microRNA-622 acts as a tumor suppressor in hepatocellular carcinoma

microRNAs (miRNAs) are important regulators of tumor development and progression. In this study, we aimed to explore the expression and role of miR-622 in hepatocellular carcinoma (HCC). We found that miR-622 was significantly downregulated in human HCC specimens compared to adjacent noncancerous liver tissues. miR-622 downregulation was significantly associated with aggressive parameters and poor prognosis in HCC. Enforced expression of miR-622 significantly decreased the proliferation and colony formation and induced apoptosis of HCC cells. In vivo studies demonstrated that miR-622 overexpression retarded the growth of HCC xenograft tumors. Bioinformatic analysis and luciferase reporter assays revealed that miR-622 directly targeted the 3′-untranslated region (UTR) of mitogen-activated protein 4 kinase 4 (MAP4K4) mRNA. Ectopic expression of miR-622 led to a significant reduction of MAP4K4 expression in HCC cells and xenograft tumors. Overexpression of MAP4K4 partially restored cell proliferation and colony formation and reversed the induction of apoptosis in miR-622-overexpressing HCC cells. Inhibition of JNK and NF-κB signaling phenocopied the anticancer effects of miR-622 on HCC cells. Taken together, miR-622 acts as a tumor suppressor in HCC and restoration of miR-622 may provide therapeutic benefits in the treatment of HCC.     

Proteomic analysis of glutathione S-transferase isoforms in mouse liver mitochondria

AIM: To survey glutathione (GSH) S-transferase (GST) isoforms in mitochondria and to reveal the isoforms’ biological significance in diabetic mice.METHODS: The presence of GSTs in mouse liver mitochondria was systematically screened by two proteomic approaches, namely, GSH affinity chromatography/two dimensional electrophoresis (2DE/MALDI TOF/TOF MS) and SDS-PAGE/LC ESI MS/MS. The proteomic results were further confirmed by Western blotting using monoclonal antibodies against GSTs. To evaluate the liver mitochondrial GSTs quantitatively, calibration curves were generated by the loading amounts of individual recombinant GST protein vs the relative intensities elicited from the Western blotting. An extensive comparison of the liver mitochondrial GSTs was conducted between normal and db/db diabetic mice. Student’s t test was adopted for the estimation of regression and significant difference.RESULTS: Using GSH affinity/2DE/MALDI TOF/TOF MS, three GSTs, namely, alpha3, mu1 and pi1, were identified; whereas five GSTs, alpha3, mu1, pi1, kappa1 and zeta1, were detected in mouse liver mitochondria using SDS-PAGE/LC ESI MS/MS, of these GSTs, GST kappa1 was reported as a specific mitochondrial GST. The R² values of regression ranged between values of about 0.86 and 0.98, which were acceptable for the quantification. Based on the measurement of the GST abundances in liver mitochondria of normal and diabetic mice, the four GSTs, alpha3, kappa1, mu1 and zeta1, were found to be almost comparable between the two sets of animals, whereas, lower GST pi1 was detected in the diabetic mice compared with normal ones, the signal of Western blotting in control and db/db diabetic mice liver mitochondria is 134.61 ± 53.84 vs 99.74 ± 46.2, with P < 0.05.CONCLUSION: Our results indicate that GSTs exist widely in mitochondria and its abundances of mitochondrial GSTs might be tissue-dependent and disease-related.     

Expression of interferon inducible protein-10 in pancreas of mice

AIM: To investigate the expression of interferon inducible protein-10 (IP-10) in pancreas of mice and to discuss its possible role in the pathogenesis of type 1 diabetes. METHODS: Non-obese diabetic (NOD) mice were used as experiment group and BALB/c mice as non-diabetic prone model. Immunohistochemistry method was used to evaluate the expression of IP-10 in the pancreas of NOD mice and BALB/c mice. Immunoelectron microscope was used to show the location of IP-10 in pancreatic islet (3 cells. RESULTS: Pancreatic islets were positively stained in all the NOD mice. Insulitis could be found in mice at the age of 4 wk. The weakly positive results were found in control group with no insulitis. Immunoelectron microscopy further demonstrated that IP-10 was produced by pancreatic (3 cells and stored in cytoplasm of the cells. CONCLUSION: IP-10 can be largely produced in pancreatic islets of NOD mice at the age of 2 wk when there is no significant insulitis, and may play an important part in the pathogenesis of type 1 diabetes by attracting immune cells to infiltrate the pancreatic islets.     

