1.Effect of matrine on cell proliferation, apoptosis and autophagy of human medullobIastoma D341 cells in vitro
Kaiyu ZHOU ; Hailong JL ; Tianming MAO ; Zhiqiang BAL
Chinese Journal of Pharmacology and Toxicology 2015;(2):240-246
OBJECTIVE To explore the effect of matrine induced proliferation, apoptosis and auto-phagy on human medulloblastoma cell line D341 in vitro. METHODS D341 cells in vitro were incubated with matrine 0, 0.5, 1.0, 1.5 and 2.0 g.L-1 for 24, 48 and 72 h, respectively. The proliferation of D341 cells was analyzed using Cell Counting Kit-8 assay. Apoptosis was detected by flow cytometry. The mor-phologic change of cells was observed under a transmission electron microscope. The expression of Bax, Bcl-2, caspase 3, microtubule associated protein 1 light chain 3 (LC3) and beclin1 was detected by Western blotting, and the expression of LC3 and beclin1 was detected by Western blotting with or without the autophagy inhibitor 3-methyladenine(3-MA). 3-MA was added 1 h before matrine and the final concentration of 3-MA was 5 mmol.L-1 . RESULTS Matrine significantly inhibited the proliferation of D341 cells. There was a concentration-effect relationship ( r24 h = 0.994, r48 h = 0.992, r72 h = 0.996, P<0.01). Matrine could induce the cell apoptosis (r24 h = 0.937, r48 h = 0.947, r72 h = 0.987, P<0.01). When the concentration of matrine was 2.0 g.L-1 , the inhibitory effect on D341 cell proliferation (r=0.999, P<0.01) and the induction of cell apoptosis (r=0.990, P<0.01) had a time-dependence. When the concen-tration of matrine was 2.0 g.L-1 , the ultrastructure of the D341 cells had obvious change. Cells with acoustic cavitation bubble structure, chromatin condensation, and marginalization were observed after matrine treatment for 24 h. After 48 h treatment with matrine, nuclear chromatin condensation and more vacuoles in the cytoplasm were observed. After 72 h treatment with matrine, cells exhibited apoptotic characteristics with obvious nuclear chromatin condensation, and nuclear fragmentation, significantly increased the larger cytoplasmic vacuoles. Western blotting analysis showed that matrine could increase the expression of Bax (r24 h =0.981, r48 h =0.967, r72 h =0.998, P<0.01), and decrease the expression of Bcl-2 (r24 h = -0.977, r48 h = -0.989, r72 h = -0.968, P<0.01). Matrine could increase the expression of caspase 3 when the effect time was 48 h (r48 h =0.995, P<0.01). Matrine also increased the expression of beclin1 (r24 h =0.989, r48 h =0.986, r72 h =0.966, P<0.01). The autophagy inhibitor 3-MA could reduce this effect ( P < 0.05). Matrine decreased the expression of LC3-Ⅰ but increased the expression of LC3-Ⅱ and thus the ratio of LC3-Ⅰ/ LC-Ⅱ was decreased (r24 h = -0.795, r48 h = -0.886, r72 h = -0.901, P<0.05). 3-MA could reduce the effects of matrine on LC3-Ⅰ and LC3-Ⅱ expression of D341 cells (P<0.05). CONCLUSION Matrine can inhibit proliferation, induce apoptosis and promote autophagy of D341 cells in vitro.
2.Design and Application of Computer-assisted Distal Radial Anatomic Splint
Yong LIU ; Mao WU ; Jiefeng SHEN ; Hailong NING ; Yong MA ; Jianwei WANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(5):1167-1171
This study was aimed to design the distal radial anatomic splint which was more in line with the human anatomy and treatment characteristics using the computer design software 3D-MAX. According to ergonomic design principles, the forearm and wrist plaster splint were selected from volunteer models. Then, data of the plaster was measured and input into the computer for the design of the three-dimensional model of distal radial anatomic splint with 3D-MAX software. Finally, the blueprint was drawn for the factory to make the distal radial anatomic splint. The results showed that the distal radial anatomic splint was more in line with human anatomy, which did not require shaping in clinical using. It did not affect the biomechanical properties. And the patients never complain the squeez-ing discomfort of the splint. It was concluded that the three-dimensional model of the distal radial anatomic splint, which designed with 3D-MAX software, provided key parameters of the important part of the distal radial anatomic splint. Therefore, the produced splint was more in accordance with the human anatomy and the clinic treatment re-quirements. It avoided the loss of biomechanical properties after shaping, which was more convenient and effective in the clinical using. Its clinical promotion has a broad prospect.
