HIF1α-siRNA and gemcitabine combination-based GE-11 peptide antibody-targeted nanomedicine for enhanced therapeutic efficacy in pancreatic cancers

Abstract

Pancreatic cancer is one of the deadliest cancers across the world with an average 5-year survival rate of less than <6%. In this study, gemcitabine (GEM) and HIF1α-siRNA loaded GE-11 peptide conjugated liposome was successfully prepared and evaluated for its antitumor efficacy in pancreatic cancer cells. The GE11 increased the targeting specificity of liposome carrier and increased the intracellular concentrations in the cancer cells. Furthermore, synergistic combination of GEM and HIF1a-siRNA exhibited remarkable improvement in the declining of cancer cell proliferations. siRNA could effectively decrease the expression of HIF1a gene in the cancer cells. Importantly, GE-11 peptide-conjugated GEM/siRNA-loaded liposomes (GE-GML/siRNA) increased the total amount of apoptosis cells with higher proportion of cells in late apoptosis phase. GE-GML induced remarkable apoptosis of cancer cells and induced chromatin condensation and nuclear fragmentation which are considered to be typical features of apoptosis and cell death. GE-GML/siRNA showed a significant reduction in the tumour burden suggesting the superior anticancer efficacy of this formulation. GE-GML/siRNA showed four-fold reduction in tumour compared to control and two-fold reduction compared to GE-GML, respectively. Overall, present work lays foundation for the combination of GEM and HIF1a-siRNA loaded in a targeted nanocarrier system as a unique therapeutic option in pancreatic cancer treatment.     

门诊高血压病人健康教育的效果观察

护士门诊作为一种护理工作模式,旨在通过科学化、系统化、专业化的护理指导,使慢性病病人得到专业的护理知识.高血压是导致心脑血管病、肾脏病发生的最重要的危险因素[1],是全球人类最常见的慢性病.高血压病的预后与血压是否控制良好有关[2],而血压控制又与病人的规范用药和良好的生活方式有关.我院门诊部对高血压病病人实施健康教育综合护理取得了较好效果.现报道如下. 1资料与方法 1.1一般资料 2010年1月-2011年1月我院收治的高血压病病人320例,均符合《中国高血压防治指南》2005修订版规定的高血压诊断标准[3].其中男214例,女106例;年龄50岁～81岁(60岁±12岁);病程5年～31年(13.2年±7.9年);Ⅰ级115例,Ⅱ级135例,Ⅲ级70例;合并糖尿病58例;未服用降压药物47例.     

High-Resolution Melting Assay (HRMA) is a Simple and Sensitive Stool-Based DNA Test for the Detection of Mutations in Colorectal Neoplasms

BackgroundStool-based DNA testing for colorectal cancer is becoming a favored alternative to existing DNA screening tests. However, current methods of analysis often become more complicated and costly with increased sensitivity. The high-resolution melting assay (HRMA) is a simple and rapid mutation scanning method with low cost and superb accuracy. In this study, we verified the accuracy of HRMA for screening KRAS/TP53 mutations in stool-isolated DNA from patients with colorectal cancer.Materials and MethodsComparing to direct DNA sequencing, the accuracy of HRMA was verified by detecting KRAS/TP53 mutations in 2 independent stages. In study stage I, both tissue and stool samples from colorectal neoplasm patients were analyzed. In study stage II, stool samples from patients with colorectal neoplasms, and normal controls in clinical screening settings were examined.ResultsIn study stage I, the HRMA identified 14 of 17 target mutations (82.4%) in stools from cancer patients, and 4 of 5 (80.0%) target mutations in stools from advanced adenoma patients. The mutation detection rate in fecal samples (45.0%; 18/40) and referred tissue samples (55.0%; 22/40) was highly consistent (κ = 0.79). The HRMA detected 1% mutant DNA in a background of wild type DNA. In study stage II, the HRMA assay detected 58.8% (20/34) mutations in tumor samples, 41.5% (17/41) in advanced adenomas samples, and 3.33% (2/60) in age-matched normal control samples. The results from HRMA and DNA sequencing revealed 100% sensitivity and specificity in both tissue and stool samples.ConclusionHRMA is a simple, reliable, and sensitive method for detecting DNA mutations in the stool samples from patients with colorectal neoplasms.     

