[摘 要] 目的：探讨胶原三螺旋重复蛋白1（CTHRC1）在膀胱癌组织和细胞中的表达及其对膀胱癌5637细胞迁移和侵袭的影响及其机制。方法：利用TCGA和Arrayexpress数据库中膀胱癌基因表达数据，分析CTHRC1转录和翻译水平。收集2014年9月至2020年12月重庆医科大学附属第一医院手术切除的144例膀胱癌组织和25例全膀胱切除的癌旁组织标本，以及人膀胱癌细胞RT4、5637、T24、UMUC-3、TCCSUP和输尿管上皮永生化细胞SV-HUC-1。采用免疫组织化学染色法、qPCR法和WB法检测膀胱癌组织和细胞中CTHRC1的表达水平，通过Kaplan-Meier曲线分析CTHRC1表达对总生存期（OS）的影响。运用RNAi技术，敲降5637细胞CTHRC1表达后，通过细胞划痕实验和Transwell实验检测CTHRC1表达下调对5637细胞迁移和侵袭的影响。利用基因集富集分析（GSEA）预测CTHRC1相关的潜在信号通路，WB法检测敲降CTHRC1表达对FAK-ERK1/2通路相关蛋白表达的影响。结果：CTHRC1的转录和翻译水平在肌层浸润性膀胱癌（MIBC）组织和细胞中表达显著上调（均P<0.05），CTHRC1高表达组患者5年OS较低表达患者缩短（P<0.05）。干扰CTHRC1表达后，膀胱癌5637细胞迁移及侵袭能力均显著降低（均P<0.01）。GSEA预测显示，CTHRC1高表达组主要富集在黏着斑激酶（FAK）、肌动蛋白细胞骨架调节、FAK和ERK1/2信号通路。WB法实验结果表明，重组CTHRC1蛋白促进膀胱癌5637细胞FAK-ERK1/2信号通路活化（P<0.05或P<0.01）。结论：CTHRC1在MIBC中表达上调，且与膀胱癌患者不良预后密切相关；CTHRC1促进膀胱癌细胞迁移和侵袭，该过程可能与FAK-ERK1/2信号通路的激活有关。

Adolescent ; Female ; Humans ; Sacrum ; pathology ; surgery ; Spondylolysis ; diagnosis ; pathology ; surgery

[摘 要] 目的：探讨CD68+ 肿瘤相关巨噬细胞（TAM）、CD8+ T细胞、Foxp3+ Treg细胞等在肺腺癌（LUAD）组织中浸润分布及其与患者预后的关系。方法：收集2004年9月至2009年4月间在苏州大学附属第三医院手术切除的93例LUAD组织及78例癌旁组织，采用组织芯片（TMA）及多重免疫荧光（mIF）技术检测其中的免疫细胞浸润与分布，Wilcoxon秩和检验比较癌与癌旁组织、癌巢与间质中浸润水平的差异，χ2检验分析其浸润水平及CD8+/CD68+细胞比值与临床病理特征的关系，Kaplan-Meier法和COX模型分析影响患者OS的潜在危险因素。结果：与癌旁组织比较，癌组织中CD68+ TAM、CD8+ T细胞、Foxp3+ Treg细胞浸润水平均显著增加（均P<0.01），间质CD68+ TAM、CD8+ T细胞的浸润水平均显著高于癌巢（均P<0.01）。总CD68+ TAM、癌巢及间质CD68+ TAM浸润水平与淋巴结转移呈正向关联（均P<0.05），癌巢CD68+ TAM浸润水平与T分期呈正向关联（P<0.05），间质CD68+ TAM浸润水平与病理分级呈正向关联（P<0.05）；癌组织中CD8+/CD68+细胞比值与病理分级、淋巴结转移均呈负向关联（均P<0.05）。Kaplan-Meier生存分析显示，LUAD组织中总CD68+ TAM、癌巢及间质CD68+ TAM高浸润患者OS均短于低浸润患者（P<0.05或P<0.01）、癌组织中CD8+/CD68+细胞比值高患者OS显著长于低比值患者（P<0.05）。多因素COX模型分析示，LUAD患者年龄、TNM分期及癌组织中CD8+/CD68+ 细胞比值是影响患者预后的独立风险因素（P<0.05或P<0.01）。结论:高度浸润的CD68+ TAM与LUAD的进展、侵袭、转移和不良预后显著关联，而高CD8+/CD68+ 细胞比值是影响LUAD患者OS的独立保护因素。

