目的:探究食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)患者血清程序性死亡配体-1(programmed death-ligand 1,PD-L1)含量与T淋巴细胞免疫功能的相关性.方法:收集2021年08月至2022年01月我院初诊的62例ESCC患者作为ESCC组,对照组29例来自相对健康人群,采用两独立样本Wilcoxon秩和检验比较两组血清PD-L1和T淋巴细胞亚群水平的差异及这些指标在ESCC组不同临床分期中的差异,绘制受试者工作特征曲线(receiver operating characteristic curve,ROC)评价在临床分期中有差异的指标对ESCC临床进展的预测价值;采用χ2检验分析血清PD-L1与ESCC患者临床病理特征之间的相关性,两独立样本Wilcoxon秩和检验比较血清PD-L1高值组与低值组T淋巴细胞亚群水平的差异.结果:ESCC组PD-L1水平显著高于对照组,且Ⅲ-Ⅳ期的ESCC患者血清PD-L 1含量明显高于Ⅰ-Ⅱ期(P<0.05).ESCC组CD4+PD-1+T细胞百分比显著高于对照组(P<0.05);而CD3+T细胞绝对值计数、CD4+T细胞绝对值计数、CD4+CD28+T细胞绝对值计数、CD8+CD28+T细胞百分比和绝对值计数明显低于对照组(P<0.05).ESCC组Ⅲ-Ⅳ期CD4+PD-1+T细胞百分比明显高于Ⅰ-Ⅱ期;而CD4+CD28+T细胞百分比和绝对值计数明显低于Ⅰ-Ⅱ期(P<0.05).血清PD-L1的含量与ESCC患者的临床进展密切相关,且具有较高预测价值(P<0.05).PD-L1高值组CD4+PD-1+T细胞百分比明显高于低值组(P<0.05),而CD4+CD28+T细胞百分比和绝对值计数、CD8+CD28+T细胞百分比和绝对值计数明显低于低值组(P<0.05).结论:ESCC患者血清PD-L1水平与T淋巴细胞表面抗原PD-1和CD28表达水平密切相关,这可能是其抑制T淋巴细胞免疫功能、促进ESCC细胞免疫逃逸的重要因素之一.

目的:探讨纤维蛋白原(FIB)、纤维蛋白原与白蛋白比值(FAR)和血清肿瘤标记物联合检测在食管鳞状细胞癌(ESCC)患者诊断及其临床进展中的意义.方法:选取2018年4月—2021年10月河北医科大学第四医院收治的66例ESCC患者,作为ESCC组.收集ESCC患者初诊时的临床病理资料及血常规、凝血功能、生化、血清肿瘤标记物包括癌胚抗原(CEA)和鳞状细胞癌抗原(SCC-Ag)等相关实验室检测指标.另选取无良恶性肿瘤、心血管等疾病的相对健康志愿者30例,作为对照组.比较两组患者各指标的差异,并分析不同TNM分期中各指标的表达水平,制作ROC曲线评价FIB、FAR、CEA和SCC-Ag对ESCC临床进展的价值;分析其与ESCC患者临床病理指标之间的相关性.结果:ESCC组FIB、FAR、纤维蛋白原与前白蛋白比值(FPR)、中性粒细胞与淋巴细胞比值(NLR)、血小板与淋巴细胞比值(PLR)等指标的水平明显高于对照组(P<0.05);III～IV期FIB、FAR、CEA和SCC-Ag水平明显高于I～II期,同时,ROC曲线显示其对ESCC临床进展均具有预测价值(P<0.05),且四者联合检测对ESCC临床进展具有更高的诊断价值;FIB、FAR、SCC-Ag高值组(以ROC曲线得到的cut off值为界值)患者N分期和TNM分期均明显高于低值组(P<0.05);FAR高值组患者的肿瘤最大直径和T分期也明显高于低值组(P<0.05);CEA高值组患者TNM分期明显高于低值组,男性多于女性(P<0.05).结论:血浆FIB、FAR和血清CEA、SCC-Ag水平与ESCC诊断和临床进展密切相关,与单独检测血清肿瘤标记物相比,四者联合检测用于预测ESCC的临床进展效能更好,值得临床推广使用.

