1.Protective Effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside on PC12 Cells Injury Induced by H2O2
Haihan MA ; Yang SHAO ; Lixue CHEN ; Qidong YANG
Chinese Journal of Rehabilitation Theory and Practice 2010;16(4):322-324
ObjectiveTo study the protective effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside(TSG) on the PC12 cells injury induced by H2O2.MethodsAll cells were divided into 5 groups: the saline control group(control group), Oxidative damage PC12 cells model group(H2O2 group) which was induced by H2O2, and TSG treated group, which oxidative damage PC12 cells model which was induced by H2O2 after given TSG 2 h at TSG 120 μg/L(TSG 120 μg/L+H2O2 group), 60 μg/L(TSG 60 μg/L+H2O2 group) and 30 μg/L(TSG 30 μg/L+H2O2 group) once. The survival rate of the cells was determined by MTT method, the content of lactate dehydrogenase (LDH) was determined by ultraviolet spectrophometry and the content of malonyldialdehyde (MDA) and superoxide dismutase (SOD) activity was measured respectively by thiobarbituric aicd and xanthine oxidese method. Immunohistochemitry method were used to detect the expression of bcl-2.ResultsTSG reduced obviously cells injury induced by H2O2. 30~120 μg/L TSG improved the cells survival rate, reduced LDH releasing and MDA content, increased SOD activity, and decreased the expression of bcl-2 in the PC12 cells injury induced by H2O2(P<0-05, P<0-01). ConclusionTSG has significantly potective effect on the PC12 cells injury induced by H2O2.
2.The clinical study on early minimal feeding of very low birth weight infant
Zhen CHEN ; Jing CHEN ; Zhenying YANG ; Xianhua SHAO
Chinese Journal of General Practitioners 2005;0(12):-
0.05). The amount of daily body weight gain in early minimal feeding group [(20.8?7.0)g/d] was higher than other groups (P0.05); The incidence of complications (apnea, vomit, abdominal distension, gastrorrhagia) was lower than in gastrogavage group (P0.05). Following all the patients when they were eighteen months old, there was respectively one cerebral palsy in the gastrogavage group and TPN group, and one mental retardation in TPN group. Conclusions Early minimal feeding and early sucking up could reduce the occurrence of feeding complications,shorten the length of hospitalization and reduce the incidence of sequelae in VLBWI.
3.Mini-invasive multichannel drainage in the treatment of hypertensive cerebral hemorrhage combined with severe ventricular hemorrhage
Gang YANG ; Gaofeng SHAO ; Jiangli CHEN ; Fei CHEN ; Donghai YUAN
Chinese Journal of Postgraduates of Medicine 2016;39(4):318-321
Objective To evaluate the clinical efficacy of mini-invasive multichannel drainage in the treatment of hypertensive cerebral hemorrhage combined with severe ventricular hemorrhage. Methods The clinical data of 76 patients with hypertensive cerebral hemorrhage combined with severe ventricular hemorrhage were analyzed. They were divided into observation group and control group by random digits table method with 38 cases each. The patients in observation group were performed intracranial hematoma catheterized drainage combined with ventricle drainage under CT positioning, and the patients in control group were performed small bone window craniotomy combined with ventricle drainage. The conditions related to surgery and prognosis were compared between 2 groups. Results The operation time, blood loss and hospital stay in observation group were (46.2 ± 25.2) min, (35.4 ± 18.1) ml and (15.2 ± 2.5) d, and those in control group were (108.5±32.5) min, (112.5 ± 35.2) ml and (18.5 ± 3.2) d, there were statistical differences between 2 groups (P<0.01). Two cases died perioperatively in each group. The Glasgow outcome score (GOS) 1 month after operative in observation:9 patients were 5 scores, 19 patients were 4 scores, and the rate of better prognosis was 73.68%(28/38);in control group: 8 patients were 5 scores, 18 patients were 4 scores, and the rate of better prognosis was 68.42% (26/38). There was no statistical difference between 2 groups (χ2 = 0.256, P = 0.613). Conclusions Mini-invasive multichannel drainage is a safe and effective method for hypertensive cerebral hemorrhage combined with severe ventricular hemorrhage, and has the minimal invision.
