1.Study on the differentiation and apoptosis of HL-60 cell line induced by Puerarin
Xuyang PENG ; Zhenghua QI ; Huangpin CHEN
Journal of Central South University(Medical Sciences) 2001;26(2):126-128
Objective To investigate the differentiation of HL-60 cells induced by different doses of Purerarin(PR) comparing with all-trans retinoic acid(ATRA)and to see if PR can induce apoptosis of HL-60 cells. Methods Cell differentiation was analyzed by NBT reduction, the ratio of NBT/MTT and CD11b ,apoptosis by morphology, DNA electrophoresis, and flow cytometry(FCM). Results 80 μg.ml-1, 160 μg.ml-1, 320 μg.ml-1 PR could induce differentiation of HL-60 cells, no significant difference was observed between the cells treated with 1 μmol.L-1 ATRA and 320 μg.ml-1 PR. Treated with 320 μg.ml-1, 640 μg.ml-1PR, HL-60 cells exhibited a morphological characteristic of apoptosis and typical DNA ladder on gel electrophoresis. FCM analysis showed that PR could interfere with cell cycle in HL-60 cells, with a increased ratio of sub-G1 in HL-60 cells. Conclusion PR exerts effect on differentiation and induces apoptosis in HL-60 cells.
2.Study on Quality Standard of Chushi Pill
Zuping ZENG ; Hong WANG ; Shan QIAN ; Bing PENG ; Xuyang HAN ; Xiaoping CHE ; Wei HE
China Pharmacy 2015;(24):3395-3398
OBJECTIVE:To establish the quality standard of Chushi pill. METHODS:Microscopic identification and TLC were adopted for the qualitative identification of Cortex moutan,C. dictamni,Angelica sinensis,Rubia cordifolia and Gardeniae fructus in Chushi pill;HPLC was performed to determine the contents of paeonol and baicalin. It was performed on column of Kro-masil 100-5 C18 with mobile phase of methanol-water-phosphoric acid(47∶53∶0.2,V/V/V)at the flow rate of 1.0 ml/min,the detec-tion wavelength was 280 nm,the temperature was 25 ℃ and the volume was 10 μl. RESULTS:The microscopic identification showed microscopic characteristics of C. moutan and C. dictamni,and characteristics of A. sinensis,R. cordifolia and G. fructus were identified by TLC;the linear range of paeonol was 0.106 24-2.124 8 μg(r=0.999 9)and baicalin was 0.059 04-1.180 8 μg (r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 2.06%;average recoveries were respective-ly 101.56%(RSD=1.68%,n=9)and 100.16%(RSD=1.13%,n=9). CONCLUSIONS:The method is simple,accurate and re-producible,and can be used for the quantity control of Chushi pill.
3.MRI diagnosis and clinical analysis for autosomal dominant polycystic kidney disease related male infertility
Huanjun WANG ; Yang PENG ; Fan ZHANG ; Haoqin ZHANG ; Xuyang XIAO ; Yan GUO ; Jian GUAN
Chinese Journal of Radiology 2019;53(2):115-120
Objective To improve the recognition and knowledge of autosomal dominant polycystic kidney disease (ADPKD) related male infertility through investigation for MRI characteristics of this disease. Methods Fourteen patients confirmed with ADPKD related obstructive azoospermia were retrospectively analyzed. All patients referred to clinic with male infertility, and obstructive azoospermia were additionally confirmed by laboratory tests and clinical examination. Subsequent abdominopelvic MR examinations were performed to comfirm obstructive factors and obstructive location. All patients were performed an abdominopelvic MR examination including non-enhanced and enhanced MR. MR imaging characteristics were analyzed and summarized by two experienced radiologists. Results MRI results for all cases were classified into 4 groups:10 cases with bilateral polycystic kidneys and bilateral seminal vesicle cysts, 2 cases with bilateral polycystic kidneys, polycystic liver and bilateral seminal vesicle cysts, 1 case with bilateral polycystic kidneys, polycystic liver and absence of bilateral seminal vesicles, 1 case with bilateral cystic kidneys, bilateral seminal vesicle cysts as well as Müllerian duct cyst. A wide range of coronal T2WI scan was necessary to observe cystic lesions in both liver and bilateral kidneys as well as abnormal changes in pelvis. The obstructive sites in all cases were located in level from ejaculatory duct to seminal vesicle. Bilateral seminal vesicle cysts presented as significantly dilated glandular ducts of seminal vesicles, in which flocculence or nodular sediment can be found. Conclusion Male infertility caused by ADPKD-related deferential duct obstrution is characterized by bilateral polycystic kidney disease and Seminal vesicle ejaculatory duct obstruction in MRI, which can be combined with other abnormalities.
4.The effect of rapamycin and underlying mechanism on hepatic ischemia-reperfusion injury in Sprague Dawley rats
Xuyang ZHANG ; Ningbo PAN ; Yu ZHANG ; Longcan YANG ; Xi YU ; Jiwei LI ; Ping HUANG ; Xueqin PENG ; Ying ZHANG
Chinese Journal of Hepatobiliary Surgery 2020;26(5):378-382
Objective:To investigate the effect of rapamycin on hepatic ischemia-reperfusion injury (HIRI) in Sprague Dawley (SD) rats and its underlying mechanism.Methods:Forty-eight specific pathogen-free SD male rats with the body weight of 180-200 g and the age of 4-8 weeks were randomly divided into 3 groups, 16 rats each group. In the rapamycin group, the rats were injected with rapamycin intraperitoneally everyday lasting for 3 days before the surgery, and in the model group and the sham group, the rats were injected with normal saline intraperitoneally. The HIRI model was performed in the rapamycin group and the model group. Serum of 8 rats was randomly harvested from each group at 2 h and 24 h after the surgery and was used to detect level of alanine aminotransferase(ALT), total bilirubin, and lactate dehydrogenase. At the meantime, liver tissues were collected for HE staining, and enzyme-linked immunosorbent assay of superoxide dismutase(SOD), glutathione, hexokinase 2, phosphofructokinase 1(PFK1), and adenosine triphosphate. Polymerase chain reaction and Western blots were used to determine the levels of mammalian target of rapamycin(mTOR), ribosomal protein S6 kinase 1(S6K1), and protein kinase B and their phosphorylation levels respectively.Results:Two hours post the surgery, the serum level of ALT(150.9±18.7) U/L, total bilirubin(5.15±0.69) μmol/L, and lactate dehydrogenase(9 547±365) U/L were higher in the model group than sham group (42.4±10.7) U/L, (2.48±0.24) μmol/L, (4 424±376) U/L and rapamycin group (87.7±11.2) U/L, (3.09±0.12) μmol/L, (8 268±264) U/L, and all differences were statistically significant (all P<0.05). HE staining and serum assay showed that the lesion of liver tissuesand of liver function were damaged in the model group, and mitigated in the rapamycin group at 2h and 24h after the surgery. At 2h and 24h after the surgery, liver SOD, glutathione, hexokinase 2, PFK1, and adenosine triphosphate in the model group were lower than those in the sham group and the rapamycin group, and all differences were statistically significant (all P<0.05). The relative levels of mTOR, S6K1, and their phosphorylation level in the model group were higher than those in the sham group and the rapamycin group at 2 h and 24 h after the surgery, but the relative levels of protein kinase B and phosphorylated protein kinase B were lower than those in the sham group and the rapamycin group, and all differences were statistically significant (all P<0.05). Conclusions:Rapamycin improves glucose metabolism and reduces oxidative stress via upregulating the phosphorylated protein kinase B through inhibition of mTOR signaling pathway, thus alleviates HIRI in rats.