<b>Objectiveb> To evaluate the curative effect ofeliminating flatulence and laxative cream on patients with advanced hepatocellular carcinoma and opioid-associated constipation（OIC）. <b>Methodsb> 120 patients with advanced liver cancer complicated with OIC who were treated at the Third Affiliated Hospital of Naval Medical University from June 2021 to December 2022 were selected as the study subjects. The patients were divided into a control group（lactulose+conventional treatment）and an experimental group（eliminating flatulence and laxative cream + conventional treatment）using a randomized numerical table method. Two groups were compared in terms of defecation, quality of life, and comprehensive post-treatment evaluation（economic cost, number of occurrences of diarrhea, and whether or not there was a change in the dosage of opioids used）. <b>Resultsb> After 28 d of intervention, both groups showed better results in relieving OIC（P<0.05）, and the experimental group was significantly better than the control group in terms of the quality of life of patients with advanced hepatocellular carcinoma, the economic cost and the number of diarrhea（P<0.05）. <b>Conclusionb> In the treatment of OIC in patients with advanced hepatocellular carcinoma, constipation could be relieved by using both topicaleliminating flatulence and laxative cream and oral lactulose solution. Among them, antieliminating flatulence and laxative cream was more acceptable to patients and superior in terms of quality of life and economic cost, which could be a better choice for improving patient satisfaction and safety.

[摘 要] 目的：探究炎症标志物对晚期非小细胞肺癌（NSCLC）患者抗PD-1治疗疗效及安全性的预测价值。方法：动态监测2018年1月至2020年12月在海军军医大学第一附属医院接受抗PD-1治疗的222例晚期NSCLC患者的血清炎症标志物，应用ROC曲线计算最佳截断值并将炎症指标分为高和低水平两组。以Log-Rank检验和Kaplan-Meier法分析患者临床病理特征及各炎症标志物水平与患者预后的关系，单因素和多因素Cox回归分析估算PFS和OS的风险比。Fisher精确检验分析各炎症标志物基线水平高和低两组与免疫相关不良反应（irAE）的相关性，Wilcoxon秩和检验比较治疗前（基线）与首次PR、PD时及发生irAE时各炎症标志物水平的差异。结果：血清炎症标志物NLR、MLR、PLR、LDH、CRP和IL-6基线水平升高与患者的PFS和OS显著缩短有关（圴P<0.01）。多因素分析结果显示，基线升高的PLR、MLR和LDH是PFS和OS的独立危险因素（P<0.05或P<0.01）。NLR、LDH、CRP和IL-6水平在患者首次获得PR时显著降低（P<0.05或P<0.01），LDH、CRP、IL-6和TNF-α水平在出现PD时显著升高（P<0.05或P<0.01）。发生irAE时，LDH、CRP、IL-6、IL-10和TNF-α水平与基线水平相比显著升高（P<0.05或P<0.01）。结论：晚期NSCLC抗PD-1治疗中炎症标志物水平的下降提示患者病情改善，其水平的上升则提示病情进展且与irAE的发生相关，动态监测炎症标志物可以预测抗PD-1治疗的疗效及安全性，有助于筛选抗PD-1治疗的最佳晚期NSCLC人群。

[摘 要] 目的：探讨过氧化物酶体膜蛋白4（PXMP4）对肝细胞癌（HCC）细胞迁移和侵袭及上皮间质转化（EMT）进程的影响。方法:采用生物信息学和免疫组织化学方法分析HCC组织中PXMP4的表达，分析其与患者临床病理特征的相关性。在HCCLM3和MHCC97H细胞中干扰PXMP4，在Huh7和MHCC97L细胞中过表达PXMP4，利用WB和qPCR法验证干扰/过表达效率。利用CCK-8、划痕愈合实验和Transwell侵袭实验检测干扰/过表达PXMP4对HCC细胞增殖、迁移和侵袭能力的影响，采用WB法检测干扰/过表达PXMP4或0、5、10和20 μmol/L U0126处理对HCC细胞中N-cadherin、E-cadherin、vimentin、细胞外信号调节激酶（ERK）、p-ERK表达的影响。结果：生物信息学和免疫组织化学分析显示，HCC组织中PXMP4呈高表达（P<0.05）。临床病理分析发现，PXMP4的表达与肿瘤分化程度有关联（P<0.05）。在HCC细胞中，干扰PXMP4后抑制细胞增殖、侵袭和EMT进程（P<0.05）；反之，过表达PXMP4后促进HCC细胞增殖、侵袭及EMT进程（P<0.05）。此外，PXMP4通过激活ERK1/2信号通路促进HCC细胞EMT进程，U0126处理同样能够抑制HCC细胞EMT进程。结论：PXMP4在HCC组织中呈高表达且与HCC细胞分化有关联，PXMP4通过激活ERK1/2信号通路促进HCC细胞EMT进程。

