肝细胞癌组织miR-124-3p、miR-212-5p表达与PI3K/Akt信号通路和预后的关系研究

摘要 目的：探讨肝细胞癌组织中微小核糖核酸（miR）-124-3p、miR-212-5p表达与磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）信号通路和预后的关系。方法：选择2017年9月至2019年9月徐州医科大学附属医院肝胆外科收治的93例肝细胞癌患者,采用实时荧光定量聚合酶链反应（qRT-PCR）检测肝细胞癌组织中miR-124-3p、miR-212-5p、PI3K 信使RNA（mRNA）、Akt mRNA的表达。Pearson相关性分析miR-124-3p、miR-212-5p表达与PI3K/Akt信号通路相关mRNA表达的相关性。绘制Kaplan-Meier生存曲线,Log-Rank检验不同miR-124-3p、miR-212-5p表达肝细胞癌患者3年总生存率（OS）的差异。结果：肝细胞癌组织中miR-124-3p表达低于癌旁组织（P＜0.05）,miR-212-5p、PI3K mRNA、Akt mRNA表达高于癌旁组织（P＜0.05）。肝细胞癌组织中miR-124-3p表达与PI3K mRNA、Akt mRNA表达呈负相关（P＜0.05）,miR-212-5p表达与PI3K mRNA、Akt mRNA表达呈正相关（P＜0.05）。Ⅱa期、低分化患者肝细胞癌组织中miR-124-3p表达低于Ⅰa-Ⅰb期、中高分化患者（P＜0.05）,miR-212-5p表达高于Ⅰa-Ⅰb期、中高分化患者（P＜0.05）。随访期间死亡37例, miR-124-3p低表达组3年OS为42.55%,低于miR-124-3p高表达组的78.26%（P＜0.05）,miR-212-5p高表达组3年OS为50.00%,低于miR-212-5p低表达组的71.11%（P＜0.05）。结论：肝细胞癌组织中miR-124-3p表达下调,miR-212-5p表达上调,且与肝细胞癌PI3K/Akt信号通路激活,PI3K mRNA和Akt mRNA高表达,低分化,CNLC分期Ⅱa期以及低OS有关。     

超声造影定量分析联合血清AFP、VEGFR-2、sTim-3对原发性肝癌TACE治疗疗效的预测价值

摘要 目的：探讨超声造影定量分析联合血清甲胎蛋白（AFP）、血管内皮生长因子受体-2（VEGFR-2）、可溶性T细胞免疫球蛋白粘蛋白分子3（sTim-3）对原发性肝癌肝动脉化疗栓塞（TACE）治疗疗效的预测价值。方法：选择2020年1月至2022年10月海军军医大学第二附属医院收治的原发性肝癌患者94例。行TACE治疗2个月,采用改良的实体瘤疗效评价标准评估患者疗效,根据不同疗效分为疗效不良组（n=32）和疗效良好组（n=62）。所有患者均行超声造影检查,比较两组超声造影定量分析参数、治疗前血清AFP、VEGFR-2、sTim-3水平,采用受试者工作特征（ROC）曲线分析超声造影定量分析参数联合血清AFP、VEGFR-2、sTim-3对原发性肝癌TACE治疗疗效的预测价值。结果：经TACE治疗,62例患者疗效良好、32例患者疗效不良,治疗有效率为65.96%。疗效良好组术前超声造影达峰时间、等增强开始时间显著长于疗效不良组（P<0.05）。疗效良好组术前血清AFP、VEGFR-2、sTim-3水平显著低于疗效不良组（P<0.05）。ROC曲线分析结果显示,超声造影定量分析联合血清AFP、VEGFR-2、sTim-3对原发性肝癌TACE治疗疗效预测的曲线下面积（AUC）为0.950,灵敏度为84.85%,特异度为82.12%,高于各指标单独检测。结论：超声造影定量分析参数、血清AFP、VEGFR-2、sTim-3水平可预测原发性肝癌患者TACE治疗的疗效,且联合诊断的预测效能更高。     

结直肠癌P53蛋白定量分析及其与肿瘤生物学行为的关系

应用MD-20显微图象分析仪对30例结直肠癌的P53蛋白免疫组织化学反应产物进行定量测定,以观察P53蛋白相对含量与肿瘤生物学行为的关系.结果显示:结直肠癌P53蛋白MOD值与PCNA增殖指数呈正相关(P<0.05).P53蛋白MOD值大的肿瘤多里浸润性生长方式,且浸润至浆膜外者多(PM<0.01,P<0.05).淋巴结转移与MOD值无明显关系(P>0.05).结果提示:结直肠癌P53蛋白相对含量对肿瘤的生物学行为有重要影响.     

放射治疗对鼻咽癌患者细胞免疫功能的影响

应用抗人淋巴细胞单克隆抗体检测３２例鼻咽癌患者放疗前、后外周血Ｔ淋巴细胞亚群,并与１２例非癌人群（对照组）进行对比。结果：鼻咽癌患者ＯＫＴ３、ＯＫＴ４及ＯＫＴ４／ＯＫＴ８值与对照组相比明显降低,ＯＫＴ８显著上升（Ｐ＜０．０５）;放疗后ＯＫＴ３、ＯＫＴ４及ＯＫＴ４／ＯＫＴ８值进一步下降,ＯＫＴ８进一步升高（Ｐ＜０．０１）。提示鼻咽癌患者细胞免疫功能低下,放射治疗可进一步抑制鼻咽癌患者的细胞免疫功能。     

