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Objective To explore the feasibility of interventional therapy in rat models of Budd-Chiari syndrome (BCS).Methods A total of 50 male clean SD rats were divided into model group and control group using random number table method,with 25 rats in each group.In the model group,BCS rat model was constructed by adopting partial ligation of the inferior vena cava (IVC),and the control group only had separation of surrounding tissues of IVC.Liver function was studied 12 weeks after postoperative raising,and digital subtraction angiography (DSA) and interventional therapy were performed,coupled with liver histopathology staining.Results Twenty BCS rats survived till the 12th week of raising,with the survival rate reaching 80.0％,and 22 survived in the control group.Compared with the control group,ALT [(43.1±5.5) U/L vs.(62.6±4.6) U/L] and AST [(84.7±26.5) U/L vs.(161.7±25.8) U/L)] in serum of rats in the model group increased,the differences were statistically significant (P＜0.05).HE staining showed that obvious hepatocyte necrosis and inflammatory cell infiltration were observed in BCS rats,and liver fibrosis was spotted via Masson staining.DSA examination found IVC obstruction in the model group,among which 14 (70.0％,14/20) received interventional therapy after successful probing of IVC obstructed segment,and 7 had balloon dilatation with a diameter of 3.5 mm,with 6 (85.7％,6/7) successfully dilatatedand the other 1 (14.3％,1/7) failed;the remaining 7 had balloon dilatation with a diameter of 4.5 mm,with 2 (28.6％,2/7) successfully dilatated,and the other 5 (71.4％,5/7) died of IVC rupture.Conclusion The BCS rat models by partial ligation of IVC can well simulate the pathophysiological changes and angiopathy characteristics of IVC obstructive BCS patients,which provide a platform for the basic research of interventional therapy of BCS.
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Objective To compare the repair effect on renal function between different times of bone marrow mesenchymal stem cells (BMSCs) transplant via renal artery route in experimental rats with adriamycininduced nephropathy.Methods Adriamycin-induced nephropathy model was established in 32 rats through injection of adriamycin though the caudal vein.Based on the scheduled times of BMSCs transplant,the experimental rats were randomly and equally divided into M0 group (zero time),M1 group (one time),M2group (2 times) and M3 group (3 times) with 8 rats in each group.Other 8 SD rats were used as normal control group (N group).Single dose of 0.5 rnl BMSC suspension (2×106 cells/ml) was transplanted to the rats of M0 group (zero time),M1 group (one time),M2 group (2 times) and M3 group (3 times),for the rats of the groups not receiving BMSC transplant a single dose of 0.5 ml L-DMEM culture medium,used as a placebo,was adopted to replace BMSC suspension.The transplant interval was one week.Before transplant as well as one and two weeks after last time of transplant,the serum urea nitrogen,serum creatinine,24 h urine protein and 24 h urine microprotein were tested,and one week after last time of transplant pathological sections were made for laser focusing microscope examination to observe renal pathological changes and the distribution of BMSC cells in the kidney.Results The values of serum urea nitrogen,serum creatinine,24 h urine protein and 24 h urine microprotein determined at each observation time point in M0 group,M1 group,M2 group and M3 group were significantly higher than those in N group (P＜0.001).The values of 24 h urine protein and 24 h urine microprotein determined at one week after last time of transplant in M2 group and M3 group were strikingly lower than those in M1 group (P＜0.05),but these differences between M2 group and M3 group were not statistically significant (P=0.063).Conclusion For the treatment of adriamycin-induced nephropathy in experimental rats,two times of using BMSCs transplant via renal artery route can achieve optimal curative effect.
