Objective:To investigate the effect of oxidized low-density lipoprotein(ox-LDL)on expression of lectin-like oxidized low-density lipoprotein receptor-1(LOX-1),transforming growth factor beta 1(TGF-?_1),and secretion of extracelluar matrix(ECM)in cultured rat glomerular mesangial cells(GMCs),and to investigate the influence of LOX-1 inhibitor polyinosinic acid(PIA)on the effect of ox-LDL.Methods:Rat glomerular masengial cells were cultured in vitro.RT-PCR was employed to determine the LOX-1 mRNA expression in GMCs incubated with different concentrations of ox-LDL(0,25,50, 100?g/ml).The expression of LOX-1 and TGF-?_1 mRNA was also determined by RT-PCR in the blank control group,ox-LDL (50?g/ml)group and PIA(50?g/ml ox-LDL+250?g/ml PIA)group.The contents of TGF-?_1,fibronectin(FN),and collagenⅣ(ColⅣ)in the supernatants of the above 3 groups were determined by ELISA.Results:RT-PCR showed that LOX-1 mRNA expression in 25,50 and 100?g/ml ox-LDL groups was significantly higher than that of blank control group(P

Matrix metalloproteinase (MMP) are a family of zinc-dependent endoproteinases whose enzymatic activity is directed against components of the extracelluar matrix(ECM). Their activities are inhibited by the tissue inhibitor of metalloproteinase (TIMP). MMP exerted an important role in invasion and metastasis of gastrointestinal cancer through degrading the ECM. With further research on inhibition of MMP, synthetic MMP inhibitor will have good prospects in the treatment of tumor invasion and metastasis.

Objective: To investigate the effect of Tangshentang on the expression of LOX-1 and secretion of TGF-?1,FN,ColⅣ in cultured rat masengial cells(RMCs) induced by ox-LDL and to explore its molecular mechanism of preventing and treating diabetic nephropathy.Methods: The cultured RMCs were divided into control group,ox-LDL group,Tangshentang group(respectively high,medium and low group)and rosiglitazone group.The mRNA expression was tested by RT-PCR.Synthesis of TGF-?1,FN,ColⅣ in cultured RMCs were determined by ELISA methods.Results: Tangshentang drug serum significantly attenuated up-regulation of TGF-?1,LOX-1 mRNA expression and synthesis of TGF-?1,FN,ColⅣ in a dose-dependent manner,with the peak at middle concentration,in RMCs stimulated by ox-LDL(50?g/ml).Conclusion: Tangshentang drug serum may protect kidney from injury by ox-LDL via the decreased expression of LOX-1 to reduce the uptake of ox-LDL and subsequently inhibiting TGF-?1 secretion,as well as the deposition of FN,ColⅣ.

AIM:To investigate the effects of MMP-7asODN on MMP-7 expression in human stomach cancer cell line KATOIII and explore the effect of MMP-7 on cancer invasion and metastasis. METHODS: A 15mer PS-asODNs targeted against the MMP-7 mRNA were introduced into KATOIII cells by FuGENE TM 6. The levels of MMP-7 mRNA was observed by reverse transcription polymerase chain reaction (RT-PCR). The ability of the invasion was observed by Boyden Chamber invasion assay. RESULTS: The MMP-7/?-actin ration was 0.31?0.02 for the PS-asODNs treated cells which is significantly lower compared with the control and PS-sODN and PS-mODN( 1.59?0.01, 1.14?0.03,1.51?0.02, respectivly.) P

Objective] To investigate the relationship between the AGE-RAGE and Qi and Yin deficiency and blood stasis of pathogenesis in diabetic nephropathy(DN).[Methods] The model was established by intravenous injection low-dose streptozotocin(STZ, 30 mg·kg-1) after having the high fat/high glucose diets for one month. And then that were divided into DN group, Tangshenfang low, medium, high groups, Valsartan group. Fasting blood-glucose (FBG) and Creatinine clearance(Ccr) were monitored. The morphological changes of renal tissue were examined under microscopy on sections stained with PAS. The RAGE mRNA and protein expression were determined by Real-time PCR and Western blot respectively;the urine album(U-alb) and content of AGE in serum were determined by ELISA. [Results] The model group rats acted obviously as deficiency of both Qi and Yin and obstruction of collaterals. compared with normal group, FBG, urinary albumin excretion rate and the content of AGE, RAGE expression were markedly increased; Ccr was markedly decreased in DN model group;blood viscosity and whole blood reduction viscosity were increased. FBG, Urinary albumin excretion rate and the content of AGE, RAGE expression were markedly decreased, Ccr was markedly increased by the intervention of Tangshenfang and valsartan. Blood viscosity and whole blood reduction viscosity were decreased; it was obviously and positively correlated between AGE with RAGE expression and U-alb, excretion rate and negatively correlated with the Ccr, r value was-0.456. [Conclusion] The early DN model rats were in the pathogenesis of Qi and Yin deficiency and blood stasis. And the AGE-RAGE may have close relationship with the pathogenesis of Qi and Yin deficiency and blood stasis.