Esophageal cancer is a common malignant disease that is generally treated with radiotherapy. High mobility group box 1 (HMGB1) plays an essential role in tumor cell proliferation, migration, and cell cycle progression. Here, we aimed to clarify the effects of HMGB1 on radioresistance in esophageal squamous cell carcinoma (ESCC) cell lines and patient survival. We performed immunohistochemistry for HMGB1 in biopsy samples of 39 stage I-III ESCC patients grouped by HMGB1 expression status. Then, 1-, 3-, 5-, and 10-year overall survival outcomes were calculated by Kaplan-Meier survival analysis. The cellular localization of HMGB1 was examined before and after irradiation by Immunofluorescence staining. Stable cell lines (KYSE30 and KYSE510) with differential HMGB1 expression were constructed using lentiviruses. Furthermore, we examined phosphorylated histone H2AX (γ-H2AX) expression in both HMGB1 overexpression and negative control groups by western blotting. HMGB1-negative expression was associated with superior ESCC patient 10-year survival (P=0.016). HMGB1 overexpression promoted cell migration, proliferation, and radioresistance and mitigated cell cycle arrest at the G0/G1 phase induced by irradiation. This demonstrates that HMGB1-positive expression is correlated with unfavorable clinical outcomes, and HMGB1 overexpression may promote the malignant phenotype of ESCC cells and induce radioresistance by regulating cell cycle distribution in ESCC.

目的:分析miR-221在肝癌患者癌组织和正常组织中以及血浆中的表达水平及其表达水平与临床病理特征、诊断和预后之间的相关性.并进一步分析探讨其在肝癌发生发展中的作用机制.方法:利用基因表达数据库(Gene Expression Omnibus database,GEO)中的芯片数据集分析miR-221在肝患者癌组织及癌旁组织中的表达水平.利用实时荧光定量PCR法验证miR-221在74例临床肝癌患者肿瘤组织和对应癌旁组织中的表达水平,进一步检测miR-221在25例肝癌患者血浆和正常人血浆中的表达水平.进一步分析miR-221在肝癌诊断及预后方面的价值.进一步进行miR-221的靶基因进行功能富集分析,及其对肝癌细胞增殖、侵袭以及EMT进程的影响.结果:GEO数据库分析结果显示miR-221在肝癌组织中的表达水平显著高于癌旁组织(P<0.05),同样经实时荧光定量PCR验证结果证明miR-221在肝癌患者的肿瘤组织和血浆中显著高表达,且其表达水平与肝癌患者的TNM分期和肿瘤大小显著相关.MiR-221的表达水平降低与肝癌患者的总生存时间和无病生存时间延长显著相关.功能富集分析发现miR-221所参与的信号通路主要为p53信号通路、ErbB信号通路、RNA转运和mTOR信号通路等信号通路,且体外实验表明低表达miR-221显著抑制肝癌细胞的增殖和侵袭能力,同时肝癌HepG2和MHCC97H细胞的E-cadherin mRNA和蛋白表达水平显著上调(P<0.01),而N-cadherin和vimentin mRNA和蛋白的表达水平明显降低.结论:MiR-221在肝癌组织和血浆中显著高表达,且与肝癌患者的预后不良显著相关,主要在肝癌中通过EMT进程而发挥作用.

Metastasis-associated protein 2 (MTA2) is frequently amplified in many types of cancers; however, the role and underlying molecular mechanism of MTA2 in esophageal squamous cell carcinoma (ESCC) remain unknown. Here, we reported that MTA2 is highly expressed in ESCC tissue and cells, and is closely related to the malignant characteristics and poor prognosis of patients with ESCC. Through in vitro and in vivo experiments, we demonstrated that MTA2 significantly promoted ESCC growth, metastasis, and epithelial-mesenchymal transition (EMT) progression. This integrative analysis combined with expression microarray showed that MTA2 could interact with eukaryotic initiation factor 4E (EIF4E), which positively regulates the expression of Twist, known as a master regulator of EMT. Moreover, the results of chromatin immunoprecipitation revealed that MTA2 was recruited to the E-cadherin promoter by Twist, which reduced the acetylation level of the promoter region and thus inhibited expression of E-cadherin, and subsequently promoted the aggressive progression of ESCC. Collectively, our study provided novel evidence that MTA2 plays an aggressive role in ESCC metastasis by a novel EIF4E-Twist positive feedback loop, which may provide a potential therapeutic target for the management of ESCC.

