Atomic force microscopy-based cell nanostructure for ligand-conjugated quantum dot endocytosis

While it has been well demonstrated that quantum dots (QDs) play an important role inbiological labeling both in vitro and in vivo,there is no report describing the cellular nanostructure basis ofreceptor-mediated endocytosis.Here,nanostructure evolution responses to the endocytosis of transferrin(Tf)-conjugated QDs were characterized by atomic force microscopy (AFM).AFM-based nanostructureanalysis demonstrated that the Tf-conjugated QDs were specifically and tightly bound to the cell receptorsand the nanostructure evolution is highly correlated with the cell membrane receptor-mediated transduction.Consistently,confocal microscopic and flow cytometry results have demonstrated the specificity anddynamic property of Tf-QD binding and internalization.We found that the internalization of Tf-QD is linearlyrelated to time.Moreover,while the nanoparticles on the cell membrane increased,the endocytosis was stillvery active,suggesting that QD nanoparticles did not interfere sterically with the binding and function ofreceptors.Therefore,ligand-conjugated QDs are potentially useful in biological labeling of cells at a nanometerscale.     

Quality control system of the endoplasmic reticulum and related diseases

The quality control (QC) system of the endoplasmic reticulum (ER) is an important monitoringmechanism in the protein maturation process,which ensures export of properly folded proteins from the ER.Incorrectly or incompletely folded proteins are retained in the ER for refolding or degradation by the ER-residing proteasome.The calnexin/calreticulin cycle and ER-associated degradation are the key elements inQC.These two mechanisms work together to allow incorrectly folded proteins have additional opportunitiesto achieve their native conformations.The QC dysfunction is involved in many diseases caused by mutantproteins,many of which are causes of neurodegenerative disorders.A better understanding of molecularregulation in the QC system will uncover the molecular pathogenic mechanisms of many diseases caused byprotein misfolding and help discover novel strategies for preventing or treating these diseases.     

The functional roles,cross-talk and clinical implications of m6A modification and circRNA in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide. HCC has high rates of death and recurrence, as well as very low survival rates. N6-methyladenosine (m6A) is the most abundant modification in eukaryotic RNAs, and circRNAs are a class of circular noncoding RNAs that are generated by back-splicing and they modulate multiple functions in a variety of cellular processes. Although the carcinogenesis of HCC is complex, emerging evidence has indicated that m6A modification and circRNA play vital roles in HCC development and progression. However, the underlying mechanisms governing HCC, their cross-talk, and clinical implications have not been fully elucidated. Therefore, in this paper, we elucidated the biological functions and molecular mechanisms of m6A modification in the carcinogenesis of HCC by illustrating three different regulatory factors ("writer", "eraser", and "reader") of the m6A modification process. Additionally, we dissected the functional roles of circRNAs in various malignant behaviors of HCC, thereby contributing to HCC initiation, progression and relapse. Furthermore, we demonstrated the cross-talk and interplay between m6A modification and circRNA by revealing the effects of the collaboration of circRNA and m6A modification on HCC progression. Finally, we proposed the clinical potential and implications of m6A modifiers and circRNAs as diagnostic biomarkers and therapeutic targets for HCC diagnosis, treatment and prognosis evaluation.     

Assessment of the pre-clinical immunogenicity of a new VEGF receptor Fc-fusion protein FP3 with ELISA and BIACORE

Purpose  

A new VEGF receptor fusion protein FP3 was shown to have promising antitumor potency better than Bevacizumab. Characterization of its immune response is essential to the safe and effective administration in clinical trials. In this study, both BIACORE and ELISA assays were employed to assess pre-clinical immunogenicity of FP3 in monkeys.     

Asian ginseng extract inhibits in vitro and in vivo growth of mouse lewis lung carcinoma via modulation of ERK‐p53 and NF‐κB signaling

Asian ginseng (AG) is the most commonly used medicinal herb in Asian countries. It is often prescribed for cancer patients as a complementary remedy. However, whether AG in fact benefits cancer patients remains unknown because some studies reported that AG facilitates tumor growth, which contradicts its usage as a dietary remedy to cancer patients. In addition, most of research works on ginseng for anti‐cancer were using single ginsenoside rather than whole root extracts used in clinics. Thus, intensive studies using the type of ginseng as its clinical form are necessary to validate its benefits to cancer patients. In this study, anti‐tumor potency and underlying molecular mechanisms of the ethanol extract of AG (EAG) were examined in mice with Lewis lung carcinoma (LLC‐1). We showed that EAG significantly suppressed tumor growth in LLC‐1‐bearing mice with concomitant down‐regulation of PCNA proliferative marker, and it exhibited specific cytotoxicity to cancer cells. EAG also induced MAPK and p53 signaling in LLC‐1 cells, which suppressed cyclin B–cdc2 complex and in turn induced G2–M arrest and apoptosis. Although EAG could activate NF‐κB signaling, the proteasome inhibitor of MG‐132 could effectively prevent NF‐κB targeted gene expression induced by EAG and then sensitize LLC‐1 cells to induce EAG‐mediated apoptosis. Collectively, EAG in a relatively high dose significantly suppressed tumor growth in LLC‐1‐bearing mice, indicating that AG may benefit lung cancer patients as a dietary supplement. This is the first report demonstrating possible combination of EAG with proteasome inhibitors could be a novel strategy in anti‐cancer treatment. J. Cell. Biochem. 111: 899–910, 2010. © 2010 Wiley‐Liss, Inc.     