3.Balloon dilatation plus support tube for treatment of biliary stricture after orthotopic liver transplantation by using the endoscope technique
Yulong YANG ; Hailong CHEN ; Wenxiang TAN ; Zhongyi FENG ; Xiaoguang WANG ; Ligang XI ; Hongwei GUO ; Wei MAO ; Wencai Lü
Chinese Journal of Tissue Engineering Research 2008;12(31):6181-6186
BACKGROUND:Biliary stricture following liver transplantation is mainly focus on biliary stoma stricture; while, balloon dilatation temporarily keeps biliary tract open but not works out a solution at all.OBJECTIVE: To discuss the diagnosis and treatment of postoperative biliary stricture after orthotopie liver transplantation by the endoscope technique.DESIGN, TIME AND SETTING: A case analysis, which was performed at Dalian Liver and Gall Surgical Institute. Ten patients hospitalized from the Department of Liver and Gall Surgery of Dalian Friendship Hospital and four patients hospitalized from the Department of Organ Transplantation of Tianjin First Central Hospital were diagnosed as biliary stricture after orthotopic liver transplantation.PARTICIPANTS: Among 14 patients, 10 males and 4 females with mean age of 46 years provided end-to-end biliary anastomose.METHODS: Fourteen cases of postoperative biliary stricture after orthotopic liver transplantation were analyzed and diagnosed by endoscope technique. And by endoscope technique, the stricture was supported with tube after balloon dilatation.MAIN OUTCOME MEASURES: Bile duct mucous membrane under T-tube radiography and endoscope; calculary distribution and bile duct mucous membrane at stoma; healing of biliary stoma of donors and recipients; inflammatory edema and stricture; recheck of above-mentioned parameters after stricture expansion by endoscopic stone extraction technique.RESULTS: Thirteen cases of postoperative biliary stricture after orthotopic liver transplantation were analyzed and diagnosed by endoscope technique, including one was induced by calculus, and one non-stoma stricture. One case was treated with balloon dilation; biliary infection and jaundice occurred in 2 cases after endoscopic sphincterotomy (EST) + basket lithotripsy + endoscopic nasobiliary drainage (ENBD), so operations or fibrocholedochoscope treatments had to be carried out. By T tube radiography, in 1 case there was strip-like negative simulacrum or no stricture, well-healed anastomosis and good mucous membranel transition; poor or no intrahepatic visualization were found in 2 cases, so anastomosis dilation was processed after the calculi removal by fibrocholedochoscope, stricture disappeared in 3 or 4 months; in 8 cases there were blur extrahepatic or intrahepatic biliary visualization, cord-like, column or branch-like negative simulacrum in biliary ducts and sign of non-anastomosis stricture, after removal of calculi, anastomosis stricture and congestion, edema were found, all these disappeared after average 2.5 months of dilation; the other 1 case was found stricture by T the radiography, but no calculi was found with fibrocholedochoscope, finally the Ttube was removed after 2 months of stricture dilation.CONCLUSION: Endoscopy is significant to directly reflect and reliably diagnose postoperative biliary stricture and effectively treat biliary stricture by anastomosis dilation.
