1.Expression, purification and application of staphylococcal protein A fused to SUMO
Xuehui GAO ; Jiechao YIN ; Guopeng SUN ; Wenfei WANG ; Deshan LI
Chinese Journal of Microbiology and Immunology 2010;30(7):597-602
Objective To clone the full length staphylococcal protein A(SPA) gene from Staphylococcus aureus (ATCC6538), and subsequently study the gene structure and antibody binding ability.Methods The full length and the functional region of the SPA gene were cloned into pHisSUMO vector respectively, and expressed in E. coli. The full length and the functional fragment of the SPA protein were detected for antibody binding ability and stability. The functional fragment of the SPA protein fused with SUMO was coupled to the CNBr-activated agarose for antibody purification from rabbit serum. Results A variant of the full length SPA gene was cloned, which has been submitted to GenBank (the accession number is EU695225). Two fusion proteins had the same antibody binding ability as the untagged SPA protein. However, the formers was more stable than the latter at the tested conditions. SUMO-SPA conjugated-agarose kept high efficiency for antibody binding. Conclusion To our knowledge, the full length SPA gene of S.aureus(ATCC6538) is a novel variant. The SUMO tag can improve the stability of the functional region of the SPA protein without damaging the antibody binding ability. This fusion protein has been used for antibody purification successfully.
2.The correlations of single photon emission computed tomography joints scan and bone metabolic markers in active rheumatoid arthritis
Haiyan GAO ; Xiaofeng LI ; Baoniu ZHANG ; Minghui DING ; Deshan ZHAO
Chinese Journal of Internal Medicine 2016;55(11):845-848
Objective To explore the correlation between quantitative value of joint bone scan by single photon emission computed tomography (SPECT) and serum bone metabolic markers in patients with active rheumatoid arthritis (RA).Method Clinical data of 60 newly diagnosed RA patients were retrospectively collected in Department of Rheumatology and Immunology from May 2013 to December 2014,including 28 cases with medical history less than 6 months and 32 from 6 months to 2 years.Serum Cterminal telopeptide of type Ⅰ collgen(CTX-Ⅰ)and N-terminal propeptide of type Ⅰ collagen(PINP) were tested by chemiluminesence.SPECT whole body bone and target joint scan before treatment was done.Thirty-nine healthy subjects as control group received ultrasound,electrocardiogram,X-ray,and whole body bone imaging examination.Quantitative values of joint imaging were calculated for the statistical analysis.Result Demographic data between RA group and control group were comparable (P > 0.05),including sample,sex,age and BMI.The joint SPECT value,CTX-Ⅰ and PINP levels were all significantly higher than those of control group (P < 0.01),which were 6.48 ± 1.98 versus 3.73 ± 1.t6;(0.66 ± 0.37) mg/L versus (0.58 ±0.21) mg/L;(46.35 ±28.15) mg/L versus (30.47 ± 13.75) mg/L respectively.Joint SPECT values had positive correlations with serum CTX-Ⅰ levels in all RA patients,as well as PINP in patients with disease duration 6mon-2years.And the according correlation coefficients were 0.513,0.495,0.402 (P < 0.05).But SPECT value had no correlation with CTX-Ⅰ (P =0.081) in patients with disease duration less than 6 mon.The correlation coefficient was 0.336.Conclusion SPECT imaging quantitative values were positively correlated with serum bone metabolic parameters.Thus SPECT imaging alone or combined with bone markers are helpful in diagnosing active RA.
3.Comparison of SPECT/CT fusion imaging and MRI in benign hip lesions
Huixia GENG ; Deshan ZHAO ; Guizhu GAO ; Fei FENG ; Sijin LI
Chinese Journal of Medical Imaging Technology 2010;26(2):337-339
Objective To compare SPECT/CT fusion imaging and MRI in the diagnosis of benign hip lesions. Methods Twenty-two patients suspected avascular necrosis of femoral head with hip discomfort, pain or action limited were analyzed retrospectively. All patients underwent radionuclide bone scan and MR examination within 5 days, and the diagnosis was proved with clinical follow-up. Results Eighteen necrosis of the femoral head and 4 hip arthritis including 1 patient with ankylosing spondylitis were found in 44 hip joints of 22 patients. MRI detected 17 femoral head necrosis and 4 hip arthritis, while SPECT/CT fusion image found out 18 femoral head necrosis and 4 hip arthritis. There was corresponding relationship in signs of hip lesions between MRI and SPECT/CT fusion imaging. Conclusion SPECT/CT fusion imaging and MRI has no markedly difference in the diagnosis of hip benign lesions, and is complementary to each other. SPECT/CT fusion image can distinguish the hip lesions from the femoral head lesions, and has a higher accuracy of diagnosing hip lesions than whole body bone scanning.
