1.Early islet graft failure due to hypoxia: review and our experimental experience
Gang MIAO ; Jingyong XU ; Yanyang ZHAO ; Junmin WEI
Chinese Journal of Hepatobiliary Surgery 2011;17(1):73-76
Pancreas and islet transplantation is the only treatment that can cure type 1 diabetes mellitus. Less invasive and more targeted surgical and immunosuppressive regimens make islet transplantation a more attractive treatment for type 1 diabetes. Current methods of islet isolation and purification cause hypoxic stress to which β cells are extremely vulnerable. Transplanted islets need to re-establish their vascular system in order to obtain sufficient oxygen and nutrient supply for stable engraftment. However, this process takes at least 7- 14 days to complete. Massive (>50%) β cells are dead before revascularization due to hypoxia, especially the core of the islets. Therefore, the obvious critical problem is the circulatory deficit to which the islets are susceptible in the immediate post-transplant period.In the current study, we reviewed various hypoxic-related insults to islets before complete engraftment, and feasible strategies to reduce hypoxic-induced apoptosis based on our experimental experiences together with that of others and investigated the possibility of revascularization in islet transplantation.
2.Extracellular matrix improves islet cell survival by α3-integrin regulation
Gang MIAO ; Yanyang ZHAO ; Yao LI ; Jianping CAI ; Jian LI ; Junmin WEI
Chinese Journal of Hepatobiliary Surgery 2012;18(3):220-225
Objective To determine the molecular pathway of reconstituted basement membrane extract(BME)embedment in the context of promoting islet cell survival.Methods Mouse islet cells were isolated and embedded in BME for in vitro culture.Caspase-3,integrin-α1 and 5,PDX-1,Akt,FAK and phospho Erk were detected using Western blot.Results Islet cells embedded with BME were partially protected from apoptosis indicated by a lower caspase-3 level and an increased phosphoAkt activity compared with untreated control.In addition,an increase of α3-integrin,FAK protein level and FAK activity were observed as well.Furthermore,the expression of PDX-1 and phosphoErk at the 48 h mark were preserved,suggesting the positive effect of BME to islet activity.Conclusion These results indicate that the embedment of BME construction can up-regulate α3 integrin and its signal transduction,which may improve viability and function of islet cells.
3.Advantages of mini-incision to remove bile-duct stones
Gang MIAO ; Yao LI ; Jian CHEN ; Xiuwen HE ; Xiaohua YE ; Min CHEN ; Junmin WEI
Chinese Journal of Hepatobiliary Surgery 2012;18(9):668-670
Objective To explore the advantages of the technique of bile duct mini-incision (BDM) for stone removal in choledocholithiasis,and to further clarify the indications for T tube insertion during surgery.Methods 85 BDM operations were performed.The use of MRCP (Magnetic resonance cholangiopancreatography),Advantage Workstation AW4.2 0.7sdc software,choledochoscopic imaging system and illustrations were used to study the applicability of the BDM technique in stone removal.Results The width of common bile duct (CBD) was usually over 8 mm in the patients with choledocholithiasis,and the width was related to the number and size of the stones.In the patients who had a CBD width of over 11mm (n 16),the MRCP stone imaging area (MRCP-SIA) was significantly larger when compared with that [(148±67)mm2 vs.(47±31)mm2,P<0.05] in the patients (n=14) with CBD width of less than 11 mm.T tube insertion for secondary choledochoscopic examination should be performed when the stones were non drifting,multiple,and closely related to the lower part of CBD indicating difficulty in complete stone removal.Conclusions Unnecessary T tube insertion could be avoided by the BDM technique for stone removal which was especially suitable for patients with early diagnosis of choledocholithiasis.T tube insertion should only be performed in patients with difficulty in complete stone removal and in other complicated situations.
4.Recognition of experimental animal model with kidney disease.
