Effect of Tongbiantang on PKA/MPKA Signal Pathway in Colon Tissue of STC Rat Model / 中国实验方剂学杂志

Objective: To explore the effect of Tongbiantang on protein kinase A(PKA) and mitogen-activated protein kinase(MAPK) signal pathway in colon tissue of slow transit constipation(STC) rats and its related mechanism. Method: Eighty SD rats were randomly divided into blank group and model group, 20 rats in blank group, 60 rats in model group, half male and half female; blank group was fed with common diet, model group was fed with compound phenylethylpiperidine, after 120 days of modeling, 10 rats in blank group and 20 rats in model group were randomly selected, and 2 rats were determined. Four-hour stool volume, water content and small intestinal charcoal powder propelling rate were observed to observe the number of stool particles retained in colon and evaluate the success of STC rat modeling. After 1 week of drug withdrawal, 40 rats in model group were randomly divided into model group(33 g·kg-1), Tongbiantang group, Tongbiantang+H89 group (PKA signaling pathway blocker,5 mg·kg-1), Tongbiantang+U0126 group (MPKA signaling pathway blocker,0.1 mg·kg-1) each. After 4 weeks of intervention with Tongbiantang, the amount of stool excretion, water content and small intestinal charcoal powder propelling rate were measured in 10 rats, and the number of stool grains in colon was observed. The protein content and mRNA expression in aquaporins 3(AQP3), AQP4, PKA and MAPKs signaling pathways in colon was determined by immunohistochemical staining (IHC), Western blot and Real-time fluorescence quantitative PCR (Real-time PCR). Result: Compared with the blank group, the 24-hour stool volume, fecal water content, small intestinal charcoal propelling rate and the number of fecal particles in colon of rats in the model group were significantly decreased (PPPPPConclusion: Tongbiantang can inhibit the PKA and MPKA signal pathways, thus down-regulate the expression of AQP3 and AQP4, increase intestinal peristalsis and intestinal water, and effectively treat STC.

Icaritin delays CCl4-induced hepatic cirrhosis in rats by protecting hepatocytes from oxidative injury / 第二军医大学学报

Objective To investigate the role of icaritin in delaying the progression of liver cirrhotic process in rats and the related mechanism. Methods For invitro study, primary rat hepatocytes were obtained by perfusing the liver of male Wistar rats; the cultured cells we reexposed to the fresh medium containing CCl4, and then treated with various concentrations of icaritin. The activities of alanine transaninase(ALT) and glutamic-oxaloacetic transaminase(AST) inculture medium were measured with an automatic biochemical analyzer. The intracellular contents of malondialdehyde (MDA) and superoxide dismutase(SOD) were determined by spectro-photometry; the apoptotic cells were detected by the TUNEL method. Forin vivostudy, CCl4-induced experimental rat hepatic fibrosis model was established and was treated with icaritin. Serum levels of ALT, AST, albumin(ALB), andglobulin(GLB) were measured; the pathological changes and collagen-expression in livers were observed by HE staining and immunohistochemistry, respectively. Results CCl4 significantly increased the activities of ALT, AST, and the contents of MDA in culture media of hepatocytes, and significantly decreased the SOD activity. More apoptotic cells were observed in CCl4 group than that in icaritin group. The ALT, AST activities in culture supernatant and the intracellular MDA contents in icaritin group were significantly lower than those in both model group and drug carrier group, while intracellular SOD activity was much higher than that in other two groups(P<0.05). Icaritin also reduced the apoptotic ratios of hepatocytes induced by CCl4 compared with model group(P<0.05 or 0.01). In the in vivo experiment, icaritin treatment significantly reduced serum levels of ALT and AST compared with model group(P<0.05) and greatly improved CCl4-induced liver histopathological injuries and collagen I deposition in the liver tissues. Conclusion Icaritin treatment can attenuate CCl4-induced cirrhosis in rats, atleast in part, by protecting the hepatocytes from peroxidation product.

The specific cytotoxic effect of tumor infiltrating lymphocytes transfected with chimeric T cell receptor on cells which express KDR / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the specific cytotoxity of tumor infiltrating lymphocytes (TIL) transfected with chimeric T cell receptor (CTCR) on cells which express KDR.</p><p><b>METHODS</b>A recombinant retroviral plasmid (pMSCVneo-Vhgamma) was constructed by cloning VEGF121-hinger-FcRgamma (Vhgamma) into retroviral vector pMSCVneo. After packaging by PT67, the virus with high titer was used to infect TIL isolated from liver cancer tissues, and then MSCVneo-Vhgamma-TIL was generated. TIL infected with MSCVneo was used as a control. The cytotoxicty of the transgenic TIL on NIH3T3 and HepG2 expressing no KDR and on ECV304 and A375 expressing KDR was detected with MTT colorimetric assay.</p><p><b>RESULTS</b>The sequences of VEGF121 and hinger-FcRgamma were different from those reported, but the deduced amino sequences were identical to the reported ones. The cytotoxity of TIL infected with MSCVneo on target cell was similar to that of the control TIL; both only had mild cytotoxity on cancer cell line. No significant cytotoxity was found in TIL infected with MSCVneo-cTCR on NIH3T3, but its cytotoxity on ECV304 was significant. The cytotoxity on HepG2 was similar to that of MSCVneo-TIL and uninfected TIL, but cytotoxity on A375 was significantly higher.</p><p><b>CONCLUSION</b>Chimeric T cell receptor permanently grafts TIL cell with predefined new specificity. TIL expressing Vhgamma can selectively recognize and kill vascular endothelial cell and tumor cells which express VEGF receptor KDR.</p>

Correlation between serum alpha-fetoprotein variant levels in patients with hepatocellular carcinoma and the presence of alpha-fetoprotein mRNA in the circulating blood / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the relationship between serum alpha-fetoprotein (AFP) variant levels in patients with hepatocellular carcinoma (HCC) and cancer cells disseminating through blood.</p><p><b>METHODS</b>Serum AFP variant levels were measured by crossed immunoaffino-electrophoresis in the presence of lectin before initial surgical treatment in HCC patients. Circulating tumor cells were simultaneously detected in pre-operative blood samples using reverse transcription-polymerase chain reaction (RT-PCR) for AFP mRNA.</p><p><b>RESULTS</b>Forty-six HCC patients with serum AFP positive were studied. Serum AFP variant level > or 20% was showed in 37 patients, among whom there were 22 (59.5%) showing AFP mRNA positive. In contrast, the positive AFP mRNA expression was only observed in 2 out of 9 patients (22.2%) with AFP variant level<20% (x(2)=4.02, P<0.05).</p><p><b>CONCLUSION</b>In hepatocellular carcinoma patients, increased AFP variant levels are associated with a haematogenous spread of tumor cells.</p>