4.The hemangioma of alimentary tract
Zulin CHEN ; Xuequan HUANG ; Zhitang SHAN ; Yunsheng LUO ; Deshan GAO ;
Chinese Journal of General Surgery 1994;0(05):-
Eighteen patients with hemangioma of alimentary tract were treated in our hospital from 1980 to 1994 There were 11 males and 7 females,with average age of 48?18 years (ranged from 19 to 76 years) Of them,14 patients were treated with operation,2 with hemangioma of rectum were treated with ligation with snare.All of the 16 patients were cured. The etiology,pathology,diagnosis and management are discussed in this paper The experiences of finding small hemangiomas of jejunum and ileum during operation are also introduced in the paper
5.Alteration of AQP2 expression in kidney tissues of emphysema model rats
Zhe WANG ; Chunying LIU ; Yuan GAO ; Ying WANG ; Huan JING ; Jiyan CHAI ; Shouyan WANG ; Deshan WANG
Chinese Journal of Pathophysiology 2010;26(2):318-321
AIM: To observe the expressions of aquaporin 2 (AQP2) in kidney tissues and the contents of endotoxin (ET), interleukin-1 β (IL-1β), tumor necrosis factor-α (TNF-α) in serum in emphysema model rats, and to investigate the relationship between lungs and kidney in humoral metabolism. METHODS: The rats of emphysema were treated by injecting lipopolysaccharide into the trachea with cigarette smoking. Immunohistochemistry and Western blotting analysis were used to observe the expression of AQP2 in kidney tissues. RT-PCR was applied to detect the expression of AQP2 mRNA in kidney tissues. Blood sample and lung tissue were taken and the levels of ET, IL-1β and TNF-α were measured by radioimmunoassay. RESULTS: AQP2 expression in the kidney tissue in model group was greater than that in control group, and the expression of AQP2 mRNA showed the same results (P<0.01). ET, IL-1β and TNF-α levels in serum and lung tissue in model group were markedly higher than those in control group (P<0.01). CONCLUSION: In the emphysema model rats, AQP2 expression is up-regulated in the kidney tissue. The mechanism of emphysema may be related to increasing the levels of ET, IL-1β and TNF-α in the serum and lung tissue obviously.
6.Establishment of a novel cell model targeted on FGF-21 receptor for screening anti-diabetic drug candidates.
Hongmei GAO ; Wenfei WANG ; Qiao ZHANG ; Yang HAN ; Qi WANG ; Guiping REN ; Yunwei FU ; Deshan LI
Acta Pharmaceutica Sinica 2011;46(8):904-9
The aim of this project is to establish a fibroblast growth factor-21 (FGF-21) signaling pathway targeted cell model, for screening a class of FGF-21 receptor agonists as anti-diabetic candidates. FGF-21 requires beta klotho transmembrane proteins as co-receptor for the activation of tyrosine kinase FGF receptor (FGFR) signaling, thereby activating a series of intracellular signaling pathways and regulating gene transcription for glucose metabolism. Firstly a recombinant plasmid expressing co-receptor beta klotho and EGFP reporter genes was constructed. After introducing the recombinant plasmid into package cells, the cell culture supernatant was used to infect 3T3-L1 cells, which were then screened for stably expressing beta klotho gene. Administration of FGF-21 increased the expression of GLUT1 and stimulated GLUT1-mediated glucose uptake. This novel cell model can be conveniently used in high-throughput drug screening of FGF-21 or FGF-21 analogues.
7.Expression and pharmacological evaluation of fusion protein FGF21-L-Fc.
Wenbing YAO ; Guiping REN ; Yang HAN ; Hongwei CAO ; Hongmei GAO ; Fangming KAN ; Qi WANG ; Deshan LI
Acta Pharmaceutica Sinica 2011;46(7):787-92
FGF21 (fibroblast growth factor 21) is a recently described member of the FGF family. It has been previously demonstrated that FGF21 is a potent regulator of glucose homeostasis. To improve stability of FGF21 for better efficacy, a new form of recombinant FGF21 was generated by fusion of a full length FGF21 gene and the Fc fragment of human IgG4 with flexible linker sequence. To examine the glucose regulation activity of FGF21-L-Fc, 3T3-L1 pre-adipocytes were differentiated into adipocytes, and glucose uptake activity of FGF21-L-Fc was examined by glucose oxidase and peroxidase (GOD-POD) assay. The results showed that in comparison with wild type FGF21, FGF21-L-Fc was more potent in stimulation of glucose uptake by 3T3-L1. In vivo studies on the modified protein demonstrated that FGF-L-Fc had a better efficacy in lowering blood glucose of the STZ-induced diabetic animals and controlled glucose level for a longer time. The results provided a sound basis for further studies.