2584 Background: PD-1 inhibitors have transformed the treatment landscape for patients (pts) with many advanced malignancies. Combination therapy with PD-1 inhibitors for cancer is a trend. However, Biomarkers for the efficacy of combination therapy remains unknown. In order for the benefited population to be screened out, biomarkers need to be established. we will conduct the following study, to explore the IFN-α, IFN-γ, IL-2 combined with TNF-α for predicting efficacy of PD-1 inhibitors combination therapy. Methods: Using postoperative without lesions as control group (n=7). Pts with lesions as the experimental group (n=66). 27 of 66 pts received chemoradiotherapy (group A), 39 of 66 pts received PD-1 inhibitors combined with therapy (group B). IFN-α, IFN-γ, IL-2, TNF-α in peripheral blood of all pts were measured using flow cytometry. Results: 1) There was significant difference in proportion above normal concentrations (ANCs) of IFN-α between two groups (57.1% vs 43.5%, P<0.05), but there was no significant difference in IFN-γ, IL-2 and TNF-α between two groups (IFN-γ 57.1% vs 52.2%, IL-2 14.3% vs 5.8%, TNF-α 42.9% vs 43.5%, P>0.05). 2) The normal ratios of IFN-α, IFN-γ and TNF-α in group B was significantly higher than that in group A (IFN-α 64.1% vs 51.9%, IFN-γ 59% vs 37%, TNF-α 69.2% vs 44.4%, P<0.05). The proportion ANCs of IFN-α, IFN-γ, and TNF-α were lower in group A (IFN-α 35.9% vs 63%, P>0.05; IFN-γ 41% vs 63%, P<0.05; TNF-α 30.8% vs 55.6%, P<0.05). However, the proportion ANCs of IL-2 detection was lower (7.4% vs 5.1%). 3) In group B, 21 of 39 pts were evaluable. ORR was 52.4% (11/21) and DCR was 85.7% (18/21). The proportion ANCs of IFN-α, IFN-γ and TNF-α in the pts with PR was higher than that with SD (IFN-α 37.5% vs 28.6%, IFN-γ 37.5% vs 28.6%, TNF-α 50% vs 38.8%, P<0.05). 4) We found that the coincidence rate of IFN-α+ IFN-γ and IFN-α+ IFN-γ+TNF-α was higher in group B (Table). Conclusions: Our results suggest that the proportion ANCs of IFN-α, IFN-γ, and TNF-α in the pts with lesions were lower than that without lesions, it may be the decrease of immune function with lesions. There was positive correlation between proportion ANCs of IFN-α, IFN-γ and TNF-α and efficacy in these pts. IL-2 was not used as a routine detection indicator. The coincidence rate of IFN-α, IFN-γ combined with TNF-α was higher, it may help predict the outcome of PD-1 inhibitors combination therapy in pts with solid cancers, and helpful to screen the benefit population. Further study is needed.[Table: see text]

目的 探讨食管鳞状细胞癌(ESCC)患者血清IL-24及IL-6的含量及其临床意义.方法 收集55例ESCC患者外周血,ELISA法检测IL-24及IL-6的含量.分析血清IL-24及IL-6含量与患者临床病理特征的关系.结果 ESCC患者血清IL-24含量明显低于正常对照组患者,而IL-6含量明显增高(P<0.01);血清IL-24及IL-6含量与临床分期及淋巴结状态有关(P<0.01),与年龄、性别、瘤组织大小及分化程度均无明显相关(P>0.05).ESCC患者血清IL-24及IL-6含量与患者淋巴结侵犯密切相关.结论 血清 IL-24 及 IL-6 含量测定可作为 ESCC 的辅助诊断指标,为临床治疗提供科学依据.