5.Evaluation of ELISA Method for Detection of Neopterin
Songjin CHEN ; Songhua JIN ; Xiaochao YANG ; Pingyang SHAO
Chinese Journal of Nosocomiology 2009;0(20):-
OBJECTIVE To evaluate the detect method of neopterin(Npt) by enzyme-linked immunosorbent assay(ELISA) and discuss the application of Npt in the viral encephalitis.METHODS The Npt was detected by ELISA.The methodology was investigated by sensitivity,precision,rate of recovery,interference and reference value.The npt was detected by ELISA assay in peripheral blood of 50 healthy people,30 cases of cerebrospinal fluid in children with viral encephalitis and 12 cases of cerebrospinal fluid in the control group.RESULTS In this method,within-run CVs were 4.94% and 5.55%;between-run CVs were 5.99% and 6.57%.The sensitivity was 1.08 nmol/L;the rate of recovery was 95.8-107.8%.Various indexes of the methodology coincided with the requirements of clinical laboratory.The reference value of serum Npt was 0-7.84 nmol/L.The Npt of cerebrospinal fluid was(34.09 ? 36.34) nmol/L in the viral encephalitis group,and(4.55 ? 2.89)nmol/L in the control group,and the Npt of cerebrospinal fluid in patient was significantly higher than that in control group(P
6.Cellular uptake characteristics of salvianolic acid B in myocardial cells and blood vessel endothelial cells
Jingping SHAO ; Shanfei ZHANG ; Yuancheng CHEN ; Wenliang YANG ; Xiaoquan LIU
Journal of China Pharmaceutical University 2009;40(3):263-268
Aim: To determine the uptake characteristics of salvianolic acid B(Sal B) in myocardial cells and blood vessel endothelial cells. Method: The effects of various factors, such as time, temperature, drug concentration, pH of the medium, on the uptake of Sal B in myocardial cells and aorta endothelial cells were investigated. LC/MS was employed to determine the intracellular concentration of Sal B. Results: Uptake kinetics of Sal B in the myocardial cells and aorta endothelial cells fitted well to the logarithmic model at 37 ℃ and 4 ℃. The a-mount of uptake was in direct proportion to the extracellular concentration of Sal B in the experimental concentration range. Uptake of Sal B both in the myocardial cells and blood vessel endothelial cells would significantly increase while the medium pH decreased, and some water-soluble components extracted from danshen would also facilitate the uptake of Sal B both in the myocardial cells and blood vessel endothelial cells obviously. The energy metabolism inhibitors would significantly inhibit the uptake of Sal B in the myocardial cells and blood vessel endothelial cells. When lactic acid and fatty acid were added to the incubation solution, the uptake of Sal B both in the myocardial cells and aorta endothelial cells increased more than 20%. Conclusion: pH is the most important factor influencing the cellular uptake of Sal B, and the amount of uptake tends to increase in acidic medium. Results suggest that the uptake of Sal B would increase in the acidified internal environment induced by myocardial ischemia, thus exerting better cardiovascular activities.
