Clinical Efficacy of Gandou Fumu Granules on Wilson Disease with Liver-kidney Deficiency and Phlegm-blood Stasis / 中国实验方剂学杂志

ObjectiveTo observe the clinical efficacy of Gandou Fumu granules （GDFM） in the treatment of Wilson disease （WD） with liver-kidney deficiency and phlegm-blood stasis. MethodNinety WD patients in The First Affiliated Hospital of Anhui University of Chinese Medicine were randomly divided into a control group （45 cases） and a treatment group （45 cases）. All patients were treated with sodium 2，3-dimercaptopropane-1-sulfonate （DMPS）， while those in the treatment group received additional GDFM. All patients were treated for four courses （32 days）. The traditional Chinese medicine （TCM） syndrome scores，clinical effective rate，24 h urinary copper，ceruloplasmin （CER），tumor necrosis factor-α（TNF-α），interleukin-1β（IL-1β），interleukin-6 （IL-6），superoxide dismutase （SOD），glutathione peroxidase （GSH-Px） and malondialdehyde （MDA） levels of the two groups before and after treatment were observed. ResultAfter treatment， the TCM syndrome scores of the two groups decreased （P<0.01），and the score of TCM syndrome in the treatment group was lower than that of the control group （P<0.01）. The total effective rate of the treatment group was 82.22% （37/45）， higher than 57.78% （26/45） of the control group （χ2=6.402，P<0.05）. There was no significant difference in CER before and after treatment in both groups. The post-treatment 24 hour urinary copper increased （P<0.01）， which was higher in the treatment group than that in the control group （P<0.05）. The TNF-α，IL-1β， and IL-6 levels were significantly reduced in both groups after treatment（P<0.01），and the above indicators in the treatment group were significantly lower than those in the control group （P<0.01）. After treatment，the SOD level increased and the MDA level decreased in the control group （P<0.01）， while no significant difference in GSH-Px level was observed. The SOD and GSH-Px levels increased and the MDA level decreased in the treatment group （P<0.01）. After treatment， SOD and GSH-Px levels of the treatment group were higher than those in the control group， while the MDA level was lower than that in the control group（P<0.05，P<0.01）. ConclusionGDFM can improve the TCM syndrome score and clinical efficacy，enhance the copper removing effect，and inhibit the inflammatory response and antioxidative stress in the treatment of WD with liver and kidney deficiency and phlegm-blood stasis.

Protective Effect and Mechanism of Gandou Fumu Decoction on Oxidative Damage of HepG2 Cells Induced by CuCl2 / 中国实验方剂学杂志

Objective:To explore the effect of Gandou Fumu decoction （GDFMD） on the oxidative damage of HepG2 cells induced by CuCl₂based on the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway. Method:CuCl₂ （200 μmol·L^-1） was used to induce a copper-loaded HepG2 cell model. HepG2 cells were divided into a blank group （HepG2 cells + blank rat serum）， a model group （HepG2 cells + CuCl₂ + normal rat serum）， a GDFMD group （HepG2 cells + CuCl₂ + GDFMD-medicated rat serum）， an inhibitor group （HepG2 cells + NVP-BEZ235 + normal rat serum）， and a GDFMD + NVP-BEZ235 group （HepG2 cells + NVP-BEZ235 + GDFMD-medicated rat serum）. ELISA method was used to determine superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px） activity， and malondialdehyde （MDA） content. The expression of microtubule-associated protein 1 light chain 3 （LC3） was detected by immunofluorescence. Phospho-PI3K/PI3K （p-PI3K/PI3K）， p-Akt/Akt， p-mTOR/mTOR， Beclin-1， LC3Ⅱ/LC3Ⅰ， and p62/Actin were determined by Western blot. PI3K， Akt， mTOR， Beclin-1， LC3Ⅰ， LC3Ⅱ， p62 mRNA expression was measured by real-time polymerase chain reaction （PCR）. Result:Compared with the blank group， the model group displayed decreased activities of SOD and GSH-Px and increased content of MDA （<italic>P</italic><0.01）. Compared with the model group， the GDFMD group showed elevated activities of SOD and GSH-Px and reduced content of MDA （<italic>P</italic><0.05， <italic>P</italic><0.01）， while the inhibitor group exhibited weakened GSH-Px activity and up-regulated content of MDA （<italic>P</italic><0.05）. Compared with the blank group， the model group showed diminished expression of p-PI3K/PI3K， p-Akt/Akt， p-mTOR/mTOR， and p62， and increased expression of Beclin-1 and LC3Ⅱ/LC3Ⅰ （<italic>P</italic><0.01）. The expression of p-PI3K/PI3K， p-Akt/Akt， p-mTOR/mTOR， and p62 was elevated， and the expression of Beclin-1 and LC3Ⅱ/LC3Ⅰ declined in the GDFMD group （<italic>P</italic><0.05，<italic> P<</italic>0.01）， while the p-PI3K/PI3K and p-mTOR/mTOR expression was down-regulated and the Beclin-1 and LC3Ⅱ/LC3 I expression was increased in the inhibitor group （<italic>P</italic><0.05， <italic>P<</italic>0.01） as compared with those in the model group. Compared with the GDFMD group， the GDFMD + NVP-BEZ235 group showed down-regulated expression of p-Akt/Akt and p-mTOR/mTOR and up-regulated expression of Beclin-1 and LC3Ⅱ/LC3Ⅰ（<italic>P</italic><0.05， <italic>P</italic><0.01）. The expression of LC3Ⅱ protein in the model group was increased （<italic>P</italic><0.01） as compared with that in the blank group. The expression of LC3Ⅱ protein was lower in the GDFMD group than in the model group， and higher in the GDFMD + NVP-BEZ235 group than in the GDFMD group. No significant difference in the expression of PI3K， Akt， and mTOR mRNA was observed among the groups. Compared with the blank group， the model group displayed lowered expression of p62 mRNA， and elevated expression of Beclin-1， LC3Ⅰ， and LC3Ⅱ mRNA （<italic>P</italic><0.01）. Compared with the model group， the GDFMD group exhibited increased expression of p62 mRNA， and declining expression of Beclin-1， LC3Ⅰ， and LC3Ⅱ mRNA （<italic>P</italic><0.01）， while the inhibitor group showed increased expression of Beclin-1 mRNA （<italic>P</italic><0.05）. The expression of Beclin-1 and LC3Ⅱ mRNA in the GDFMD + NVP-BEZ235 group was elevated （<italic>P</italic><0.01） as compared with that in the GDFMD group. Conclusion:GDFMD may inhibit the excessive autophagy and alleviate the oxidative damage of HepG2 cells induced by CuCl₂， with the underlying mechanism related to the activation of PI3K/Akt/mTOR signalling pathway.

Professor 's experience of four elements of acupuncture manipulation practice / 中国针灸

Through collecting the 30-year medical experience of professor , the methods of acupuncture manipulation practice were summarized so as to provide the reference for the medical staffs in acupuncture teaching and clinical work. Professor proposes four key elements of acupuncture manipulation practice, i.e. the style of needling practice, the consciousness of needling practice, the strength of needling practice and of needling practice. Acupuncture manipulation is a highly operational clinical skill. The knowledge learning from the experience of the instructors is the important way to improve acupuncture techniques. Besides, beginners need to improve their techniques through learning the theories, a large amount of exercises as well as the self-perception.