运用丰田生产方式缩短急性缺血性卒中静脉溶栓时间临床研究

目的 研究丰田生产方式（Toyota production system,TPS）缩短急性缺血性卒中患者入院至溶栓时间 （door to needle time,DNT）和提高DNT≤60 min的比例。 方法 收集2012年6月-2013年12月重组组织型纤溶酶原激活剂（recombinant tissue plasminogen activator,rt-PA）静脉溶栓的急性缺血性卒中患者为对照组,2014年1月-2015年6月启用TPS改善溶栓 流程后rt-PA静脉溶栓的急性缺血性卒中患者为实验组,比较两组患者DNT时间及DNT≤60 min比例。 结果 研究共纳入对照组68例,实验组87例。对照组DNT平均（92.27±16.98）min,实验组DNT平均 （63.52±11.86）mi n,两组有显著差异（P =0.002）。改善流程后DNT≤60 min的比例由对照组的6.11%提 高到实验组的51.09%（P =0.001）。 结论 通过组建TPS团队多学科合作进行溶栓流程改造,能够显著降低急性缺血性卒中rt-PA静脉 溶栓的院内延误时间,缩短DNT,提高DNT≤60 min的比例。     

慢性心力衰竭合并肌肉衰减综合征的研究进展

肌肉衰减综合征是一种与年龄增长相关,以进行性全身骨骼肌质量减少、肌肉强度下降和肌肉功能减退为主要特征的临床综合征。慢性心力衰竭是心血管疾病中影响患者生存率的重要疾病之一。越来越多的研究发现慢性心力衰竭可能造成或加重肌肉衰减综合征的进展,其机制涉及营养不良、运动量下降、炎症反应、氧化应激及激素变化等。目前的治疗策略主要包括运动干预、营养支持和药物治疗。本文就近年慢性心力衰竭合并肌肉衰减综合征的发病机制及治疗方案等方面的研究进展进行综述。     

乔松素对脂多糖诱导的人脐静脉内皮细胞凋亡的影响

目的探讨蜂胶乔松素对脂多糖诱导的人脐静脉内皮细胞凋亡的影响。方法用消化灌注法收集人脐静脉内皮细胞进行体外原代培养,于培养液中给予10 mg/L脂多糖处理以建立细胞凋亡模型,乔松素处理组于模型条件培养液中分别加入不同浓度的乔松素(50、100和200 mg/L)干预,共同孵育24 h。光镜下观察细胞形态,MTT法测定细胞存活率以观察细胞增殖变化,缺口末端标记技术TUNEL法检测细胞凋亡率,免疫细胞化学法检测信号转导系统核因子κB亚基p65核易位变化。结果模型组内皮细胞凋亡率明显高于阴性对照组,不同浓度乔松素组细胞凋亡率均低于模型组,同时细胞存活率增高(P0.01)。加入不同浓度乔松素能显著降低脂多糖诱导的人脐静脉内皮细胞核因子κB p65核易位活性(P0.01)。结论乔松素可通过抑制脂多糖诱导的人脐静脉内皮细胞凋亡而保护内皮细胞功能,其机制可能与抑制核因子κB p65活化有关。     

Cytotoxic ent-kaurane diterpenoids from Isodon weisiensis C. Y. Wu

A new ent-kaurane diterpenoid, weisiensin B (1), was isolated from the leaves of Isodon weisiensis C. Y. Wu, along with four known ones, kamebanin (2), kamebacetal A (3), macrocalyxin D (4) and excisanin D (5). Their structures were determined by spectroscopic means. Compound 1-4 showed significant cytotoxic activity against Bel-7402 and HO-8910 cells.     

