Immunohistochemical and molecular analysis of PI3K/AKT/mTOR pathway in esophageal carcinoma

Among the numerous signaling pathways involved in tumorigenesis, PI3K‐AKT‐mTOR is a key one that regulates diverse cellular functions. However, its prognostic value in esophageal carcinoma remains unclear. In our study, we examined the immunohistochemical expression of phosphorylated (p‐) AKT, mTOR, p70S6K and 4E‐BP1 along with the mutational status of PIK3CA and AKT1 genes by High Resolution Melting Analysis and Pyrosequencing in 44 esophageal carcinomas. The results were correlated with the clinicopathological characteristics of the patients in an effort to define their possible prognostic significance. Total p‐mTOR cytoplasmic expression, assessed in 10 random areas, was positively correlated with tumor stage (Kruskal–Wallis ANOVA, I/II vs III/IV, p = 0.0500). Μoreover, maximum p‐mTOR cytoplasmic immunoexpression, estimated in hot spot areas, was positively associated with tumor grade (Mann–Whitney U test, I/II vs III, p = 0.0565). Interestingly, p‐4E‐BP1 immunoreactivity was negatively correlated with tumor histological grade (Mann–Whitney U test, I/II vs III, p = 0.0427). No mutation was observed in exons 9 and 20 of PIK3CA gene and in exon 4 of AKT1 gene. In conclusion, our findings depict the presence of activated PI3K/AKT/mTOR pathway in esophageal cancer bringing forward p‐mTOR and p‐4E‐BP1 for their potential role in esophageal carcinogenesis. Additional studies are warranted to validate our findings.     

临床护理路径对脑出血偏瘫患者康复效果的影响

目的 探究与分析临床护理路径对脑出血偏瘫患者康复效果的影响.方法 选取本院自2013年7月至2015年7月收治的90例脑出血偏瘫患者,按照就诊时间顺序分为常规护理组与临床护理路径组,每组45例.常规护理组给予基础护理对策,包括入院时宣教、介绍病情、观察病情变化等;临床护理路径组指派至少由1名护士长及3名以上护理人员建立临床护理路径小组,由护士长组织组内人员系统性的学习并加强脑出血偏瘫后康复护理的相关知识,根据每位患者的个体情况给予评估,并对每个阶段后的康复护理效果给予评价.对比两组患者康复护理效果、活动能力、肢体功能及负面情绪评分.结果 临床护理路径组的护理效果总有效率明显高于常规护理组,差异具有统计学意义(P＜0.05).两组患者治疗后较治疗前相比Barthel评分及Harris评分均明显升高,临床护理路径组较常规护理组相比Barthel评分及Harris评分升高更加明显,差异具有统计学意义(P＜0.05).两组患者治疗后较治疗前相比HAMA评分及HAMD评分评分均明显降低,临床护理路径组较常规护理组相比HAMA评分及HAMD评分降低更加明显,差异具有统计学意义(P＜0.05).结论 临床护理路径可有效提高脑出血偏瘫患者的护理效果,促进改善日常活动能力及肢体功能,降低负面情绪.     

Clinicopathological features,diagnosis, and treatment of sessile serrated adenoma/polyp with dysplasia/carcinoma

Sessile serrated adenoma/polyps (SSA/Ps) are early precursor lesions in the serrated neoplasia pathway, which results in BRAF‐mutated colorectal carcinomas with not only high levels of microsatellite instability but also microsatellite stable. SSA/Ps with advanced histology, including cytological dysplasia or minimally invasive carcinomas, are important lesions because SSA/Ps are considered major contributors to “interval cancers” and these lesions can rapidly become dysplastic or invasive carcinomas. Clinicopathologically, SSA/Ps with dysplasia or invasive carcinoma were associated with advanced age, female sex, and proximal colon. Although SSA/Ps with submucosal invasive carcinoma were smaller and invaded less deeply into the submucosal layer than conventional tubular adenomas with submucosal invasive carcinoma, SSA/Ps with submucosal invasive carcinoma frequently had a mucinous component and exhibited a higher potential for lymphatic invasion and lymph node metastasis. In an SSA/P series, endoscopic characteristics, including (semi)pedunculated morphology, double elevation, central depression, and reddishness, may help accurately diagnose SSA/Ps with advanced histology. Removal of SSA/Ps with dysplasia or invasive carcinoma was recommended. Endoscopic treatment such as endoscopic mucosal resection or endoscopic submucosal dissection is useful for those lesions. However, surgical resection with lymph node dissection might be indicated when SSA/Ps with invasive carcinoma are endoscopically suspected, because these have the high risk of lymph node metastasis. Greater awareness may promote further research into improving the detection, recognition, and complete resection rates of SSA/Ps with dysplasia or invasive carcinoma and reduce the interval cancer rates.     

