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.     

Influence of SNPs in cytokine-related genes on the severity of food allergy and atopic eczema in children

Although many single nucleotide polymorphism (SNP) studies have reported an association of atopy, allergic diseases and total serum immunoglobulin E (IgE) levels, almost all of these studies sought risk factors for the onset of these allergic diseases. Furthermore, many studies have analyzed a single gene and hardly any have analyzed environmental factors. In these analyses, the results could be masked and the effects of other genes and environmental factors may be decreased. Here, we described the correlation between four genes [interleukin (IL)-4 (C-590T), IL-4 receptor (A1652G), FCER1B (G6842A) and STAT6 (G2964A)] in connection with IgE production; the role of IL-10 (C-627A) as a regulatory cytokine of allergy; and the severity of food allergy (FA) and atopic eczema (AE) in 220 Japanese allergic children. In addition to these SNPs, environmental factors, i.e., patient's attitude, indoor environment, and so on, were also investigated in this study. Our study was retrospective, and the correlation was analyzed by our defined clinical scores divided into three terms: worst symptoms, recent symptoms and general amelioration at the most recent examination during the disease course. Our results indicated that IL-10 AA, the genotype with lower IL-10 production, is associated with higher IgE levels in the serum (p < 0.0001, estimate; 0.912). Marginal liver abnormalities were observed in the subject group with both FA and AE (p < 0.1191, estimate; 0.1490). Our defined clinical scores enabled evaluation of various aspects of disease severity. Based on the scores, while no single SNP selected in this study determined severity, the combination of the SNP with laboratory data and environmental factors appeared to determine severity.     

A facile synthesis of [14C]pyrithiobac‐sodium

Condensation of thiourea 1 with diethyl malonate 2 in the presence of sodium methoxide furnished 4,6‐dihydroxy‐2‐mercaptopyrimidine 3 . Compound 3 on methylation with diazomethane followed by oxidation with H₅IO₆/CrO₃ in ethyl acetate gave 4,6‐dimethoxy‐2‐methylsulphonylpyrimidine 5 . Compound 5 on condensation with 2‐mercapto‐6‐chlorobenzoic acid in the presence of a phase transfer catalyst, tetrabutylammonium bromide and sodium carbonate gave the title compound – pyrithiobac‐sodium 6 with an overall yield of > 35% starting from thiourea. Following the above standardized procedure, using [¹⁴C]‐thiourea in lieu of thiourea, ¹⁴C labelled product 6 , was synthesized with an overall radiochemical yield > 30% (with respect to [¹⁴C]‐thiourea) for further evaluations of environmental fate of 6 , in soils and plants. Copyright © 2006 John Wiley & Sons, Ltd.     

Measurement of xenon diffusing capacity in the rat lung by hyperpolarized 129Xe MRI and dynamic spectroscopy in a single breath-hold.

We used the dual capability of hyperpolarized 129Xe for spectroscopy and imaging to develop new measures of xenon diffusing capacity in the rat lung that (analogously to the diffusing capacity of carbon monoxide or DLCO) are calculated as a product of total lung volume and gas transfer rate constants divided by the pressure gradient. Under conditions of known constant pressure breath-hold, the volume is measured by hyperpolarized 129Xe MRI, and the transfer rate is measured by dynamic spectroscopy. The new quantities (xenon diffusing capacity in lung parenchyma (DLXeLP)), xenon diffusing capacity in RBCs (DLXeRBC), and total lung xenon diffusing capacity (DLXe)) were measured in six normal rats and six rats with lung inflammation induced by instillation of fungal spores of Stachybotrys chartarum. DLXeLP, DLXeRBC, and DLXe were 56 +/- 10 ml/min/mmHg, 64 +/- 35 ml/min/mmHg, and 29 +/- 9 ml/min/mmHg, respectively, for normal rats, and 27 +/- 9 ml/min/mmHg, 42 +/- 27 ml/min/mmHg, and 16 +/- 7 ml/min/mmHg, respectively, for diseased rats. Lung volumes and gas transfer times for LP (TtrLP) were 16 +/- 2 ml and 22 +/- 3 ms, respectively, for normal rats and 12 +/- 2 ml and 35 +/- 8 ms, respectively, for diseased rats. Xenon diffusing capacities may be useful for measuring changes in gas exchange associated with inflammation and other lung diseases.     

