1.Separation of polyphyllin from Rhizoma Paridis by foam fractionation technique
Jie LAN ; Rui LI ; Li HAN ; Ming YANG ; Jin WANG
Chinese Traditional and Herbal Drugs 1994;0(01):-
Objective To study the process of foam fractionation of polyphyllin in semi-batch mode. Methods Taking enrichment ratio, recovery rate of polyphyllin, and purity ratio as the performance criteria and using single examining method to examine the operational parameters, i.e. operation mode, air flow rate, initial feed concentration, solution pH value, initial feed height and temperature on separation performance. The optimal conditions of the process were obtained finally. Results The separation performance is good when gas flow rate is 400 mL/min, initial feed concentration (polyphyllins content) is 0.3 mg/mL, pH value is 7, feed height is 30 cm, and feed temperature is 40 ℃. The enrichment ratio is 25.7, recovery ratio is 42.1%, and the foam liquids purity of total polyphyllin is 41.4%, which is 4.5 times higher than that in feed purity. Conclusion Foam fractionation technique could be applied to separate polyphyllin.
2.Development of mass spectrometry technique for quality assessment of monoclonal antibodies
Wen-wen ZHU ; Meng-lin LI ; Jin-lan ZHANG
Acta Pharmaceutica Sinica 2020;55(12):2843-2853
The research and development of monoclonal antibodies (mAbs) is a rapidly developing field. From the first generation of murine mAbs to the fourth generation of fully human mAbs, the efficacy and safety of mAbs in the treatment of various diseases have been continuously improved. In order to regulate the development and evaluation of mAbs, drug regulatory agencies and pharmacopeias of America and China have tried to issue feasible test procedures and acceptance criteria for quality evaluation of mAbs and biosimilars. Mass spectrometry (MS) technique with high sensitivity, resolution, selectivity, and specificity has become an important tool to evaluate the quality characteristics of monoclonal antibody-related products or specify mAb quality. The research of MS-based monoclonal antibody study involves structure characterization, impurity analysis, pharmacokinetics/pharmacodynamics (PK/PD), etc. This review focuses on the current quality control requirements of mAb related products and the development of MS technique for mAb quality characterization and specification. It is expected to provide information and references for evaluating the quality of monoclonal antibodies under research and development.
3.N-Glycans and intact glycopeptide-based characterization of N-glycosylation of monoclonal antibody drugs
Meng-lin LI ; Wen-wen ZHU ; Jin-lan ZHANG
Acta Pharmaceutica Sinica 2021;56(9):2360-2366
In recent years, the biopharmaceutical industry has grown rapidly, and the market size of monoclonal antibody drugs has increased significantly. Accurate structural characterization and quality control are the supporting technologies for the development of monoclonal antibody drugs. As a significant post-translational modification of antibody drugs, glycosylation has an important influence on its efficacy, stability, and immunogenicity. The existing literature usually uses liquid chromatography-mass spectrometry to perform major glycosylation modifications of monoclonal antibody drugs. Characterization, there are few studies on low-abundance glycosylation, but the characterization and control of low-abundance glycosylation cannot be ignored. In this study, we have established a qualitative and quantitative analysis technology for N-glycans based on RapiFluor-MS reagent-labeled monoclonal antibody drugs. This method has a short sample processing time and high sensitivity. It can not only characterize the main glycoforms of three monoclonal antibody drugs (adalimumab, bevacizumab, and trastuzumab) but also can quantify low-abundance N-glycans. The results of the study showed that the main glycoforms specified in the Pharmacopoeia could be detected in different batches of monoclonal antibody drugs, but the content of N-glycans in different batches of samples is not identical. After that, we analyzed the N-glycans connection sites and glycoforms at the intact glycopeptide level, further enriching the N-glycans structure information of the monoclonal antibody. The qualitative and quantitative analysis technology of N-glycans based on RapiFluor-MS reagent-labeled monoclonal antibody drugs can realize the in-depth characterization and control of glycosylation modification of monoclonal antibody drugs.
