression. Conclusion Over-expression of NAIF1 in gastric cancer cells can promote the expression of some immune system-related IFIT family proteins.

A proportion of up to 10% of breast cancer resulted from hereditary germline pathogenic variants (GPVs) in cancer predisposition genes (CPGs), which been demonstrated distinct clinical features and imaging manifestations. However, the performance of imaging modalities for breast cancer surveillance in CPG mutation-carriers is still unclear, especially in Asian women. A population of 3002 breast cancer patients who received germline genetic testing of CPGs was enrolled from three hospitals in China. In total, 343 (11.6%) patients were found to harbor GPVs in CPGs, including 137 (4.6%) in BRCA1 and 135 (4.6%) in BRCA2. We compared the performances of ultrasound, mammograms, MRI, and the combining strategies in CPG mutation carriers and non-carriers. As a result, the ultrasound showed a higher detection rate compared with mammograms regardless of the mutation status. However, its detection rate was lower in CPG mutation carriers than in non-carriers (93.2% vs 98.0%, P=2.1×10-4), especially in the BRCA1 mutation carriers (90.9% vs 98.0%, P=2.0×10-4). MRI presented the highest sensitivity (98.5%) and the lowest underestimation rate (14.5%) in CPG mutation carriers among ultrasound, mammograms, and their combination. Supplemental ultrasound or mammograms would add no significant value to MRI for detecting breast cancer (P>0.05). In multivariate logistic regression analysis, the family or personal cancer history could not replace the mutation status as the impact factor for the false-negative result and underestimation. In summary, clinicians and radiologists should be aware of the atypical imaging presentation of breast cancer in patients with GPVs in CPGs.

长非编码RNA SLC25A25-AS1在结直肠癌的发展中具有肿瘤抑制作用,然而,其在宫颈癌中作用机制有待深入研究.本文研究了宫颈癌和宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)病人血清中SLC25A25-AS1的异常表达,并探讨了SLC25A25-AS1在宫颈癌发展中可能的作用机制.采用RT-qPCR检测正常对照组,宫颈癌病人和CIN病人血清中SLC25A25-AS1的表达水平.在体外和体内研究中分析了SLC25A25-AS1在宫颈癌系HeLa细胞中的重要作用.与健康对照组相比,宫颈癌病人血清中的SLC25A25-AS1含量降低.在体外,SLC25A25-AS1的过表达抑制HeLa细胞的增殖、迁移和侵袭.体内裸鼠成瘤实验表明,注射过SLC25A25-AS1过表达的HeLa细胞的裸鼠皮下肿瘤的重量和体积明显小于注射对照细胞的裸鼠(P＜0.05).SLC25A25-AS1在宫颈癌的发生发展中可能发挥重要作用,有望成为宫颈癌的新治疗靶点.

Background: Ubiquilin-4 (UBQLN4) is a component of the ubiquitin-proteasome system and regulates the degradation of many proteins implicated in pathological conditions. The aim of this study was to determine the role of UBQLN4 in regulating the proliferation and survival of the normal gastric epithelial cell line GES-1. Material/Methods: We constructed GES-1 lines stably overexpressing UBQLN4 by lentiviral infection. Cell proliferation, apoptosis, and the cell cycle were analyzed using the MTT assay and flow cytometric assays. Phosphorylation of ERK, JNK, p38, and expression of cyclin D1 were detected by western blot analysis. Results: Overexpression of UBQLN4 significantly reduced proliferation and induced G2/M phase arrest and apoptosis in GES-1 cells. Moreover, upregulation of UBQLN4 increased the expression of cyclin D1 and phosphorylated ERK, but not JNK or p38. Conclusions: These data suggest that UBQLN4 may induce cell cycle arrest and apoptosis via activation of the ERK pathway and upregulation of cyclin D1 in GES-1 cells.

