目的 构建抑癌基因SEMA3B真核表达载体pcDNA3.1-SEMA3B,并检测其对肺癌A549细胞恶性生物学行为的影响。方法 应用PCR扩增SEMA3B全长cDNA片段,构建真核表达载体pcDNA3.1-SEMA3B。克隆PCR、双酶切法、基因测序验证过表达载体构建成功。将pcDNA3.1-SEMA3B真核表达载体和空载体pcDNA3.1分别转染入A549细胞中,应用qRT-PCR、Western blot检测SEMA3B mRNA、蛋白表达水平的变化;MTS法检测细胞增殖;流式细胞仪检测细胞凋亡、细胞周期;克隆形成实验检测细胞集落形成能力。结果 SEMA3B基因扩增片段与预测片段一致,克隆成功,且测序鉴定证实真核表达载体构建成功。转染pcDNA3.1-SEMA3B真核表达载体可上调SEMA3B mRNA、蛋白表达水平,且可抑制A549细胞的增殖,诱导凋细胞亡,细胞被阻滞在G1期,抑制细胞集落形成能力。结论 成功构建了SEMA3B基因真核表达载体,抑癌基因SEMA3B在肺癌恶性生物学进程中可能发挥重要作用。

Objective To construct the eukaryotic expression vector of the cancer suppressor gene, SEMA3B, and research the effects on malignant biological behavior of lung cancer A549 cells. Methods By reverse transcriptase-polymerase chain reaction (RT-PCR), the full length SEMA3B gene was amplified and then was inserted into pcDNA3.1. The recombinant plasmid pcDNA3.1-SEMA3B was confirmed correctly through double enzyme digestion and PCR identification, which was transfected into lung cancer A549 cells by lipid media transfection. The untransfected A549 and A549 transfected with pcDNA3.1 were used as controls. SMEA3B gene was detected by qRT-PCR and western blot. MTS assay, flow cytometry, and colony formation test were performed to evaluate the effect of overexpression of SEMA3B gene on A549 cell proliferation, apoptosis, cell cycle, and colony forming ability. Results The amplied fragment of SEMA3B gene by PCR was consistent with the anticipated result, the SEMA3B gene was cloned successfully. And the recombinant plasmid pcDNA3.1-SMEA3B was constructed successfully through gene sequence identification. After transfection of pcDNA3.1-SEMA3B, SEMA3B mRNA and protein expression levels were raised, and overexpression of SEMA3B gene in A549 cells significantly inhibited the proliferation of A549 cells, induced apoptotic cell death, blocked cell cycle in the G1 phase, and suppressed cell colony-forming ability. Conclusion The recombinant pcDNA3.1-SEMA3B is constructed successfully. SEMA3B gene can significantly inhibit the malignant biological behavior of lung cancer A549 cells.

目的 分析乳腺癌细胞中Snail与MTDH基因的作用,明确Snail是否通过结合于MTDH的启动子区域促进乳腺癌转移。方法 克隆、转染Snail基因至乳腺癌细胞,观察过表达Snail的乳腺癌细胞中MTDHmRNA及蛋白表达的变化;再使用免疫共沉淀法检测Snail与MTDH基因的共作用。结果 转染Snail基因进入乳腺癌MDA-MB-435细胞后,转染组、空白组和对照组中MTDHmRNA的表达水平分别为1.61±0.22、1.02±0.18、0.99±0.20,转染组高于空白组和对照组,差异有统计学意义(P<0.05),而后两组表达无差异(P>0.05);Westren blot检测结果显示,Snail可促进MTDH蛋白的表达;免疫共沉淀显示,Snail与MTDH在细胞内存在相互结合作用。结论 Snail在乳腺癌细胞中可通过结合于MTDH基因的启动子区域,促进MTDHmRNA转录及相关蛋白的表达,从而导致乳腺癌转移。