4.Practice of “one week with one case”teaching method with college-enterprise cooperation in the theoretical classes of contact lens courses
Yang, YANG ; Li, WANG ; Wen-Lan, LIU ; Jin, YAN
International Eye Science 2016;16(6):1140-1143
?AIM: To explore the effects of “one week with one case” teaching method with college - enterprise cooperation in the theoretical classes of contact lens courses, which provide the basis for teaching reform.?METHODS: Fifty-six students in optometry major of Grade 2012 from Xi’an Medical College were divided into 2 groups randomly. The experimental group of 28 students used “one week with one case” teaching method with college-enterprise cooperation; the control group of 28 people used traditional “one week with one case”teaching method. The examination scores and questionnaire were used to evaluate the teaching effects.?RESULTS:The students of experimental group acquired higher test scores in short-answer questions and the case analysis questions compared with students of control group (P<0. 05). The questionnaire survey showed the“one week with one case” teaching method with college-enterprise cooperation acquired higher scores in 6 items which include the intensity of learning interest, information acquisition ability, team cooperation ability, communication skills, oral communication ability and the satisfaction of teaching method (P<0. 05). Four items included participation, preparation, communication ability and team spirit of experimental group students were significantly superior to those of the control group students (P<0. 05).?CONCLUSION:“One week with one case” teaching method with college-enterprise cooperation can improve comprehensive ability of students. It is an effective teaching method with the characteristics of the contact lens courses.
5.Effect of ecdysterone on apoptosis induced by hypoxia in HUVECs
Jun JIN ; Lan HUANG ; Shanjun ZHU ; Changqing XIANG ; Hong LI ;
Chinese Pharmacological Bulletin 1986;0(05):-
AIM To explore the effect of hypoxia on apoptosis in human umbilical vein endothelial cells(HUVECs) and the role of ecdysterone(EDS)in inhibition of apoptosis. METHODS ① Culture and identification of HUVECs;② Hypoxic model(0,12,24,48 h) in HUVECs was established. While HUVECs was incubated 24 h with EDS(100 mg?L -1 ) under hypoxic condition. Apoptosis in HUVECs was detected by TUNEL;③ HUVECs received EDS 100 mg?L -1 in normal and hypoxic condition. After 24 h, vascular endothelial growth factor (VEGF) was detected with immunohistochemical technique. RESULTS The apoptosis percentages are different under different hypoxic condition. The longer hypotic the time was, the higher the apoptosis percentage was. EDS reduced apoptosis of HUVECs. EDS could enhance expression of VEGF protein in cardiac myocytes of rat in normal and hypoxic condition. CONCLUSION The role of hypoxia in HUVECs apoptosis is more significant along with prolonging of hypoxic time. VEGF play an important role in protective effect of EDS on endothelial cell apoptosis.
6.Parthenolide enhances the apoptosis induced by PKC inhibitor in human gastrointestinal stromal tumors cell lines
Xiangdan LI ; Lan LIU ; Xuesen FANG ; Toufeng JIN
Basic & Clinical Medicine 2017;37(2):202-205
Objective To investigate the effect of parthenolide ( PTL) and PKC inhibitor on human gastrointestinal stromal tumor (GIST) cell proliferation and apoptosis and the mechanism involved .Methods Human GIST cell lines were cultured in vitro, and the cell proliferation rate of GIST , was determinate by MTT;flow cytometry was used to test the early apoptosis rate of GIST;Western blot assay was applied to detect the expression of endoplasmic reticulum stress-related proteins , GRP78 and GADD153.There were four groups: control group , PTL group, PKC inhibitor group , combine PTL and PKC inhibitor group .Results PTL and PKC inhibitor combination therapy for GIST was sig-nificantly more effective than a single-drug therapy (P<0.05);as for the early apoptosis rate , the combination ther-apy for GIST cells was significantly higher than that medication alone group (P<0.05).the expression of endoplas-mic reticulum stress-associated protein GRP 78 and GADD153 was obviously higher in PTL and PKC inhibitor combi-nation group than that in medication alone group (P<0.05).Conclusions PTL and PKC inhibitor combination therapy for GIST cells can induce apoptosis , which is possibly mediated via endoplasmic reticulum stress pathway .