硫唑嘌呤在胰腺癌动物模型制作中的应用

目的筛选建立胰腺癌动物模型最适合成瘤的硫唑嘌呤(AZA)浓度。方法将40只SD大鼠按照AZA浓度的不同分为5组:空白对照组(A组)、2.6mg/kg AZA组(B组)、2.2mg/kg AZA组(C组)、1.8mg/kg AZA组(D组)、1.4mg/kgAZA组(E组),每组8只。A组每天灌服2mL生理盐水,B、C、D、E组每天灌服相应浓度的AZA生理盐水液2mL。第30天后进行SD大鼠皮下移植Mia PACAⅡ胰腺癌细胞悬液,通过检测大鼠血浆CA24-2和制作胰腺组织病理切片的方法来判断其是否成瘤。结果接种胰腺癌细胞后的第15、20、25、30天,各造模组大鼠血中的CA24-2水平均有不同程度的升高,其中B组在移植后的第15、20、25、30天,体内CA24-2水平分别为(51.56±2.14)、(51.61±1.21)、(54.66±1.02)、(56.00±0.38)U/mL,与C、D、E组比较,差异有统计学意义(P<0.05),且成瘤率最高,为83.3%。此外,胰腺病理切片镜下显示细胞核畸形、细胞大小不等,可见双核细胞及1～2个核仁,细胞核大且深染等病理特征,同时发现在移植部位的淋巴结和胰腺周围的淋巴结肿大。结论当AZA浓度为2.6mg/kg时,最有利于制作大鼠胰腺癌模型。     

p-Cymene Modulates In Vitro and In Vivo Cytokine Production by Inhibiting MAPK and NF-κB Activation

The present study was designed to investigate the effects of p-cymene on lipopolysaccharide (LPS)-induced inflammatory cytokine production both in vitro and in vivo. The production of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and interleukin-10 (IL-10) in LPS-stimulated RAW 264.7 cells and C57BL/6 mice was evaluated by sandwich ELISA. Meanwhile, the mRNA levels of cytokine genes were examined in vitro by semiquantitative RT-PCR. In a further study, we analyzed the activation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways by western blotting. We found that p-cymene significantly regulated TNF-α, IL-1β, and IL-6 production in LPS-stimulated RAW 264.7 cells. Furthermore, the levels of relative mRNAs were also found to be downregulated. In in vivo trail, p-cymene markedly suppressed the production of TNF-α and IL-1β and increased IL-10 secretion. We also found that p-cymene inhibited LPS-induced activation of extracellular signal receptor-activated kinase 1/2, p38, c-Jun N-terminal kinase, and IκBα. These results suggest that p-cymene may have a potential anti-inflammatory action on cytokine production by blocking NF-κB and MAPK signaling pathways.     

537例经产妇分娩方式及难产因素分析

目的:分析经产妇分娩方式及难产因素,探讨经产妇各项剖宫产指征的合理性,寻求降低经产妇剖宫产率的对策,提高经产妇母儿安全水平。方法:将1999年3月～2009年2月无妊娠合并症的537例经产妇按分娩时间分为前5年组和后5年组,对其基本特征、分娩方式、难产主要因素进行分析比较。结果:后5年组与前5年组比较,经产妇年龄呈明显增加趋势;自然分娩率、阴道助产率、臀位助产率降低,剖宫产率增高;臀位妊娠发生率无变化,瘢痕子宫比例明显增加,其剖宫产率均明显增高。结论:应加强经产妇的围产期保健及高危妊娠管理,重视有阴道助产史和胎儿体重较前次分娩新生儿体重有较大增加的孕妇的产程,避免发生母儿并发症;通过降低臀位妊娠和瘢痕子宫经产妇的剖宫产率,可以合理降低经产妇的剖宫产率。     

Microbial transformation of asiatic acid by Alternaria longipes

Asiatic acid is a major pentacyclic triterpene isolated from Centella asiatica. It shows a variety of bioactivities. In order to obtain its derivatives, potentially useful for detailed pharmacological studies, the substrate was subjected to incubations with selected micro-organisms. In this work, asiatic acid was converted into three new compounds: 2α,3β,23,30-tetrahydroxyurs-12-ene-28-oic acid (1), 2α,3β,22β,23-tetrahydroxyurs-12-ene-28-oic acid (2), and 2α,3β,22β,23,30-pentahydroxyurs-12-ene-28-oic acid (3) by the fungus Alternaria longipes AS 3.2875. The structures of the three metabolites were determined by 1D and 2D NMR spectral data.