Oxycodone inhibits myocardial cell apoptosis after myocardial ischemia-reperfusion injury in rats via RhoA/ROCK1 signaling pathway.

OBJECTIVE: The purpose of this study was to investigate the effect of oxycodone on myocardial ischemia-reperfusion injury in rats through the Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil containing protein kinase 1 (ROCK1) signaling pathway. MATERIALS AND METHODS: A total of 48 Sprague-Dawley (SD) rats were randomly divided into sham operation group, model group, oxycodone group, and inhibitor group, with 12 rats in each group. The rats in the sham operation group only underwent thoracotomy without ischemia-reperfusion injury, those in the model group were used to prepare the myocardial ischemia-reperfusion model with normal saline intervention, those in the oxycodone group were used to prepare the myocardial ischemia-reperfusion model with oxycodone intervention, and those in the inhibitor group were utilized to prepare the myocardial ischemia-reperfusion model with AG490 intervention. Then, the expressions of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (BAX) were detected by immunohistochemistry, the relative protein expressions of RhoA and ROCK1 were examined via Western blotting, and the messenger ribonucleic acid (mRNA) expressions of Bcl-2 and BAX were measured by quantitative Polymerase Chain Reaction (qPCR). Thereafter, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was adopted for apoptosis detection, and the levels of creatine kinase-muscle/brain (CK-MB), and cardiac Troponin I (cTnI) in serum were detected using an automatic biochemical analyzer. RESULTS: Immunohistochemistry results showed that compared with those in the sham operation group, the positive expression of BAX was remarkably increased (p<0.05), while that of Bcl-2 was significantly decreased (p<0.05) in the model group, oxycodone group, and inhibitor group. Compared with the model group, oxycodone group and inhibitor group had an evidently reduced positive expression of BAX (p<0.05) and an evidently raised positive expression of Bcl-2 (p<0.05). No differences were found in the positive expressions of BAX and Bcl-2 between oxycodone group and inhibitor group (p>0.05). According to Western blotting results, the relative protein expressions of RhoA and ROCK1 in the model group, oxycodone group, and inhibitor group were notably increased compared with those in the sham operation group (p<0.05). In comparison with those in the model group, the relative protein expressions of RhoA and ROCK1 in the oxycodone group and inhibitor group were predominantly reduced (p<0.05). There were no differences in the relative protein expressions of RhoA and ROCK1 between oxycodone group and inhibitor group (p>0.05). Moreover, it was discovered from qRT-PCR results that compared with those in the sham operation group, the mRNA expression of BAX was markedly raised (p<0.05), whereas that of Bcl-2 was decreased predominantly (p<0.05) in the model group, oxycodone group, and inhibitor group. Compared with the model group, oxycodone group, and inhibitor group had an evidently reduced mRNA expression of BAX (p<0.05) and a significantly raised mRNA expression of Bcl-2 (p<0.05). No differences were found in the mRNA expressions of BAX and Bcl-2 between oxycodone group and inhibitor group (p>0.05). In addition, TUNEL assay results manifested that compared with sham operation group, model group, oxycodone group, and inhibitor group had a markedly elevated apoptosis rate (p<0.05). In comparison with the model group, the apoptosis rate in oxycodone group and inhibitor group was remarkably reduced (p<0.05). There was no difference in the apoptosis rate between oxycodone group and inhibitor group (p>0.05). According to biochemical analysis results, the serum levels of CK-MB and cTnI in model group, oxycodone group, and inhibitor group were significantly increased compared with those in the sham operation group, with statistically significant differences (p<0.05). The levels of serum CK-MB and cTnI in the oxycodone group and inhibitor group were substantially lowered in comparison with those in the model group, displaying statistically significant differences (p<0.05). Besides, the levels of serum CK-MB and cTnI in the oxycodone group were not different from those in the inhibitor group (p>0.05). CONCLUSIONS: Oxycodone inhibits myocardial cell apoptosis after myocardial ischemia-reperfusion injury by suppressing the RhoA/ROCK1 signaling pathway.

[Inhibition of Sirolimus on the growth of pancreatic carcinoma and its effect on the expression of glucose transporter and hexokinase â ¡].

OBJECTIVE: To investigate whether Sirolimus could affect the glycolytic catabolism pathways of pancreatic carcinoma through the control of hypoxia induced factor (HIF-1α) to inhibit the growth of tumor, and explore the potential mechanism of targeting the signaling pathways of mTOR for the treatment of pancreatic carcinoma. METHODS: Sirolimus was applied to treat the pancreatic carcinoma in nude mice orthotopic transplantation model, its difference with the control group was compared; RT-PCR and Western blot were used to measure the mRNA and protein expression of mTOR, HIF-1α, Glucose carrier protein 1 (GLUT-1) and Hexokinase Ⅱ (HK-Ⅱ), respectively; the changes of activity of HK-Ⅱ in the tumor was determined. RESULTS: The tumor mass of the control group (1.97±0.21)g was significantly larger than that of the Sirolimus group (0.38±0.10)g (P<0.01), and the volume of the control group (1.40±0.15) mm(3) was significantly larger than that of the Sirolimus group (0.27±0.07) mm(3) (P<0.01). The expressions of mTOR, GLUT-1 and HK-Ⅱ mRNA in the control group were higher than those of the Sirolimus group (P<0.05), while no significant change was observed in the expression of HIF-1α (P>0.05); the expressions of p-mTOR, HIF-1α, GLUT-1 and HK-Ⅱ proteins in the control group were higher than those of the Sirolimus group (P<0.05). The activity of HK-Ⅱ in the control group was higher than that of the Sirolimus group (P<0.05). CONCLUSION: Sirolimus could affect the expression of GLUT-1 and HK-Ⅱ in pancreatic carcinoma through the effects of HIF-1α to inhibit tumor growth, indicating that blocking the mTOR pathway could control the glycolytic metabolism pathways of pancreatic carcinoma, which may become the new strategy for the treatment of pancreatic carcinoma.

