Effect of Qingyi Decoction on gene expression profiles of severe acute pancreatitis rats by gene chip technique / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of Qingyi Decoction (QYD) on pancreatic gene expression profiles in rats with severe acute pancreatitis (SAP).</p><p><b>METHODS</b>Totally 60 Sprague-Dawley (SD) rats were randomly divided into the sham-operation group (SO group), the SAP group, and the QYD group, 20 in each group. SAP model was replicated by the pancreatic duct retrograde injection with 4% sodium taurocholate. Rats in the QYD group was intragastrically intervened by QYD (0.75 mL/100 g) for 3 times. Pancreatic RNA expression was analyzed using Illuminate whole genome expression profiles. Changes of mRNA and protein in specific genes [heat shock proteins a8 (Hspa8) and heat shock proteins b1 (Hspb1)] were verified by real-time quantitative PCR and Western blot analysis.</p><p><b>RESULTS</b>Compared with the SAP group, 575 differential genes were screened in the QYD group, including 92 up-regulated genes and 483 down-regulated genes. Gene Ontology (GO) categories indicated the genes are associated with negative regulation of transcription regulator activity, oxidoreductase activity and enzyme inhibitor activity. Effects of QYD on the SAP rats were major related to mitogen-activated protein kinase (MAPK), NOD like receptors (NLR) receptor-like signaling pathway, cell cycle, metabolic pathways, oxidoreductase activity. Protein and mRNA changes of Hspa8 and Hspb1 in microarray were verified [relative mRNA expression for Hspa8 and Hspb1 was increased by (13.24 +/- 1.22) times and (7.55 +/- 1.09) times respectively, P < 0.01].</p><p><b>CONCLUSION</b>QYD was effective in treating experimental SAP involved the MAPK and NLR signaling pathways, cell cycle, metabolic pathways, and oxide reductase activities.</p>

Pioglitazone ameliorates nonalcoholic steatohepatitis by down-regulating hepatic nuclear factor-kappa B and cyclooxygenases-2 expression in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Pioglitazone is effective in nonalcoholic steatohepatitis (NASH), but the mechanisms of action are not completely understood. This study was designed to investigate the effects of pioglitazone on hepatic nuclear factor-kappa B (NF-κB) and cyclooxygenases-2 (COX-2) expression in NASH rats.</p><p><b>METHODS</b>Thirty Sprague-Dawley male rats were randomly assigned to a control group (n = 10), NASH group (n = 10), and pioglitazone treatment group (n = 10). Liver tissues were processed for histology by hematoxylin & eosin and Masson stained. Serum alanine aminotransferase (ALT), cholesterol, triglyceride, fasting blood glucose (FBG), fasting insulin (FINS) levels and biochemical parameters of antioxidant enzyme activities, tumor necrosis factor alpha (TNF-α), prostaglandin E(2) (PGE(2)) levels in serum and liver were measured. The mRNA and protein expression of peroxisome proliferator-activated receptor gamma (PPARγ), NF-κB and COX-2 were determined by real-time polymerase chain reaction, Western blotting and immunohistochemistry. One-way analysis of variance (ANOVA) and Wilcoxon's signed-rank test was used for the statistical analysis.</p><p><b>RESULTS</b>There were severe steatosis, moderate inflammatory cellular infiltration and fibrosis in NASH rats. After pioglitazone treatment, steatosis, inflammation and fibrosis were significantly improved compared with the NASH group (χ(2) = 20.40, P < 0.001; χ(2) = 20.17, P < 0.001; χ(2) = 13.98, P = 0.002). Serum ALT, cholesterol, triglyceride, FBG, FINS levels were significantly elevated in the NASH group (P < 0.05). In the NASH group, total anti-oxidation competence (T-AOC), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX) and malondialdehyde (MDA) levels in serum and liver were conspicuous disordered than those parameters in the control group. Meanwhile, TNF-α and PGE(2) levels in serum and liver were significantly increased compared with the control group. Immunohistochemistry showed NF-κB and COX-2 expression in liver was significantly elevated. However, PPAR? level was decreased in the NASH group. Real-time PCR and Western blotting revealed mRNA and protein expression of COX-2 were increased in the NASH group compared with the control group (0.57 ± 0.08 vs. 2.83 ± 0.24; 0.38 ± 0.03 vs. 1.00 ± 0.03, P < 0.001 and P = 0.004, respectively). After pioglitazone intervention, all of those parameters markedly improved (P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>Down-regulating hepatic NF-κB and COX-2 expression, at least in part, is one of the possible therapeutic mechanisms of pioglitazone in NASH rats.</p>

