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OBJECTIVE: To establish a New Zealand rabbit model of acute kidney injury induced by cisplatin. METHODS: A total of 24 male New Zealand rabbits were randomly divided into control group,low-,medium-and high-dose cisplatin group according to the body mass. Rabbits were injected with cisplatin at 0. 0,1. 0,2. 0,4. 0 mg/kg body weight by auricular vein. Rabbits in low-dose group was continuously injected for 5 days,medium-dose group was continuously injected for 3 days,and the high-dose group was injected for once per day. Rabbits in the control group did not receive any treatment. Blood was collected from the middle ear artery and 24 h urine was taken before exposure and on day 1,day 3,day 5 and day 7 of injection. The serum creatinine( Scr) and urea nitrogen( BUN) were detected by colorimetric method,and 24 h urine kidney injury molecule 1( KIM-1) was measured by enzyme-linked immunosorbent assay. Plasma platinum,24 h urinary platinum and renal platinum level were detected by inductively coupled plasma mass spectrometry.At the end of the experiment,rabbits were sacrificed and the left kidney was taken for histopathological examination.RESULTS: The body mass of rabbits of the low-,medium-and high-dose groups on day 7 after cisplatin exposure was lower than that of the control group( P < 0. 05),and lower than that of the same group before exposure( P < 0. 05). After 3 days of exposure,the Scr level in each dose group was higher than that of the control group( P < 0. 05),the Scr level on day 3and day 5 in medium-and high-dose groups were higher than that of the low-dose group( P < 0. 05). The BUN levels on day 3 and day 5 in medium-and high-dose group were higher than that of the control group and low-dose group( P <0. 05),the BUN levels on day 7 in three dose groups were higher than that of the control group( P < 0. 05). The levels of plasma platinum and 24 h urinary platinum in the three doses groups of New Zealand rabbits on day 1,day 3,day 5 and day 7 after exposure were higher than that of the control group( P < 0. 05),and were higher than the pre-treatment levels of the same group( P < 0. 05). The level of 24 h urinary KIM-1 in the meclium-dose group of New Zealand rabbits was higher than that of the control group on day 3 of exposure( P < 0. 05). The level of 24 h urinary KIM-1 in the mediumdose group of New Zealand rabbits on the 5th day after exposure was higher than that of the control group( P < 0. 05). The renal platinum levels in the three groups of New Zealand rabbits were higher than that in the control group( P < 0. 05).The pathological changes of rabbit kidney caused by cisplatin are mainly tubular dilatation,protein cast,alkalophilic and interstitial nephritis. CONCLUSION: Cisplatin can induce acute kidney injury in rabbits,and the degree of injury is dosedependent. The dose of 1. 0 mg/kg body weight continuous injection for 5 days is closely related to clinical use of cisplatin,which is recommended for model establishment.
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Objective To detect the blood parameter, blood biochemical and electrolyte indices of the F1 generation of Rongshui miniature pig ( RMP) .Methods The blood of 43 female and 42 male RMPs of 4 th month old, and 36 RMPs of 12th month old ( half male and female) were extracted from jugular vein.And the blood parameter, blood biochemical and electrolyte indices were detected by blood analyzer and automatic biochemical analyzer.Results In the same month-old RMP, no significant difference between male and female were found in most indices of blood parameter, blood biochemical and electrolyte indices.On the other hand, many indices were difference between 4th month old and 12th month old RMPs of same gender.Compared with the 4th month old RMP, the 12th month old RMP decreased significantly in WBC and PLT, increased in HGB ( P 0.05 ) .Serum ALT, AST, ALP, CK (male), LDH(male), A/G, BUN, GLU (female), CHOL (male) and K+decreased significantly (P <0.05), while serum TP, TBIL, CR and Ca2+increased significantly (P <0.05),but serum CHOL, TG, HDL-C and LDL-C were not different.86.4%(19/22) biochemical and electrolyte indices in RMP were in/or close to the range of normal value of human.Conclusion Most of the blood parameter, blood biochemical and electrolyte indices of RMP were close to human’ s normal value.
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Objective To construct a recombinant Ientiviral vector of mFVII/Fc and investigate its transfective efficiency into human bone mesenchymal stem cells (hBMSCs),and to detect the expression of mFVII/Fc fusion gene in vitro. Methods Coagulation factor VII (FVII) was cloned in vitro,with a point mutation from Lys to Ala in the position of 341 in the gene level.The cDNA fragments of mutational FVII (mFVII) and those of IgG1Fc were fused together with DNA ligase.After digestion,integration and sequencing,the fusion DNA was identified and transfected human embryonic kidney 293T cell packaging for re-mFVII/Fc lentiviral vector.After successful identification of vectors,detect the Ientiviral titer determination,bulk transfer after the determination of best MOI value of the third generation of hBMSCs,obseve the GFP expression with fluorescence microscope,have relative quantitative analyse of mRNA and protein expression of mFVII/Fc with RT-PCR and ELISA at different time points. Results In contrast with GenBank ID: AF 272774,the fusion gene matches exactly except the synonymous mutation,and the titer of packaging lentivirus was 2×108 TU/ml.Analyzed by Flow cytometry, indentification results of hBMSCs were as follows,CD+29(98.08%),CD+44 (97.63%),CD+34(0.31%) and CD+45(0.58%),respectively.The transfection efficiency of hBMSCs after 72 hours was (84±3)%,and the hBMSCs with mFVII/FC transfcetion have a large number of mRNA transcription and protein expression levels. Conclusions In this experiment we obtained a stable genetic vector with hBMSCs fusion gene expression successfully,which lay a foundation for the tissue factor study of prostate cancer targeting therapy and cancer gene therapy research.
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Objective To establish a spontaneous abortion rat model for a syndrome in traditional chinese medicine, kidney deficiency, and observe the changes of physiological indicators and related cytokine expression in the model. Methods 40 female and 20 male rats were used in this study. The female and male rats were mated (mating ratio 2:1). The day of vaginal smear with a large number of sperm was considered as the first day of pregnancy. The rats were randomly divided into control group and model group. The model group received 450 mg/kg hydroxyurea every day. Mifepristone was given on the eighth day in a dose of 3.75 mg/kg. The diet amount, the diameter index of kidney, ovary and embryos were analyzed. The mRNA expression of Th1/Th2 cytokine was detected by RT-PCR, and the expression of co-stimulating factors CD80, CD86, CD28, CTLA-4 were determined by flow cytometry.Results Comparing the model group with control group on the eighth day, there were significant differences between the model and control groups in quantity of food and water intake, and weight increase (P<0.05), and also in the embryonic diameter index, average of abortion rate, Th1/Th2 type cytokines, co-stimulating factor CD80, CD86, CD28, and CTLA-4 (P<0.05). Conclusion A rat model of spontaneous abortion due to kidney deficiency can be successfully established with hydroxyurea and mifepristone. The high expression of Th1 (TNF-α, IFN-γ) may cause abortion and be harmful to pregnancy. Th2 type (IL-4, IL-10) may facilitate pregnancy. The expression co-stimulating factor CD80, CD86, CD28, CTLA-4 may be relevant to the spontaneous abortion.