目的:探讨干扰HMGB1基因对X线照射后食管鳞癌细胞增殖活性、存活能力及侵袭、迁移能力的影响及其机制.方法:构建含有HMGB1 shRNA的慢病毒载体并转染人食管鳞癌EC9706细胞(HMGB1 shRNA组),并以转染阴性序列的细胞为对照组,两组均设置X射线照射和未照射2个亚组.采用Western blot法评估HMGB1基因的干扰效果;MTS法和集落形成实验分别检测食管鳞癌EC9706细胞的增殖活性和存活能力;划痕实验和Transwell小室实验分别检测干扰HMGB1基因对EC9706细胞迁移和侵袭能力的影响;流式细胞术和Western blot分别检测照射后各组的细胞凋亡率及上皮间质转化(EMT)相关蛋白的表达.结果:与阴性对照组比较,Western blot结果显示干扰HMGB1基因明显降低了HMGB1蛋白的表达(P<0.01),MTS法检测结果显示X射线照射后干扰HMGB1显著抑制了食管鳞癌细胞的增殖水平(P<0.05),集落形成实验结果显示HMGB1 shRNA组细胞的放射敏感性明显增加(P<0.01).划痕实验结果显示,X射线照射后HMGB1 shRNA组细胞24 h的划痕愈合率为(20.78±4.38)％,低于阴性对照组的(39.02±3.25)％(P<0.01).Transwell实验结果显示X射线照射后HMGB1 shRNA组细胞在3.5 h的迁移细胞数为(67.00±16.56)个,低于阴性对照组的(194.00±19.74)个(P<0.01).流式细胞术结果显示,X射线照射后阴性对照组细胞凋亡率为(13.64±1.24)％,HMGB1 shRNA组细胞凋亡率为(20.67±1.38)％;与阴性对照组比较,X射线照射后HMGB1 shRNA组细胞E-cadherin表达显著增加,而N-cadherin、Vimentin、MMP-2、MMP-9的表达明显降低(均为P<0.01).结论:干扰食管鳞癌细胞中HMGB1基因的表达后经X射线照射,降低了肿瘤细胞的增殖、侵袭、迁移水平和存活能力,诱导了细胞凋亡,增加了放射敏感性,并调控EMT相关蛋白的表达.

目的:探讨miR-93-5p和miR-106b-5p在酒精性肝硬化肝癌(酒精相关性肝癌)患者中的表达水平及其临床意义.方法:选取酒精相关性肝癌患者穿刺组织蜡块10例,根据性别、年龄配对原则选取10例酒精性肝硬化患者穿刺组织蜡块,采用实时荧光定量PCR(qPCR)检测组织中的miR-93-5p和miR-106b-5p表达水平.另收集河北医科大学第四医院2014年1月1日—2016年12月31日住院治疗的酒精相关性肝癌患者资料71例,治疗前抽取患者外周血,qPCR法检测血清中miR-93-5p和miR-106b-5p的表达水平,分析miR-93-5p和miR-106b-5p表达水平与患者临床病理指标和预后的关系.结果:酒精相关性肝癌患者癌组织中miR-93-5p和miR-106b-5p的表达水平显著高于酒精性肝硬化患者(均P<0.01).miR-93-5p或miR-106b-5p高表达组肿瘤最大径较低表达组显著增加(P<0.01),且临床分期较晚(P<0.01).全组患者中,年龄<60岁组患者生存率显著优于≥60岁组患者(P=0.03);肿瘤最大径<5 cm组患者生存率显著优于≥5 cm组患者(P=0.01);TNM分期Ⅰ+Ⅱ期患者生存率高于Ⅲ期患者(P=0.02);miR-93-5p或miR-106b-5p高表达组患者生存率较低表达组患者显著降低(P≤0.01);年龄和miR-106b-5p表达水平是患者预后的独立影响因素(P值分别为0.02和0.03).结论:miR-93-5p和miR-106b-5p有作为酒精相关性肝癌肿瘤标志物的潜力,治疗前检测其表达水平可以预测患者临床病理指标和预后.

目的:分析胃癌骨髓侵犯患者临床实验室检测结果的特点,筛选对胃癌细胞骨髓侵犯具有提示意义的实验室指标.方法:回顾性分析2013年1月至2021年3月河北医科大学第四医院收治的30例胃癌发生骨髓侵犯患者,收集血常规、凝血功能、免疫、生化及骨髓等临床指标检测结果资料.分析上述患者与未发生骨髓侵犯的Ⅳ期胃癌患者临床实验室检测结果的差异,并制作受试者工作特征(receiver operating characteristic,ROC)曲线评价各指标在提示胃癌细胞骨髓侵犯中的意义.结果:与未发生骨髓侵犯的Ⅳ期胃癌患者相比较,发生骨髓侵犯的患者血小板(Plt)计数、凝血酶原时间(PT)、凝血酶时间(TT)、乳酸脱氢酶(LDH)、D二聚体(D-DIMER)、纤维蛋白原降解产物(FDP)、CEA及CA72-4等指标的检测结果差异均具有统计学意义(均P<0.05).其中FDP的ROC曲线下面积最大(AUC=0.988),且阳性似然比最高.同时,两组患者外周血幼红、幼粒细胞检出率差异具有统计学意义(P<0.01).结论:胃癌骨髓侵犯患者的部分临床实验室检测指标较未发生骨髓侵犯患者异常检出率更高且变化更为明显,上述指标对骨髓侵犯具有提示意义.

