OBJECTIVE To investigate the induction of apoptosis in hepatocellular carcinoma cells by polyphyllin 9 （PP9） through the regulation of the Fas/Fas ligand （FasL） signaling pathway， and its inhibitory effect on the growth of transplanted tumor in nude mice. METHODS Based on the screening of cell lines and intervention conditions， HepG2 cells were selected as the experimental subject to investigate the effects of 2 μmol/L and 4 μmol/L PP9 treatment on cell colony formation activity， apoptosis rate， as well as the protein expressions of Fas， FasL， cleaved caspase-8 and cleaved caspase-3. Additionally， Fas inhibitor KR- 33493 was introduced to investigate the underlying mechanism of PP9’s anti-hepatocellular carcinoma activity. Using HepG2 cell tumor-bearing nude mice model as the object， and 5-fluorouracil （20 mg/kg） as the positive control， the effects of 10 mg/kg PP9 on tumor volume， tumor mass， and the protein expressions of the nuclear proliferation-associated antigen Ki-67 and cleaved caspase-3 in tumor-bearing nude mice were investigated. RESULTS Compared with the control group， 2， 4 μmol/L PP9 significantly decreased the number of clones and the clone formation rate of cells， but significantly increased the apoptosis rate， the protein expressions of Fas， FasL， cleaved caspase-8 and cleaved caspase-3 （P＜0.05 or P＜0.01）. However， the combination of Fas inhibitor KR-33493 could significantly reverse the effect of PP9 on the up-regulation of proteins related to the Fas/FasL signaling pathway （P＜0.01）. Compared with the control group， the tumor volume （on day 27）， mass and protein expression of Ki- 67 in nude mice of the PP9 group were significantly decreased， while the protein expression of cleaved caspase-3 was significantly increased （P＜0.01）. CONCLUSIONS PP9 can induce apoptosis of HepG2 cells by activating the Fas/FasL signaling pathway. Meanwhile， PP9 can also effectively inhibit the growth of transplanted tumors in nude mice.

OBJECTIVE To investigate the effects of polyphyllin Ⅵ（PPⅥ） on the proliferation and apoptosis of glioma cells and potential mechanism. METHODS Using human glioma LN229 cells as objects， MTT assay was used to detect the survival rate after treated with different concentrations of PPⅥ [0 （control group）， 1， 2， 4， 8， 16， 32， 64 μmol/L] for different time （24， 48， 72 h）. The clone formation experiments were adopted to detect the number of cell clones and clone formation rate after being treated with different concentrations of PPⅥ [0 （control group）， 2， 4， 8 μmol/L] for 14 days. The flow cytometry and Western blot assay were used to detect the apoptotic rate of cells， the expressions of apoptosis-related protein [B cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， cleaved caspase-3]， and the expressions of related proteins of Fas/Fas ligand （FasL） death receptor pathway and protein kinase B （Akt）/glycogen synthesis kinase-3β （GSK-3β） pathway after being treated with different concentrations of PPⅥ [0（control group）， 4， 8 μmol/L] for 24 h. RESULTS Compared with the control group， the survival rate of cells， the number of clones and clone formation rate， the protein expression of Bcl-2， and the phosphorylation levels of Akt and GSK-3β protein were decreased significantly in different concentration groups of PPⅥ （P＜0.05 or P＜0.01）. The apoptotic rate， the protein expressions of Bax， cleaved caspase-3， Fas， FasL and cleaved caspase-8 were increased significantly （P＜0.05 or P＜ 0.01）. CONCLUSIONS PPⅥ can inhibit the proliferation and induce the apoptosis of human glioma LN229 cells， which may be related to the activation of the Fas/FasL death receptor pathway and the inhibition of the Akt/GSK-3β pathway.

Objective: To establish the quality standard for Fuzheng capsules. Methods: TLC was adopted to identify Astragali Radix and Glycyrrhiza uralensis. The method validation for Fuzheng capsules was conducted by microbial limit test as described in the appendixes of Chinese Pharmacopeia (2015 edition). The content of epimedii in Epimedium brevicornu was determined by HPLC. The chromatographic separation was carried out on an Agilent TC-C18(2) (250 mm×4. 6 mm, 5 μm) column. The mobile phase consis-ted of acetonitrile-water( 30: 70) with gradient elution at a flow rate of 1. 0 ml·min-1,and the injection volume was 5 μl. The detec-tion wavelength was 270 nm. Results: The spots in TLC were clear without any interference. The methods of plating and direct inocu-lation could be used for the microbial limit test. The linear range was 0.101-1.008 μg (r =0.999 7). The average recovery was 99. 36% with the RSD of 0. 81% (n=6). Conclusion: The method is simple with high specificity and good repeatability, which can be used as the quality control method for Fuzheng capsules.

Objective: To establish the quality standard for Shenzhi Xiaoping capsules. Methods: The characteristics of Astragali Radix were identified by microscopy. TLC was adopted to identify Astragali Radix, Fructus Aurantii and Glycyrrhiza uralensis. The con-tent of ginsenoside Rb1in Panax ginseng was determined by HPLC. Results: The microscopic characteristics were obvious. The TLC spots were clear with highly specific identification and good separation. The linear range of ginsenoside Rb1was 47. 06-376. 5 μg· ml-1(r=0. 999 9), and the average recovery was 102. 7% with the RSD of 1. 1% (n=6). Conclusion: The method is simple and specific, which can be used for the quality control of Shenzhi Xiaoping capsules.

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 analyze the rationality of proton pump inhibitors (PPIs) in the prevention of stress ulcer in perioperative orthopedic patients of our hospitals,and to provide reference for ratioual use of PPIs in clinic.METHODS:The medical records of discharged orthopedic patients were selected during Jan.1st-30th in 2016.Those records were analyzed statistically in respects of the use of PPIs,general information of patients,operation situation,the stressors and risk factors of stress ulcer.The rationality of the use of PPIs was analyzed comprehensively.RESULTS:Among 664 surgical patients,PPIs were used in 210 cases,and H2 receptor antagonist (H2RA) were used in 74 cases,with prophylactic rate of 42.77％.The main types of operation were fracture fixation (74 cases,35.24％),discectomy and discectomy (38 cases,18.11％).Among 210 patients receiving PPIs,all of them used PPIs,among which 68 cases (32.38％) had no medication indications,128 cases (60.95％) were medication timing error,74 cases (35.24％) were administration frequency error and 44 cases (20.95％) were more than 3 d prophylactic drug use.CONCLUSION:When orthopedic patients use PPIs for the prevention of stress ulcer,there are serious irrational phenomena in indications,species selection,usage and dosage,treatment course and so on.Clinicians and regulators should pay great attention to it so as to avoid the excessive application of PPIs.