目的 探讨化学发光微粒子免疫分析法(CMIA)检测梅毒抗体用于梅毒感染筛查的效能.方法 用1份低值血清和1份高值血清对CMIA检测血清梅毒抗体进行精密度评价;将梅毒抗体标准品稀释至临界值附近,对CMIA法检出限进行验证;阴性、阳性样本以不同比例混合,对cut-off值附近的重复性进行验证.对CMIA法检测阳性标本进行TP-PA确证,同时进行RPR试验,此外对CMIA法检测阴性的384例标本进行TP-PA检测,评价CMIA检测梅毒抗体的检测效能.结果 CMIA检测梅毒抗体的批内和批间精密度CV均＜8％,检出限为4 NCU/ml,cut-off值附近样本重复性结果显示低于cut-off值20％的20份样本均为阴性,高于cut-off值20％的20份样本均为阳性.38 902例血清样本经CMIA检测615例梅毒抗体阳性,其中TP-PA阳性为401例,RPR阳性为110例.384例CMIA检测阴性样本经TP-PA验证后均为阴性.结论 CMIA法可用于有效筛查梅毒感染,但对于弱阳性结果应进一步验证.

目的 探讨三阴性乳腺癌患者血清长链非编码RNA(lncRNA)HOX转录反义RNA(HOTAIR)、生长停滞特异性RNA5(GAS5)、肺腺癌转移相关转录子1(MALAT1)的表达变化,并探讨其临床意义.方法 选择35例三阴性乳腺癌患者为观察组、29例健康体检者为对照组,观察组患者入院时、对照组患者体检时收集血清,采用实时荧光PCR法检测HOTAIR、GAS5、MALAT1水平.分析血清HOTAIR、GAS5、MALAT1水平与患者临床病理参数的关系.结果 两组血清HOTAIR 相对表达量相比,P>0.05.观察组血清GAS5相对表达量低于对照组(P<0.05),MALAT1相对表达量高于对照组(P<0.05).观察组患者血清GAS5相对表达量与患者TNM分期和淋巴结转移有关(P<0.05),与患者年龄无关(P>0.05).MALAT1与GAS5相同.结论 三阴性乳腺癌患者血清GAS5表达量低于健康人,血清MALAT1表达量高于健康人,血清HOTAIR表达量与健康人无差异.检测三阴性乳腺癌患者血清GAS5、MALAT1表达量有助于三阴性乳腺癌的诊断和病情判断.

恶性肿瘤通常伴有机体的高凝状态,是血栓形成的高危因素,而血栓形成也是恶性肿瘤严重的并发症之一[1].但凝血指标中与凝固时间相关的指标,如凝血酶时间(thrombin time, TT)、凝血酶原时间(prothrombin time,PT)和活化部分凝血活酶时间(activated partial thromboplastin time,APTT)在肿瘤患者中多表现为正常或缩短,TT明显延长或"不凝固"报警的病例较为罕见.本研究报道1例肺下叶癌伴淋巴结肿大并且进行了放疗、化疗的患者凝血功能检验结果 中出现单纯TT延长的情况.

Objective To screen and identify the interacting proteins with ubiquilin 4 and explore its mechanism in tu-mor development and progression. Method Cell immunofluorescence technique was used to determine the location of exogenous and endogenous ubiquilin 4 in MKN45 cells; a recombinant prokaryotic expression vector pGEX-UBQLN4 containing UBQLN4 and GST tags was constructed and then transformed into Escherichia coli, and the protein was ob-tained after 6 h. Ubiquilin 4 was purified using affinity tag and the interacting proteins were detected by GST-pull down. The candidate proteins were identified by mass spectrometry, and the interaction was verified by co-immunoprecipitation assay. Result Immunofluorescence technique results showed that the exogenous and endogenous ubiquilin 4 were main-ly expressed in the nucleus and cytoplasm. After purification and mass spectrometry analysis, it was found that MYH9 might be the possible interacting protein with ubiquilin 4. Conclusion MYH9 protein is identified to interact with ubiqui-lin 4. These results may provide a new clue for the role of ubiquilin 4 in the development and progression of cancer.

