2.Limited Septoplasty Under Nasal Endoscopy
yun-hai, FENG ; shan-kai, YIN ; yu-jun, ZHANG
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(01):-
Objective To compare the outcomes of powered-assisted septoplasty with CO_ 2 laser-assisted septoplasty. Methods Thitry patients with limited deviation of nasal septum were analyzed retrospectively. Among 30 patients, 18 underwent powered-assisted septoplasty and the rest underwent CO_ 2 laser-assisted septoplasty. The surgical results were assessed by the visual analogue scale (VSA) and acoustic rhinometry. Results VSA scores significantly improved in both groups after surgery (P
3.Isolation, culture and surface markers detection of human umbilical cord mesenchymal stem cells
Kai FENG ; Li XIAO ; Xihui MA ; Yu GAO ; Xiangrui KONG
Journal of Leukemia & Lymphoma 2013;22(6):354-356
Objective To investigate the methods of isolation and culture in vitro and detect the surface markers of human umbilical cord mesenchymal stem cells.Methods Human umbilical cord Wharton' s jelly was separated and cut up as small as possible,and then cultured with α-MEM.Human umbilical cord mesenchymal stem cells could be obtained by culturing the tissue block adhered the bottle wall.And the cells were passaged at a certain density.The surface markers of human umbilical cord mesenchymal stem cells were detected by FACS when the cells were in Generation Three.Results Human umbilical cord mesenchymal stem cells were obtained from Wharton' s jelly conveniently,with fibroblast shape and stable proliferation and passage.CD29,CD44,CD105 were strongly expressed on human umbilical cord mesenchymal stem cells.But CD45,CD34,HLA-DR,HLA-G,CD80,CDs6 were not expressed.Conclusion Human umbilical cord mesenchymal stem cells can be obtained effectively from the culture of the tissue block,which provides a rich source of cells for tissue engineering.
5.Development of a novel reporter gene method for determination of ADCC potency of anti-CD20 monoclonal antibody.
Chunyu LIU ; Lan WANG ; Wei GUO ; Chuanfei YU ; Feng ZHANG ; Wenbo WANG ; Meng LI ; Kai GAO
Acta Pharmaceutica Sinica 2015;50(1):94-8
The biological activity of ADCC by anti-CD20 monoclonal antibody was determined by BioGlo™ Luciferase Assay System using Jurkat/NFAT-luc+FcγRIIIa cell line as effector cell and WIL2-S cell line as target cell. The developed method was verified for specificity, precision and accuracy. Anti-CD20 monoclonal antibody showed a dose-response mode by the developed method, and the determination result complied with the following four-parameter equation: y = (A-D)/[1 + (X/C)(B)] + D. The optimized parameters of the method were determined including the antibodies diluted concentration (18,000 ng·mL(-1)), dilution rate (1:5), the ratio of effector cell and target cell (6:1), and induction time (6 h). The values of eight independent tests have passed a statistical test for curve regression analysis, linear or parallelism, which showed the method possessed good specificity. Four different dilute groups of recovery rates sample were determined for 3 times, and the result showed mean relative potencies of (44.39±3.93)%, (72.74±2.78)%, (128.28±7.01)% and (168.19±2.70)% respectively, with a variation coefficient of less than 10%, and the recoveries of (88.78±7.85)%, (96.99±3.70)%, (102.63±5.61)% and (112.12±1.80)% respectively. A novel reporter gene method for determination of biological activity of ADCC by anti-CD20 monoclonal antibody was successfully developed, which showed strong specificity, good reproducibility and high accuracy, and might be used routinely.
6.Application value of laparoscopic ultrasonography in laparoscopic common bile duct exploration
Xiaomei CHEN ; Mingxing LI ; Zhijian LUO ; Yu ZHANG ; Chunhong FENG ; Kai HE
Chongqing Medicine 2014;(3):281-282
Objective To explore the value of the laparoscopic ultrasonography (LUS) in laparoscopic common bile duct explora-tion(LCBDE) .Methods 47 cases with cholecystolithiasis combined with bile duct dilatation or abnormal liver function were tested by conventional ultrasound ,magnetic resonance cholangiopancreatgraphy (MRCP) CT and LUS after LCBDE .All results were com-pared with the surgical findings and the intraoperative biliary endoscopy results .Results The diagnosis coincidence rate of conven-tional ultrasonography ,CT ,MRCP and LUS in 47 cases were 85 .1% ,72 .3% ,95 .7% ,100 .0% respectively .6 cases completed the surgery under the guidance of LUS ,1 case was converted to open surgery .Conclusion LUS is an examination method to avoid bile duct injury effectively and reduce postoperative residual stone in LCBDE .
7.Comparative study of size and charge heterogeneities of anti-TNF-αantibodies by high performance liquid chromatography
Wei GUO ; Wenbo WANG ; Chuanfei YU ; Feng ZHANG ; Lan WANG ; Chunyu LIU ; Meng LI ; Kai GAO
Chinese Journal of Microbiology and Immunology 2014;(9):723-726
Objective To analyze the differences of size and charge heterogeneities between origi-nal humanized anti-TNF-αantibody and four similar biotherapeutic products ( SBP ) .Methods The size exclusion chromatography ( SEC-HPLC ) and weak cation exchange chromatography ( WCX-HPLC ) were used to analyze the size and charge heterogeneities , respectively.Carboxypeptidase B (CpB) treatment was employed to analyze the source of charge heterogeneity of the antibody products .Results Four SBPs showed the same pattern with the originator in SEC-HPLC, and no significant difference with the percentage of mono-mer was observed .The percentages of the aggregates of SBP-3 and SBP-4 were a little higher than those of the originator .The charge distribution of SBPs was significantly different from the originator ′s, especially in the basic region .The results from the samples treated with CpB indicated that the difference of charge distri -bution in the basic region might be caused by the C-terminal lysine variants .Conclusion Four SBPs showed similar size heterogeneity with the originator , but significant differences with charge heterogeneity were observed among them .The study suggested that more attention should be paid to the charge heterogene -ity analysis of the biosimilar products .