扁塑藤素调控Notch信号通路诱导人肺癌条件重编程细胞死亡的作用机制研究

目的：应用条件性重编程细胞（conditionally reprogrammed cells，CRCs）技术建立人肺癌细胞体外长期培养体系，研究扁塑藤素对人肺癌CRCs增殖和迁移能力的影响，并探讨相关作用机制。方法：免疫组化法检测Notch 1、HES1和Cyclin D3在肿瘤组织和癌旁组织中的表达水平；使用CRCs技术分离培养非小细胞肺癌原代细胞；使用2，4，8，16 μmol·L^-1的扁塑藤素处理人肺癌CRCs，MTS法检测细胞活力，Annexin V/PI流式细胞术检测细胞凋亡，Transwell法检测细胞迁移能力，Western Blot检测细胞中Notch信号通路相关蛋白Notch 1、HES1和Cyclin D3的表达水平。结果：在非小细胞肺癌患者肿瘤组织中Notch 1、HES1和Cyclin D3的表达水平高于癌旁组织；扁塑藤素能够诱导人肺癌CRCs死亡，并在一定范围内呈现时间和浓度依赖性（P<0.05）；随着扁塑藤素作用浓度的增高，人肺癌CRCs凋亡率明显增高（P<0.05），细胞迁移能力下降（P<0.05）；扁塑藤素能够下调人肺癌CRCs中Notch 1、HES1和Cyclin D3的蛋白表达水平。结论：扁塑藤素可能通过调控Notch信号通路相关蛋白的表达水平，抑制人肺癌CRCs的增殖和迁移，并诱导其凋亡，为肺癌的治疗提供新的实验数据和理论依据。     

A basal‐enriched microRNA is required for prostate tumorigenesis in a Pten knockout mouse model

MicroRNAs (miRNAs) play important roles in prostate cancer development. However, it remains unclear how individual miRNAs contribute to the initiation and progression of prostate cancer. Here we show that a basal layer‐enriched miRNA is required for prostate tumorigenesis. We identify miR‐205 as the most highly expressed miRNA and enriched in the basal cells of the prostate. Although miR‐205 is not required for normal prostate development and homeostasis, genetic deletion of miR‐205 in a Pten null tumor model significantly compromises tumor progression and does not promote metastasis. In Pten null basal cells, loss of miR‐205 attenuates pAkt levels and promotes cellular senescence. Furthermore, although overexpression of miR‐205 in prostate cancer cells with luminal phenotypes inhibits cell growth in both human and mouse, miR‐205 has a minimal effect on the growth of a normal human prostate cell line. Taken together, we have provided genetic evidence for a requirement of miR‐205 in the progression of Pten null‐induced prostate cancer.     

Causal Diagram Techniques for Urologic Oncology Research

Large quantities of data are now available to medical researchers; however, observational studies are plagued by bias and confounding. Additionally, much of this research only speculates on variable associations, leaving prospective randomized clinical trials as the sole purveyors of claims about causal relations between variables. There has been a growing movement of causal inference that uses new techniques to investigate causality using observational data. These techniques include the implementation of directed acyclic graphs, which allow researchers to explicitly and reproducibly define the causal relationships between study variables, thus making statistical analysis more robust. Directed acyclic graphs further allow researchers to identify confounding and other sources of bias and to discover causal effects among complex networks of variables. This review aims to introduce these techniques to the general urology and urologic oncology research communities in order to provide a basic understanding of causal inference and analysis and call for integration of these practices more generally in research methodology.     

铜绿假单胞菌注射液对肺癌A549细胞自噬及PI3K/Akt通路的影响

目的探究铜绿假单胞菌注射液(PA-MSHA)对肺癌A549细胞自噬及磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B(Akt)通路的影响。方法体外培养人肺癌A549细胞,随机分为空白对照组、LY294002组(加入20μmol/L LY294002),PA-MSHA干预组(加入0.5×10^9/mL、1.0×10^9/mL、2.0×10^9/mL PA-MSHA)。干预培养48 h后,MTT法检测各组细胞活性,平板克隆形成实验检测各组细胞增殖能力,透射电子显微镜观察各组细胞自噬情况,Western blot检测各组细胞中自噬相关蛋白p62、LC3Ⅰ、LC3Ⅱ及PI3K/Akt通路相关蛋白PI3K、Akt、p-PI3K、p-Akt表达。结果与空白对照组相比,LY294002组、0.5×10^9/mL、1.0×10^9/mL、2.0×10^9/mL PA-MSHA组细胞增殖抑制率、LC3Ⅱ/LC3Ⅰ蛋白表达显著升高(P<0.05),自噬体个数增加,细胞克隆形成率、p62、p-PI3K/PI3K、p-Akt/Akt蛋白表达显著降低(P<0.05);与LY294002组相比,0.5×10^9/mL、1.0×10^9/mL PA-MSHA组细胞增殖抑制率、LC3Ⅱ/LC3Ⅰ蛋白表达显著降低(P<0.05),自噬体个数减少,细胞克隆形成率、p62、p-PI3K/PI3K、p-Akt/Akt蛋白表达显著升高(P<0.05);与0.5×10^9/mL、1.0×10^9/mL PA-MSHA组相比,2.0×10^9/mL PA-MSHA组细胞增殖抑制率、LC3Ⅱ/LC3Ⅰ蛋白表达显著升高(P<0.05),自噬体个数增加,细胞克隆形成率、p62、p-PI3K/PI3K、p-Akt/Akt蛋白表达显著降低(P<0.05)。结论PA-MSHA可能通过调控PI3K/Akt信号通路,促进肺癌A549细胞自噬,抑制细胞增殖。     

