Objective To investigate the effect and mechanism of emodin (EM) in renal interstitial fibrosis of unilateral ureteral obstruction (UUO) mice.Methods Male C57BL/6J mice were randomly divided into 4 groups,including sham operation group (n=8),UUO operation group (n=8),UUO operation+losartan (LST) group (n=8) and UUO operation+EM group (n=8).The mice in each group were ingested the suspensions by gavage for 14 days after surgery.Mice in UUO+LST and UUO+ EM groups were given 10 mg· kg-1· d-1 LST and 20 mg· kg-1 · d-1 EM,respectively.LST and EM were mixed with 0.5％ sodium carboxymethyl cellulose.Mice in sham group and UUO group were given 0.5％ sodium carboxymethyl cellulose.The mice were sacrificed at the 14th day.Interstitial fibrosis was observed by HE,Masson and PAS stain.Real-time PCR was used to detect LC3,Beclin-1 and mTOR mRNA.Protein expressions of TGF-β1,α-SMA,E-cadherin,LC3,Beclin-1,PI3K,p-Akt and mTOR were detected by Western blotting.The autophagy was observed with transmission electron microscopy in the renal tissue.Results Compared with sham mice,UUO mice at the 14th day displayed obvious renal fibrosis.Meanwhile,UUO mice had increased expressions of TGF-β1 and α-SMA (all P ＜ 0.01),and decreased expressions of E-cadherin (P ＜ 0.01).Their renal expressions of PI3K,p-Akt and mTOR were also raised (all P ＜ 0.01).Compared with those in UUO group,in UUO+LST group and UUO+EM group,expressions of autophagy protein LC3 and Beclin-1 were increased (all P ＜ 0.01),and the number of autophagic was increased.Additionally,expressions of TGF-β1 and α-SMA were reduced in UUO+LST group and UUO+EM group (all P ＜ 0.01),while the expression of E-cadherin was increased by emodin treatment (P＜ 0.05).And expressions of PI3K,p-Akt and mTOR were decreased in UUO + LST group and UUO + EM group (all P ＜ 0.05),meanwhile renal tissue fibrosis significantly reduced.Conclusions Emodin can promote autophagy,ameliorate renal interstitial fibrosis and protect renal function through PI3K/Akt/mTOR signaling pathway.

Objective: To investigate the mechanism of emodin ( EM) in the expression of related protein for the fibrosis of the transforming growth factor-β1(TGF-β1)-stimulated human renal tubular epithelial (HK-2) cells. Methods: HK-2 cells were randomly divided into the normal control group, TGF-β1-stimulated model control group and emodin ( TGF-β1 +EM) group. The contents of Collage Ⅰ and fibronectin in the culture supernatant were determined by ELISA. After HK-2 cells were stimulated with TGF-β1 for 24 h, the cells were collected for immunofluorescence, Western blot and RT-PCR analysis. RT-PCR was used to detect PI3K, p-Akt and mTOR. The protein expressions of PI3K, p-Akt and mTOR were detected by Western blot. Immunofluorescence was used to detect PI3K. Results: Compared with those in the model control group, the contents of CollageⅠand fibronectin in the supernatant of emod-in group significantly decreased (P<0. 05), the expression of PI3K protein was inhibited, the expression of downstream p-Akt protein decreased, and the downstream mTOR decreased (P<0. 05), the expression levels of PI3K, p-Akt and mTOR mRNA decreased, the differences were statistically significant (P<0. 05), and the expression of PI3K decreased. Conclusion: Emodin can alleviate fibrosis of HK-2 cells stimulated by TGF-β1 through the classical Akt/mTOR pathway of autophagy.

OBJECTIVE Based on the retrieval and analysis of cases related to the invalidation of drug patents in recent years in China, this paper puts forward relevant suggestions on the operation and management of pharmaceutical enterprises in China. METHODS According to analyzes a total of 503 invalidation request cases filed with the National Intellectual Property Administration from January 1, 2005 to July 12, 2020, analyzed them from the dimensions of the number of cases, involved parties, legal basis, related litigation, etc., which aimed to systematically and comprehensively understand the general situation of patent invalidation cases in the field of medicine in China. RESULTS The number of cases was increasing rapidly year by year, gradually forming a complete closed loop of patent challenges with litigation; the average closing period of administrative litigation after an invalidation case declared was 31.1 months. Judging from the judgment results, the success of patent invalidation rate over 50%. CONCLUSION Pharmaceutical enterprises should pay attention to patent evaluation, analysis and management in the whole process of drug R&D and marketing, improve their independent R&D capabilities and patent management quality.