Objective Salidroside is a major active component of integripetal rhodiola herbal medicine, which has a significant activity against hypoxia and ischemia.This study was to investigate the effects of side chain carbon losssalidroside analogues (N04) on the expressions of HIF-1α-related factors in the hypoxia-injured EAhy926 human vascular endothelial cells.Methods EAhy926 human umbilical vein endothelial cells in the logarithmic growth phase were randomly divided into a normal control, a hypoxia model control, a salidroside, a high-dose N04, a medium-dose N04, and a low-dose N04 group.The hypoxia model was established by depriving the culture medium of sugar and serum and culturing the EAhy926 cells in an environment of 95%N2+5%CO2 for 2 hours, followed by intervention with salidroside at 1×10-6 mol/L and N04 at 1×10-6, 1×10-7, and 1×10-8 mol/L, respectively.Then, the activity of the cells was detected by MTT assay, their LDH activity examined by spectrophotometry, the mRNA expressions of HIF-1α and VEGF measured by RT-PCR, the protein expressions of HIF-1α, VEGF and pVHL determined by Western blot, and the activity of eNOS measured by ELISA.Results Compared with the normal control group, the hypoxia model cells showed significantly reduced activity (0.51±0.05 vs 0.27±0.02, P<0.01), an elevated LDH level ([6.65±1.43] vs [78.82±2.33] U/L, P<0.01), and decreased eNOS activity ([1.56±0.23] vs [1.16±0.20] U/100 mL, P<0.01).In comparison with the hypoxia model group, the cells treated with high-, medium-, and low-dose N04 exhibited remarkably increased activity (0.27±0.02 vs 0.0.42±0.05, 0.40±0.03 and 0.37±0.04, P<0.01), a reduced LDH level ([78.82±2.33] vs [53.05±3.90], [58.42±4.45] and [62.73±3.63] U/L, P<0.01), and increased eNOS activity ([1.16±0.20] vs [3.01±0.47], [2.60±0.26] and [2.32±0.29] U/100 mL, P<0.01).The activity of eNOS was also increased in the salidroside group ([2.32±0.29] U/100 mL, P<0.01).The cell activity in the high-and medium-dose N04 groups was markedly higher than that in the salidroside group (P<0.05), and so was the eNOS activity in the high-dose N04 group and the LDH level in the medium-and low-dose N04 groups (P<0.05).In comparison with the normal control group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were significantly up-regulated in the hypoxia model group (P<0.01) while that of the pVHL protein markedly down-regulated (P<0.01).Compared with the hypoxia model group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were remarkably reduced (P<0.05), while that of the pVHL protein markedly elevated (P<0.05).Both the expressions of VEGF mRNA and HIF-1α protein were significantly lower in the medium-and low-dose N04 groups than in the salidroside group (P<0.05).Conclusion N04 can protect vascular endothelial cells against hypoxia-induced injury by regulating the expression of HIF-1α-related factors and eNOS.

OBJECTIVETo explore the effects on cell proliferation and invasion as well as molecular basis after suppressing EZH2 expression in endometrial carcinoma cells by using siRNAs.

METHODSRT-PCR was used to examine the expression of EZH2 in endometrial carcinoma and their paracancerous tissues. SiRNAs targeting to EZH2 were transfected to endometrial carcinoma cells, and MTT, FACS, and boyden assays were utilized to examine cell proliferation, cell cycle change, and cell invasion. Finally, the molecular mechanisms of EZH2 on cell function alteration were investigated.

RESULTSCompared with paracancerous tissues, increased expression trend of EZH2 mRNA was showed in endometrial carcinoma tissues. Further, knocking down EZH2 expression inhibited cell growth, cell cycle transition from G1 to S phase, and cell invasion ability. Molecular basis indicated that suppression of EZH2 downregulated the expression of E2F1 and MMP9 and upregulated tumor suppressor p21 expression.

CONCLUSIONEZH2 expression is increased in endometrial carcinoma tissues. Knocking down EZH2 expression suppresses the cell growth, cell cycle transition and cell invasion by downregulated E2F1 and MMP9, and upregulated tumor suppressor p21 expression.

Cell Line, Tumor ; Cell Proliferation ; E2F1 Transcription Factor ; metabolism ; Endometrial Neoplasms ; genetics ; pathology ; Enhancer of Zeste Homolog 2 Protein ; Female ; Genes, Tumor Suppressor ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; Polycomb Repressive Complex 2 ; genetics ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Transfection

Objective To study the method of preparation and blood sugar lowering effect of oral chitosan-insulin nanoparticles (INS-NPs) in streptozotocin-induced diabetic Wistar rats. Methods The INS-NPs were prepared by an ionic gelation method. The changes in the morphology and size of the INS-NPs were observed with transmission electron microscope and Zetasizer 3000HS, respectively. The blood sugar lowering effect of the INS-NPs was evaluated by monitoring the blood glucose levels in healthy and streptozotocin-induced diabetic rats. Results INS-NPs were spherical in shape with a mean size of 220.6±15.9nm. Entrapment efficiency of INS-NPs was 75.4%±3.2% and the loading efficiency of INS-NPs was 19.5%±2.6%. In vivo blood sugar lowering study showed that the levels of blood glucose of healthy Wistar rats were significantly reduced from 6h to 12h after oral administration of INS-NPs(25U/kg). The blood glucose level of diabetic rats were significantly reduced at 6h after oral administration of INS-NPs (25U/kg), and this effect was maintained for more than 9h, and the levels of blood glucose were kept in normal range for 7h. Conclusion The INS-NPs prepared by ionic gelation method has the blood glucose lowering effect in streptozotocin-induced diabetic Wistar rats.

The early histological myocardial ischemic changes in both experimental rabbitsand forensic caseworks of sudden death were demonstrated by the hematoxylin basicfuchsin-picric acid staining method.After the ligation of the coronary artery,therewere discrete patchy fuchsin-stained fibers which in contrast with the normal fibersof light-brown color.The number of fuchsin-stained fibers increased with the pro-longation of the intervals after ischemia.The results were not affected by the post-mortem autolysis.We consider that this method can be used as a routine test ofearly myocardial ischemia in both clinico-pathology and forensic pathology.

Proteolysis and amino acid metabolism in autolyzing rabbit left ventricular myocardium were studied. Tissue slices were incubated in a moist chamber at 37℃ from 15 min to 480 min. Their free acidic, neutral and basic amino acids and ammonia were determined by ion exchange chromatography. It was found thatmost of the free tissue amino acids in the course of autolys is increased significantlg, Increasing of alanine, lysine and histidine was noted at 15min of autolysis while the tissue glutamic acid content decreased,and the tissue ammonia content increased. The results suggest that the protcelysis was accelerated in early autolyzing heart muscles. It is helpful for the timing of death.