Objective To investigate the function of Snail to MTDH gene in breast cancer cells. Methods We observed the changement of MTDHmRNA and protein expression in breast cancer cell line MDA-MB-435 after transfected with Snail gene. Then, we used co-immunoprecipitation to determine the domain of Snail and MTDH binding in vitro. Results After transfected with Snail gene into MDA-MB-435 cell, the expression levels of MTDHmRNA in transfection group, blank group and control group were 1.61±0.22,1.02±0.18,0.99±0.20. The level of transfection group was significantly higher than the other groups(P< 0.05). Western blot showed that the expression of MTDH protein can be promoted by Snail. Co-immunoprecipitation showed that Snail and MTDH are binding interactions in breast cancer cell line MDA-MB-43. Conclusion Snail can promote transcription and expression of MTDH in breast cancer cells by binding to the promoter region of the MTDH gene resulting in metastasis of breast cancer.

       目的  探讨TRIB2在结肠癌中的表达水平及与预后及免疫浸润之间的关系。方法  TIMER数据库分析TRIB2在泛癌种中的表达；TCGA、GSE17538下载结肠癌患者RNA-seq数据和临床信息，评估其与临床病理特征的相关性；生存曲线、单因素和多因素Cox分析探讨TRIB2与预后的相关性，并构建列线图；对TRIB2进行差异基因的富集分析；分析TRIB2表达水平与免疫细胞浸润、免疫检查点、肿瘤突变负荷（TMB）以及免疫治疗敏感性之间的相关性。结果  TRIB2在结肠癌组织中高表达（P＜0.05）；CMS1结肠癌患者TRIB2 mRNA表达水平最高；TRIB2是结肠癌患者的独立预后因素（单因素Cox回归分析：HR=1.397，95%CI：1.100~1.774，P=0.006；多因素Cox回归分析：HR=1.502，95%CI：1.158~1.947，P=0.002）；TRIB2与免疫细胞的浸润密切相关，并且与免疫检查点分子表达水平以及TMB正相关（r=0.39，P＜0.001）；TRIB2的表达水平与免疫检查点抑制剂的疗效相关。结论  TRIB2在结肠癌中高表达且与结肠癌患者预后差和免疫微环境密切相关。

       Objective  To explore the expression of TRIB2 in colon cancer and its relationship with prognosis and immune cell infiltration．Methods  TIMER database was used to analyse the expression of TRIB2 in pan-cancer．RNA-seq  data and clinical information of colon cancer patients were downloaded from TCGA and GSE17538 to assess the correlation between TRIB2 with clinicopathological features．Survival curves，univariate and multivariate COX regression analysis were performed to explore the correlation between TRIB2 and prognosis，and a nomogram was constructed．Gene enrichment analyses were performed for TRIB2．Correlations between TRIB2 expression and immune cell infiltration，immune checkpoints，tumor mutation burden（TMB），and immunotherapy sensitivity were analyzed．Results  TRIB2 was highly expressed in colon cancer tissues（P＜0.05）．The highest level of TRIB2 mRNA expression was found in CMS1．TRIB2 was an independent prognostic factor for colon cancer patients（univariate Cox regression analysis：HR=1.397，95%CI：1.100-1.774，P=0.006；multivariate Cox regression analysis：HR=1.502，95%CI：1.158-1.947，P=0.002）．TRIB2 was closely associated with immune cell infiltration and positively correlated with the expression level of immune checkpoint molecules as well as TMB（r=0.39，P＜0.001）．The expression of TRIB2 was correlated with the efficacy of immune checkpoint inhibitors．Conclusions  TRIB2 is highly expressed in colon cancer and is closely associated with poor prognosis and the immune microenvironment of colon cancer patients．