7.Application of trauma assessing method to a emergency triage
Jingfen JIN ; Shuihong CHEN ; Muying YANG ; Linling SHAO
Chinese Journal of Emergency Medicine 2008;17(6):631-633
Objective To study a quick and effective assessing method used in the emergency trauma patients in order to prevent delaying treatment for patients from overlook or misdiagnosis during triage. Method From 1 January 2006 to 31 September 2006, there were 4023 patients evaluated with trauma assessment method, which was composed of CRAMS scale and trauma assessment program. Results Of 4023 patients, there were 2430 cases with CRAMS below 1( < 7) ,of whom 1979 cases assessed with trauma assessment program were found to be injuried severely to threaten their lives. And more noticeably,of 1593 patients with CRAMS ≥7 there were 46 patients found to have life-threatening injuries assessed with trauma assessment method. Conclutions CRAMS score can be used for triage, and trauma assessment program can be used to evaluate the injuries anatomically. Trauma assessment program and CRAMS employed conjointly can increase the sensitivity and specificity for the stratification of the severity in the injuried patients in order to get a successful triage and lo take advantage of the resources of e-mergency care appropriately.
8.Construction of cell model targeted on the damage by α-synuclein
Wenfen YAN ; Yang HENG ; Qianhang SHAO ; Naihong CHEN ; Yuhe YUAN
Chinese Pharmacological Bulletin 2015;(4):586-590
Aim To construct the cell model targeted on the damage by α-synuclein for screening anti-Parkinson’s Disease (PD)compounds.Methods The cDNA fragment of α-synucle-in gene was obtained by PCR methods and inserted into the re-combinant prokaryotic plasmid by molecular cloning technique. The recombinant plasmid was transformed into Escherichia coli, and subsequently induced to express α-synuclein protein.The recombinant α-synuclein was purified and identified by affinity chromatography,immunoblotting and mass spectrometry.The cells damage by α-synuclein was evaluated through cell viability measured by 3-(4,5-dimethyl-2-thiazolyl )-2,5-diphenyl-2-H-tetrazolium bromide.Results The obtained cDNA fragment ofα-synuclein in accordance with its theoretic molecular weight was cloned into pET30a plasmid and verified by sequencing.The re-combinant plasmid was transformed into bacteria E.Coli.BL21 (DE3)and induced to express α-synuclein by isopropyl β-D-1 -thiogalactopyranoside (IPTG).The expression condition was op-timized according to the culture temperature,the concentration of IPTG and the proliferation state of bacteria.The purified α-synu-clein was proved to be a 1 5.3 ku molecule weight protein,and could be immunoblotted with anti-α-synuclein antibody.The pu-rified α-synuclein could decrease the viability of PC1 2 cells and primary neurons significantly,and its effect was in a concentra-tion-dependent manner.Conclusion We have succeeded in constructing the cell model targeted on the damage by α-synucle-in.
9.Expression,purification and preparation of monoclonal antibody of HCA518 protein
Xiaoang YANG ; Xiaoping QIAN ; Xiuyuan SUN ; Qixiang SHAO ; Weifeng CHEN
Chinese Journal of Immunology 1985;0(01):-
Objective; To express and purify HCA518 protein and prepare its monoclonal antibody ( McAb). Methods: The HCA518 protein was expressed with gene recombinant technique in prokaryotic system and purified with nickel chelate nitrilotriacetic acid(Ni-NTA) affinity chromatography column. Hybridoma cell lines that secreted anti-HCA518 McAb were established by cells fusion and screened by enzyme linked immunosorbent assay( ELISA). The specificity of anti-HCA518 McAb wa3 identified by Western blot assay. The HCA518 protein in tumor cells was stained by immunoflourescence assay. Results: Rcombinant HCA518 protein was expressed with a purity of 98%. Two hybridoma cell lines was selected and anti-HCA518 McAb was purified from mice ascites. The titers of anti HCA518 McAb in ascites were 1?10-4 and 5?10-4 respectively. The antibody belonged to IgG2b subtype and IgM. Anti-HCA518 McAb specifically reacted with recombinant HCA518 protein and tumor cells'nuclear protein (P100). The HCA518 protein was mainly located in cell nucleus. Conclusion: Stable hybridoma cell lines that secreted anti-HCA518 McAb have been established and anti HCA518 McAb was prepared with high specificity. It has important significance for detecting HCA518 protein in tumor tissues and determining malignant proliferation status of tumor cells and predicting its prognosis.