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Background Visual adaptive mechanism of mammalian is close responsible for the development of visual cortex.The various genes with different biological functions in different developing stages of visual cortex participate in regulation of visual development.To investigate the differential expression profiles of various genes in different ages of rat cortex can offer basis and evidence for the study of visual development.Objective Present study aimed to investigate the genes that changed continuously in the postnatal developmental process of rat visual cortex by microarray analysis of visual cortex RNA.Methods Sixty clean SD rats were grouped numbered and randomized into the postnatal day 0 group (P0,n =20),before eye opening group (postnatal day 10,P10,n =15),before the critical period of visual cortex growth group (postnatal day 20,P20,n =15) and the end of development of visual cortex group(postnatal day 45,P45,n=15).The rats were sacrificed at corresponding time point respectively,and the fresh visual cortex were obtained for the extraction of total RNA and microarray analysis.Genes exhibiting changes in expression by≥2.0 folds were further confirmed using real-time PCR(RT-PCR).In order to evaluate the association of differential gene expression with growth,the postnatal stages were paired as 36 groups with the 3 pairs for each target gene (P45/P0,P20/P0,P10/P0).Results Microarray analysis showed that the gene with differential ratio ≥ 2.0 folds in rat visual cortex included Akap7,Asam,Casp3,Cxcr4,Egr1,Ennp2,Fabp7,Gpr88,Inpp5d,Rpsa,Stk32c and Vamp1.Real-time PCR verified that 24 genes form 26 probe sets had the same-phase regulating tendency,including 20 up-regulating probe sets and 6 down-regulating probe sets.The homodromous expressing tendency was seen in Akap7,Asam,Casp3,Cxcr4,Egr1,Ennp2,Fabp7,Inpp5d,Rpsa and Vamp1 genes between microarray analysis and RT-PCR.However,reverse expressions were found in the P45/P0 of Gpr88 and Stk32c genes,showing the up-regulation in the microarray analysis and down-regulation in RT-PCR.The concordant rate of gene expression between microarray analysis and RT-PCR was 94.44％.The expressing genes mainly functioned nervous system development,(metal) ion binding/transport,metabolism,regulation of neuronal synaptic plasticity.Conclusions New relevant candidate genes of age-associated rat visual cortex can be identified by microarray analysis,which provide a clue for the research of visual plasticity.
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Background Streptozotocin (STZ)-induced type 1 diabetic model is an acceptable model and is often used to the study of diabetic retinopathy (DR).However,the model is often established using retinal digest preparation in vitro in albino rats,and therefore it is difficult to evaluate the change of retinal vessels by fundus fluorescein angiography (FFA) in vivo.Recently,pigmented rats are being used as the DR model.But the study on the comparison between albino rat model and pigmented rat model is seldom.Objective This study was to observe and compare the manifestations of FFA and retinal digest preparation in early diabetic pigmented diabetic rat and albino diabetic rat,and to select the right model of DR using FFA.Methods Type 1 diabetic models were induced in 15 six-week-old health male BN rats and 15 six-week-old health male SD rats by the injection of STZ (55 mg/kg) via tail vein.The models were thought to be successful in the rats with the blood glucose level ≥ 16.7 mmol/L.The right eyes of the rats were extracted 6 weeks after injection of STZ.Lens were examined by slit lamp microscope.Retinal vascular changes were examined by fundus photography,FFA and periodic acid Schiff staining of retinal digest preparation.The manifestations of FFA and retinal digest preparation were contrasted between BN rats and SD rats.The number of eyes with lens opacification was compared by Chi-square test and the ratio of vascular endothelial cells and perithelial cells (E/P) was compared between BN rats and SD rats.The use and care of experimental animals complied the Statement of Ethic Committee of Tianjin Medicine University.Results Six weeks after injection of STZ,11 BN rats and 10SD rats were included in this study.The blood levels were (24.73±2.98) mmol/L and (22.36±3.65) mmol/L in BN rats and SD rats,respectively,without significant difference between the 2 types of rats (t =7.873,P＞0.05).Lens opacification occurred in 6 BN rats and in 5 SD rats (P=0.717).FFA showed the clear retinal vascular under the brown background.Evident vessel disorder and fluorescence leakage like background stage of DR were seen in 9 eyes.However,in the SD rats,retinal vessel abnormality could not exhibited owing to the interference of choroid fluorescent light from choroidal vessels and vortex vein.Retinal digest preparation exhibited that unevenness of vessel diameter,decrease of perithelial cells and increase of endothelial cells in both types of rats.The E/P values were 11.50±3.68in the BN rats and 12.86±3.94 in the SD rats,without significant difference between them (t=9.785,P＞0.05).Conclusions The abnormality of fundus vascular morphology can be better displayed in pigmented diabetic rats than albino rats by FFA in vivo.