[Purpose] To investigate the effect of cathepsin D(CTSD) on the chemosensitivity of gastric cancer cells to cisplatin.[Methods] CTSD and its mutation 231N were transfected into gastric cancer MKN45 cells by lentivirus.The cell proliferation was determined by MTT and the expression of apoptosis related proteins was tested by Western blot.The apoptosis rate was detected by flow cytometry.[Results] Compared to cisplatin alone,CTSD or 231N combined with cisplatin inhibited cell proliferation and enhanced cell apoptosis more markedly.Western blotting results indicated that the expression levels of cleaved caspase 3,cleaved caspase 9 in CTSD or 231N combined with cisplatin were increased,however,there were no significant changes in expression of procaspase 8.[Conclusion] CTSD and 231N can increase the inhibition effect of cisplatin on gastric cancer MKN45 cells and enhance cell apoptosis,which may be associated with mitochondrial pathway.

Objective: To investigate the clinical value of combined detection of serum miR-378 and miR-21 in gastric cancer (GC). Methods: Eighty-seven patients with GC and 78 patients with colorectal cancer(CRC) from National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences were selected, 83 individuals undergoing healthy physical examination were selected as the healthy controls. The levels of serum miR-378 and miR-21 were detected by quantitative real-time PCR (RT-qPCR) (result data were transformed as log2 for analysis). Results: Relative expression levels of miR-378 in the serum were -1.24, -3.25 and -2.73 in healthy controls, GC and CRC patients, respectively. Compared with the healthy controls, the levels of serum miR-378 were significantly decreased in GC and CRC patients (both P<0.05). Relative expression levels of miR-21 in the serum were 0.11, 2.34 and 2.47 in healthy controls, GC and CRC patients, respectively. Compared with the healthy controls, the levels of serum miR-21 were significantly up-regulated in GC and CRC patients (both P<0.05). Moreover, the serum level of miR-378 in GC patients was inversely associated with tumor clinical stage (P<0.05). However, the level of miR-21 showed no significant differences among patients with different clinical and pathological characteristics (all P>0.05). The area under the receiver operating characteristic curve (AUC), sensitivity and specificity of miRNA-378 to diagnose GC was 0.770, 82.0% and 66.0%, respectively, and were 0.900, 85.0%, and 88.0% of miR-21, respectively. The AUC, sensitivity and specificity of combined detection of serum miR-378 and miR-21 to diagnose GC were 0.930, 92.0% and 87.0%, respectively, while the AUC of combined detection of serum CEA and CA-199 was 0.767, the AUC of combined all of the four factors was 0.946. Conclusion: The combined detection of serum miR-378 and miR-21 have a certain effect on diagnosis of GC.

PURPOSEPreviously, we identified six miRNAs that are differentially expressed in colorectal cancer compared with healthy controls. Here, we tested them in gastric cancer GC.METHODSWe performed quantitative RT-PCR on serum samples from 92 patients with gastric cancer and 89 controls for the six miRNAs, and analyzed their risk scores to evaluate the diagnostic value of the serum miRNA profiling system.RESULTSAfter a two-phase selection and validation process, five miRNAs were found to significantly differ in expression between gastric cancer samples and control samples, including miR-21, miR-31, miR-92a, miR-181b, and miR-203. Risk score analysis showed that this miRNA panel could distinguish gastric cancer cases from controls with high sensitivity and specificity. Under receiver operating characteristic curves, areas under the curve for tumor identification were 0.933 (95% confidence interval [CI]: 0.86-1.007) for the training set and 0.919 (95% CI: 0.863-0.975) for the validation set-markedly higher than those of carcinoembryonic antigen (0.624) and carbohydrate antigen 19-9 (0.603).CONCLUSIONSThe signature of these five miRNAs is a novel and noninvasive biomarker for gastric cancer, and could facilitate and simplify its diagnosis.