4.Relationship between homocysteine and sleep disorders in patients with Parkinson's disease
Hao WANG ; Hailong ZHAO ; Yuqing FANG ; Fei MAO ; Shanshan LU ; Xiangqi TIAN ; Zhangning ZHAO ; Jiyou TANG ; Xiuhua LI
Chinese Journal of Behavioral Medicine and Brain Science 2022;31(2):141-147
Objective:To explore the characteristics of sleep disorders in patients with Parkinson's disease (PD) and its correlation with homocysteine.Methods:Totally 75 PD patients hospitalized in the department of neurology from January 2017 to June 2021 were selected and divided into sleep disorder group ( n=39) and non-sleep disorder group ( n=36)according to polysomnography, Parkinson's disease sleep scale(PDSS) and Epworth sleepiness scale(ESS). The basic clinical data, hematological examination results, scale evaluation data and polysomnography monitoring data of the above patients were collected during hospitalization to analyze the sleep characteristics of patients with Parkinson's disease and its correlation with homocysteine.SPSS 26.0 statistical analysis software was used for t test, Mann-Whitney U test, Pearson analysis, Spearman analysis and multivariate Logistic analysis. Results:The sleep efficiency (56.82±19.07)%, N2 phase ratio(48.67±17.70)%, N3 phase ratio(9.20%(19.00%)) and the leg movement micro-arousal index(0(1.20)) in the sleep disorder group were lower than those in the non-sleep disorder group (sleep efficiency (82.15±5.55)%, N2 phase ratio(57.02±2.80)%, N3 phase ratio(20.01%(3.93%)), the leg movement micro-arousal index(1.15(1.80)). The differences were statistically significant ( t/ Z=-6.087, -2.905, -3.773, -3.683, all P<0.05). The proportion of AHI (0.90(14.60)), N1 stage (19.50%(15.70%)), and periodic limb index (0(24.80)) in sleep disorder group were higher than those in non-sleep disorder group (AHI (0.60(0.30)), N1 stage (12.15%(3.15%)), and periodic limb index (0(0)). The difference was statistically significant ( Z=2.154, 5.250, 3.559, all P<0.05). The homocysteine (15.80(3.90) μmol/L), NMSS-insomnia correlation score (3.00(5.00)), MDS-UPDRS-Ⅰ(7.00 (10.00)), MDS-UPDRS-Ⅲ (23.00 (16.00)) in the sleep disorder group were higher than those in the non-sleep disorder group (homocysteine (14.10 (4.20)μmol/L), NMSS-insomnia correlation score (0(1.00)), MDS-UPDRS-Ⅰ(3.00 (2.00)), MDS-UPDRS-Ⅲ (17.00 (4.00)), and the differences were statistically significant( Z=2.557, 4.487, 2.952, 2.180, all P<0.05). The NMSS-olfactory correlation scores (2.00(4.00)) and PDSS (99.00 (40.00)) were lower than those in the non-sleep disorder group (NMSS-olfactory correlation scores (4.50 (7.00)) and PDSS (122.00 (28.00)), and the differences were statistically significant ( Z=2.450, 4.126, both P<0.05). Hcy was positively correlated with sleep disorder in PD patients ( r=0.297, P<0.05). Binariate logistic regression analysis showed that elevated homocysteine level might be a risk factor for sleep disorder in PD patients ( β=0.193, OR=1.213, 95% CI=1.029-1.430). Conclusion:Parkinson's disease patients with sleep disorder have the characteristics of sleep structure disorder, often accompanied by more serious motor disorders, and the olfactory function impairment is relatively mild. Elevated homocysteine levels may be a risk factor for sleep disorder in Parkinson's disease.
5.Content Determination of Alkaloids and Catalpol in Different Proportions of Qianjin Huanglian Pills
Hailong MAO ; Rui BAO ; Xiao WAN ; Hongli YAN
Herald of Medicine 2024;43(4):601-606
Objective To establish an HPLC method for the determination of alkaloids(epiberberine,coptisine,palma-tine,berberine)and catalpol in different ratios(1∶1,1∶10)of ancient and modern Qianjin Huanglian Pills,and to compare the differences in their contents.The content differences were compared to preliminarily evaluate the differences in the efficacy of Qianjin Huanglian Pills in the treatment of diabetes under different preparation processes and different ratios.Methods The alkaloid solvent was methanol∶ hydrochloric acid(100∶1).The detection conditions were as follows:C18 column,acetonitrile-0.05 mol·L-1 potassium dihydrogen phosphate solution(50∶50),detection wavelength 345 nm,column temperature 30℃,flow rate 1 mL·min-1,injection volume 10 μL.The catalpol solution was methanol∶ water(20∶80).The detection conditions were as follows:chromatographic column C18 column,methanol-0.1%phosphoric acid solution(1∶ 99),detection wavelength 210 nm,column temperature 30℃,flow rate 1 mL·min-1,injection volume 10 μL.Results The established method was spe-cific,and the separation effect of the five components was good.It exhibited a good linear relationship(R2>0.999)in their respec-tive linear ranges.The repeatability,precision,stability,and sample recovery rate all met the requirements.The content of four alka-loids in the ancient method 1∶1 was the highest,and the content of catalpol was the lowest.The content of four alkaloids in the ancient method 1∶10 was the lowest;the content of 1∶1 in the present method was higher than that in the ancient method 1∶10,and the content of berberine in the present method 1∶10 was slightly lower than that in the present method 1∶1,and the rest were higher than that in the present method 1∶1.The PCA results showed that the chemical composition contents of the four kinds of Qianjin Huanglian pills were very different.Conclusion The method is simple,accurate,and reproducible,making it suitable for the quality control of Qianjin Huanglian Pills.It provides a theoretical basis for exploring the difference in efficacy of Qianjin Huanglian Pills.