AIM:To investigate the effects of silent information regulator 1 (SIRT1) on high glucose-induced acetylation of NF-κB p65 subunit and its protective role in rat mesangial cells .METHODS:Rat mesangial cells were cul-tured in DMEM supplemented with 10% FBS and were divided into control group , mannitol group , high glucose group , resveratrol group and SIRT1 RNAi group.The cell viability was determined by MTT assay .The mRNA expression of SIRT1, monocyte chemoattratant protein 1 (MCP-1), vascular cell adhesion molecule 1 (VCAM-1-1), tumor necrosis fac-torα( TNF-α) , transforming growth factor β1 ( TGF-β1 ) was analyzed by real-time quantitative PCR .The protein expres-sion of SIRT1 and the acetylation of NF-κB p65 subunit were determined by Western blotting .The protein concentrations of MCP-1, VCAM-1, TNF-α, TGF-β1 and malondialdehyde (MDA) were detected by ELISA.RESULTS:The cell viabili-ty, superoxide dismutase (SOD) activity, and the expression of SIRT1 at mRNA and protein levels were decreased by high glucose treatment as compared with control group .The acetylation of NF-κB p65 subunit was significantly increased after interfered with high glucose , resulting in the increase in the secretion of MCP-1, VCAM-1, TNF-αand TGF-β1 .Resvera-trol decreased high glucose-induced acetylation of NF-κB p65 subunit.However, silencing SIRT1 significantly enhanced the acetylation of NF-κB p65 subunit and the expression of MCP-1, VCAM-1, TNF-αand TGF-β1 .CONCLUSION:SIRT1 remarkably inhibits the inflammatory reactions by deacetylating NF-κB p65, suggesting that SIRT1 is a possible tar-get for preventing diabetic nephropathy .

Aim To investigate the effect of bufalin on proliferation and expression of WT1 in K562 cells. Methods The colony number of K562 cell was detec-ted with semi-solid culture assay. The cell cycle was measured by flowcytometry, and the expression of WT1 was observed with immunocytochemistry. Subcutaneous tumor models established by K562 cells in BALB/C nu/nu mice were divided into three groups, including model group, bufalin group and positive control group. After 21 days, the subcutaneous tumors were removed for calculating the inhibitory rate of tumor growth. HE staining and immunohistochemistry were used to ob-serve the morphological changes and the expression of WT1 . Results ① Bufalin could significantly decrease the colony number of K562 cell, arrest it at G0/G1 phase and down-regulate its expression of WT1 in a dose-dependent manner. ② Compared with the model group, the tumor inhibitory rate was much higher, while the volume and the weight were obviously lower in the other two groups. ③Bufalin could induce apop-tosis, necrosis, hemorrhage and fibrosis with HE stai-ning, and down-regulate the expression of WT1. Con-clusion Bufalin could inhibit the proliferation, arrest the cell cycle at G0/G1 phase and down-regulate the expression of WT1 in vitro. Bufalin could inhibit the tumor inhibitory rate, the volume and the weight of the subcutaneous tumors, induce apoptosis, necrosis, hemorrhage and fibrosis with HE staining and down-regulate the expression of WT1 .

Objective To supply large amount of seed cells for the cartilaginous tissue engineering,adult adipose-derived stem cells(ADSCs) were differentiated directly into cartilaginous cells.Methods Adipose tissue donated voluntarily by the adult patients after liposuction was isolated and subcultured.The cell activity was detected by methyl thiazolyl tetrazolium(MTT) assay and the proliferation curve line was drawn.The cultured stem cells were identified with flow cytometry.After the subculturing,the fourth generation of ADSCs were differentiated directly into cartilaginous cells,and then were marked by Alcian blue.Type II collagen expression in the ADSCs after cartilaginous differentiation was examined by immunohistochemical assay.Results ADSCs proliferated in vitro fast,and kept a stable multiplication till the 13th~15th generation.The results of flow cytometry showed that the cultured ADSCs had the specific features of the stem cells,and then were differentiated into cartilaginous cells.Positive Alcian blue staining and type II collagen expression indicated that the obtained cells had the function of normal cartilaginous cells.Conclusion ADSCs are differentiated directly into cartilaginous cells successfully,which will supply seed cells for the cartilaginous tissue engineering.

Objective To evaluate the biochemical function of rat nature three-dimensional acellular pancreatic bioscaffold (3D-APB) in promoting cell proliferation and differentiation in vitro.Methods The fresh pancreas from 10 rats were perfused to prepare 3D-APB.The biocompatibility of 3D-APB was eva luated.The experiment was divided into 4 groups based on 4 kinds of three-dimensional media for AR42J culture,including blank control group,ECM group,PLGA group and 3D-APB group.We compared the proliferation and differentiation of AR42J pancreatic acinar cell at 3 days,5 days,7 days and 10 days after seeding among 4 groups.Results The 3D-APB could represent a biocompatible scaffold capable of integrating within host tissue.The proliferation rate of AR42J cells by MTT in 3D-APB was higher,while the apoptosis rate by flow cytometry was lower than those in other 3 media,which were all significantly different (P ＜ 0.05),respectively.The protein expression of PDX-1 and PTF-1 by western blot in 3D-APB group was greatly higher than those in other 3 groups (both P ＜ 0.05).The mRNA expression of PDX-1 and PTF-1 through qRT-PCR was significantly higher than those in other 3 groups (both P ＜ 0.05).Conclusion Compared with the commonly used chemical and natural scaffold at present,3D-APB could promote cell proliferation and differentiation,which was more appropriate for regenerative medicine.

Purpose:To study the expression of MMP 7 gene in gastric cancer tissue and different stomach cancer cell line and explore its clinical significance.Methods:Reverse transcription polymerase chain reaction (RT PCR) was used to detect the expression of MMP 7 mRNA in 35 gastric cancer samples and the surrounding normal tissues and stomach cancer cell line. The ability of the invasion was observed by Boyden Chamber invasion assay. Results:The expression rate of MMP 7 mRNA in 35 gastric cancer samples and the surrounding normal tissues was 60% and 11%, respectively. The expression level of MMP 7 gene in gastric cancer was much higher than that in the surrounding normal tissues[(0 627?0 276)vs(0 237?0 12), P