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Background Studies showed that some members of matrix metalloproteinases (MMPs) play an important role in the pathogenesis of diabetic retinopathy (DR).However,whether MMPs inhibitor can deter blood retinal barrier (BRB) from damage is below understood.Objective This study was to investigate the effects of GM6001,a MMPs inhibitor,on BRB permeability.Methods Twenty-four adult clean SD rats were randomized to the control group,diabetic group and diabetes+GM6001 group according to randomized number table.Diabetic models were induced by the intraperitoneal injection of streptozotocin in the rats of the diabetic group and the diabetes + GM6001 group,and equal volume of citrate buffer was used in the same way in the rats of the control group.GM6001 10 μ1 (100 μ mol/L) was intravitreously injected in the third and fourteenth day after modeling in the diabetes+ GM6001 group,and equal volume of normal saline solution was injected in the same way in the control group and the diabetic group.The rats were sacrificed and eyeballs were extracted 1 month after injection,and the relative expressions of MMP-2 mRNA and MMP-9 mRNA in rat retinas were detected by reverse transcription PCR (RT-PCR).Evens blue (EB) was infused via the right jugular vein,and paraformaldehyde solution 1％ was then infused via left ventricle at the perfusion pressure 120 mmHg.The eyeballs were extracted 2 minutes later,and the leakage of EB in rat retinas was examined.Results RT-PCR electrophoresis exhibited the response bands of MMP-2 mRNA,and MMP-9 mRNA and GAPDH,with the gene size of 436,536 and 484 bp,respectively.The difference of the MMP2 mRNA and MMP-9 mRNA was statisticaly significant (F =20.336,P =0.000 ; F =8.742,P =0.002) ; and the relative expressions of MMP-2 mRNA and MMP-9 mRNA were significantly higher in the diabetic group and diabetes +GM6001 group than those in the control group (all at P＜0.01),and the relative expressions of MMP-2 mRNA and MMP-9 mRNA in the diabetes+GM6001 group was significantly reduced in comparison with the diabetic group(both P=0.01,P=0.02).The standardized EB content in the retinas of the control group,diabetic group and diabetes+ GM6001 group was (12.60±3.50) ng/mg,(26.52±7.14) ng/mg and (17.55±2.65) ng/mg,showing a significant difference (F=17.032,P＜0.01),and EB content in rat retinas in the diabetic group was higher than that in the control group (P=0.003),and that in the diabetes+GM6001 group was lower in comparison with the diabetic group (P=0.020).Conclusions Intravitreal injection of GM6001 can down-regulate the expression of MMP-2 mRNA and MMP-9 mRNA in diabetic rats and therefore protect BRB.
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Objective To discuss the impacts of different doses of alcohol on the level of microparticles (EMPs) of rat endothelial cells. Methods Sixty male SD rats were randomly allocated into four groups:high dose group (group A), me-dium dose group (group B), low dose group (group C) and the blank contrast group (group D). There were15 rats in each group. Rats were fed alcohol for 8 weeks. Flow cytometer was used to measure the level of circulating CD31+/CD42-EMPs in four groups, and which was compared with the alcohol dosage. Results Compared with group D, the level of circulating EMPs was significantly increased in group A and B (P＜0.05). There was no significant change in the level of circulating EMPs in group C (P>0.05). Conclusion The moderate and high doses of alcohol are harmful to the function of vascular en-dothelial cells in rats, which show a significant dose-effect relationship. The low dose of alcohol shows no effect on the func-tion of vascular endothelial cells in rats. The protective effect of alcohol needs further investigation.
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Insulin receptor substrate(IRS)-1 and IRS-2 expression levels of liver tissues and skeletal muscle in intrauterine growth retarded(IUGR)rats were investigated by RT-PCR and immunohistochemistry.An IUGR animal model was established by maternal nutrition restriction during pregnancy.IRS-2 expression level of liver tissue and IRS-1 expression level of skeletal muscle in IUGR rats at 0 and 3 weeks old were significantly lower than those in normal rats at the same age respectively,and insulin resistance was induced in IUGR,and these findings might be the molecular mechanisms susceptible to metabolic syndrome in IUGR rats.
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Diabetic model of SD rats was induced by streptozotocin injected intraperitoneally. Transforming growth factor (TCF)-?1 mRNA level was significantly decreased in renal cortex of diabetic rats by treatment of Pioglitazone. The result suggests that the protection of Pioglitazone against diabetic nephropathy seems to be related to the decrease of TGF-?1 gene expression in renal cortex.