目的:探讨micro RNA-21、micro RNAlet-7a在三阴性乳腺癌与Luminal A型乳腺癌中的差异及与临床参数间相关性,为三阴性乳腺癌的治疗提供分子靶点。方法:收集三阴性、Luminal A型乳腺癌各33例以及健康者血清29例,通过q PCR检测血清中micro RNA-21、let-7a的表达情况。结果:micro RNA-21在三阴组血清中表达高于Luminal A组（P＜0.05）及正常组（P＜0.05）。Let-7a在三阴组血清中表达低于正常组（P＜0.05）。三阴组血清中micro RNA-21高表达与临床分期晚呈正相关（P＞0.05）。结论:micro RNA-21在Luminal A型与三阴性乳腺癌中的表达差异可作为三阴性乳腺癌特异性治疗的候选靶点。

目的:在肝癌细胞HepG2中过表达外源NAIF1(核凋亡诱导因子1),探讨NAIF1的亚细胞定位以及对HepG2增殖和迁移能力的影响.方法:以真核表达质粒pEGFP-N1为对照组,pEGFP-N1-NAIF1为实验组,瞬时转染肝癌细胞HepG2,利用免疫印迹方法检测NAIF蛋白表达效率;以DAPI染核,荧光显微镜下观察绿色荧光蛋白定位,确定NAIF1的亚细胞定位;通过MTT方法绘制细胞增殖曲线;通过transwell小室法检测NAIF1对HepG2迁移能力的影响.结果:在肝癌细胞HepG2中,外源表达NAIF1主要定位于细胞核;与对照组HepG2/pEGFP-N1相比,HepG2/pEGFP-N1-NAIF1的细胞增殖、迁移能力下降(P＜0.05).结论:外源表达NAlF1蛋白定位于HepG2细胞核,过表达NAIF1抑制HepG2的细胞增殖与迁移能力,NAIF1可能作为肝癌治疗的潜在靶点.

Nuclear apoptosis-inducing factor 1 (NAIF1) could induce apoptosis in gastric cancer cells. Previously, we have reported that the expression of NAIF1 protein is down-regulated in gastric cancer tissues compared with the adjacent normal tissues. However, the role of NAIF1 in gastric cancer cells is not fully understood.

Prognosis of patients with colorectal cancer (CRC) is generally poor because of the lack of simple, convenient, and noninvasive tools for CRC detection at the early stage. The discovery of microRNAs (miRNAs) and their different expression profiles among different kinds of diseases has opened a new avenue for tumor diagnosis. We built a serum microRNA expression profile signature and tested its specificity and sensitivity as a biomarker in the diagnosis of CRC. We also studied its possible role in monitoring the progression of CRC. We conducted a two phase case-control test to identify serum miRNAs as biomarkers for CRC diagnosis. Using quantitative reverse transcription polymerase chain reactions, we tested ten candidate miRNAs in a training set (30 CRCs vs 30 controls). Risk score analysis was used to evaluate the diagnostic value of the serum miRNA profiling system. Other independent samples, including 83 CRCs and 59 controls, were used to validate the diagnostic model. In the training set, six serum miRNAs (miR-21, let-7g, miR-31, miR-92a, miR-181b, and miR-203) had significantly different expression levels between the CRCs and healthy controls. Risk score analysis demonstrated that the six-miRNA-based biomarker signature had high sensitivity and specificity for distinguishing the CRC samples from cancer-free controls. The areas under the receiver operating characteristic (ROC) curve of the six-miRNA signature profiles were 0.900 and 0.923 for the two sets of serum samples, respectively. However, for the same serum samples, the areas under the ROC curve used by the tumor markers carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) were only 0.649 and 0.598, respectively. The expression levels of the six serum miRNAs were also correlated with CRC progression. Thus, the identified six-miRNA signature can be used as a noninvasive biomarker for the diagnosis of CRC, with relatively high sensitivity and specificity.