Anti-inflammatory diterpenes from the seeds of Vitex negundo

Phytochemical investigation of a dichloromethane-soluble extract of Vitex negundo seeds led to the isolation of five labdane diterpenes, negundoins A–E (1–5), a 9,10-seco-abietane diterpene, negundoin F (6), a sandaracopimara-7,15-diene diterpene, negundoin G (7), and two known diterpene derivatives (8, 9). Their chemical structures were elucidated by detailed spectroscopic analyses on the basis of NMR, IR, and MS data. The anti-inflammatory effects of metabolites 1–7 were also evaluated in vitro. Compounds 3 and 5 were among the most potent inhibitors on nitric oxide production by LPS-stimulated RAW 264.7 macrophages, with IC₅₀ values of 0.12 and 0.23 μM, respectively. Further studies revealed that compounds 3 and 5 (5 μM) significantly reduced the levels of the iNOS protein to 0.40 ± 0.13% and 41.02 ± 6.02%, respectively, and COX-2 protein to 2.06 ± 0.53% and 26.40 ± 7.43%, respectively.     

喀斯特木论自然保护区土壤养分的空间变异特征

基于网格(20m×20m)采样法采集土壤样品,利用经典统计学和地统计方法分析了典型喀斯特峰丛洼地(200m×100m)土壤养分的空间变异特征.结果表明:研究区土壤pH值表现为弱变异,其他各养分指标均为中等程度变异,大小顺序为速效磷(AP)速效钾(AK)碱解氮(AN)土壤有机质(SOM)全钾(TK)全磷(TP)全氮(TN);pH半变异函数的最佳拟合模型为球状模型,TK和AK的最佳拟合模型为指数模型,其他养分指标的最佳拟合模型均为高斯模型;pH、AK的变异尺度(变程)较小,分别为58.1和41.1m,SOM、TN、TP、AN、AP的变异尺度相近,在100～150m,TK的变异尺度最大(463.5m);除研究区土壤TK、TN表现为中等的空间自相关性外,其他土壤养分指标均表现为强烈的空间自相关性.pH、AK呈零星斑块状分布,表现为高异质性;SOM、TP、TK的变化趋势较平缓,呈中间高、两边低的分布格局;AN、AP的空间分布具有显著的相似性,均随坡度的增加而呈片状上升趋势;TN的分布较特殊,呈中间低、两边高的趋势.植被、地形和高异质性的微生境是造成喀斯特木论自然保护区土壤养分格局差异的主要因素.     

乳源活性肽对抗猪瘟病毒抗体IgG、IgA和IgM水平的影响

不同剂量乳源活性肽与2头份猪瘟活疫苗同时分点肌肉注射仔猪,每周一次耳静脉采血,间接ELISA法检测抗猪瘟病毒抗体IgG、IgA和IgM水平.发现乳源活性肽能够明显提高抗猪瘟病毒抗体水平.当乳源活性肽为15 g/L时,与对照组相比,IgG抗体水平在注射后28 d时提高了21.0%,IgA和IgM抗体水平在注射后14 d时分别提高了13.8%和7.8%.实验组腹泻率、发病率和死亡率明显比对照组低,同时发现乳源活性肽具有促进生长的作用.     

C57-ras致癌性转基因小鼠模型的脏器及血液学参数

目的测定自主建立的致癌性转基因动物模型C57-ras小鼠的血液生理生化值和主要脏器重量,计算脏器系数并作统计学分析。方法选取同窝C57-ras转基因阳性和阴性小鼠,雌雄各半,采血,测量血液生理指标和血清生化指标,并称主要脏器重量。结果 C57-ras转基因阳性雌鼠和阴性雌鼠间比较,NEUT、NEUT%存在显著性差异（P〈0.05）,PCT存在极显著性差异（P〈0.01）。C57-ras转基因阳性雄鼠和阴性雄鼠间比较,RBC、HCT、PLT、PCT存在显著性差异（P〈0.05）,MON%存在极显著性差异（P〈0.01）。血清生化指标中,ALT和TG存在显著性差异（P〈0.05）。主要脏器重量和脏器系数比较结果显示,除C57-ras转基因阳性雌鼠和阴性雌鼠在肺重量存在极显著性差异（P〈0.01）外,其余均无显著性差异（P〉0.05）。结论新建C57-ras致癌性转基因小鼠模型和正常C57BL/6小鼠的主要生物学特性基本一致,利于该模型在致癌性安全性评价等领域的实际应用。