       目的   探究血清多配体蛋白聚糖-1（SCD-1）与可溶性血管内皮生长因子受体-2（sVEGFR-2）表达水平在老年慢性心力衰竭患者预后评估的判定价值。方法   选取2023年1月—2024年3月珠海市第五人民医院检验科收治的110例老年慢性心力衰竭患者，检测其血清SCD-1和sVEGFR-2水平，对患者进行随访调查，了解其再次由于心力衰竭住院、心源性死亡的情况。运用多因素Logistic回归分析，探究老年慢性心力衰竭患者预后影响因素。结果  Logistic回归分析显示，心功能分级（OR=3.433，95%CI：0.934~6.431）、B型脑钠肽升高（OR=2.462，95%CI：0.861~4.765）、血清SCD-1升高（OR=3.795，95%CI：0.972~6.894）、血清sVEGFR-2升高（OR=3.842，95%CI：0.942~6.912）为影响老年慢性心力衰竭患者预后不良的重要因素（P＜0.05）；联合血清SCD-1和sVEGFR-2曲线下面积0.962与B型脑肽钠曲线下面积0.844，相较于单一SCD-1曲线下面积0.658、sVEGFR-2曲线下面积0.712明显偏高（P＜0.05）。结论   经研究证实，老年慢性心力衰竭患者预后效果不理想，其血清SCD-1和sVEGFR-2监测水平异常升高，和老年慢性心力衰竭预后不佳存在关联性，可视为老年慢性心力衰竭患者判定预后效果的主要标志物。

      Objective  To investigate the prognostic value of serum syndecan-1（SCD-1）and soluble vascular endothelial growth factor receptor-2（sVEGFR-2）expression levels in elderly patients with chronic heart failure．Methods  A total of 110 elderly patients with chronic heart failure admitted to our hospital were selected，with a time interval of January 2023 to March 2024．Serum SCD-1 and sVEGFR-2 levels were detected and follow-up investigations were conducted to understand their re hospitalization and cardiogenic death due to heart failure．Multiple logistic  regression analysis was used to explore the prognostic factors affecting elderly patients with chronic heart failure．Results  According to logistic retrospective analysis，heart function grading（OR=3.433，95%CI：0.934-6.431），elevated B-type brain natriuretic peptide（OR=2.462，95%CI：0.861-4.765），elevated serum SCD-1（OR=3.795，95%CI：0.972-6.894），and elevated serum sVEGFR-2（OR=3.842，95%CI：0.942-6.912）were important factors affecting the poor prognosis of elderly patients with chronic heart failure，with differences P＜0.05．The area under the curve of combined serum SCD-1 and sVEGFR-2 was 0.962，and the area under the curve of B-type brain peptide sodium was 0.844，which was significantly higher than that of a single SCD-1 curve of 0.658 and sVEGFR-2 curve of 0.712，with a difference of P＜0.05．Conclusions  Research has confirmed that the prognosis of elderly patients with chronic heart failure is not satisfied，and their serum SCD-1 and sVEGFR-2 monitoring levels are abnormally elevated，which is related to the poor prognosis of elderly patients with chronic heart failure．It can be regarded as the main biomarker for defining the prognosis of elderly patients with chronic heart failure．