比较经颅超声造影和数字减影血管造影对大脑中动脉狭窄的诊断价值研究

目的 比较经颅超声造影（transcranial contrast-enhanced ultrasound,t-CEUS）和DSA在大脑中动脉 （middle cerebral artery,MCA）狭窄性疾病中的诊断价值。 方法 收集2017年12月-2019年4月疑似MCA狭窄或闭塞的患者197例,记录其225条狭窄或闭塞 的大脑中动脉的t-CEUS和DSA检查结果。根据MCA狭窄程度分为轻度狭窄（30%～50%）,中度狭窄 （50%～69%）,重度狭窄（70%～99%）和闭塞（100%）,以DSA为金标准,分析t-CEUS对轻、中和重度 MCA狭窄的诊断价值。 结果 197例疑似MCA狭窄闭塞患者中,男132例,女65例,年龄43～72岁,平均（58.4±4.6）岁。经 DSA确诊的狭窄和闭塞的225条MCA中,轻度狭窄16条,中度狭窄70条,重度狭窄121条,闭塞18条。 t-CEUS诊断狭窄和闭塞的共224条MCA,其中,轻度狭窄18条,中度狭窄76条,重度狭窄112条,闭塞18条。 t-CEUS和DSA诊断MCA狭窄和闭塞高度吻合（Kappa=0.91）。t-CEUS诊断轻度MCA狭窄的敏感度、特异 度和准确度分别为80%、80%和91%;诊断中度MCA狭窄的敏感度、特异度和准确度分别为83%、93% 和97%;诊断重度MCA狭窄的敏感度、特异度和准确度分别为83%、96%和96%。 结论 t-CEUS能够清楚地显示轻、中、重度MCA狭窄和闭塞,适用于MCA狭窄和闭塞类疾病患者的确 诊,尤其是适用于不适合DSA检查的疑似MCA狭窄和闭塞患者。     

肝X受体的激活对小鼠糖尿病肝脏脂肪酸合成酶表达的调节

目的 探讨肝X受体(LXR)对糖尿病肝脏脂肪酸合成酶(FAS)表达的影响及机制.方法 将16周龄、雄性、C57BL/6背景下瘦素受体基因缺陷的db/db小鼠和对照的db/m小鼠,分别予以LXR激动剂TO901317(TO)(3 mg·kg-1·d-1)或DMSO灌胃处理7 d;TO(10 μmol/L)刺激人肝癌细胞系HepG2细胞24 h.此外,HepG2细胞转染小鼠FAS启动子报告基因表达质粒,同时转染pcDNA3.1表达载体或LXR或活化型固醇调节元件结合蛋白(SREBP-1c)表达质粒12 h;采用免疫组织化学方法检测FAS蛋白在肝脏上的表达,实时荧光定量PCR和蛋白印迹方法分别在mRNA和蛋白水平检测FAS和SREBP-1的表达及TO对其表达的影响,荧光素酶报告基因方法检测LXR激动剂和SREBP-1c对小鼠FAS启动子活性的影响.结果 免疫组化结果显示FAS蛋白在肝脏广泛表达,主要位于肝细胞胞质内.TO可显著降低db/db小鼠空腹血糖水平[由(12.00±1.06)mmol/L下降到(7.73±0.69)mmool/L,P＜0.01]和糖化血红蛋白水平(由5.67%±0.10%下降到4.87%±0.08%,P＜0.01).db/db小鼠肝脏FAS mRNA水平明显高于db/m小鼠,约为5.5倍(P＜0.01);在蛋白水平,db/db小鼠肝脏上的FAS也明显高于db/m小鼠.TO处理可使db/m小鼠肝脏FAS mRNA表达升高约3.5倍(P＜0.05),可使db/db小鼠肝脏FAS mRNA升高约1.7倍(P＜0.05);TO处理可使db/m小鼠肝脏SREBP-1 mRNA升高约2.4倍(P＜0.05),使db/db小鼠肝脏SREBP-1 mRNA升高约2.1倍(P＜0.01).TO能够上调HepG2细胞FAS的mRNA表达水平,升高约1.9倍(P＜0.05);LXR的激活还能显著增加HepG2细胞中FAS基因启动子活性,约为对照组的1.5倍;过表达LXR或SREBP-1c也能增加HepG2细胞FAS基因启动子活性,分别为对照组的1.9倍和1.6倍(均P＜0.01).结论 LXR可能通过其本身的直接作用和SREBP-1C的间接作用上调肝脏FAS的表达,LXR可能介导了糖尿病肝脏的脂质生成过程.