Integrated regulatory network involving differently expressed genes and protein-protein interaction on pancreatic cancer.

OBJECTIVE: Pancreatic cancer is a deadly disease with poor prognosis. However, comprehensive understanding about its pathogenesis remains insufficient. In this study, we aimed to find potential novel approaches for the treatment of pancreatic cancer and explore the regulatory mechanisms underlying pancreatic cancer progression. MATERIALS AND METHODS: The gene expression profile data GSE32688 were downloaded from Gene Expression Omnibus database followed by background correction and normalization through GCRMA (GC Robust Multi-array Average) method. Then DEGs (differentially expressed genes) were identified using t-test method and DEGs-related PPIs (protein-protein interaction) were extracted from STRING database. The PPI networks were constructed by calculating the pearson correlation coefficient under different conditions. Moreover, the network was divided into a number of unit modules, and KEGG pathway and GO analysis were performed for genes in module networks using clusterProfiler. RESULTS: In total, 199 DEGs (165 up-regulated genes and 34 down-regulated genes) were screened between tumor and normal samples. The integrated DEG. PPI network was established by comparing two different networks under tumor and normal conditions respectively. The top ten genes with high degrees such as ANLN, PSRC1 and ECT2 were identified in the integrated network, and they were mainly enriched in cell cycle pathway. CONCLUSIONS: ECT2 and PSRC1 might be used as two novel biomarkers for diagnosis and management of pancreatic cancer.

Inorganic phosphorus fractionation and its translocation dynamics in a low-P soil.

The translocation of different inorganic phosphorus (Pi) forms in a low-P soil (Langfang experimental station, Hebei province, China) over time was investigated using P fractionation extraction and a (32)P tracer technique. The L-value and P availability of the soil was assessed using 5 different maize genotype (Zea mays L.) cultivars. The results showed that the different Pi fractions in the soil increased in the order of H(2)SO(4)-extractable P (Ca(10)-P) > Na(3)C(6)H(5)O(7)-Na(2)S(2)O(4)-extractable P (O-P) > NH(4)Ac-extractable P (Ca(8)-P) > NaHCO(3)-extractable P (Ca(2)-P), NH(4)F-extractable P (Al-P), NaOH-Na(2)CO(3)-extractable P (Fe-P), and the content of plant-unavailable P (Ca(10)-P + O-P) was high, up to 79.1%, which might be an important reason for P deficiency in this low-P soil. The (32)P tracer results showed that after the addition of (32)P-Pi to the soil with no P fertilizer applied for 25 d, 29.0% of (32)P was quickly transformed into Ca(2)-P (rapidly available P), and 66.1% of (32)P was transformed into Al-P, Fe-P and Ca(8)-P (slowly available P). Only 5.0% of (32)P was transformed into O-P and Ca(10)-P (plant-unavailable P). Moreover, in the soil with P fertilizer applied, (32)P transformation into Ca(2)-P increased, and the transformation into Ca(8)-P + Fe-P + AL-P and O-P, Ca(10)-P significantly decreased compared to the soil with no P fertilizer applied (p < 0.05). This result suggested a higher rate for water-soluble P transformation to slowly available and plant-unavailable P in P deficient soil than in soil with sufficient P. The results of maize L-value determination showed that different genotype maize cultivars had different soil P-use efficiency and low-P tolerance mechanisms. Low-P tolerant cultivar DSY-32 regulated soil P-use efficiency and plant P content according to exogenous P fertilizer application. However, another low-P tolerant cultivar, DSY-2, used soil P more efficiently, regardless of the application of exogenous P.

Cryosurgery for treatment of subcutaneously xenotransplanted tumors in rats and its effect on cellular immunity.

Cryosurgery has shown encouraging therapeutic effects on some solid tumors but its effect on the cellular immunity remains unclear. We developed a subcutaneously xenotransplanted tumor model in SD rats to directly evaluate the immune response by detecting the serum cytokine levels, T-cell responses to tumor derived antigens, and the cytolytic activity of peripheral blood mononuclear cells against the W256 cancer line with apoptosis of cells being detected using TUNEL method. 66 SD rats were divided into 2 groups with group A having 36 rats and group B having 30 rats. 30 rats in group B were equally divided into 3 groups, tumor group B, cryosurgery group B and surgery group B, with 10 rats each. 36 rats in group A were equally divided into 2 groups, cryosurgery group A and tumor group A and treatment was done accordingly. The results showed that cryosurgery induced not only destruction of the tumor cells but also cell apoptosis around the cryosurgery foci. The apoptosis ratio reached the peak 12 h after cryosurgery, with an apoptosis rate of (68.28 ± 7.85)% .Compared with surgical resection that caused significant reduction in CD31 and CD41 cell percentages, cryosurgery resulted in significantly increased percentages of CD31 and CD41 cells (P < 0.05) with a relative increase of the CD41/CD81 cell ratio. However, sIL-2R level of peripheral blood of rats in cryosurgery group which decreased more rapidly than that in surgery group over time was significantly different 3 and 5 weeks after treatment compared to surgery group (P < 0.01). Moreover, cytotoxicity of mononuclear cell was significantly enhanced after cryosurgery, which is significantly higher in cryosurgery group (P = 0.05). These results demonstrate that in addition to tumor cell destruction, cryosurgery also results in enhanced cellular immunity and antitumor immune response of the rats with subcutaneously xenotransplanted tumor, suggesting the great potential of argon-helium cryosurgery in clinical management.