Yi-gang WAN ; Yan-ru HUANG ; Wei SUN ; Zhi-min MAO ; Xi-miao SHI ; Jian YAO
China Journal of Chinese Materia Medica 2014;39(21):4075-4081
Animal models with kidney disease are generally divided into two types. One belongs to the models which imitate human kidney disease by the artificial operations, such as anti-glomerular basement membrane antibody nephritis, Heymann nephritis, anti-Thyl. 1 antibody nephritis, BSA nephritis and puromycin nephropathy. The other one pertains to the models which make themselves kidney disease, and appear the pathological characteristics naturally as like as human, such as HIGA mice with IgA nephropathy and NZB/WF1 and MRL/1pr mice with lupus nephritis. In addition,the transgenic animal models with kidney disease can also be established by the modern molecular biologic techniques including gene knockout and siRNA transfection. As for the studies related with kidney disease in pharmacodynamics and pharmacology of Chinese herbal medicine (CHM), it is important to understand deeply the features of each animal model with kidney disease, and select accurately the proper models according to the different experimental objectives, and then, build the special models provided with the combination of disease with syndrome in traditional Chinese medicine (TCM). Therefore,it is the developmental direction for the further study to establish animal models with kidney disease, which should possess the characteristics of syndrome in TCM.
Animals
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Diabetic Nephropathies
;
etiology
;
Disease Models, Animal
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Humans
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Kidney Diseases
;
etiology
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Medicine, Chinese Traditional
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Mice
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Streptozocin
5.Influence of TIEG1 on apoptosis of HL-60 cells and expression of Bcl-2/Bax.
Kun YAO ; Ying YANG ; Rong HU ; Miao MIAO ; Ai-Jun LIAO ; Wei YANG ; Zhuo-Gang LIU
Journal of Experimental Hematology 2013;21(3):587-590
This study was aimed to investigate the influence of TIEG1 on apoptosis of HL-60 cells and the expression of Bcl-2/Bax. Different concentration of TIEG1 were used to treat HL-60 cells, the cell growth inhibition rate was detected by MTT method. After treating HL-60 cells with 12.03 ng/ml TIEG1, cell apoptosis was detected with flow cytometry. Bcl-2 and Bax was detected with RT-PCR. The results showed that TIEG1 had inhibitory effect on HL-60 cell proliferation, and in time-and dose-dependent manners. The more obvious inhibitory effect was observed in HL-60 cells treated with TIEG1 of 12.03 ng/ml. During the course of cell apoptosis, Bax expression increased, but Bcl-2 expression decreased (P < 0.05). It is concluded that TIEG1 inhibits HL-60 cell proliferation and induces apoptosis in time and dose-dependent manners. During the course of HL-60 cells apoptosis induced by TIEG1, Bcl-2/Bax are associated with HL-60 cell apoptosis induced by TIEG1.
Apoptosis
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drug effects
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Cell Proliferation
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drug effects
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Early Growth Response Transcription Factors
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pharmacology
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Gene Expression Regulation, Leukemic
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HL-60 Cells
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Humans
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Kruppel-Like Transcription Factors
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pharmacology
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Proto-Oncogene Proteins c-bcl-2
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metabolism
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bcl-2-Associated X Protein
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metabolism
6.Effect of valproic acid sodium on proliferation and apoptosis of acute T-lymphoblastic leukemia Jurkat cells.
Miao MIAO ; Bing DU ; Rong HU ; Ying YANG ; Wei YANG ; Ai-Jun LIAO ; Zhuo-Gang LIU
Journal of Experimental Hematology 2013;21(2):343-346
This study was aimed to investigate the effects of valproic acid sodium (VPA) on the proliferation and apoptosis of acute T-lymphoblastic leukemia Jurkat cells. Jurkat cells were treated with different concentration of VPA. Proliferation-inhibition curve was assayed and plotted by CCK-8 method and the cell apoptosis was detected by flow cytometry with Annexin V/PI double staining. The expression level of anti-apoptotic gene BCL-2 and pro-apoptosis gene Bak1 were detected by semi-quantitative RT-PCR. The results showed that the VPA inhibited the proliferation of Jurkat cells in concentration-dependent manner. As compared with the control group, the apoptosis of cells increased along with adding concentration of VPA; VPA could decrease the expression of BCL-2 gene, but did not show obvious effect on the expression of Bak1. It is concluded that the VPA can inhibit proliferation of Jurkat cells which possibly associates with the decrease of BCL-2 expression.
Apoptosis
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drug effects
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Cell Proliferation
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drug effects
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Humans
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Jurkat Cells
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Proto-Oncogene Proteins c-bcl-2
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metabolism
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Sodium
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pharmacology
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Valproic Acid
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pharmacology
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bcl-2 Homologous Antagonist-Killer Protein
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metabolism
7.Construction of recombinant gene adenovirus encoding enhanced green fluorecence protein-peroxisome proliferator-activated receptor gamma2 fusion protein and its expression in bone marrow mesenchymal stem cells.