8.Optimization and characterization of a novel FGF21 mutant.
Xianlong YE ; Huashan GAO ; Wenfei WANG ; Guiping REN ; Mingyao LIU ; Kun HE ; Yakun ZHANG ; Jingzhuang ZHAO ; Dan YU ; Deshan LI
Acta Pharmaceutica Sinica 2012;47(7):897-903
Fibroblast growth factor 21 (FGF21) is a member of FGF family. It has been demonstrated that FGF21 is an independent, safe and effective regulator of blood glucose levels in vivo. In order to improve the activity of FGF21, we exchanged the beta10-beta12 domain of the human FGF21 with that of the mouse FGF21 to construct a novel FGF21 gene (named hmFGF21), and then subcloned hmFGF21 gene into the SUMO expression vector to create pSUMO-hmFGF21 and transformed it into E. coli Rosetta for expression of the fusion protein SUMO-hmFGF21. Both in vitro and in vivo glucose regulation activity of hmFGF21 was evaluated. The SDS-PAGE result showed that compared with wild-type hFGF21, the soluble expression of hmFGF21 increased about 2-fold. HmFGF21 was more potent in stimulation of glucose uptake in HepG2 cells in vitro. The results of anti-diabetic effect on db/db mice demonstrated that hmFGF21 had better efficacy on controlling the blood glucose of the db/db diabetic animals than wild-type hFGF21. These results suggest that the biological properties of FGF21 are significantly improved by optimization.
9. A survey of the status of care quality evaluation in the institution that integrating pension and health care service:a qualitative research
Man LI ; Xinyu ZHOU ; Yuqin LI ; Hong LI ; Deshan LIU ; Minghao PAN ; Shushu ZHU ; Wei GAO
Chinese Journal of Practical Nursing 2018;34(19):1493-1497
Objective:
To explore the status of care quality evaluation in the built-in type institution that integrating pension and health care service and analyze the focus on nursing service and management,it aims to provide theoretical basis for nurse managers to implement scientific nursing management.
Methods:
Descriptive phenomenological methodology was adopted in this study,a semi-structured interview was conducted with 8 nurse managers from 7 built-in type institutions that integrating pension and health care service.Data were analyzed based on Colaizzi′ s phenomenological research method.
Results:
Two themes were extracted, including the status of care quality evaluation in the built-in type institution that integrating pension and health care service; key indicators of nursing quality in the built-in type institution that integrating pension and health care service.
Conclusions
Through in-depth interviews with 8 nurse managers, we suggest that state should attach importance to the quality inspection of nursing care in the built-in type institution that integrating pension and health care service, and explore the evaluation criteria for nursing quality,in order to ensure the safety of elderly care, and it also can provide a basis for the comparison between inter-agency nursing quality and promote healthy competition among institutions.
10.The work experiences in nursing homes among nurses:a qualitative research
Man LI ; Xinyu ZHOU ; Lianrong GUO ; Lingyun LI ; Deshan LIU ; Yang ZHANG ; Wei GAO
Chinese Journal of Practical Nursing 2018;34(6):450-454
Objective To explore the work experiences in nursing homes among nurses and to provide the referenced basis for managers to understand the needs of nurses and develop humanistic nursing management measures. Methods Descriptive phenomenological methodology was adopted in this study,a semi-structured interview was conducted with 9 nurses working in nursing homes.Data were analyzed based on Colaizzi′s phenomenological research method. Results Four themes were extracted including:heavy work tasks;high psychological pressure;strong demand in training;emotional experience of work. Conclusions Through in-depth interviews with 9 nurses working in nursing homes,we suggest that in order to promote the professional development of geriatric nursing, managers should change and update the concept of geriatric nursing and attach importance on professional study of nurse, and take appropriate intervention measures to lighten nurses' work pressure and adjust their bad mood, so as to arouse the enthusiasm of nurses.Finally,we can achieve the goal of stabilizing the nursing personnel and improving the quality of care in the nursing homes.