3.Gene construction and screening of humanized single chain antibody library against VSTM1-v2 cytokine.
Xiaojin FU ; Yongxia ZHANG ; Yunjian DAI ; Mingrong WANG
Acta Pharmaceutica Sinica 2013;48(11):1651-6
To rapidly select potent anti-VSTM1-v2 scFv (single-chain antibody fragment) by construction and screening of a humanized scFv library in which a murine VH-CDR3 library was grafted onto a human scFv framework. A murine VH-CDR3 library was amplified from anti-VSTM1-v2 murine cDNA and grafted on human scFv (VH3-VK1) framework. Anti-VSTM1-v2 scFv templates were selected and enriched through ribosome display, TA-cloned into expression vector, and transformed into BL21 (DE3) for soluble expression of target scFv. A total of 1000 clones were randomly picked. Positive ones were first identified using colony PCR, indirect ELISA, Western blotting and then verified with sequencing and dose response ELISA. At last an anti-VSTM1-v2 humanized scFv with good binding affinity (EC50 = 21.35 nmol x L(-1)) was selected from the humanized library of 10(12) members generated in this study. This scFv antibody might have potential applications. This study provides a new approach for rapid screening of humanized antibodies.
4.Clinical effect of small dose of glucocorticoid in the treatment of AECOPD patients with severe disease associated with low levels of cortisol
Haiwei FU ; Xiaobo LI ; Yiming KONG ; Tianwei ZHANG ; Yuanrong DAI
Chinese Journal of Biochemical Pharmaceutics 2017;37(6):58-59,62
Objective To explore clinical effect of small dose of glucocorticoid in the treatment of AECOPD patients with severe disease associated with low levels of cortisol.Methods72 patients with AECOPD in our hospital were randomly divided into control group and treatment group.The control group was treated with conventional AECOPD method,the treatment group was treated on the base of the treatment group with a small dose of corticosteroids,CRP, IL-6, TNF-α, FEV1, PCT and other indicators, as well as patients with ventilation, shock and mortality were compared after the end of treatment.ResultsThe time of non mechanical ventilation, the time of non-mechanical ventilation, and the time of non-shock were increased, the shock and the mortality was lower in the treatment group,(P<0.05);The CRP level of patients in the treatment group was lower(P<0.05);IL-6, TNF-α, PCT levels of patients in the treatment group was lower (P<0.05);FEV1, FVC and FEV1/FVC level in the treatment group were higher(P<0.05).ConclusionSmall dose glucocorticoid can improve the pulmonary function of patients with AECOPD complicated by severe disease, and alleviate the disease, significantly reduce the levels of inflammatory factors, promote the improvement of the lung function.
5.The effect of the shrinkage of thermoplastic mask on patient positioning
Yin ZHANG ; Jianrong DAI ; Minghui LI ; Guishan FU
Chinese Journal of Radiation Oncology 2015;(4):454-456
Objective This study analyzes the effect of the shrinkage of thermoplastic mask on patient positioning. Methods Design of the two test. Test 1:thermoplastic film shrinkage test. Get some thermoplastic film by the size of 10 cm×5 cm, extrude it at a certain rate after heated. Measure the length of thermoplastic film on different time, and calculate the contraction. Test 2:phantom test. Take advantage of head and neck phantom, and simulate the procedure that from making mask for patients to radiation therapy. Measure the off set of isocenter which caused by the contraction of thermoplastic mask. Results The largest shrinkage of thermoplastic had happened in 20 minutes. Different tensile ratio had little effect on the shrinkage. The offset of isocenter which caused by the shrinkage of thermoplastic mask were:LR ( -0?? 1± 0?? 3) mm,SI (-0?? 2±0?? 2) mm, AP (0.6±0?? 4) mm,respectively. There was little change in the course of six weeks ( P= 0.185?0?? 961). Conclusions The cooling time should be more than 20 minutes, when making a mask for the patient. The setup errors which caused by the shrinkage of thermoplastic mask is at an acceptable level on this premise.
6.Kinetics of Fed-batch Cultivation on Recombinant Escherichia coli Containing Human-like-Collagen cDNA
Yu MI ; Xiu-Fu HUA ; Dai-Di FAN ; Xi ZHANG ;
China Biotechnology 2006;0(04):-
The kinetics of batch and fed-batch cultures of recombinant Escherichia coli to produce humanlike collagen were investigated. Through examining the density of substrate and the amount of mushrooms and the density of products during the process of fermenting, a set of kinetic models are set up. The influence of cell without plasmid was considered. The results show that the kinetic model may well simulate the fermenting process.
7.Volatile Oil Constituents of Two Acremonium Endophyte Isolates from Aquilaria sinensis
Xiu-Huan ZHANG ; Wen-Li MEI ; Hao-Fu DAI ;
Microbiology 1992;0(01):-
The volatile oils of endophyte isolates R1 (Acremonium sp.) and R2 (Acremonium sp.) from Aqui-laria sinensis were extracted by solvent method. Twenty and sixteen ingredients were identified from the volatile oils of R1 and R2 by GC-MS, respectively. Fatty acids, such as oleic acid, linoleic acid, hexa-decanoic acid etc., were the main components, whereas terpenoids like 1,8-cineole, zingiberene, ar-curcumene were also found in both oils. It’s the first report about the volatile oil constituents of endophyte isolates from A. sinensis.