褪黑素和6-羟褪黑素保护神经细胞抗缺血再灌注损伤比较研究

目的研究褪黑素(Mel)和6-羟褪黑素(6-OHMel)神经保护作用及作用机理。方法体外培养N2a细胞,模拟缺血再灌注(OGSD),加入Mel和6-OHMel,检测以下指标:①细胞生存能力:MTT法、乳酸脱氢酶释放;②细胞凋亡分析:DNA片断化,细胞色素C,Caspase3活性;③活性氧(ROS)和线粒体跨膜电位。结果①Mel和6-OHMel都能减轻OGSD诱导的N2a细胞损伤,Mel的作用强于6-OHMel。②Mel和6-OHMel均能抑制细胞色素C释放,但6-OHMel强于Mel。③Mel和6-OHMel都能稳定线粒体跨膜电位,但Mel作用时间比6-OHMel长。④Mel和6-OHMel能清除ROS,6-OHMel表现为直接作用,Mel表现为间接作用。⑤Mel和6-OHMel均能抑制caspase3的活性,但是作用时间不同。6-OHMel表现在OGSD后12h,Mel在OGSD后24h。结论Mel和6-OHMel的神经保护作用与其抗氧化、稳定线粒体功能相关,Mel的作用机制更复杂。     

大黄素和小剂量雌激素合用对去卵巢大鼠骨质疏松的预防作用

目的观察大黄素和小剂量雌激素联合应用对去卵巢大鼠骨质疏松的预防作用.方法3月龄大鼠双侧卵巢去除术后预防用药90 d.用骨组织形态计量学方法,测定大鼠胫骨近端松质骨静态参数和动态参数,观察骨物理生长指标、血清生化指标和器官指数.结果大黄素90 mg·kg-1·d-1和己烯雌酚5 μg·kg-1·d-1联合应用可抑制去卵巢大鼠的破骨细胞活性,完全对抗其骨转化率增高和骨量丢失的骨质疏松症状,与己烯雌酚30 μg·kg-1·d-1的作用相当,且降低己烯雌酚对子宫和肝脏的刺激作用.结论大黄素和小剂量己烯雌酚联合应用可预防去卵巢大鼠骨质疏松.     

抑癌基因KLF-6和APC在结直肠癌组织中的表达及临床意义

目的 研究抑癌基因KLF-6和APC在结直肠癌中的表达规律及其临床意义。方法 应用RT-PCR和免疫组织化学法对32例结直肠癌组织和正常黏膜组织中KLF-6和APC基因mRNA及蛋白的表达进行检测和分析。结果 KLF-6与APC的mRNA阳性表达率在结直肠癌组织中分别为37、5％和34，3％，明显低于正常结直肠黏膜组织的96．9％和93．8％（P〈0．05）。KLF-6与APC的免疫组化染色阳性表达率在结直肠癌组织中分别为28．1％和25．0％，明显低于正常结直肠黏膜组织的81．3％和84．4％（P〈0．05）。KLF-6与APC在结直肠癌中的表达存在相关性（P〈0．05），并与肿瘤组织的分化程度、浸润深度、淋巴结转移及临床分期均有明显相关性（P〈0．05）。结论 抑癌基因KLF-6和APC的低表达可能与结直肠癌的发生、发展、转移及预后有关，且两者存在相关性。     

运动疲劳对大鼠十二指肠电活动的影响

本文报道了大鼠十二指肠电活动在运动疲劳前后的数值。结果表明：在运动疲劳后，快波、慢波振幅明显增加，而其频率却减少；Ⅰ相缩短，Ⅱ相延长，Ⅲ相、Ⅳ相及ＩＤＭＥＣ周期无显著变化；并在运动疲劳后９．７±２．４ｍｉｎ内出现不规律的食后型的电活动。这些改变可了解运动疲劳对消化系统的影响。     

体外循环对TXB2／6—keto—PGF1α的影响

对１３例体外循环病人进行了观察，发现体外循环后ＴＸＢ２／６－ｋｅｔｏ－ＰＧＦ１α显著增高，表明体外循环使血小板受损，而血小板受损是体外循环后失血的主要原因之一。