7.Analysis of the clinical features and electroencephalogram characteristics in 24 patients with agyria-pachygyria
Xinghui LAN ; Li JIANG ; Yue HU ; Jin CHEN ; Na XU
Chinese Journal of Applied Clinical Pediatrics 2015;30(9):702-706
Objective To analyze the clinical manifestations and electroencephalogram (EEG)characteristics of agyria-pachygyria for its early diagnosis,treatment and prognosis judgment in clinical practice.Methods The clinical manifestations and EEG features of twenty-four patients with agyria-pachygyria who were diagnosed by CT or magnetic resonance imaging(MRI) at Pediatric Neurology of Children's Hospital of Chongqing Medical University from July 2004 to July 2013 were retrospectively analyzed.Results Of twenty-four patients,eighteen cases were diagnosed as diffuse agyria-pachygyria and six cases were diagnosed as partial agyria-pachygyria.The clinical features were mainly manifested as mental retardation (twenty-four patients),and motor retardation (twenty-four patients),and epilepsy (eighteen patients).All of the twenty-four patients had abnormal EEG pattern which were mainly three tapes.Type Ⅰ had diffused high amplitude alpha and beta activity in all cortical regions,frontal-central,or parietal-occipital region (fourteen patients).Type Ⅱ showed alternating high amplitude bursts with sharp and slow waves (seven patients).Type Ⅲ was characterized by high amplitude spike or sharp wave activity generalized or multifocal distribution and δ,θ wave mixing graphics (twelve patients).Nine of twenty-four patients showed two or three EEG characteristic patterns in an awake-asleep EEG recording.During the follow-up of 1-8 years old,twelve of the thirteen patients who were diagnosed as epilepsy in diffuse agyria-pachygyria had refractory epilepsy,mainly with infantile spasms or Lennox-Gastaut syndrome.One of the five patients who was diagnosed as epilepsy in focal agyria-pachygyria had refractory epilepsy,mainly for partial epilepsy secondary generalized seizures.There was a significant difference between them (P =0.008).Eighteen of twenty patients who had moderate-severe mental retardation or dyskinesia were diagnosed as diffuse a gyria-pachygyria,while two were focal agyria-pachygyria.Both of them had a significant difference (P =0.005).Conclusions Agyria-pachygyria is a brain malformation caused by neuronal migration abnormality.Diffuse agyria-pachygyria is presented with serious clinical manifestations and poor outcome while the clinical manifestation of focal agyria-pachygyria is relatively mild and epilepsy could be controlled by antiepileptic drugs or epilepsy surgery.These characteristics of EEG patterns along with clinical findings could provide important evidence for early diagnosis,timely treatment and prognosis judgment of agyria-pachygyria.
8.Quantification of tryptase and TIM-3 double positive mast cells in human chronic periodontitis tissue
Juan LI ; Hua JIN ; Tian LAN ; Shiguang HUANG
Journal of Practical Stomatology 2014;(4):482-486
Objective:To quantify the tryptase and T cell immunoglobulin mucin domain-3(TIM-3)double positive mast cells in hu-man chronic periodontitis tissue using double immunofluorescence staining.Methods:25 healthy controls,28 chronic mild periodontitis and 30 chronic advanced periodontitis patients were included.The gingival specimens were stained with HE for histology,and with double immunofluorescence staining for the identification of tryptase and TIM-3 double positive mast cells in gingival tissue.Results:In chronic periodontitis tissue the degree of gingival inflammation was significantly increased,the densities(cells/mm2 )of tryptase and TIM-3 double positive mast cells were significantly increased(P<0.05),in addition,that in chronic advanced periodontitis group was significantly higher than in the chronic mild periodontitis group(P<0.05).Conclusion:Tryptase and TIM-3 double positive mast cells has the similar tendency as the severity of periodontitis inflammation in human periodontitis tissue.Tryptase and TIM-3 double positive mast cells may play an important role in human chronic periodontitis.
10.A clinical study on two methods of common bile duct exploration in open surgery for secondary common bile duct stone
Guoqing WU ; Wei HAN ; Lan JIN ; Jianshe LI ; Zhongtao ZHANG
International Journal of Surgery 2009;36(1):40-42
Objective To compare the clinical efficiency of two methods in the open surgery treatment of seconda-ry common bile duet stones: transeytie common bile duet exploration with eholedoehoscope (TCBDE),or traditional eholedoehotomy with T-tube drainage.Methods Retrospectively analysis the treatment of 113 eases with secondary bile duet stones (TCBDE group 58 cases ,T-tube group 45 cases) ; Compare the achievement ratio,operating time; complications,average hospital stay and cost between two groups.Results All procedure is successful,without any severe complications.There are no significant differences of operating time.The TCBDE group has sig-nificantly shorter hospital stay and lower cost.Conclusion TCBDE has significant advantages than the traditional choledochotomy,and should be the predominant method in the open surgery of secondary common bile duet stones.