Li-zi LIAO ; Jin-gang XIAO ; Miao-miao YANG ; Zi-ren KONG ; Qin-ce SUN ; Wei-dong TIAN
West China Journal of Stomatology 2010;28(4):430-434
OBJECTIVETo construct mouse enhanced green fluorecence protein (EGFP) -peroxisome proliferator-activated receptor (PPAR)gamma2, and to detect EGFP-PPARgamma2 expression in infected mouse bone marrow mesenchymal stem cells (BMSC).
METHODSCut the fragment of PPARgamma2 from the expression plasmid pcDNA flag PPARgamma2, then cloned the gene fragment into pEGFP-C1 and pEGFP-N1 vector. Subsequently, subclone the fragment EGFP-PPARgamma2 from pEGFP-C1-PPARgamma2 into the shuttle plasmid DC315. HEK293 cells were co-transfected with the constructed recombinant shuttle plasmid DC315-EGFP-PPARgamma2 and large adenovirus helper plasmid pBHGlox deltaE1, 3Cre in mediation of liposome. The obtained replication-defective recombinant adenovirus Ad-EGFP-PPARgamma2 was confirmed. Then it was propagated in HEK293 cells. After the BMSC were transfected for 72 h, adipogenic differentiation was demonstrated.
RESULTSHEK293 cells were transfected with the pEGFP-C1-PPARgamma2 or pEGFP-N1-PPARgamma2 in mediation of liposome. The former green fluorescence protein was better than the latter by fluorescence microscope. The recombinant plasmids were digested and identified. Western blot analysis showed the expression of EGFP-PPARgamma2 in vitro. EGFP-PPARgamma2 protein was detectable in the nucleus of BMSC.
CONCLUSIONThe recombinant adenovirus encoding EGFP-PPARgamma2 fusion protein was successfully constructed, which provided a basis for application of EGFP-PPARgamma2 gene to adenovirus-mediated gene therapy.
Adenoviridae ; Animals ; Bone Marrow Cells ; metabolism ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; HEK293 Cells ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Mice ; PPAR gamma ; metabolism ; Recombinant Proteins ; metabolism ; Transfection
8.Effects and mechanisms of huangkui capsule ameliorating renal fibrosis in diabetic nephropathy rats via inhibiting oxidative stress and p38MAPK signaling pathway activity in kidney.
Zhi-min MAO ; Yi-gang WAN ; Wei SUN ; Hao-li CHEN ; Yan-ru HUANG ; Xi-miao SHI ; Jian YAO
China Journal of Chinese Materia Medica 2014;39(21):4110-4117
OBJECTIVETo demonstrate the effects and mechanisms of Huangkui capsule (HKC) on renal fibrosis in rats with diabetic nephropathy (DN).
METHODRats were randomly divided into 5 groups, the sham-operated group (Sham group, n = 5), the vehicle-given group (Vehicle group, n = 7), the low dose of HKC-treated group (L-HKC group, n = 7), the high dose of HKC-treated group (H-HKC group, n = 7) and the lipoic acid (LA)-treated group (LA group, n = 7). DN models were induced by intraperitoneal injection of streptozotocin (STZ,35 mg x kg(-1)) twice and unilateral nephrectomy. After models were successfully established, the rats in HKC and LA groups were daily administrated with HKC suspensions (0.75, 2 g x kg(-1)) or LA suspensions (60 mg x kg(-1)) respectively, and at the same time, the rats in Vehicle group were daily administrated with distilled water (2 mL) for 8 weeks. All rats were sacrificed at the end of week 8 to collect blood and renal tissues. UAlb, renal function, renal fibrotic morphologic characteristics, as well as oxidative stress (OS)-related markers, the protein expressions of the key signaling molecules in p38 mitogen-activated protein kinase (p38MAPK) signaling pathway, fibrogenic cytokines and inflammatory factors were examined respectively.
