1.Expression and purification of rat brain NT-3 fusion protein and its antibody preparation
Zhiquan LI ; Yunyu HU ; Qingsheng ZHU ; Jinyu ZHU ; Fang LIU
Basic & Clinical Medicine 2006;0(03):-
Objective To clone NT-3 gene from normal rat brain and to purify its fusion protein and to prepare specific high titer antibody so that to provide a foundation for further study for peripheral nerve injury.MethodsWe amplified target gene by RT-PCR and cloned it into the vector of pMD-18T,then analyzed its sequence and compared it with the sequence from GenBank.We subcloned it into pRSET-A vector and introduced it into Escherichia coli BL21.The expression was induced by IPTG,and identified by SDS-PAGE.The fusion protein was purified by niccolum purify kit.We immuned rabbits with immunological adjuvant for specificity antibody preparation.Results We got a 777 bp gene segment by RT-PCR.The DNA sequence was identical to rat NT-3 gene sequence in GenBank.It proved that the target gene was correctly inserted into the vector.A new protein band of about 34 ku appeared on SDS-PAGE after induction of IPTG.A specific high titer antibody of 1∶64000 was gained by immunizing rabbits with adjuvant.
2.Effects of Recreational Therapy on Social Communication of Stroke Patients
Jiaqi FAN ; Yunyu ZHU ; Wenhua CHEN ; Yaping GU
Chinese Journal of Rehabilitation Theory and Practice 2010;16(3):262-263
ObjectiveTo investigate the effect of the recreational therapy on the ability of social communication in stroke patients. Methods58 cases of stroke patients were divided into recreational group (28 cases) and control group (30 cases). All of them received routine rehabilitation therapy, while the recreational group received the recreational therapy in addition. All patients were evaluated with Functional Independence Measure (FIM) before and after treatment. ResultsThe scores of FIM significantly improved after treatment (P<0.05) in both groups, and the scores in social communication improved more (P<0.01) in recreational group than in control group. ConclusionRecreational therapy may improve the ability of social communication more effectively for stroke patients.
3.Application of Recreational Therapy in Stroke Rehabilitation (review)
Yunyu ZHU ; Wenhua CHEN ; Bo YU ; Yanyan YU ; Qi WI
Chinese Journal of Rehabilitation Theory and Practice 2009;15(2):152-154
Stroke patients usually are complicated with degeneration of body function as well as emotional and cognitive problems; consequently it will affect capacities of daily lives and recreation in most cases. Recreational therapy (RT) being favorable as far as stroke rehabilitation is concerned. The authors reviewed current developments of applications of RT to carry on a brief talk about present and future utilization of RT in stroke rehabilitation and it holds the point that when practicing RT we should introduce relevant evaluation Methods and techniques adopted in physical therapy (PT) and occupational therapy (OT) area into comprehensive rehabilitation program for some better Results . Evidence based principal should also play a positive role to make a systematic and specific intervention plan in further procedure of RT treating stroke patients.
4.Optimizations of an ELISA-like high-throughput screening assay for the discovery of β-catenin/TCF4 interaction antagonists.
Zhenghao FU ; Gangan YAN ; Xiaohong ZHU ; Xiaoping LIU ; Yunyu CHEN
Chinese Journal of Biotechnology 2021;37(8):2878-2889
In canonical Wnt/β-catenin signaling pathway, β-catenin/TCF4 (T-cell factor 4) interaction plays an important role in the pathogenesis and development of non-small cell lung cancer (NSCLC), and it is tightly associated with the proliferation, chemoresistance, recurrence and metastasis of NSCLC. Therefore, suppressing β-catenin/TCF4 interaction in Wnt/β-catenin signaling pathway would be a new therapeutic avenue against NSCLC metastasis. In this study, considering the principle of enzyme-linked immunosorbent assay (ELISA), an optimized high-throughput screening (HTS) assay was developed for the discovery of β-catenin/TCF4 interaction antagonists. Subsequently, this ELISA-like screening assay was performed using 2 μg/mL GST-TCF4 βBD and 0.5 μg/mL β-catenin, then a high Z' factor of 0.83 was achieved. A pilot screening of a natural product library using this ELISA-like screening assay identified plumbagin as a potential β-catenin/TCF4 interaction antagonist. Plumbagin remarkably inhibited the proliferation of A549, H1299, MCF7 and SW480 cell lines. More importantly, plumbagin significantly suppressed the β-catenin-responsive transcription in TOPFlash assay. In short, this newly developed ELISA-like screening assay will be vital for the rapid screening of novel Wnt inhibitors targeting β-catenin/TCF4 interaction, and this interaction is a potential anticancer target of plumbagin in vitro.
Carcinoma, Non-Small-Cell Lung
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Cell Line, Tumor
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Enzyme-Linked Immunosorbent Assay
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High-Throughput Screening Assays
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Humans
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Lung Neoplasms
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Transcription Factor 4/genetics*
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beta Catenin/genetics*