Correlation between magnetic resonance imaging characteristics and BRAF alteration status in individuals with optic pathway/hypothalamic pilocytic astrocytomas

Background and purposeMost individuals with optic pathway/hypothalamic pilocytic astrocytoma (OPHPA) harbor either the BRAF V600E mutation or KIAA1549-BRAF fusion (K-B). This study aimed to investigate the imaging characteristics of OPHPA in relation to BRAF alteration status.Materials and methodsSeven cases of OPHPA harboring either the BRAF V600E mutation or K-B fusion were included in the study. Preoperative magnetic resonance imaging (MRI) was assessed for degree of T2 hyperintensity on T2-weighted images (T2WI) and the ratio of nonenhancing T2 or fluid-attenuated inversion recovery (FLAIR) hyperintense area to the contrast enhanced area (CE) on gadolinium-enhanced-T1 weighted images (T2/FLAIR-CE mismatch). The T2 signal intensity was normalized to cerebrospinal fluid (T2/CSF) for both the V600E and K-B group and compared. T2/FLAIR-CE mismatch was assessed by calculating the proportion of the tumor volume of nonenhancing high T2 signal intensity to the whole lesion (nonenhancing and enhancing components).ResultsFour and three cases of OPHPA harboring the BRAF V600E mutation and K-B, respectively, were analyzed. The T2/CSF value was higher in the K-B group than in the V600E group. Moreover, the V600E group had a larger T2/FLAIR-CE mismatch than the K-B group.ConclusionsThe BRAF alteration status in individuals with OPHPA was associated with preoperative MRI by focusing on T2 signal intensity and T2/FLAIR-CE mismatch. The BRAF V600E mutation was associated with a lower T2/CSF value and larger T2/FLAIR-CE mismatch, whereas K-B fusion was associated with a higher T2/CSF value and smaller T2/FLAIR-CE mismatch.     

PARP1: A potential biomarker for gastric cancer

Gastric cancer (GC) is the third leading cause of cancer mortality worldwide, with an overall 5-y survival rate of 25%. The majority of GCs are caused by infectious agents, including the bacterium Helicobacter pylori (H. pylori) and Epstein–Barr virus (EBV). Furthermore, inappropriate repair of DNA damage can also result in genomic instability, which has shown to be a key factor in carcinogenesis of different regions including gastric region. Present study was designed to explore the association between base excision repair pathway genes, PARP1 and APEX1 and gastric pathology and H. pylori infection. Two hundred gastric cancer tissue samples (114 H. pylori positive and 86 H. pylori negative) and adjacent uninvolved area taken as controls was used for expression analysis of BER pathway genes at mRNA level and protein levels using quantitative PCR (qPCR) and immunohistochemistry (IHC) respectively. Oxidative stress and DNA damage was also determined by measuring the level of antioxidant enzymes and comet assay respectively. Significant upregulation in PARP1 (p < 0.001) and APEX1 (p < 0.02) was observed in GC tissue samples compared to controls and this upregulation was more pronounced in H. pylori positive cases (HPGC) (PARP1, p < 0.02: APEX1, p < 0.04) than H. pylori negative cases (HNGC). Upregulation of BER pathway genes in HPGC was found correlated with smoking status (p < 0.0001), T stage (p < 0.01) and lymph node metastasis (p < 0.03). Moreover, immunohistochemical staining of BER pathway genes was found correlated with a number of clinicopathological characteristics such as tumor type (p < 0.03), tumor size (p < 0.01) and lymph node metastasis (p < 0.01). Expression levels of APEX1 and PARP1 gene also correlated with increased oxidative burden (p < 0.0001) and DNA damage (p < 0.001) in GC patients. Survival analysis showed that upregulation of PARP1 gene was associated with poor overall survival outcome of gastric cancer patients (HR = 2.04 (95% CI = 1.10–3.76; p < 0.02). Univariate and multivariate cox regression analysis showed the upregulated PARP1 gene (HR = 5.03; 95%CI (2.22–11.35); p = 0.0001), positive smoking status (HR = 3.58; 95%CI (1.67–7.65); p = 0.001), positive status for H pylori infection (HR = 4.38; 95%CI (1.82–10.56); p = 0.001) and advance N-stage (HR = 5.29; 95%CI (2.28–12.24); p = 0.0001) were independent prognostic factors for gastric cancer and may serve as a valuable biomarker for the diagnosis and progression of GC and can be helpful in developing individualized treatment strategies for treating GC.