目的:比较老年局部晚期食管癌患者采用同步放化疗(CRT)和单纯放疗(RT)的疗效和急性不良反应.方法:回顾性分析2010年1月-2016年1月于河北医科大学第四医院就诊,年龄＞70岁的82例老年局部晚期食管癌患者.比较CRT和RT两个治疗组间的完全缓解率(CRR)、部分缓解率(PRR)、疾病缓解率(DCR)、无进展生存期(PFS)、总生存期(OS)和急性不良反应,并分析食管癌预后的影响因素.结果:CRT组患者的CRR(33.3％)明显高于RT组(14.5％,P=0.049),两组之间PRR和DCR的差异无统计学意义(P=0.058,0.064).CRT组患者的中位PFS及PFS≥1、2、3年的患者比例均显著高于RT组(P=0.007).CRT组患者的中位OS及OS≥1、2、3年的患者比例亦显著高于RT组(P=0.012).单因素分析显示不同治疗方案、CRR是PFS和OS的影响因素.多因素分析显示CRR为PFS的独立预后影响因素,不同治疗方案、CRR是OS的独立预后影响因素(均为P＜0.05).CRT组患者的部分急性不良反应有恶心、呕吐、白细胞减少症、血小板减少症,发生率均高于RT组(均为P＜0.05).结论:同步放化疗可作为老年局部晚期食管鳞癌患者的治疗方案,且疗效优于单纯放疗,但仍需密切关注患者的不良反应.

目的:探讨异黏蛋白(MTDH)和叉头框蛋白M1(FoxM1)在肝细胞癌中的表达及其临床意义.方法:收集156例肝细胞癌(HCC)手术切除标本,免疫组织化学和反转录PCR(RT-PCR)分别检测MTDH、FoxM1蛋白和mRNA在HCC中的表达情况,并分析其表达与临床病理指标及预后的关系.结果:免疫组织化学结果显示,MTDH在HCC中的阳性表达率为71.2％(111/156),FoxM1在HCC中的阳性表达率为64.7％(101/156),其表达与肿瘤数目、TNM分期、微血管侵犯、卫星结节等相关(P<0.05).RT-PCR方法检测显示,MTDH和FoxM1在肝细胞癌中的表达较对照组明显升高(P<0.05).Spearman相关性分析结果显示,MTDH与FoxM1的表达存在明显的相关性(r=0.507,P<0.01).Kaplan-Meier生存分析结果显示肝细胞癌患者生存时间与TNM分期、微血管侵犯、卫星结节、MTDH和FoxM1有关,差异均有统计学意义(P<0.05);多因素Cox回归分析结果显示TNM分期和MTDH的表达是影响患者生存时间的主要因素.结论:对于HCC患者,MTDH和FoxM1与诸多临床因素有关,MTDH为患者预后影响因素,且MTDH和FoxM1的表达存在相关性.

