1.Effect of mulberry anthocyanin suppressing autophagy on morphology and function of vascular endothelial cells through activating PI3K/Akt/mTOR pathway
Kun WANG ; Chuancai DAN ; Xiang HU ; Zhaoyang PI ; Huanlu WANG ; Dandan ZHU ; Yumeng ZHOU ; Shaofeng XIONG
Chongqing Medicine 2025;54(2):289-296
Objective To investigate the effect of mulberry anthocyanin on the morphology and func-tion of vascular endothelial cells(VEC)and its possible mechanism at the level of autophagy.Methods VEC were cultured in vitro and divided into 5.5/0 mmol/L control group,11/0.125,22/0.250,33/0.500,44/1.000,55/2.000 mmol/L high glucose/high fat groups.The cell functional levels and autophage levels in each group were detected.The optimal high glucose/high lipid concentration was selected as the VEC function inju-ry model group.10,25,50 μmol/L mulberry anthocyanins were added into the model group for processing and divided into the low,middle and high concentrations of mulberry anthocyanin groups.The effects of mulberry anthocyanin on the morphology,function and autophagy of VEC in high sugar and high fat stress were ob-served.Then the autophagy regulator(rapamycin)was added to study its action mechanism.HE staining was used to observe the number and morphology of cells,the CCK-8 assay was used to detect the viability of VEC,the flow cytometry was used to detect the level of reactive oxygen species(ROS)in VEC,the Western blot was used to detect the expression level of Beclin-1 and p62 proteins in VEC,and the kit method was used to detect the level of nitric oxide(NO)and endothelin-1(ET-1)in VEC.Results Compared with the control group,the number and morphology of cells in the middle and high concentrations of high glucose/high fat groups were changed,the cell viability,p62,NO levels were significantly decreased(P<0.05),while the ROS,ET-1,and Beclin-1 expression levels were increased significantly(P<0.05).Compared with the model group,the number of cells in the mulberry anthocyanin groups was increased,morphology was improved,the cellular viability,p62 and No levels were significantly increased,and ROS,Beclin-1 and ET-1 levels were significantly decreased(P<0.05).Compared with the high concentration mulberry anthocyanin group,the cells number was decreased af-ter adding rapamycin,the morphology changed to be irregular,the NO level was decreased,and the ET level was increased(P<0.05).Compared with the control group,the p-phosphatidylinositol 3-kinase(PI3K)and p-protein kinase B(Akt)relative expression levels in the model group were significantly decreased(P<0.05);compared with the model group,the p-PI3K and p-Akt relative expression levels in the high concentration mulberry anthocyanin group were significantly increased(P<0.05).Conclusion Mulberry anthocyanin could in-hibit autophagy by activating the PI3K/Akt/mammalian target of rapamycin(mTOR)signaling pathway and improve the morphology and function of VEC under high glucose and lipid stress.
2.Determination of 17 Characteristic Ingredients of Plant Extracts in Hair Growth Cosmetics by Ultra High Performance Liquid Chromatography
Jianhua TAN ; Huiyong LI ; Shaofeng XI ; Changhong GUO ; Jicai WANG ; Xiaoting XIONG ; Yanping XIAN ; Xindong GUO
Chinese Journal of Analytical Chemistry 2015;(1):110-114
A method was developed for the simultaneous determination of 17 characteristic ingredients of plant extracts, including paeoniflorin, hydroxysafflor yellow A, calycosin_7_glucoside ferulic acid, etc. , in hair growth cosmetics using ultra high performance liquid chromatography ( UPLC ) . Different cosmetic samples were extracted by ultrasonic_assisted extraction with the solvent of methanol/water (4∶1, V/V) solution. After demulsified by the addition of appropriate amount of NaCl and high speed centrifugation, the supernatant was transferred and analyzed with UPLC. The separation was conducted on a Waters reversed phase column of ACQUITY UPLC CSH C18(50 mm×2. 1 mm, 1. 7μm), and the mobile phases were methanol and the solution of 0. 05% phosphate in water. The detection was performed with a photodiode_array ( PDA) detector. The linear range was 0 . 2-25 mg/L with correlation coefficients higher than 0 . 999 . The limits of detection were within 0. 3-1. 5 mg/kg, and the limits of quantification were from 1. 0 to 4. 0 mg/kg. The average recoveries of 17 characteristic ingredients were within 93 . 5%-105 . 0%, with the intra_and inter_day precision ( n=6 ) less than 4. 6%. This method was simple, rapid, with good_repeatability, and had been applied to the analysis of real samples.
3.The study of the expression of heat shock protein 70 in lung tissue of rats with acute paraquat poisoning and the invention with ulinastatin
Congyang ZHOU ; Zhijian ZHANG ; Shaofeng GONG ; Huawei XIONG
Chinese Journal of Emergency Medicine 2010;19(9):953-956
Objective To observe the expression of heat shock protein 70 (HSP70) in lung of rats after paraquat (PQ) poisoning and to investigate the therapeutic effects of ulinastatin. Method Seventy-two adult healthy SD rats were randomly(random number) divided into control group (group A, n = 24), poisoning group (group B, n =24) and ulinastatin group (group C, n =24). The rat models of acute PQ poisoning were established by intra-gastric administration of 80 mg/kg PQ to rats of group B and group C, and the rats of group C were intra-peritoneally injected with 100 000 IU/kg ulinastatin 30 minutes after poisoning. The expression of HSP70 in lung tissue was observed, and W/D and histopathological changes in lung tissue were compared 12 h,24 h,48 h and 72 hours after poisoning. The expression of HSP70 in lung tissue was assayed by using RT-PCR. All quantitative data were processed with one-way analysis of variance to compare multiple sample means. Results Compared with group A, the expression of HSP70 in the lung of rats in group B and group C increased significantly at all intervals ( P < 0.05). The pathological changes in lung tissue of rats with PQ poisoning showed mainly congestion,leukocytes infiltration and local hemorrhage, and the pathological changes in lung tissue of group C were significantly lessened. Conclusions Ulinastatin may ameliorate the acute lung injury to a certain extent after PQ poisoning in rats by enhancing the expression of HSP70.
4.Study on surgical approaches for small hepatocellular carcinoma with biliary tumor thrombi
Keke YU ; Baimeng ZHANG ; Pingguo LIU ; Shaofeng WU ; Yong LI ; Yu XIONG
Chinese Journal of Postgraduates of Medicine 2006;0(08):-
Objective To investigate the surgical approaches and effects for small hepatocellular carcinoma(HCC) with biliary tumor thrombi.Mothods The clinical data of 14 small HCC patients with biliary tumor thrombi who underwent surgical intervention were analyzed retrospectively.Results The postoperative mean survival time of patient with removal of biliary tumor thrombi(n=4) was 7 months.Three patients underwent limited hepatectomy and removal of biliary tumor thrombi,2 of them survived 4 and 21 months respectively,the other cases underwent second operation 2 months later for recurred tumor in liver.Eight patients underwent left or right hemihepatectomy and removal of biliary tumor thrombi,3 of them combined with extrahepatic bile duct resection and reconstruction.Survival times of these patients were 55 months,53 months,44 months(still alive),31 months(still alive),29 months,18 months,11 months(still alive) and 1 month respectively.Conclusions Regular hemihepatectomy,meanwhile combined with the invaded extrahepatic bile duct resection,might help to prolong the survival time of patients of small HCC with biliary tumor thrombi.

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