Effects and molecular mechanism of benserazide hydrochloride on LPS-induced inflammation in human umbilical vein endothelial cells / 中国药科大学学报

@#In this article, human umbilical vein endothelial cells(HUVECs)were induced by lipopolysaccharides(LPS)to establish an in vitro inflammation model to further verify the anti-inflammation effects of benserazide hydrochloride and to explore the molecular mechanisms involving in the anti-inflammation and anti-atherosclerosis of benserazide hydrochloride. The experiments were divided into blank groups(PBS+0. 5% FBS DMEM medium), model group ［LPS(500 μg/mL)+ 0. 5% FBS DMEM medium］ and drug group ［LPS(500 μg/mL)+benserazide hydrochloride+0. 5% FBS DMEM medium］. Western blot, ELISA and qPCR were used to detect the protein and mRNA expression levels of inflammatory cytokines SAP, TNF-α and MCP-1 in HUVECs cells. The expression levels of p65/p-p65, p38/p-p38, IκBα/p-IκBα, AKT/p-AKT and the nuclear translocations of p65, p38 and IκBα were detected by Western blot. The results showed that benserazide hydrochloride(1×10-9, 1×10-10, 1×10-11 mol/L)could significantly inhibit the protein and mRNA expression of pro-inflammatory cytokines SAP, TNF-α and MCP-1. Besides, it could down-regulate the protein expression of p65/p-p65, p38/p-p38, IκBα/p-IκBα and AKT/p-AKT in the signal pathway while inhibiting the nuclear translocation of p65, p38 and IκBα, thereby inhibiting the transcriptional activity of the related genes.

Effect of heparin-derived oligosaccharide on lipopolysaccharides-induced inflammation in HUVECs and its molecular mechanisms / 中国药科大学学报

@#In this study, the effect of heparin-derived oligosaccharide(HDO)on lipopolysaccharides(LPS)-induced inflammation in human umbilical vein endothelial cells(HUVECs)and the molecular mechanisms were investigated. The generation of intracellular reactive oxygen species(ROS)was detected by 20, 70-dichlorofluorescein diacetate(DCFH-DA). Experiment is divided into blank group(0. 5% serum medium), model group(LPS+0. 5% serum medium)and HDO dosing group(LPS+0. 5% serum medium +0. 01, 0. 1, 1mol/L HDO). The intracellular reactive oxygen species level was detected by reactive oxygen species experiment, the level of key regulatory proteins p38 and p-p38 in MAPK pathways and VCAM-1 were determined by Western blot. The results showed that HDO at 0. 01, 0. 1 and 1 μmol/L could inhibit the expression of VCAM-1 in HUVECs induced by 100 μg/mL LPS, and reduce the expression of key regulatory proteins p38 and p-p38, but could not obviously affect NF-κB nuclear translocation. The results all above showed that HDO could decrease the key regulatory proteins expression, and suppress the transcription of VCAM-1, resulting in inhibiting inflammation.

Clinical and pathologic features of small gastrointestinal stromal tumor / 中华消化内镜杂志

Objective To investigate the clinical and pathologic features of small gastrointestinal stromal tumors (small GISTs,d ＜ 2.0 cm).Methods Medical records of 95 patients undergoing surgery (endoscopic surgery,thoracoscopic/laparoscropic surgery and open surgery)and diagnosed as having GISTs by pathology and immunohistochemistry in Nanfang hospital from October 2003 to June 2014 were retrospectively analyzed.Based on clinical and pathological results,correlation analyses between risk factors for endoscopic ultrasonography(EUS) and Mitotic count(MI),clinicopathologic parameter and NIH risk classification were performed.Results Among 95 cases (104 lesions),88 were single,while 7 were multiple;81.7％ (85/104) small GISTs arose from stomach,including 87.1％ (74/85)in middle-upper stomach;5 cases (5.3％) presented calcification of different degrees,3 cases(3.2％) presented local necrosis and 2 cases (2.1％) with arrangement of epithelioid cells;88 cases (92.6％) were very low grade of NIH risk classification,6 cases (6.3％) were intermediate risk and 1 case(1.1％) was high risk.Positive rates of CD34 and CD117 were 95.8％ (91/95) and 96.8％ (92/95) respectively.The risk factors (border,mucosal surface,echo and heterogeneity) of EUS had no correlation with mitotic count(P＞0.05).The correlation analysis between clinicopathologic features and NIH risk classification revealed tumors more than 1.5 cm had a striking correlation with NIH risk classification (P＜ 0.05).Conclusion Most small GISTs,single or multiple,located at middle-upper stomach,were of very low or low risk,and have a favorable prognosis.But it has worse biological behavior and a higher grade risk when the diameter is more than 1.5 cm,intervention should be recommended.