Objective: To investigate the effect and mechanism of p-hydroxycinnamaldehyde(CMSP) momomer compounds extract from the Cochinchina momordica seeds on the growth and metastasis of melanoma transplanted tumors in mice with in vivo and in vitro experiments. Methods: A tumor-bearing mouse model was established, and 18 C57 BL/6 mice were randomly divided into 3 groups(6 mice in each group): a control group(intraperitoneal injection of 0.1 ml normal saline) and two CMSP treatment groups(intraperitoneal injection of 1 and 2 mg/ml CMSP of 0.1 ml). From the 5 thday of drug administration, the transplanted tumor in mice was measured with calipers and the volume was calculated before each administration. At the end of the experiment, the mass of transplanted tumor was weighed. The pathological changes of liver tissues were observed under light microscope after H-E staining. The expressions of E-cadherin and vimentin in the transplanted tumor tissues were detected by SP immunohistochemical method. The metastatic ability of melanoma B16 cells in the CMSP group(10, 20 μg/ml) was detected by cell scratch test and Transwell test after treatment for 24 and 48 h. The expressions of EMT-related m RNAs in B16 cells after treatment with CMSP for 24 h were detected by qPCR. The protein expression levels of β-catenin, p-β-catenin(Ser675), vimentin and E-cadherin in B16 cells treated with CMSP for 48 h were detected by Western blotting(WB). Results: The mean tumor volume and tumor mass of the mice in CMSP treatment group were significantly decreased(all P 0.05), but it was significantly decreased in the 20 μg/ml treatment group(P<0.01). Compared with the control group, the protein expressions of β-catenin, p-β-catenin and vimentin in B16 cells of CMSP groups(10 and 20 μg/ml) were significantly decreased(all P<0.01), while the protein expression of E-cadherin was significantly increased(all P<0.01). Conclusion: CMSP can inhibit the growth and metastasis of melanoma transplanted tumors in mice, and its mechanism may be related to the activity of wnt/β-catenin pathway.

Objective:To examine the effect of FAM83D knockdown on proliferation, survival ability and invasion of human esophageal squamous cell carcinoma after X-ray radiation, and explore the mechanism.Methods:The expression of FAM83D, E-cadherin and vimentin in tumor tissues was detected in 69 cases of esophageal squamous cell cancer by using immunohistochemical method. The siRNA based on the sequences of the FAM83D mRNA were synthesized to transfect into the cultured ECA109 cells as FAM83D shRNA group. The effect of silencing FAM83D gene was evaluated to determine the protein levels of FAM83D in the human oesophageal squamous cell carcinoma ECA109 and KYSE30 cells using western blotting. MTS, clone formation, and Transwell assay were employed to examine the proliferation, survival ability and invasion of ECA109 and KYSE30 cells in vitro, respectively. We used flow cytometry assay to analyze distribution of cell apoptosis in different groups. Western blotting was used to examine the expression of cell metastasis-related molecules and apoptosis-related protein. Results:The strong expression rates of FAM83D, E-cadherin, and vimentin were 55%(38/69), 36%(25/69) and 61%(42/69) in the tumor tissues, respectively. FAM83D protein expression was significantly and negatively correlated with the expression of E-cadherin ( r=-0.350, P<0.01), and positively with the expression of vimentin ( r=0.470, P<0.01). Western blotting results demonstrated that silencing FAM83D gene significantly reduced the FAM83D protein expression ( P<0.01). MTS data demonstrated that FAM83D knockdown after irradiation significantly inhibited the proliferation of esophageal squamous cell carcinoma ECA109 and KYSE30 cells ( P<0.05). The data from the clone formation assay revealed that the radiosensitivity was increased after downragulation of FAM83D expression ( P<0.01). In addition, the invasive abilities of oesophageal carcinoma cells transfected with FAM83D shRNA after irradiation were significantly inhibited compared with those of the NC group ( P<0.01), followed by the downregulation of N-cadherin, vimentin, Snail, p-Akt and p-GSK-3β expression, and the upregulation of E-cadherin expression ( P<0.01). The apoptosis rate of tumor cells in FAM83D shRNA group after irradiation was markedly increased ( P<0.01), followed by a decrease of Bcl-2 and Mcl-1 expression and an increase of Cleaved caspase-3 expression ( P<0.01). Conclusions:FAM83D expressions was found to be closely related to the invasion and development of ESCC. Furthermore, siRNA interference technology inhibited the expression of FAM83D gene in oesophageal squamous cell carcinoma cells, reduced the proliferation, invasion of cells, induced cell apoptosis, and increased radiosensitivity, which may be associated with regulating the epithelial-mesenchymaltransition via Snail/Akt/GSK-3β signaling pathways.