8.Size heterogeneity analysis of monoclonal antibody products
Chuanfei YU ; Wenbo WANG ; Meng LI ; Lan WANG ; Feng ZHANG ; Chunyu LIU ; Kai GAO
Chinese Journal of Microbiology and Immunology 2014;(9):718-722
Objective To compare the capability of capillary electrophoresis-sodium dodecyl sul-fate ( CE-SDS) and size exclusion-high performance liquid chromatography ( SE-HPLC) for analysis of size heterogeneity of monoclonal antibody products .Methods The size heterogeneity of one humanized anti-VEGF monoclonal antibody was analyzed by using non-reduced and reduced CE-SDS, and conventional , de-natured and denatured reduced SE-HPLC.Results The percentage of aggregates detected by non-reduced CE-SDS (0.82%±0.01%) was equal to that by using denatured SE-HPLC (1.05%±0.02%), but it was significantly lower than that by using conventional SE-HPLC analysis (5.08%±0.10%).With regard to fragments analyzed with non-reduced antibodies, its percentage was (7.12±0.04)% measured by non-re-duced CE-SDS analysis that was significantly higher than that by conventional SE -HPLC analysis (0.02%± 0.01%) and denatured SE-HPLC analysis (0.62%±0.01%).Using reduced antibodies , the percentage of fragments was (3.19±0.50)%tested by reduced CE-SDS analysis that was significantly higher than that by using denatured reduced SE-HPLC analysis (0.07%±0.01%).Conclusion Conventional SE-HPLC was more objective than CE-SDS for content analysis of aggregates , as both the covalent and non-covalent forms of aggregates could be detected .Non-reduced CE-SDS could demonstrate the content of clips , while reduced CE-SDS showed the degraded fragments .Therefore, CE-SDS had an advantage over conventional SE-HPLC for content analysis of fragments .The use of the two analytical methods in combination provided solid techni-cal supports for the quality control of size heterogeneity of monoclonal antibodies .
9.Empirical Study on the Effect of Osmotic Opening of Blood Brain Barrier on the Facilitation of Nerve Growth Factor Passing
tian-ming, JIA ; ru, BAO ; yu-feng, LIU ; kai-xian, DU ; bin, GAN
Journal of Applied Clinical Pediatrics 1992;0(06):-
Objective To explore the change of nerve growth factors(NGF) through the blood brain barrier(BBB) after distal intravenous injection of mannitol into the experimental rats and the effect of exogenous NGF on the expression of growth associated protein-43 in hypoxic-ischemic brain.Methods One hundred cases of 7 days rats were divided into 2 units.One unit was divided into 3 groups:treatment group,control group and sham operated group,20 rats in each group.The other unit was divided into 4 groups:mannitol and NGF treated group,NGF treated group,control group,and sham operated group,there were 10 rats in each group.The model rats with perinatal hypoxic-ischemic brain damage(HIBD) rats were prepared by ligation of left common carotid artery with a temporary systemic hypoxia(inhaling 80 mL/L O2 and 920 mL/L N2).The sections of brains were processed by immunochemistry with antibodies against GAP-43,and the study and memory ability of rats were tested by maze test.The effect of osmotic opening of BBB on the facilitation of NGF′s passage was tested by ELISA.Results The expression of GAP-43 increased after NGF treatment,and the differences were remarkable(P
10.Investigation of inductive effect of triptolide on cytochrome P450 s in rat hepatocytes and HepG2 cells and possible mechanism
Xi YU ; Zijin WAN ; Yanping ZHU ; Hairong XU ; Kai LIAO ; Feng LIU ; Wei LI
Chinese Pharmacological Bulletin 2017;33(3):366-372
Aim Toinvestigatingtheinductionof CYPs in hepatocytes or HepG2 cells by triptolide(TP) andthepossiblemechanism.Methods AfterTPtreat-ment,the expression of CYPs in rat primary hepato-cytes or human HepG2 cells was detected by real-time PCR and Western blot assays.Specific inhibitors or gene knockdown method were employed to analyze the possiblemechanism.Results Theexpressionof CYP1A2,2C7,2C11,2C12,2D2,2E1 and 3A1 in rat primary hepatocytes was induced by TP.The fold was 19,2,31,3,21,88 and 34 at 50 nmol·L-1, respectively while at 100 nmol·L-1 it was 20,5,30,23,61,83 and 38,respectively.In HepG2 cells,the expression of human CYP1A1,2B6,2C9,2C19, 2D6,2E1 and 3A4 was also induced by TP.The ac-tivities of nuclear receptor PXR and CAR were inhibi-ted.TP upregulated p53 expression,and the induction of several CYPs caused by TP was blocked when p53 wasinhibited.Conclusions TPinducesCYPsexpres-sion in rat hepatocytes and HepG2 cells.Nuclear re-ceptors may not be involved in TP induced CYPs, while the mechanism may partly attribute to p53.