The mechanisms of heparin-derived oligosaccharide on the inhibition of smooth muscle cells proliferation induced by platelet-derived growth factor / 药学学报

In this study, the effect of heparin-derived oligosaccharide (HDO) on platelet-derived growth factor (PDGF) induced vascular smooth muscle cells (VSMCs) proliferation and the related signal transduction mechanisms were investigated. MTT assays were used to measure VSMCs proliferation. Cell cycle distribution was analyzed by flow cytometry. The level of key regulatory proteins in PKC, MAPK and Akt/PI3K pathways were determined by RT-PCR, Western blot and immunocytochemical methods. Meanwhile, mRNA expressions of some proto-oncogenes were assayed by RT-PCR method. Our data showed that HDO (0.01, 0.1 and 1 μmol · L(-1)) inhibited 30 ng · mL(-1) PDGF-induced VSMCs proliferation in a dose-dependent manner, blocked the G1/S transition and inhibited the level of key regulatory proteins and some proto-oncogenes (P < 0.05). The results showed that HDO may decrease the key regulatory proteins expression, hence suppress the transcription of proto-oncogene and G1/S transition, finally inhibiting VSMCs proliferation.

Molecular mechanisms of combretastatin A-4 amino sugar derivative CPU-XT-008 on inhibiting proliferation of vascular endothelial cells / 中国药科大学学报

Taking human umbilical vein endothelial cells (HUVEC)as experimental model which can simulate tumor-derived vascular endothelial cells;the effects of CPU-XT-008;an amino sugar derivative of combretastatin A-4 (CA-4);on HUVEC proliferation;cell cycle distribution;tubulin polymerization and the key regulatory pro-tein of cell cycle were studied.The effect of CPU-XT-008 on the proliferation of HUVEC was determined by MTT assay.The cell cycle distribution caused by CPU-XT-008 was detected by flow cytometry.Immunofluorescence was used to detect apoptosis and tubulin polymerization.The expressions of Cyclin B1;Cdc2 and β-tubulin were detected by Western blotting.Results demonstrated that CPU-XT-008 could target tubulin and inhibit the poly-merization ofβ-tubulin;and it could lead to G2/M cell cycle arrest in HUVEC by up-regulating Cyclin B1 expres-sion and down-regulating Cdc2 and p-Cdc2 expression;which resulted in inhibiting the proliferation of HUVEC and inducing its apoptosis.

Molecular cloning, expression and identification of recombinant hyaluronan synthase from Pasteurella multocida / 中国药科大学学报

Aim: To set a suitable route for efficient expression and purification of recombinant hyaluronan syn-thase from Pasteurella multocida (rPmH AS). Methods: Coding sequences were cloned and expressed in Esche-richia coli strain of BL-21( DE3). Then rPmHAS was purified through a simple Ni-affinity chromatography and identified by hyaluronic acid binding protein ( HABP) based ELISA assay. Results: High yield expression of functional rPmHAS exceeding the highest production reported hitherto was achieved, and the purity of rPmHAS was as high as 90%. Conclusion: In this study, an optimized system of the expression, purification and identifica-tion route for large scale preparation of rPmHAS was established, which will be helpful for accelerating further study of PmHAS and facilitating fine researches on peculiar hyaluronic acid with special properties.

EXPRESSION OF Bcl-2 AND Bax PROTEINS IN THE DEVELOPING SMALL INTESTINE OF HUMAN FETUS / 解剖学报

Objective To explore the role of cell proliferation and apoptosis, and the expression significance of involved protein as Bcl-2(B-cell lymphoma/leukemia-2) and Bax (Bcl-associate X protein) in the developing small intestine of human fetus. Methods The expression product of Bcl-2 and Bax was investigated with immunohistochemical methods in the 2nd, 3rd and 4th months of gestation respectively. Results In the second, the third and the fourth month of gestation, the Bcl-2 immunoreactive positive signals were found in the ganglion cells of intermuscular and submucous nerve plexus. Bax positive cells were observed in the cytoplasm of the simple columnar epithelium cells of the small intestinal mucous layer.Conclusion Bcl-2 and Bax proteins regulate the developing small intestine of human fetus.