目的:采用siRNA技术干扰人食管鳞癌细胞中高迁移率族蛋白B1(HMGB1)基因表达,观察经X射线照射后细胞增殖活性、存活能力及细胞凋亡率的变化.方法:针对HMGB1 mRNA序列,设计合成有效的干扰序列HMGB1 siRNA,并转染食管鳞癌KYSE30细胞,同时设置转染阴性对照序列的阴性对照组以及未进行转染的空白对照组.采用实时荧光定量PCR(qPCR)和Western blot法检测各组细胞HMGB1 mRNA和蛋白的表达;分别用MTS比色、克隆形成实验、流式细胞术和Western blot法检测HMGB1 siRNA干扰对食管鳞癌细胞KYSE30细胞的增殖活力、存活能力、细胞凋亡率及凋亡相关蛋白表达的影响.结果:HMGB1 siRNA干扰后,KYSE30细胞HMGB1 mRNA和蛋白的表达水平明显降低(P均<0.01);MTS数据显示HMGB1 siRNA干扰后经X射线照射显著抑制了食管鳞癌细胞的增殖水平(与空白对照组和阴性对照组比较,P<0.05);克隆形成实验结果显示空白对照组、阴性对照组、HMGB1 siRNA组的D0值分别为2.57、2.54、1.55 Gy;Dq值分别为1.69、1.65、1.30 Gy;SF2值分别为0.27、0.27、0.13;外推数N值分别为1.93、1.91、2.31;X射线照射后HMGB1 siRNA组肿瘤细胞的凋亡率明显增加,同时bcl-2蛋白表达显著降低,而bax、caspase-3蛋白的表达明显增加(与空白对照组和阴性对照组比较,P<0.01).结论:siRNA干扰技术可用来抑制食管鳞癌细胞中HMGB1基因的表达,联合X射线照射降低了肿瘤细胞的增殖水平和存活能力,诱导了细胞凋亡,增加了放射敏感性,该作用可能与调控bcl-2、bax和caspase-3基因表达有关.

目的 探讨弥漫大B细胞淋巴瘤(DLBCL)骨髓侵犯形态学及免疫表型特点,为临床DLBCL侵犯骨髓的准确诊断提供一定的科学依据.方法 收集2014年6月—2019年6月我院收治的59例DLBCL患者的骨髓组织和外周血,分析DLBCL患者的骨髓细胞形态学、骨髓病理学、流式细胞学、荧光原位免疫杂交及其他相关检测的结果与临床特征.结果 59例发生骨髓侵犯的DLBCL患者多数表现为白细胞增高、贫血、血小板减少,有43例(72.88％)外周血中可见数量不等的瘤细胞.59例患者骨髓涂片中瘤细胞占有核细胞比例≥10％的有31例(52.54％).瘤细胞侵犯骨髓方式以弥漫性增生为主,造血组织明显减少或缺乏.免疫组化染色及流式细胞学检查示,淋巴瘤细胞CD20、CD19、CD79b及cCD79a均呈阳性表达,部分患者表达HLA-DR、CD22、sIgM、CD43、CD10、FMC7及CD123,均不表达CD38、TdT、CD103、CD25、CD3、CD2、MPO、CD33、CD13、CD7及CD56.12例患者进行FISH检测均未见C-myc基因重排.结论 骨髓形态学、病理学并结合免疫表型检测是DLBCL患者准确诊断的重要依据.

目的:观察多发性骨髓瘤(MM)患者骨髓中浆细胞免疫表型特点,及其与患者临床预后的相关性.方法:收集35例MM患者资料,骨髓涂片及切片分别经瑞氏-吉姆萨和HE染色后,观察分析骨髓细胞形态及病理学特点.同时,取患者骨髓或外周血分别进行流式细胞术检测及相关实验室检查.结果:MM患者骨髓浆细胞不仅数量明显增多,且形态明显异常,切片中浆细胞分布类型包括塞实型16例(45.71％),结节型14例(40.00％)和间质型5例(14.29％).MM患者中出现血清LDH、β2-MG、Cr和BUN等检测指标升高的比例分别占80％、85.71％、62.86％和60％.MM患者浆细胞CD38、CD138均呈阳性表达,CD56、CD200、CD117和CD28阳性患者比例分别为65.71％、94.29％、60％和54.29％,CD20、CD81、CD33、CD27及CD13阳性患者比例分别为37.14％、42.86％、28.57％、57.14％和34.29％,而CD19及CD45阳性患者比例分别为22.86％和31.42％,且Igκ或Igλ链表达缺失呈单克隆增生.其中CD27和CD28的表达情况与MM患者预后密切相关,浆细胞CD27表达缺失及CD28阳性表达的患者5年总生存率较低(P<0.05).结论:MM患者骨髓中浆细胞免疫表型明显异常,其中CD27和CD28的表达与患者病情进展和临床预后具有显著相关性,因此免疫表型检测在MM患者临床诊断、治疗监测中具有重要意义.