       目的  探讨转录因子E盒结合锌指蛋白1（ZEB1）、溶酶体相关膜蛋白5（LAMP5）在结直肠癌组织中的表达水平分析及预后预测价值。方法  选取驻马店市中心医院2018年1月—2020年1月收治的120例结直肠癌患者，分别采取所有患者的结直肠癌组织及癌旁组织进行免疫组化染色，对比ZEB1、LAMP5阳性率。对比不同病理特征结直肠癌患者ZEB1、LAMP5表达水平差异。对所有患者进行4年随访，依照随访结果将患者分为2个亚组，即预后不良组（n=35）和预后良好组（n=85），对比两组患者一般临床特征及ZEB1、LAMP5表达水平，应用Logistic回归分析ZEB1、LAMP5对结直肠癌预后的预测价值。结果 结直肠癌组织ZEB1、LAMP5相对表达量（38.26±5.49、26.77±3.85）与ZEB1、LAMP5阳性率（86.67%、72.22%）高于癌旁组织（15.46±2.54、8.04±1.59、23.33%、15.56%］，对比差异有统计学意义（t=41.280，χ²=25.437；t=49.255，χ ² =16.071；P＜0.05）。不同TNM分期［Ⅰ~Ⅱ期（35.55±4.13）、Ⅲ~Ⅳ期（42.32±4.75）］、淋巴结转移患者［是（44.37±4.28）、否（35.84±3.77）］、肿瘤分化程度［低分化（35.27±4.57）、中高分化（41.34±4.60）］ZEB1相对表达量对比差异有统计学意义（t=-8.281，P＜0.001；t=10.746，P＜0.001；t=-7.253，P＜0.001）；不同TNM分期［Ⅱ期（24.88±3.37）、Ⅲ~Ⅳ期（29.61±2.57）］、淋巴结转移［是（30.72±2.19）、否（25.21±3.19）］、肿瘤分化程度［低分化（24.57±3.62）、中高分化（29.04±2.55）］患者LAMP5相对表达量对比差异有统计学意义（t=-8.254，P＜0.001；t=9.227，P＜0.001；t=-7.797，P＜0.001）；预后良好组与预后不良组患者性别、年龄、大体类型、肿瘤大小对比差异无统计学意义（P＞0.05），预后良好组与预后不良组患者TNM分期、淋巴结转移、肿瘤分化程度、ZEB1、LAMP5阳性比例对比差异有统计学意义（P＜0.05）；Logistic回归分析显示：淋巴结转移、ZEB1阳性、LAMP5阳性为结直肠癌预后不良独立预测因素（P＜0.05）。结论  ZEB1、LAMP5在结直肠癌组织中呈现高表达状态，且与结直肠癌的发生有关，同时ZEB1、LAMP5是结直肠癌预后的独立预测因素，两者有希望成为结直肠癌的治疗靶点。

       Objective  To investigate the expression levels and prognostic value of transcription factor E-box binding to zinc finger protein 1（ZEB1）and lysosomal associated membrane protein 5（LAMP5）in colorectal cancer tissues．Methods  A total of 120 colorectal cancer patients admitted to a hospital from January 2018 to January 2020 were selected．Immunohistochemical staining was performed on the colorectal cancer tissues and adjacent tissues of all patients，and the positivity rates of ZEB1 and LAMP5 were compared．The expression levels of ZEB1 and LAMP5 in colorectal cancer patients with different pathological characteristics were compared．All patients were followed up for 4 years and divided into two subgroups based on the follow-up results，namely the poor prognosis group（n=35）and the good prognosis group（n=85）．The general clinical characteristics and expression levels of ZEB1 and LAMP5 were compared between the two groups．Logistic  regression analysis was used to evaluate the predictive value of ZEB1 and LAMP5 for the prognosis of colorectal cancer．Results The relative expression level of ZEB 1 and LAMP 5 in colorectal cancer tissues ［（38.26±5.49），（26.77±3.85）］ and the positive rate of ZEB 1 and LAMP 5（86.67%，72.22%）were significantly higher than that of adjacent tissues ［（15.46±2.54），（8.04±1.59），23.33%，15.56%］，the contrast  difference was statistically significant（t=41.280，χ²=25.437；t=49.255，χ ² =16.071；P＜0.05）．Relative ZEBI expression levels in different TNM stages ［I-Ⅱstage（35.55±4.13），Ⅲ-Ⅳstage（42.32±4.75）］，lymph node metastasis［Yes（44.37±4.28），No（35.84±3.77）］，degree of tumor differentiation ［hypodifferentiated（35.27±4.57），and middle or high differentiated （29.04±2.55）］，those differences were statistically significant（t=-8.254，P＜0.001；t=9.227，P＜0.001；t=-7.797，P＜0.001）．The relative expression of LAMP 5 between different TNM stages ［I-Ⅱstage（24.88±3.37），Ⅲ-Ⅳstage（29.61±2.57）］，lymph node metastasis ［yes（30.72±2.19），no（25.21±3.19）］，degree of tumor  differentiation ［hypodifferentiated（24.57±3.62），and middle or high differentiated（29.04±2.55）］，the contrast was statistically significant（t=-8.254，P＜0.001；t=9.227，P＜0.001；t=-7.797，P＜0.001）．There were no differences in gender，age，gross type，and tumor size between the good prognosis group and the poor prognosis group（P＞0.05），while there were differences in TNM stages，lymph node metastasis，tumor differentiation degrees，ratio of ZEB 1 and LAMP 5（P＜0.05）．Logistic regression analysis showed that TNM stage，lymph node metastasis，ZEB 1 positive，and LAMP 5 positive were independent predictive factors of poor prognosis in colorectal cancer（P＜0.05）．Conclusions  ZEB1 and LAMP5 are highly expressed in colorectal cancer tissues and closely related to the occurrence and development of colorectal cancer．ZEB1 and LAMP5 are independent prognostic factors for colorectal cancer，and they have the potential to become therapeutic targets for colorectal cancer．