Preparation, identification and application of polyclonal antibodies against human PIWIL2 protein / 第二军医大学学报

Objective: To prepare rabbit polyclonal antibody against human P-element-induced wimpy testis like 2 (HIWI2) protein, identify its properties and investigate its distribution in normal and tumor tissues by means of tissue chip. Methods: PIWIL2 peptide was synthesized chemically and conjugated to Keyhole limpet hemocyanin (KLH) as an immunogen. Then the PIWIL2-KLH conjugation was injected into rabbits subcutaneously to produce polyclonal antibodies. The specificity and sensitivity of antibodies were identified by ELISA and Western blotting after purification by affinity chromatography. PIWIL2 was then immuno-stained on the tissue chip to study its distribution in the normal and tumor tissues. Results: Rabbit's antibodies against human PIWIL2 were prepared after the injection of PIWIL2-KLH conjugation subcutaneously. These antibodies were identified as PIWIL2 peptides by ELISA and Western blotting assay. PIWIL2 protein expression was tissue-specific in tumor tissues, with PIWIL2 protein found in the cytoplasm of smooth muscle cells of most normal and tumor tissues. Conclusion: The polyclonal antibodies against human PIWIL2 protein have been successfully prepared, which provides a basis for further study on the role of PIWIL2 in the pathway of miRNA/RNA.

Antitumor effects of specific cyclooxygenase inhibitors combined with chemotherapeutic agents on gastric cancer cells in vitro / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the effects of two specific cyclooxygenase inhibitors (SCI), rofecoxib and celecoxib, combined with chemotherapeutic drugs 5-Fu, DDP and VP-16 on gastric cancer cell line BGC-823, and to evaluate whether specific cyclooxygenase inhibitors can be used as a synergetic agent in chemotherapy.</p><p><b>METHODS</b>The gastric cancer cell line BGC-823 cells were incubated for 48 hours with rofecoxib and celecoxib, 5-Fu, DDP and VP-16 (concentration gradient of 5-Fu, DDP and VP-16:1 microg/ml, 10 microg/ml and 100 microg/ml), or in combination, respectively. MTT working solution was added to each culture and calculated the survival rates of gastric cancer cells. Median-effect principle and Professor Jin's evaluation methods were applied to detect the interaction between the specific cyclooxygenase inhibitors and chemotherapeutic agents.</p><p><b>RESULTS</b>The inhibition rates of gastric cancer cells were 42.63% +/- 1.26% and 50.67% +/- 2.35% by treatment with 0.1 micromol/L rofecoxib and 50 micromol/L celecoxib, respectively. The inhibition rates of gastric cancer cells by treatment with 5-Fu, DDP and VP-16 at different concentrations (1 microg/ml, 10 microg/ml and 100 microg/ml) were 39.75% +/- 3.14%, 49.96% +/- 2.08%, 87.93% +/- 3.66%; 48.28% +/- 2.08%, 59.46% +/- 1.69%, 88.23% +/- 4.81%; and 29.23% +/- 3.27%, 49.34% +/- 3.75%, 79.24% +/- 2.44%, respectively. However, the inhibition rates showed a synergetic role while combined the two SCI (0.1 micromol/L rofecoxib and 50 micromol/L celecoxib) with chemotherapeutic agent at different concentrations (P <0.05).</p><p><b>CONCLUSION</b>Both rofecoxib and celecoxib have an ability to suppress gastric cancer cells in vitro, and the synergetic role becomes evident when rofecoxib and celecoxib are combined with chemotherapeutic agents at different concentrations, which indicate that the two specific cyclooxygenase inhibitors may be used as a chemotherapeutic sensitizer.</p>