Role and Significance of Ikaros Family in the Development of Hematopoietic System -Review / 中国实验血液学杂志

Hematopoietic cells are regulated by many transcriptional factors during their development, among them the Ikaros family is one of the most important representatives. They have characteristic conserved structural motifs, binding with DNA specific short sequences-containing key gene promoter or enhancer, to regulate their transcription activity. Meanwhile, the Ikaros family interact with other related transcriptional regulators to regulate the development and differentiation of hematopoietic cells. The structure of the Ikaros family, which belong to the zinc finger transcription factor, including Ikaros, Helios, Aiolos, Eos and Pegasus, are encoded by IKZF1-5 genes, respectively. They are master regulators of hematopoiesis, playing important roles in the occurrence, development and function of hematopoietic cells such as lymphocytes via individual and joint regulation. When working abnormalities, they are often related with the occurrence and development of the disease. In this review, the research achievements of the Ikaros family in recent years are summarized. On the one hand, it is helpful to understand the role and significance of this family in hematopoietic system; on the other hand, it provides the possible research direction for further research work.

EGFR-ERK signaling pathway down-regulates miRNA-145 in lung cancer cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the relationship between EGFR activation and down-regulation of miRNA-145 in lung cancer.</p><p><b>METHODS</b>Normal human lung epithelia cell line (BEAS-2B), human lung adenocarcinoma cell lines with wild-type EGFR (A549 and H292) and human lung adenocarcinoma cell lines with EGFR mutation (H1975 and H1650) were chosen in this study. The levels of miRNA-145 and p-EGFR were determined by quantitative real-time PCR (qRT-PCR) and Western blotting, respectively, and the relationship between p-EGFR and miRNA-145 levels was analyzed. The miRNA-145 levels were determined by qRT-PCR after activating EGFR with EGF or blocking EGFR signal pathway with AG1478. In addition, ERK1/2 inhibitor U0126 was used to inhibit ERK1/2 activation and then the expression of miRNA-145 was detected.</p><p><b>RESULTS</b>The miRNA-145 levels were closely negatively related with p-EGFR in lung cancer cells (r = -0.926, P = 0.024). EGF down-regulated miRNA-145 expression, particularly in BEAS-2B cells (53.0%; t = 30.993, P = 0.001) and A549 cells (42.6%; t = 14.326, P = 0.005).The miRNA-145 was up-regulated after inhibiting p-EGFR with AG1478, and significantly enhanced by 67.5% in H1975 cells when treated with AG1478 (t = 8.269, P = 0.014). The ERK1/2 signal pathway was activated by p-EGFR. U0126 restored the miRNA-145 down-regulation induced by EGFR-activation in lung cancer cells.</p><p><b>CONCLUSION</b>The activation of EGFR down-regulates miRNA-145 expression through ERK1/2 in lung cancer cells.</p>

Protective effects of rhu TNFR: Fc against the lipopolysaccharide induced intestinal damage of rats and its underlying mechanism / 药学学报

To investigate the protective effects of recombinant human tumor necrosis factor receptor II: IgG Fc fusion protein (rhu TNFR: Fc) against the lipopolysaccharide (LPS) induced intestinal damage of rats and its underlying mechanism. SD rats were randomly divided into four groups: control group, rhuTNFR: Fc group, LPS group and rhu TNFR: Fc + LPS group. Mean arterial pressure (MAP) was continuously monitored and the mortality rates were assessed. The levels of TNF-alpha and its bioactivity in the serum were assessed by ELISA and flow cytometry respectively. Pathologic changes of intestinal tissue were observed by HE staining. The rats of control and rhu TNFR: Fc group all survived with stable MAP, and the low level and bioactivity of TNF-alpha in the serum were maintained. While 83% of the rats in LPS group died by 6 h with the levels and bioactivity of TNF-alpha increasing significantly. In rhu TNFR: Fc + LPS group, the mortality rate of rats dropped to 33%. The TNF-alpha level increased compared with control group but its bioactivity decreased significantly compared with LPS group. The MPO activity and content of MDA decreased significantly. The status of pathological manifestation in the intestine was also ameliorated. These data suggest that rhu TNFR: Fc could protect rats from the acute intestine injury induced by LPS through ablating the rise in serum TNF-alpha level and bioactivity as well as anti-oxidation.

Effect of ryanodine receptor 2 gene silencing on ischemia-reperfusion injury of rat myocardial cells / 中华病理学杂志

<p><b>OBJECTIVES</b>To block the synthesis of ryanodine receptor 2 (RyR2) in myocardial cells by RNA interference and to investigate its biological impact on ischemia-reperfusion (I/R) in rat myocardial cells.</p><p><b>METHODS</b>Rat myocardial cells were isolated and cultured for an I/R model in vitro. RNA interference technique was used to block the synthesis of RyR2 in myocardial cells. Changes of LDH level, apoptosis, RyR2 mRNA expression and cytosolic Ca(2+) concentration were analyzed accordingly.</p><p><b>RESULTS</b>Myocardial cells after I/R manipolation were severely injuried (LDH leakage, 125 IU/L vs 12 IU/L, P < 0.05), apoptosis (60.1% vs 5.5%, P < 0.05), significant cytosolic Ca(2+) overload (21.2 vs 7.6, P < 0.05) and remarkable mitochondrial membrane potential loss (37.2 vs 85.1, P < 0.05). However, no visible change of RyR2 was observed (20.1 vs 22.7, P > 0.05). Pre-treatment with RyR2 specified siRNA demonstrated suppressed expression of RyR2 (6.8 vs 20.1, P < 0.05), increased mitochondrial membrane potential (55.8 vs 37.2, P < 0.05), attenuated cytosolic Ca(2+) overload (8.6 vs 21.2) and cellular apoptosis (31.2% vs 60.1%, P < 0.05).</p><p><b>CONCLUSION</b>RyR2 gene silencing enables to protect myocardial cells from I/R injury in vitro.</p>

Construction of lentivirus vector containing human ?-catenin-EGFP and its expression in human hair follicle stem cells / 上海第二医科大学学报

Objective To construct the lentivirus carrying human ?-catenin-EGFP(enhanced green fluorescent protein)and observe its expression in human follicle stem cells.Methods The ?-catenin gene sequence was amplified by RT-PCR from extraction of total RNA of human vascular endothelial cells.TA cloning technique was utilized to acquire gene subcloned pUCm-T-?-catenin.After transformation reaction,candidate clone was further analyzed by PCR and gene sequencing.Then the plasmid was transfected into FT293 cells.After identification by Western blotting,the plasmid was transfected into FT293 cells again for packaging.Infection titer was monitored by green EGFP expression.The expression of ?-catenin-lentivirus in human follicle stem cells were observed under inverted fluorescence microscope.Results The ?-catenin gene was cloned into the lentivirus successfully.The high expression of green fluorescence protein in FT293 cell line was found under fluorescent microscope.Viral titer checked by real-time PCR was about 2.0?108 TU/mL.When the multiplicity of infection(MOI)was 10,the infection efficiency of ?-catenin-lentivirus in human follicle stem cells was nearly 80% after infection 48 h around.After 3 weeks of continuous observation,we found the infection efficiency still keeping in the range of 80%-90%.Conclusion The lentivirus expression vector for ?-catenin was successfully constructed.It can steadily infect human follicle stem cells and the infection efficiency is considerable high.