[摘 要] 目的：探究敲减间质表皮转化因子（MET）表达对人喉鳞状细胞癌（LSCC）Hep-2细胞生长、迁移及5-氟尿嘧啶（5-FU）和顺铂敏感性的影响。方法：通过基因表达综合数据库（GEO）及癌症基因组图谱（TCGA）数据库数据分析人LSCC组织中MET mRNA的表达水平；常规培养正常人支气管上皮细胞16HBE与人LSCC细胞Hep-2、KBV200和TU212，采用qPCR法和WB法检测16HBE、Hep-2、KBV200和TU212细胞中MET基因和蛋白的表达水平。用LipofectamineTM 3000将MET敲减质粒（si-Met）和对照质粒（si-NC）转染至Hep-2细胞中，分为空白对照组，si-NC组和si-Met组。采用MTT法、流式细胞术、划痕愈合实验分别检测各组Hep-2细胞的增殖、迁移能力、周期分布以及对5-FU和顺铂的敏感性。结果：数据库数据分析显示LSCC组织中MET mRNA呈高表达（P < 0.05），Hep-2、KBV200和TU212细胞中MET mRNA和蛋白的表达水平也均明显高于16HBE细胞（均P < 0.01）。敲减MET表达后，Hep-2细胞中MET mRNA及蛋白表达水平均明显降低（P < 0.01或P < 0.001）、细胞增殖活力显著下降（P < 0.000 1）、G0/G1期细胞数量明显升高（P < 0.000 1）、S期细胞数量明显降低（P < 0.000 1）。敲减MET表达后，不同浓度5-FU或顺铂对细胞增殖的抑制率均显著升高、药物半数抑制浓度（IC50）均降低（均P < 0.000 1），划痕愈合率明显降低、迁移能力下降（均P < 0.05）。结论：MET在人LSCC组织和细胞中呈高表达，敲减MET可有效抑制Hep-2细胞中MET的表达，抑制细胞增殖、迁移能力，使其周期阻滞于G1期，增强Hep-2细胞对5-FU和顺铂的敏感性。

<b>OBJECTIVEb>This study was aimed to characterize the Helicobacter pylori strains isolated from different geographic regions in China and different ethnic groups in Yunnan province in terms of cagA, iceA, vacA and HP0519 genes which were proposed to be related to the pathogenesis.

<b>METHODSb>150 Helicobacter pylori strains were collected from Yunnan province, Fujian province and Beijing. Chromosome DNA was extracted and polymerase chain reaction (PCR) was carried out to determine the 3' region of cagA, iceA, vacA and HP0519 status with specific primers. PCR results were analyzed statistically according to their isolated original and clinical outcomes.

<b>RESULTSb>For cagA 3' region, 93% (139/150) of the Chinese Helicobacter pylori strains belonged to East Asian type according to the specific primer of TF/JR. Among the 150 strains, 75% (113/150) belonged to iceA1, and 19% (29/150) to iceA2. The dissemination of iceA was not associated with any of the geographic regions, different ethnic groups or different clinical outcomes. 96% (144/150) of the vacA s region belonged to s1. In the vacA middle region, m2, m1b, m1b-m2 were 57% (85/150), 27% (41/150) and 11% (16/150) respectively. However, m1a was only observed in two strains from Fujian. Neither vacA s1 nor m2 showed significant difference between Yunnan, Fujian and Beijing. However, the distribution of mlb-m2 in Yunnan was higher than that in Fujian and Beijing. In Yunnan province, the distribution of vacA s1 was not associated with different ethnic groups but m2 from Bai group was less than other two ethnic groups. The ratio of m1b in Bai group was higher than that in other groups. Both vacA' s region and m region alleles had no significant relationship with the clinical outcomes. With the 15 bp and 24 bp DNA insertion and deletion primers test, 93% (140/150) of the strains were positive. The distributions of the 15 bp and 24 bp DNA insertion or deletion were different according to the different ethnic groups.

