RESUMO
【Objective】 To reveal possible mechanisms of miRNA in diabetic hepatopathy through bioinformatics method. 【Methods】 Subset data of miRNA and their matched mRNAs in the liver of STZ-induced diabetic mice and the normal liver tissues of congenial mice by detecting on microarrays were collected from GEO database； information from the database and bioinformatics analysis were applied to mine a batch of miRNAs in diabetic hepatopathy and targeted mRNAs regulated. Then qRT-PCR was used to verify the expressions of miRNAs in diabetic liver from 20 STZ-treated Kunming mice and 10 normal homologous mice. 【Results】 Via detection and analysis， miRNAs differentially expressed （including 96 up-regulated and 77 down-regulated） were significantly obtained. Groups of miRNAs and their effectors （mRNAs） that may be related to the pathological process of diabetic liver disease in mice were screened by GO and KEGG enrichment analyses， combined with relevant protein annotations in the databases and references. The expressions of miR-200a-3p， miR-200b-3p and miR-222-3p in the mice’s liver tissue detected by qRT-PCR were significantly down-regulated. In addition， the expressions of related effector genes CERS6， MYBL1， SCD2， SLCO1A4 and PLK2 were up-regulated， while the expressions of ACSS2， BCL6 and SLC10A2 were down-regulated. 【Conclusion】 The variation trend of those candidate miRNAs in mouse diabetic liver compared with that in control livers was consistent with that of the previous studies and prediction， which revealed their potential molecular regulation in this disease process.
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Objective To investigate the influence of m4-1BBL on the anti-tumor effects induced by truncated human prostate specific membrane antigen (tPSMA) gene in mice. Methods A eukaryotic expression plasmid encoding tPSMA and m4-1BBL (pDC316-tPSMA-IRES-m4-1BBL), pDC316-tPSMA and pDC316 were constructed. C57BL/6 mice were vaccinated in the quadriceps femoris, respectively. The CTL activity of spleen cells from the immunized mice against prostate cancer RM-1-tPSMA was detected by CCK-8 kit in vitro. The tumor growth was then observed. Results The target cell specific cytotoxicity rate induced by pDC316-tPSMA-IRES-m4-1BBL was 42.6%, compared to 24.8% in the pDC316-tPSMA group and 10.8% in the pDC316 group. The difference was significant (P<0.05). The volume of tumor in the pDC316 group was 2657.4mm3 7 d after vaccination, compared to 1334.5 mm3 in the pDC316-tPSMA group, 9 d after vaccination. In the pDC316-tPSMA-IRES-m4-1BBL group, the tumor volume was 445.8 mm3, 12d after vaccination. The difference was significant (P<0.05). Conclusion Gene vaccines co-expressing tPSMA gene and m4-1BBL gene could significantly enhance anti-prostate cancer effects in mice.
RESUMO
Objective To investigate the effects of isehemic postconditioning (IPO) on the acute renal ischemia/reperfusion (I/R) injury in dogs. Methods Fifteen adult male mongrel dogs were randomly divided into three groups with 5 animals in each group. In sham operation group (S), after the dogs were anesthetized, the midline laparotomy was made and right nephrectomy was performed;In I/R group, animals were subjected to the similar surgical procedures, except that the left renal vessels were clamped; In IPO group, the IPO was induced by 6 cycles of reperfusion (30 s) and ischemia (30 s) after 60 min renal ischemia before reperfusion completely. Blood samples were obtained for determination of blood creatinine (Cr) and urea nitrogen (BUN) concentrations before operation and at 24, 48 and 72 h after operation. The dogs were killed at the thirdday after operation and left kidneys were removed for determination of SOD activity and MDA and MPO concentrations.The apoptosis in the nephridial tissue was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and apoptotic index (AI) was calculated. The changes of renal tissue were examined by a microscope. Results Blood Cr and BUN concentrations in I/R group, IPO group and S group were decreased in turn after operation (P<0. 05). MDA and MPO concentrations were decreased significantly, SOD activity was significantly increased and AI was decreased significantly in IPO group as compared with I/R group at 72 h after operation (P<0. 05). Microscopic examination showed that there was no renal injury in S group and renal I/R resulted in tubular necrosis, medullary hemorrhage congestion and proteinaceous casts in I/R group. The renal I/R injury was significantly attenuated by IPO. In S group, IPO group and I/R group the renal AI was 2. 7 ±1.3, 28. 4 ± 6. 2 and 15.4±4. 1 respectively (P<0. 05). Conclusion IPO can attenuate renal damage induced by I/R by inhibiting oxidative stress and apoptosis and decreasing inflammation.