目的:研究miR-429和Bmi-1 mRNA在弥漫性大B细胞淋巴瘤(DLBCL)组织中的表达及其临床意义.方法:选取自2014年1月—2015年12月河北医科大学第四医院的45例DLBCL患者瘤组织和28例正常对照人群淋巴结组织,用实时荧光定量反转录PCR(RT-qPCR)技术检测组织中miR-429及Bmi-1 mRNA的表达水平,并分析二者之间的关系及其与患者临床病理指标之间的关系.结果:DLBCL组织中miR-429表达水平较正常淋巴结组织明显降低,而Bmi-1 mRNA的表达水平明显升高(P<0.01).miR-429低表达组和Bmi-1 mRNA高表达组患者临床分期、骨髓浸润程度、Ki-67阳性率和IPI积分均分别高于miR-429高表达组和Bmi-1 mRNA低表达组(P<0.05).结论:DLBCL组织中miR-429及Bmi-1 mRNA的表达水平与患者病情进展和预后密切相关.组织中低表达miR-429及高表达Bmi-1 mRNA可能是患者临床预后较差的预测指标.

目的:观察输注的异体血小板对人肺癌细胞A549侵袭、转移的影响,初步探讨其作用机制。方法:选取2017年1月至2018年12月于河北医科大学第四医院化疗科就诊输注血小板的89例晚期肺癌患者,实验分为Ctrl组（与培养液共孵育的A549细胞组）、Before组和After组（分别指与输注血小板前和后患者血浆共孵育的A549细胞组）。通过划痕实验和Transwell实验检测与输血小板前后患者血浆共孵育的A549细胞的迁移和侵袭能力,采用Western blotting法检测金属基质蛋白酶（MMPs）及其抑制剂（TIMPs）和上皮-间质转化（EMT）相关蛋白E-cadherin、N-cadherin及Vimentin,以及血管内皮生长因子VEGF及受体2（VEGFR2）的表达水平。结果:After组的A549细胞的划痕愈合率明显高于Before组及Ctrl组[（73.67±2.60）%vs（58.33±2.33）%、（35.33±2.03）%,P＜0.01或P＜0.05],Before组与Ctrl组比较也具有显著差异（P＜0.05）。细胞迁移实验结果显示,After组的穿膜细胞数明显高于Ctrl组和Before组[（69.67±7.84）vs（18±2.08）、（39.33±2.03）个,均P＜0.01]。细胞侵袭实验显示,After组的穿膜细胞数明显高于Ctrl组和Before组[（59.34±3.46）vs（18.34±1.56）、（37.58±2.79）个,均P＜0.01]。A549细胞与输注血小板前、后的血浆共孵育48 h后,MMP9、MMP2的表达均升高（P＜0.05）而其抑制剂TIMP1和TIMP2的水平均下降（P＜0.01）;EMT相关蛋白N-cadherin、Vimentin表达升高（P＜0.05）而E-cadherin表达降低（P＜0.01）;血管形成相关蛋白VEGF、VEGFR2的表达均升高（P＜0.05）。结论:输注异体血小板能促进肺癌A549细胞的侵袭和转移,其作用机制可能与调节EMT、金属基质蛋白酶及血管生长因子相关蛋白的表达有关。

目的:研究环磷腺苷效应元件结合蛋白(CREB)在食管鳞状细胞癌(ESCC)组织中的表达及其与患者临床特征、预后之间的关系.方法:选取2010年1—12月在河北医科大学第四医院就诊的60例ESCC患者癌组织及癌旁组织,采用Western blot及免疫组化技术检测组织中CREB蛋白的表达水平,并分析其与临床病理指标及患者预后之间的关系.结果:ESCC组织中CREB表达水平较癌旁组织明显升高(P<0.01).组织中CREB高表达的患者往往瘤组织较大、临床分期较晚、更易发生淋巴结转移,同时患者5年总生存率较低而累积复发率较高(P<0.05).结论:ESCC组织中CREB表达水平明显升高,且高表达CREB的患者往往提示临床进展及较差预后.因此,CREB具有成为ESCC相关肿瘤标志物的潜力.