湖北地区宫颈癌组织中人乳头瘤病毒16型E7基因的分离、克隆和序列分析

采用加端聚合酶链反应技术,从湖北地区一宫颈癌患者癌组织ＤＮＡ中分离出人乳头瘤病毒１６型（ＨＰＶ１６）Ｅ７基因,并在ｐＵＣ１８载体中克隆。经限制性核酸内切酶分析和ＤＮＡ序列分析,确认了含ＨＰＶ１６Ｅ７重组克隆质粒,命名ｐＨＰＶ１６Ｅ７─ＨＢ。ＤＮＡ序列分析表明,ＨＰＶ１６Ｅ７─ＨＢ基因全长２９４ｂｐ（与报道的标准株基因长度相同）,但其核苷酸顺序中有两处发生了Ｃ→Ｔ突变,即第４３位密码子ＣＡＡ变为ＴＡＡ,第７６位ＣＧＴ变为ＴＧＴ;前者使谷氨酰胺密码子变为终止密码,即无义奕变（ｎｏｎｓｅｎｓｅｍｕｔａｔｉｏｎ）。这种突变发生在２９４个碱基的ＤＮＡ扩增产物之中,不像是ＰＣＲ本身的错配,而很可能是湖北株与标准株之间的结构差异。     

5’核苷酸酶碱性磷酸酶双重染色法在毛细淋巴管研究中的应用

本文应用5’-N-ALP双重染色法观察了裸鼠皮肤及人胃癌组织内淋巴管的形态和细微分布.在光镜下毛细淋巴管、淋巴管呈5’-N强阳性反应,管壁显示明显的棕色或深棕色,而毛细血管、血管的ALP呈强阳性,管壁呈明显的蓝色.据此可用组化方法将毛细淋巴管和毛细血管区别开来.本法能显示呈褐色的毛细淋巴管,特别是呈实性条索状的毛细淋巴管,因而双重染色比HE染色更能客观、准确地显示毛细淋巴管的分布.     

The Expression of Two Splice Variants of Metabotropic Glutamate Receptor Subtype 5 in the Rat Brain and Neuronal Cells During Development

Abstract: We previously reported that a variant with extra amino acid residues exists in the metabotropic glutamate receptor subtype 5 (mGluR5). Either of the two isoforms, named mGluR5b and mGluR5a for the isoforms with and without the inserted sequence, respectively, generated Ca²⁺-activated Cl⁻ current when expressed in Xenopus oocytes. We herein report that these two isoforms are produced by the alternative splicing of the exon skipping type. When examined during the course of postnatal development, the major mGluR5 isotype mRNA was observed to switch from mGluR5a to mGluR5b in the rat hippocampus and the cerebral cortex. We also investigated two cell lines that could be differentiated into neuron-like cells in vitro. Whereas the mGluR5b mRNA was hardly detectable in either undifferentiated or differentiated NG108-15 cells, the relative amounts of the two variant mRNAs changed after the induction of differentiation in the P19 cells. An extracellular application of trans - d,l -1-amino-1,3-cyclopentanedicarboxylate on the neuron-like P19 cells induced intracellular Ca²⁺ mobilization, thus suggesting that the cells could express functional mGluR(s) coupled to phospholipase C and other components that could mediate the signal transduction pathway. This cell line may thus provide a model system for studying both mGluR5 expression and other mGluR-induced phenomena at the molecular level.     

Extracellular matrix components and proteolytic enzymes in uterine cervical carcinoma

The important components of mucopolysaccharides and collagen have been analyzed in tissues of control and carcinoma of uterine cervix. Among these components hyaluronic acid and chondroitin sulphate levels were found to be increased, whereas decreased level of collagen was observed in uterine cervical carcinoma. Serum cathepsin B, D and acid and alkaline phosphatases have also been analyzed in controls and carcinoma patients before and after treatments. The activities of these enzymes have been found to increase prominently in advanced stages. Among these enzymes cathepsin B and alkaline phosphatase have exhibited remarkable increase in activity in uterine cervical carcinoma. Different modes of treatment exerted reversion of the elevated activities of these enzymes. However, combined therapy type II (radiation combined with cisplatin and cyclophosphomide) seems to be more effective in reverting the activities of these enzymes to normal levels.     

Functional and genetic analysis of annexin VI

This study is concerned with the determination of the function of the 68kDa calcium-binding protein, annexin VI. Studies on the structure and regulation of the gene include a detailed analysis of annexin VI expressed heterologously in human A431 carcinoma cells. We have recently discovered that annexin VI is subject to a novel growth dependent post-translational modification. Interestingly, the protein exerts a negative effect on A431 cells. This effect was manifested as a partial reversal of the transformed phenotype. We are currently exploring the hypothesis that the post-translational modification of annexin VI is required for sub-cellular targeting, and that correct localisation within the cell is essential for function.     

Therapy of bovine ocular squamous-cell carcinoma with local doses of interleukin-2: 67% complete regressions after 20 months of follow-up

We have tested the therapeutic potency of peritumorally injected low doses of interleukin-2(IL-2). Seventy tumours of the bovine ocular squamous-cell carcinoma (BOSCC), 1–3 cm in diameter, were treated with 5000, 20 000 or 200 000 U IL-2 from Eurocetus (Chiron) to find the optimal dose for treatment. Injections were given peritumorally on Monday to Friday on 2 consecutive weeks. The size of the tumours was measured before treatment and 1, 3, 4, 9 and 20 months after treatment. After 9 months complete regression was observed in 89% of the tumours treated with 5000 U IL-2, 80% treated with 20 000 U and 67% treated with 200 000 U. After 20 months, there was complete regression of 35%, 31% and 67% of the tumours respectively. The 9-and 20-month results of the 200 000-U treatment are significantly better than those of the 5000-U and 20 000-U treatments taken together. This protocol may be useful to treat advanced inoperable tumours (e.g. of the nasopharynx or skin) of human patients.