A comparative study of using comet assay and gammaH2AX foci formation in the detection of N-methyl-N'-nitro-N-nitrosoguanidine-induced DNA damage.

Comet assay is a useful technique in the detection of DNA damages, particularly DNA strand breaks; and it has been utilized to show that a potent carcinogen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), can induce such damages. Recently, gammaH2AX foci formation has been suggested as another sensitive way to detect DNA double strand breaks (DSBs). However, there is no systematic comparison being conducted to evaluate the consistency of these two methods. Using MNNG as a model chemical, the sensitivity of neutral comet assay and gammaH2AX foci formation in detecting MNNG-induced damage was studied. It was found that at concentrations of 0.1 and 1 microg/ml, both methods can detect MNNG-induced damage in human amnion FL cells. However, at 0.1 microg/ml, comet assay revealed more percentage of cells with DNA damage than gammaH2AX fluorescence revealed. On the other hand, while gammaH2AX foci were readily formed at very early times by 10 microg/ml MNNG treatment, neutral comet assay did not detect any significant DNA damage at the same time points. In addition, 10 microg/ml MNNG induced a distinct whole nuclei staining pattern of gammaH2AX, a type of DNA damage which was not detected by neutral comet assay but could be detected by alkaline comet assay. Therefore, gammaH2AX may be used as a sensitive indicator for DNA damage.     

F-19 MR imaging of glucose metabolism in the rabbit

Noninvasive metabolic magnetic resonance (MR) imaging reflecting glucose metabolism in the aldose-reductase-sorbitol (ARS) pathway was performed in the rabbit head; after administration of the fluorinated glucose analogue 3-fluoro-3-deoxy-D-glucose (3FD-glucose), fluorine-19 images were generated. Images of 3FD-glucose showed significant 3FD-glucose uptake by adipose tissue, indicating its buffering effects in case of excess loads of glucose. Images of 3-fluoro-3-deoxy-D-sorbitol (3FD-sorbitol) demonstrated the spatial distribution of aldose reductase activities and significant sorbitol accumulation in the lens. Images of 3-fluoro-3-deoxy-D-fructose (3FD-fructose) showed preferential uptake of fructose by muscle tissue. The extremely low toxicity of 3FD-glucose indicates promise for its clinical application in metabolic imaging.     

Upregulation of CD147 protects hepatocellular carcinoma cell from apoptosis through glycolytic switch via HIF-1 and MCT-4 under hypoxia

Purpose

Hypoxia is a vital factor in supporting and directing hepatocellular carcinoma (HCC) progression. HIF-1 transactivates target genes involved in metabolic reprogramming and antiapoptosis under hypoxia. However, key molecules involved in HCC hypoxia adaptation remain to be characterized. The aim of this study was to investigate the mechanism and biological function of CD147 on HCC cells resistant to apoptosis under hypoxic conditions.

Methods

Apoptotic rates of hypoxia-treated HCC cells were investigated by flow cytometer, and the expression levels of CD147, HIF-1α, MCT-1 and MCT-4 were assayed by immune blot. The in vitro glycolytic capacity was investigated in SMMC-7721 cells and 7721-shCD147 cells (CD147 is stably knocked down). Immunohistochemical staining of CD147, Glut-1, MCT-1, MCT-4, LAT-1 and CD98 was detected in tumor tissues from a xenograft model. Immunofluorescence double-labeled staining allowed further exploration of the expression levels and localizations of CD147, MCT-1 and MCT-4.

Results

Upregulation of CD147 under hypoxia correlates with higher viability of HCC cells compared with that in HSC cells. Silencing of CD147 significantly inhibited the glycolytic rate and induced apoptosis in SMMC-7721 cells. The expression level of lactate secretion transporter MCT-1/MCT-4 is dependent on CD147, while the expression level of Glut-1, LAT-1 and CD98 remained unchanged. Particularly, MCT-4 is demonstrated to be essential for the membranal localization of CD147.

Conclusions

With the above findings, we conclude that within the HCC hypoxic microenvironment, upregulation of CD147 and MCT-4 involved in glycolytic reprogramming is decisively important for the viability of HCC cells under hypoxia adaptation.     

