1.Over-expression and purification of the recombinant p30 antigen of Toxoplasma gondii
Dianbo ZHANG ; Defu ZAI ; Maoqing GONG ; Jin LI ; Qingkuan WEI ; Yong CUI ; Bingcheng HUANG ; Keyi LIU
Chinese Journal of Zoonoses 2005;(12):1089-1093
To provide the basis for preparation of diagnostic kits and vaccines in Toxoplasma gondii infection, the gene coding for the qualified recombinant p30 protein (SAG1) of this parasite was amplified by PCR, and the amplified gene was cloned into prokaryotic expression vector pET-30a(+) to construct the recombinant plasmid, and then transformed to E.coli DH5α. The positive recombinant plasmid was screened by PCR and double enzymes digestion, and the nucleotide sequence of p30 gene was determined by automated DNA sequencing. Meanwhile, the identified recombinant plasmid was transformed to E.coli BL21(DE3) with the expression of p30 on bacteria induced by IPTG and the expressed protein was identified by SDS-PAGE. The protein obtained was then further purified and refolded, and its biological activity was checked by Western blotting. It was shown that the size of the amplified gene was 750 bp with molecular weight of 30 ku, and this protein could specifically react with monoclonal antibody against p30 protein.
2.Establishment and application of the training mode for undergraduate clinical medical talents'practical ability
Xueming GU ; Jian YIN ; Xu ZHANG ; Ying YANG ; Di JIN ; Defu ZHANG ; Hang LI ; Jun WANG ; Yingxia LI ; Guochao SUN
Chinese Journal of Medical Education Research 2017;16(6):546-550
Under the guidance of strengthening medical undergraduates' practical ability and their competence in future jobs, Dalian Medical University has established a clinical medical talents practical ability training mode. This model has implementedThree Turnsclinical skills training pattern to strengthen the three stage (before, during and after practice) clinical skills training and concentrated on humanistic quality education through overall evaluations and set up clinical skill examination system to evaluate teach-ing effectiveness comprehensively and truly , which has effectively improved the quality of education by perfecting safeguard mechanisms and guaranteeing teaching quality.
3.Expression and identification of the multiple gene ROP2-P30 of Toxoplasma gondii in E.coli BL21
Dianbo ZHANG ; Qingkuan WEI ; Defu ZAI ; Yong CUI ; Jin LI ; Honggang GAO ; Xuelian BAI ; Lijiang ZHAO ; Guangdong HAN ; Keyi LIU
Chinese Journal of Zoonoses 2006;(6):538-543
To obtain the functional fusion protein of rhoptry protein 2, compound rhoptry protein2 and surface antigen 1 of Toxoplasma gondii. the ROP2 and P30 genes from genomic DNA of T.gondii RH strain were amplified by PCR, and were inserted into pMD18-T cloning vector. Then the ROP2 fragment was subcloned to pET-30a(+) plasmid digested by EcoRⅠand Hind Ⅲ to construct plasmid pET-ROP2. Furthermore,the P30 fragment was subcloned into pET-ROP2 digested by BglⅡand EcoRⅠto create plasmid pET-ROP2-P30, the resulting recombinant plasmids , transformed into E.coli BL21 (DE3), were induced with IPTG. and the proteins identified by SDS-PAGE were further purified and refolded. The biological activity was analyzed by Western blot with specific antibody. It was found that the sizes of ROP2 and ROP2-P30 were 1212 and 1896bp with corresponding molecular weight 50- kDa and 75-kDa, respectively. The recombinant protein ROP2 (50-kDa) could specifically react with rabbit-polyclonal antiserum, and complex fusion protein ROP2-P30 (75- kDa) could react with P30 monoclonal antibody.
4.Studies on the Immuno-Protection of ROP2 Nuclei Acid Vaccine in Toxoplasma gondii Infection
Qingkuan WEI ; Jin LI ; Tingxia FU ; Xuelian BAI ; Yong CUI ; Dianbo ZHANG ; Hongfa WANG ; Yubing LIU ; Bin FU ; Defu ZAI ; Bingcheng HUANG ; Keyi LIU ; Guangdong HAN
Chinese Journal of Parasitology and Parasitic Diseases 1997;0(05):-
Objective To study the protective effect of ROP2 nuclei acid vaccine in mice.Methods Forty-two BALB/c mice were divided into three groups.Each mouse in experiment group was injected with 50 ?g recombinant plasmid pc-DNA3-ROP2 through musculus quadriceps fexoris.In control groups,each mouse was injected with 50 ?g blank plasmid pc-DNA3 and with 50 ?l PBS respectively.All mice were immunized for three times with an interval of three weeks.The volume was doubled for the final injection in the two plasmid groups.Blood,spleens and lymph nodes of 4 mice in each group were taken for the detection of CD4+,CD8+ T cells and cytokines 2 weeks after the final immunization.The rest mice in 3 groups were challenged with 500 tachyzoites of Toxoplasm gondii RH strain for further observation.Results The vaccine induced strong cellular and humoral immune response.The titer of antibody in serum was high after inoculation and recognized ROP2 protein antigen expressed in vitro.The lymphocyte phenotype was analyzed.CD4+ T cells proliferated sharply(69.5?3.4)%,and the ratio of CD4+/CD8+ increased considerably by(4.69?1.32)%(P
5.Practice and exploration of training innovative talents through tutorial system in the whole process of "5+3" integrated undergraduate education for clinical medicine specialty
Di JIN ; Guochao SUN ; Jian YIN ; Xueming GU ; Ying YANG ; Defu ZHANG ; You ZHOU ; Youfei GUAN
Chinese Journal of Medical Education Research 2020;19(8):926-929
Dalian Medical University, aiming at cultivating innovative talents, has formulated and implemented a "5+3" innovative talents training reform plan. In the whole process of medical undergraduate education, tutorial system is used as the carrier to develop a phased innovative ability training system, which covers basic scientific research ability training such as courses, lectures, experimental design, papers and so on, and strengthens the cultivation of undergraduate scientific research ability. Through statistical analysis of the results of the first "5+3" students' periodic training, it is found that the proportion of "5+3" students publishing Chinese periodicals and SCI, and hosting national innovation projects and provincial innovation projects is significantly higher than that of ordinary 5-year students ( P<0.05). Although there are some problems and deficiencies in the implementation process, the basic scientific research training with tutorial system as the core has a significant effect on improving students' scientific research thinking and innovation ability, and is feasible for training medical innovative talents.
