应用人肝细胞研究人参皂苷Rb1和Rg1、丹参素钠、冰片对CYP450药物代谢酶1A2、2B6和3A4的体外诱导作用

目的 使用原代培养的人肝细胞研究人参皂苷Rb1和Rg1、丹参素钠、冰片对细胞色素P450酶(CYP450)的诱导作用.方法 分别将3批次的冷冻原代人肝贴壁细胞进行接种培养,使用人参皂苷Rb1和Rg1、丹参素钠、冰片(30μmol/L)对CYP1A2、CYP2B6和CYP3A4进行诱导,实时荧光定量PCR(qRT-PCR...     

MiR-873-5p targets THUMPD1 to inhibit gastric cancer cell behavior and chemoresistance

BackgroundGastric cancer is one of the most common gastrointestinal tumors. Evidence has pointed to the fact that miRNAs play critical roles in the occurrence, development, and metastasis of gastric cancer by regulating cell proliferation, differentiation, apoptosis, and invasion.MethodsIn this study, first the relationship of miR-873-5p level and tissues types/LN(+/-)/metastasis(+/-)/tumor size was analysis, respectively. Second, the CCK8 and Transwell assay was used to determine the proliferation, invasion and migration of GC cells transfected with overexpression-/low expression-miR-873-5p. Third, the cell viability were analysis in the GC cells transfected with overexpression-/low expression-miR-873-5p treatment with different chemotherapy drugs. Fourth, the target gene of miR-873-5p was predicted using bioinformation methods. Fifth, the relationship of miR-873-5p with target gene-THUMPD1 were explored by using Wb and luciferase activity assay, et al.ResultsWe confirmed that miR-873-5p was negatively correlated with GC including tumor size, LN metastasis, distant metastasis. The miR-873-5p enhanced the sensitivity of Doxorubicin/Fluorouracil and cisplatin. The THUMPD1 was the target gene of miR-873-5p. Moreover, miR-873-5p could target the THUMPD1 axis so as to inhibit gastric cancer cell behavior as well as chemoresistance.ConclusionsMiR-873-5p plays a role in regulating cell behavior as well as regulating chemoresistance in gastric cancer. In addition, THUMPD1, as a downstream molecule of miR-873-5p, plays an important role in the cell behavior and chemoresistance of gastric cancer. The research first confirmed that miR-873-5p could inhibit gastric cancer cell behavior and chemoresistance by targeting the THUMPD1.     

马蓝色氨酸合成酶基因的克隆及表达分析

目的 克隆马蓝Baphicacanthus cusia吲哚类生物碱合成途径的色氨酸合成酶（tryptophan synthase,TSB）基因,命名为BcTSB（GenBank登录号AYM45644.1）,对其生物信息学和表达进行分析。方法 基于前期马蓝转录组数据库,获得BcTSB基因开放阅读框（ORF）,运用生物信息学手段对基因功能做出初步预测,构建pET32a-BcTSB原核表达载体,并转化大肠杆菌BL21（DE3）,IPTG诱导表达后进行SDS-PAGE检测。通过实时荧光定量PCR（qRT-PCR）技术检测马蓝不同组织（根、茎、叶）中TSB表达水平。结果 克隆的BcTSB基因ORF长度为1 452 bp,编码483个氨基酸,生物信息学预测为亲水性蛋白,定位于叶绿体,该蛋白相对分子质量为51 665.89,pET-32a载体包含18 000的标签。SDS-PAGE分析在70 000处出现目的蛋白条带,与理论预期值大小一致。qRT-PCR分析显示BcTSB基因在马蓝不同组织中均有表达,且在茎中的表达量最高。结论 通过对BcTSB基因的克隆及表达分析,为进一步研究该基因功能及调控奠定了实验基础。     

S-poly(T)plus荧光定量PCR技术在检测肝癌相关miR-199中的临床应用价值CSCD

目的探讨S-poly(T)plus荧光定量PCR法与普通Poly(A)加尾法检测外周血血清miR-199敏感度的优劣性。方法采用荧光定量PCR S-poly(T)plus法和Poly(A)加尾法检测肝癌患者和健康人血清miR-199,分别从敏感度、线性关系和重复性三方面进行评估其潜在临床应用价值。结果S-poly(T)plus法检测健康人miR-199的Ct值显著低于Poly(A)加尾法(30.32±0.72 vs.35.42±0.67,P<0.0001);而且S-poly(T)plus法具有更好的线性关系(R2=0.9843±0.0047 vs.R2=0.9573±0.0045,P=0.0027)和更好的的重复性(CV=2.59%vs.CV=3.52%,P=0.010)。S-poly(T)plus法在检测肝癌患者外周血miR-199时具有更好的敏感度。结论S-poly(T)plus荧光定量PCR检测肝癌血清miR-199比Poly(A)加尾法敏感度更高、重复性及稳定性更优。     

没食子软膏对二硝基氯苯致大鼠湿疹的保护作用及机制研究

目的：分析没食子软膏治疗大鼠湿疹可能的作用机制。方法：选取健康SD大鼠60只，按随机数字表法分为正常组、模型组、阳性组（地塞米松）、没食子软膏低剂量组、没食子软膏中剂量组、没食子软膏高剂量组，除正常组外，其余组均利用7%二硝基氯苯丙酮溶液涂抹背颈部皮肤构建湿疹大鼠模型，模型建立成功后各组连续给予相应药物14 d，取样。试剂盒检测大鼠血液学；苏木精-伊红（HE）染色观察湿疹大鼠病灶处皮肤组织的病理改变；免疫组织化学法测定皮肤中白细胞介素-4受体α、白细胞介素-5、JAK1、p-信号转导及转录激活因子6蛋白表达水平；荧光定量PCR（qRT-PCR）测定皮肤中PI3K、AKT基因表达水平。结果：没食子软膏低剂量组、没食子软膏高剂量组可以显著降低湿疹大鼠血液中单核细胞百分比，没食子软膏中剂量组、没食子软膏高剂量组可以显著降低湿疹大鼠血液中嗜酸性粒细胞百分比；没食子软膏可以显著改善湿疹大鼠的皮肤病理特征，下调皮肤中白细胞介素-4受体α、白细胞介素-5、JAK1、p-STAT6蛋白表达水平，PI3K、AKT基因表达水平。讨论：没食子软膏能够起到治疗湿疹大鼠的作用，其机制可能与影响JAK1/STAT6/PI3K/AKT信号通路有关。     

The prognostic signature based on glycolysis-immune related genes for acute myeloid leukemia patients

Acute myeloid leukemia (AML) is widely considered an immunoresponsive malignancy. However, potential association between glycolysis-immune related genes and AML patients’ prognosis has been seldom studied. AML-related data was downloaded from TCGA and GEO databases. We grouped patients according to Glycolysis status, Immune Score and combination analysis, basing on which overlapped differentially expressed genes (DEGs) were identified. The Risk Score model was then established. The results showed that totally 142 overlapped genes were probably correlated with glycolysis-immunity in AML patients, among which 6 optimal genes were screened to construct Risk Score. High Risk Score was an independent poor prognostic factor for AML. In conclusion, we established a relatively reliable prognostic signature of AML based on glycolysis-immunity related genes, including METTL7B, HTR7, ITGAX, TNNI2, SIX3 and PURG.