十全大补汤联合肠内营养支持对老年胃癌术后气血两虚证病人营养状况和免疫功能的影响

目的探讨十全大补汤联合肠内营养支持对老年胃癌术后气血两虚证病人营养状况和免疫功能的影响。方法选取 2016年 1月至 2017年 12月邯郸市第一医院老年胃癌手术后气血两虚证病人 76例,按照随机数字表法分为观察组 38例和对照组 38例。对照组给予常规肠内营养支持治疗,观察组在对照组基础上辅助使用中药十全大补汤,给予中西医结合肠内营养支持治疗。经治疗后,比较两组病人肠鸣音恢复时间及排气、排便时间。对比两组病人在治疗前、治疗第 8天后病人的营养状况和免疫功能指标的变化情况。结果观察组在术后肠鸣音恢复时间( 41.2±5.2)h及排气( 69.5±7.8)h、排便时间( 81.5±9.8)h上显著快于对照组( 49.5±6.5)h、(73.6±8.6)h、(88.2±10.5)h,差异有统计学意义( P＜0.05)。治疗 8d后,观察组总蛋白( TP)(64.09±5.07)g·L-1、转铁蛋白( TRF)(2.10±0.18)g·L-1及血红蛋白( Hb)(117.59±14.32)g·L-1上升幅度均显著高于对照组(61.16±5.01)、(1.99±0.19)、(110.45±16.42)g·L-1(P＜0.05);观察组血清 IgA、IgG、IgM水平升高幅度均显著高于对照组( P＜ 0.05)。两组病人治疗 8d后 CD4+均升高( P＜0.05)但观察组高于对照组( P＜0.05);两组病人 CD8+均降低( P＜0.05),但观察组低于对照组( P＜0.05);两组病人 CD4+/CD8+P＜0.05),但观察组高于对照组( P＜0.05)。结论对于老年胃癌术后气血两虚证病人,运用中西医结合理念,应用十全大补汤联合肠内营养支持不仅可在一定程度上促进肠功能恢复、改善机体营均提升(,养状况,而且还能提高机体免疫力,起到了促进病人早日康复的目的。     

一例德朗热综合征患儿的致病基因变异分析

目的对1例疑诊德朗热综合征(Cornelia de Lange syndrome,CdLS)的患儿进行致病基因变异检测,明确其发病原因。方法应用高通量捕获测序对CdLS相关致病基因(NIPBL、SMC1A、SMC3、RAD21和HDAC8)进行测序,用Sanger测序验证测序结果以及致病基因的家系分析。结果患儿NIPBL基因存在c.6109-1G>A杂合剪接变异,Sanger测序验证结果表明患儿父母均未携带此变异,提示为新发变异,该变异未在HGMD及ExAC数据库收录。根据Human Splicing Finder预测剪接软件,预测该剪接变异将改变NIPBL基因剪接位点,为致病性变异。未发现SMC1A、SMC3、RAD21和HDAC8基因致病性变异。结论NIPBL基因c.6109-1G>A剪接变异可能是该例患儿的发病原因,新变异的检出丰富了NIPBL基因变异谱。     

Impact of body mass index on survival of patients with surgically treated renal cell carcinoma

PURPOSE: Population studies link increased BMI with an increased risk of cancer and cancer mortality and in particular a greater risk of RCC. We evaluated the impact of BMI and other clinical/pathological characteristics on survival in patients with RCC treated with radical or partial nephrectomy. MATERIALS AND METHODS: Between 1995 and 2003 patients undergoing radical (760) or partial (399) nephrectomy for RCC were entered into a database. BMI data were available on 1,137 of 1,159 (98%). Demographic and clinical/pathological parameters were analyzed. World Health Organization BMI definitions (normal-less than 25 kg/m(2), overweight-25 to 29.9 kg/m(2), obese-30 kg/m(2) or more) were used. RESULTS: A total of 75% of patients had greater than normal BMI with 472 (41.5%) overweight and 387 (34.0%) obese. Median followup was 33 months with a median overall survival of 110 months and a 5-year overall survival probability of 0.79. BMI categories were similar in age, gender, smoking status, presenting symptoms, tumor size, stage, and type of surgery. Significant increases in blood loss and operative time (p <0.05) were seen with increasing BMI. Although BMI 30 kg/m(2) or greater was associated with a higher proportion of clear cell histology (p = 0.002), it did not translate into an increased pathological stage, or incidence of metastasis. Multivariate analysis revealed age older than 65 years, systemic symptoms, surgery type, and pathological stage impacted overall survival (p <0.05). CONCLUSIONS: Although an increased BMI was associated with a greater proportion of clear cell histology, comorbidity, and surgical morbidity, BMI did not adversely impact overall or progression-free survival.