6.The application scenarios study on the intervention of cognitive decline in elderly population using metaverse technology.
Defu ZHOU ; Yi JIN ; Ying CHEN
Journal of Biomedical Engineering 2023;40(3):573-581
China is facing the peak of an ageing population, and there is an increase in demand for intelligent healthcare services for the elderly. The metaverse, as a new internet social communication space, has shown infinite potential for application. This paper focuses on the application of the metaverse in medicine in the intervention of cognitive decline in the elderly population. The problems in assessment and intervention of cognitive decline in the elderly group were analyzed. The basic data required to construct the metaverse in medicine was introduced. Moreover, it is demonstrated that the elderly users can conduct self-monitoring, experience immersive self-healing and health-care through the metaverse in medicine technology. Furthermore, we proposed that it is feasible that the metaverse in medicine has obvious advantages in prediction and diagnosis, prevention and rehabilitation, as well as assisting patients with cognitive decline. Risks for its application were pointed out as well. The metaverse in medicine technology solves the problem of non-face-to-face social communication for elderly users, which may help to reconstruct the social medical system and service mode for the elderly population.
Aged
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Humans
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Cognitive Dysfunction/prevention & control*
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Aging
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China
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Internet
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Technology
7.Value of serum IgG4 level in differential diagnosis of IgG4-related pancreatic and hepatobiliary diseases and non-IgG4-related pancreatic and hepatobiliary diseases
Chang LI ; Lei YAN ; Li WANG ; Chongxu HAN ; Yunfeng YE ; Defu JIN ; Yuzhang JIANG
Journal of Clinical Hepatology 2022;38(6):1307-1310
Objective To investigate the value of serum IgG4 level in the differential diagnosis of IgG4-related pancreatic and hepatobiliary disease (IgG4-PHD) and non-IgG4-related disease (non-IgG4-RD). Methods Clinical data were collected from 491 patients who were hospitalized and 50 individuals who underwent physical examination in Huaian No. 1 People's Hospital Affiliated to Nanjing Medical University, Subei People's Hospital, and The First Affiliated Hospital of Xuzhou Medical University from August 2014 to April 2021. The 491 patients were divided into IgG4-PHD group with 20 patients, non-IgG4-RD autoimmune disease group with 431 patients (104 patients with systemic lupus erythematosus, 79 with rheumatoid arthritis, 174 with Sjogren's syndrome, 16 with ankylosing spondylitis, 11 with scleroderma, 4 with adult-onset Still's disease, 30 with myositis, 3 with psoriasis, and 10 with primary sclerosing cholangitis), and malignant pancreatic/hepatobiliary tumor group with 40 patients, and the 50 individuals undergoing physical examination were enrolled as healthy control group. Scattering immunoturbidimetric assay was used to measure serum IgG4 concentration. The two-sample Mann-Whitney U test was used for comparison of normally distributed continuous data between groups, and the Fisher's exact test was used for comparison of categorical data between groups. The receiver operating characteristic (ROC) curve was plotted to determine the optimal cut-off value of serum IgG4 in the diagnosis of IgG4-PHD. Results The IgG4-PHD group had a significantly higher serum IgG4 level than the non-IgG4-RD autoimmune disease groups, the malignant pancreatic/hepatobiliary tumor group, and the healthy control group (all P < 0.05), and the Sjogren's syndrome group had a significantly lower serum IgG4 level than the healthy control group ( Z =2.958, P < 0.05). With serum IgG4 ≥1.35 g/L and IgG4 ≥2.01 g/L as the cut-off values, the IgG4-PHD group had a significantly higher positive rate than the non-IgG4-RD autoimmune disease group and the healthy control group (all P < 0.05). The ROC curve analysis showed that IgG4 had an area under the ROC curve of 0.980 in the differential diagnosis of IgG4-PHD and non-IgG4-RD autoimmune diseases, with a sensitivity of 100.00% and a specificity of 94.00% at the optimal cut-off value of 2.21 g/L. Conclusion Serum IgG4 level may also increase in non-IgG4-RD autoimmune diseases, while the cut-off value of 2.21 g/L can improve the differential diagnosis of IgG4-PHD and non-IgG4-RD autoimmune diseases, which requires further verification in clinical practice.