2.Analysis of soluble expression,purification and antigenic characterization of P protein of GⅡ.2[P16]norovirus
Chinese Journal of Biologicals 2024;37(8):952-956
ObjectiveTo obtain the soluble expression of P protein of norovirus(NV)G Ⅱ. 2[P16]and analyze its antigenic characteristics.MethodsThe recombinant plasmid GⅡ.2-NV-pET-28a was constructed by optimizing the NV GⅡ.2[P16]P protein gene,synthesizing and cloning it into pET-28a(+)vector. The recombinant plasmid was transformed into DH5α competent cells,and the expression strain was optimized to make the P protein express in soluble form. The expressed P protein was purified by Ni-NTA affinity chromatography,of which the antigenicity was detected by Western blot,the purity was detected by size exclusion high performance liquid chromatography(SEC-HPLC),and the antigenantibody binding ability was identified by ELISA.ResultsThe target protein of recombinant BL21 Star(DE3)pLySs was highly expressed in soluble form. After purification by affinity chromatography,the purity of GⅡ.2 P protein was 95. 53%,and the antigenicity was good.ConclusionSoluble NV P protein was successfully prepared by prokaryotic expression strain,and the antigenicity of P protein was confirmed,which laid a foundation of the development of NV G Ⅱ.2[P16]detection kits and the recombinant subunit vaccines.
3.Research of relationship among C-peptide level,coagulation function and microvascular disease in pa-tients with type 2 diabetes mellitus
Chinese Journal of cardiovascular Rehabilitation Medicine 2015;24(1):116-118
Diabetic microvascular disease is a severe complication of diabetes mellitus (DM).There are many hypoth-esis for its mechanisms.This overview aimed at effect of C-peptide,coagulant factors,platelet and D-dimer in dia-betic microvascular disease,and provided evidence for its prevention and treatment in clinic.
4.Usage of left internal thoracic artery in the patients aged over 70 years old during coronary artery bypass graft
Chinese Journal of Postgraduates of Medicine 2014;37(26):14-16
Objective To review the outcome of coronary artery bypass graft (CABG) using left internal thoracic artery grafts in the patients aged over 70 years old.Methods Two hundred and eighty-eight patients aged over 70 years old from January 2009 to January 2014 who underwent CABG were selected.All of the patients were divided into two groups on base of the graft:control group:only saphenous vein graft was used in 138 cases; observation group:the left internal thoracic artery to the left anterior descending artery was used in 150 cases,which also included total arterial grafting,facilitated by the use of the radial artery,right gastroepiploic artery was operated in 7 cases,beside these artery grafts,saphenous vein graft was used to anastomosis with other vessels.Results Operative mortality was 2.9% (4/138) in control group and 2.0% (3/150) in observation group,and there was no significance (P > 0.05).The volume of blood drainage was larger in observation group [(796.2 ± 382.4) ml] than that in control group [(512.8 ± 387.5) ml],and there was significant difference (P < 0.05).At the same time,there was no significant difference in the incidence rate of second thoractomy,malignant arrhythmia,stroke,poor wound healing,and usage of intraaortic balloon counterpulsation between two groups (P > 0.05).Conclusion It does not increase the mortality and incidence of serious complications using the left internal thoracic artery to the left anterior descending artery in the patients aged over 70 years old during CABG.
5.In vitro cultivation and identification of epithelial cells of keratocystic odontogenic tumor
Journal of Peking University(Health Sciences) 2004;0(01):-
Objective:To establish a method of in vitro cultivation of epithelial cells of keratocystic odontogenic tumors(KCOT).Methods: Tissue block and enzyme digestion techniques were used for primary cultivation of KCOT cells.The cells were grown in keratinocyte-serum free medium(K-SFM).The biological characteristics of the cultivated keratinocytes were identified by phase microscopic observation and by immunohistochemistry of cytokeratin,vimentin,cytokeratin10 and 14.Results: KCOT keratinocytes could survive for 30-50 days in K-SFM after passing 2-4 generations.Cells were polygon and showed typical slabstone-like appearance.Immunostaining showed positive staining for cytokeratin antibody,and negative for vimentin.Conclusion: KCOT epithelial cells could be serially cultured in vitro in K-SFM by techniques suggested in this study.
6.Does long-term effective control of body weight is a difficult problem in clinical of diabetes control.
Chinese Journal of Practical Internal Medicine 2001;0(03):-
Weight increasing would happen with the control of glucose in diabetic patients,whether the application of insulin or that of oral medication.Long-term and effective weight control should be done in order to lower the mortality rate.A large number of studies have shown that the mortality rate has increased along with weight gain,especially death caused by cardiovascular disease.The following measures can be used:(1)Rational target should be formulated.It would be the best that weight decrease 0.5~1.0 kg every week.(2)Diet should be controlled.Low-calorie,high protein,low-fat diet can be used.(3)Exercise therapy:Aerobic exercise will be recommended.(4)Drug intervention treatment:Metformin or Thiazolidinediones drugs can be use,Sibutramine or Orlistat can be used in the patients with serious obesity.
7.Research on five stagnations in seven chapters about five evolutive phases and six climatic factors of Plain Questions
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(03):-
Five stagnations are an important content of Plain Questions.Many doctors think highly of the treating principle,while overlook the other content.Through researches on Plain Questions,the author lays out the entire system,including the prevention and treatment for mild stagnation,the presymptom,changes on phenology,climate and human body for severe stagnation.
8.Effects of 2-methoxyl estradiol on the expression of L-selectin protein and its mRNA in K562 cells
Medical Journal of Chinese People's Liberation Army 2001;0(12):-
Objective To investigate the effects of 2-methoxyl estradiol(2-ME)on the expression of L-selectin and its mRNA and the relationship between L-selectin and bcr/abl fusion gene expression in chronic myelocytic leukemia(CML)K562 cells.Methods The cultured K562 cells were divided into three groups.The control group,K562 cells were cultured without 2-ME treatment.The experimental group,K562 cells were cultured in the medium containing different concentrations of 2-ME(1,2,4,8 and 16?mol/L)for 36h.The negative control group,K562 cells were replaced by water without RNase in the medium containing different concentrations of 2-ME for 36h.The expression of L-selectin protein in K562 cells was determined by flow cytometry(FCM)method.The expression of L-selectin mRNA in K562 cells was detected by half-quantitative reverse transcription-polymerase chain reaction(RT-PCR).The bcr/abl fusion gene expression in K562 cells was detected by quantitative SYBR Green Ⅰ real-time RT-PCR.Results After being treated with 2-ME at different concentrations for 36 hours,the expression of L-selectin protein and its mRNA increased in K562 cells in a concentration-dependent manner,and the expression level in the experimental group was higher than that in the control group(P
9.On Introducing Professor Li Saimei's Clinical Experiences for Mental Diseases
Journal of Zhejiang Chinese Medical University 2016;40(2):122-124
Objective] To sum up Pro. Li Saimei’s clinical experience in treating mental diseases. [Method] By following the teacher clinic and sorting out the related medical materials,the author summarizes professor Li Saimei 's academic experience of treatment of mental diseases ,and for proven cases. [Result] In professor Li Saimei's opinion,mental diseases are due to dysfunction of shaoyang's turning,and phlegm blocking the spirit orifices. Professor Li usually uses Chaihu related formulas to turn the Shaoyang,resolve the phlegm,open the three Jiao,regulate Yin and Yang.The notable clinical result has been observed.[Conclusion] Professor Li Saimei's experience is effective and worthy of inheritance and promotion.