RESULTHKC, similar to LA, improved the general state of health, body weight, UAlb, BUN, UA and Alb in DN model rats. Of note, renal fibrosis was ameliorated in HKC groups,especially in H-HKC group which was better than that in LA group. In addition, HKC not only improved the main indexes of OS in the kidney like LA, but also down-regulated the protein expressions of phosphorylated-p38MAPK (p-p38MAPK), transforming growth factor (TGF)-β1 and tumor necrosis factor(TNF)-α in the kidney, whereas, LA only decreased the protein expression of TNF-α in the kidney in DN model rats.
CONCLUSIONHKC, similar to LA, has the actions of anti-OS in vivo. Moreover, HKC could attenuate renal fibrosis by suppressing the activation of p38MAPK signaling pathway and the protein expressions of fibrogenic cytokines and inflammatory factors in the kidney in DN model rats, which is different from LA.
Abelmoschus ; chemistry ; Animals ; Capsules ; Diabetic Nephropathies ; drug therapy ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Fibrosis ; Kidney ; drug effects ; pathology ; MAP Kinase Signaling System ; drug effects ; Male ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors
9.Effects and mechanisms of multi-glycoside of Tripterygium wilfordii improving glomerular inflammatory injury by regulating p38MAPK signaling activation in diabetic nephropathy rats.
Yan-Ru HUANG ; Yi-gang WAN ; Wei SUN ; Zhi-min MAO ; Qing ZHAO ; Xi-miao SHI ; Jian YAO
China Journal of Chinese Materia Medica 2014;39(21):4102-4109
OBJECTIVETo explore the effects and mechanisms of multi-glycoside of Tripterygium wilfordii (GTW) on improving glomerular inflammatory lesion in rats with diabetic nephropathy (DN).
METHODDN model was induced by unilateral nephrectomy and intraperitoneal injection of STZ (35 mg x kg(-1)) twice. The rats were randomly divided into 3 groups, the sham-operated group (Sham group, n = 5), the vehicle-given group (Vehicle group, n = 5 ) and GTW-treated group (GTW group, n = 5). After the model was successfully established, the rats in GTW group were daily oral administrated with GTW suspension (50 mg x kg(-1) x d(-1)), meanwhile, the rats in Vehicle group were daily oral administrated with distilled water (2 mL) for 8 weeks. From the beginning of the administration, all rats were killed 8 weeks later. Blood and renal tissues were collected,and then UAlb, renal function, glomerular morphology characteristics and glomerular macrophages (ED1 + cells) infiltration, as well as the protein expressions of inflammatory cytokines including tumor necrosis factor(TNF)-α and interleukin(IL)-lβ, and the key molecules in p38MAPK signaling pathway including p38 mitogenactivated protein kinase (MAPK), phosphorylated p38 (p-p38MAPK) and transforming growth factor(TGF)-β1 were investigated respectively.
RESULTGTW not only ameliorated the general state of health and body weight,but also attenuated UAlb, glomerulosclerosis, the infiltration of glomerular ED1 + cells and the protein expressions of TNF-α, IL-1β, p-p38MAPK and TGF-β1 in the kidney in DN model rats.
CONCLUSIONBy means of DN model rats, we demonstrated that GTW has the protective effect on renal inflammatory damage in vivo via inhibiting inflammatory cells infiltration and inflammatory cytokines expression. Furthermore, GTW could improve renal inflammatory lesion through down-regulating the expressions of the key signaling molecules in p38MAPK pathway such as p-p38MAPK and TGF-β1 ,and inhibiting the activation of p38MAPK signaling in the kidney.
Animals ; Diabetic Nephropathies ; drug therapy ; Disease Models, Animal ; Glomerulonephritis ; drug therapy ; Glycosides ; pharmacology ; MAP Kinase Signaling System ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; analysis ; Tripterygium ; chemistry ; p38 Mitogen-Activated Protein Kinases ; physiology
10.Gene expression profiling of skeletal muscle in type 2 diabetic rats
Jun-Ping WEN ; Li-Xiang LIN ; Gang CHEN ; Wei-Te ZHUANG ; Lian-Tao LI ; Miao LIN ; Qing-Ming LIN ;
Chinese Journal of Endocrinology and Metabolism 2001;0(05):-
Differences in gene expressions were compared by cDNA microarray in skeletal muscle of type 2 diabetic rats and normal rats.In diabetic rats,157 genes were down-regulated and 100 genes up-regulated. Some of these genes were related to insulin resistance,glucose and lipid metabolism.