8.MRI comparison of lumbar facet joint degeneration and intervertebral disc degeneration in patients with low back pain
Changlin FU ; Bin ZHANG ; Yuan LIU ; Min DAI ; Xin ZHOU ; Xiaoxing FU
Chinese Journal of Tissue Engineering Research 2015;(46):7401-7405
BACKGROUND:During spinal degeneration process, the intervertebral disc and facet joints are interrelated and interacted to impact the normal function and stability of the spine, thereby resulting in low back pain. Moreover, there is always a controversy on the degeneration order of the intervertebral disc and facet joint. OBJECTIVE:To explore the relationship between lumbar facet joint degeneration and intervertebral disc degeneration in patients with low back pain. METHODS: A retrospective analysis was made on the clinical data of 186 patients with low back pain. The facet joint degeneration and intervertebral disc degeneration at L2-S1 motion segments of each patient were evaluated. Enroled patients were divided into three age groups: < 40 years old, 40-60 years old and≥ 60 years old. RESULTS AND CONCLUSION:The incidence of lumbar facet joint and intervertebral disc degeneration was increased with age, and degeneration of the lumbar facet joint and intervertebral disc were the most obvious at L4-5 and L5-S1 segments. The incidence of intervertebral disc degeneration was more than that of facet joint degeneration at each segment in al age groups, except groups of < 40 years old and 40-60 years old at L2-3 segments, but there was no significant difference (P > 0.05). There was a weak correlation between facet joint degeneration and intervertebral disc degeneration (χ2=100.9,P < 0.001, gamma=0.22). These findings show that the incidence of intervertebral disc degeneration and facet joint degeneration is increased with age, and there is a weak correlation between them. However, the lumbar degenerative order is stil unclear and further research is needed.
9.Randomised clinical trial on rh-Endo combined with FOLFOX4 regimen as an adjuvant therapy for stage Ⅱ and Ⅱ colorectal cancer patients
Zhihua XIE ; Dejian DAI ; Lin ZHONG ; Yi YI ; Jun FU ; Zhijin ZHANG ; Yuhao ZHANG
Chinese Journal of General Surgery 2013;28(10):758-762
Objective To evaluate the long-term efficacy of recombinant human endostatin (rh-Endo) combined with FOLFOX4 as an adjuvant treatment for patients of stage Ⅱ and Ⅲ colorectal cancer.Methods Eligible patients were randomly assigned to receive FOLFOX4 or FOLFOX4 plus rh-Endo regimen in which patients receiving 7.5 mg/m2 Ⅳ on day 1-7,repeated every 2 weeks,to a total of 12 cycles in 6 months.Results A total of 197 eligible patients were accrued in this research with 105 patients in the control group and 92 patients in the experimental arm.Median follow-up period was 42 months.The baseline characteristics distributed were balanced by treatment.Rh-Endo combined with FOLFOX4 regimen resulted in significant improvement on DFS compared to FOLFOX4 regimen for patients with stage Ⅲ colon cancer (HR =0.19,95% CI0.05-0.75,P =0.0124),and with a 34% improvement on 3-year DFS and 81% reduced recurrence.Although rh-Endo combined with FOLFOX4 regimen failed to make significant difference on DFS in the whole (HR =0.75,95% CI 0.31-1.83,P =0.5589),it was also observed a 17% improveiment on 3-year DFS.No statistical significant difference on DFS was observed in patients with stage Ⅱ disease.Conclusions Rh-Endo combined with FOLFOX4 regimen significantly improved the disease-free survival for patients with stage Ⅲ colorectal cancer,indicating that patients with stage Ⅲ disease,but not stage Ⅱ disease,can benefit from FOLFOX4 plus rh-Endo regimen in adjuvant treatment.
10.Effects of human IL-10 gene transfection on the mRNA and protein expressions of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion injury in rats
Jun FU ; Yi ZHANG ; Chengsheng ZHANG ; Changchun YANG ; Shilei WANG ; Qishui LIN ; Haixing XUAN ; Dai ZHOU
Chinese Journal of Geriatrics 2008;27(10):775-779
ObjectiveTo observe the effects of human IL-10 gene transfection on the mRNA and protein expressions of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion injury in rats and to investigate its neuroprotective mechanism. MethodsRats were divided into four groups: normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group. The mRNA and protein expressions of IL-1β and TNF-α in the penumbra area were detected by fluorescence real-time quantitative PCR and ELISA respectively. ResultsIn normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group, the levels of protein expression of TNF-α in penumbra area were(0.66±0. 04) ,(1.16±0.26),(1. 155±0. 26)ng/g and(0. 84±0. 05)ng/g, and the levels of protein expression of IL-1βin penumbra area were(0.37±0.05), (1.25±0.39), (1.21±0.57) ng/g and(0.62+0.05)ng/g, respectively. Compared with normal control group, the levels of protein expression of TNF-α and 1L-1β were significantly higher in other three groups(all P<0. 01), and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group(all P<0. 01). In normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfectedgroup, the levels of mRNA expression of TNF-α in penumbra area were 1.00 ±0.53,9.42±1.83,9.69±1.96 and 3.53±1.09, and the levels of mRNA expression of IL-1β in penumbra area were 1.00 ±0.51,27. 81±4.84,23.96 ± 4.90 and 13.55± 4.45, respectively. Compared with normal control group, the levels of mRNA expression of TNF-α and IL-1β were significantly higher in other three groups(all P<0. 01), and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group(all P<0. 01). ConclusionsThe human IL-10 gene transfection may play an protective effect on cerebral ischemia through inhibiting mRNA and protein expression of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion in rats.