       目的   探讨Ki-67、微小染色体维持蛋白2（MCM2）、p16在宫颈鳞状上皮内病变中的表达及其临床意义。方法   采用免疫组化检测Ki-67、MCM2、p16在宫颈炎症组14例、低级别鳞状上皮内病变（LSIL）组47例、高级别鳞状上皮内病变（HSIL）组49例中的表达情况，以病理结果作为金标准，对结果进行统计分析。结果  HSIL组中Ki-67、MCM2、p16阳性率均高于炎症组和LSIL组（均P＜0.017）。HSIL组中Ki-67、MCM2过表达率均显著高于炎症组和LSIL组（均P＜0.017）。随着宫颈病变级别增加，Ki-67及MCM2阳性范围从基底层至表层逐渐扩大。MCM2及Ki-67在LSIL组中表达模式多为基底层的非过表达模式，HSIL组多为中层及以上的过表达模式。Spearman相关性分析显示，MCM2和Ki-67在宫颈鳞状上皮内病变中的表达强度之间呈正相关（r=0.801，P＜0.05）；p16与MCM2在宫颈鳞状上皮内病变中的表达呈正相关（r=0.559，P＜0.05）；p16与Ki-67在宫颈鳞状上皮内病变中的表达呈正相关（r=0.478，P＜0.05）。结论  p16阳性提示宫颈高级别鳞状上皮内病变。MCM2与Ki-67在宫颈鳞状上皮内病变中的表达具有较高一致性，MCM2可作为宫颈鳞状上皮内病变新的增殖标志物。

       Objective  To investigate the expression and significance of Ki-67，MCM2 and p16 in cervical intraepithelial lesions．Methods  The expressions of Ki-67，MCM2 and p16 in cervicitis group（14 cases），low-grade squamous intraepithelial lesion（LSIL） group（47 cases）and high-grade squamous intraepithelial lesion（HSIL） group（49 cases）were detected by immunohistochemistry．The pathological results were used as the gold standard for statistical analysis．Results  The positive  rates of Ki-67，MCM2 and p16 in HSIL group were significantly higher than those in cervicitis group and LSIL group（P＜0.017）．The over-expression rates of Ki-67，MCM2 in HSIL group were significantly higher than those in cervicitis group and LSIL grou（P＜0.017）．With the increase of cervical lesion grade，the positive range of Ki-67 and MCM2 gradually expanded from basallayer to surface layer．The expression patterns of MCM2 and Ki-67 in LSIL group were mostly non-overexpressed in the basal layers，while those in HSIL group were mostly overexpressed in the middle layer and above．Spearman correlation analysis showed that the expression intensity of MCM2 and Ki-67 in cervical squamous intraepithelial lesions was positively correlated（r=0.801，P＜0.05）．There was a positive correlation between the expression of p16 and MCM2 in cervical squamous intraepithelial lesions（r=0.559，P＜0.05）．There was a positive correlation between the expression of p16 and Ki-67 in cervical squamous intraepithelial lesions（r=0.478，P＜0.05）．Conclusions  Positive p16 indicates high-grade squamous intraepithelial lesion．The expression of MCM2 and Ki-67 in cervical intraepithelial lesions is highly consistent．MCM2 can be used as a new proliferative marker for cervical intraepithelial lesions．