<b>CONCLUSIONb>By JF/TR primer, 93% of the Chinese strains cagA's 3' region belonged to East Asian type. Most of the Chinese strains vacA's allele was s1. The distribution of vacA s1 had no relationship with the clinical outcome of the isolates. From different geographic regions and ethnic groups, the distribution of vacA m region allele was different. 93% of the Chinese strains HP0519 genes had 24 bp or 15 bp insertion or deletion character. The biological meaning of the polymorphism of HP0519 needs advanced investigation.

China ; Genes, Bacterial ; genetics ; Helicobacter Infections ; ethnology ; genetics ; Helicobacter pylori ; classification ; genetics ; isolation & purification ; Humans ; Polymerase Chain Reaction

[Abstract] Objective: To explore the expression and regulation mechanism of Dickkopf-1 (DKK1) in head and neck squamous cell carcinoma (HNSCC) tissues. Methods: Based on the TCGA database, the relationship of DKK1 expression in HNSCC tissues and its methylation site with patients’prognosis was analyzed. GO and KEGG gene enrichment method were used to analyze the signaling pathways of DKK1 enrichment. STRING was used to analyze the interaction between DKK1 protein and other proteins. TargetScan was used to analyze the miRNAs that regulate the expression of DKK1, and the transcription factors of DKK1 were analyzed with the TRRUST website. Results: DKK1 gene was highly expressed in HNSCC tissues (P<0.01), and its expression level was significantly correlated with the HPV status, age, pathological grade, and clinical stage of patients (all P<0.05); the prognosis of HNSCC patients with high DKK1 expression was poorer than those with low DKK1 expression (P<0.01). There were 19 methylation sites in DKK1, 12 of which were significantly different between cancer tissues and normal tissues (P<0.05), and 11 sites were significantly related to the prognosis of HNSCC (P<0.05). In addition, miRNA, circRNA, lincRNA and transcription factors, etc. also participated in the regulation of DKK1. A total of 5 DKK1-related PPI networks that may involve in the occurrence, development, invasion and metastasis of HNSCC were obtained. Conclusion: DKK1 is highly expressed in HNSCC tissues and is a risk factor for poor prognosis of HNSCC patients. DKK1 plays an important role in the pathogenesis of HNSCC and is expected to become a potential target for HNSCC treatment.

[Abstract] Objective： ： To explore miR-103a-3p expression in the tumor tissues and the serum of breast cancer patients, and its role and mechanism in breast cancer development. Methods: Pathologically confirmed 31 cases of tumor tissues and 21 cases of para-cancerous tissues resected at Department of Oncological Surgery of the Second Affiliated Hospital of Hainan Medical University (Haikou, China) from March 1, 2017 to August 31，2017 were collected for this study; in addition, serum samples from 38 breast cancer patients and 22 healthy subjects as well as the breast cancer cell lines MCF-7 and MDA-MB-231 were used in this study. pHBLV-U6-Luc-T2A-Puro and PLL3.7 lentivirus were applied to knock down miR-103a-3p and PDK4 in MCF-7 and MDA-MB-231 cells, respectively. qPCR and Western blotting were performed to examine the mRNA and protein expressions of miR-103a-3p and PDK4 in tissues and serums of breast cancer patients as well as the in cell lines, respectively; CCK-8 assay was applied to detect the proliferation of MCF-7 and MDAMB-231 cells; Olympus AU5400 was applied to detect the glucose consumption and lactate production in indicated cell line. Results： ： miR-103a-3p was significantly decreased in tumor tissues compared with the paracancerous tissues (P<0.01). miR-103a-3p knockdown activated the glucos consumption and lactate production (all P<0.01), increased the PKD4 expression (P<0.01) in MCF-7 and MDAMD-231 cells, and promoted the proliferation of MCF-7 and MDA-MB-231 cells (P<0.01). Furthermore, knockdown of PDK4 suppressed the glucose consumption, lactate production and proliferation in MCF-7 and MDA-MB-231 cells with miR-103a-3p silencing (all P<0.01). Conclusion： ：In the breast cancer, miR-103a-3p inhibited the proliferation of breast cancer cells through down-regulation of PDK4 and PDK4-mediated aerobic glycolysis.