1.Expression and purification of Tat-GFP fusion protein and its cell membrane penetrating activity
Xingang GUAN ; Weiheng SU ; Xin YU ; Haibin TONG ; Xin SUN
Journal of Jilin University(Medicine Edition) 2014;(4):725-728
Objective To obtain the Tat-GFP fusion proteins with penetrating activity and labeled with green fluorescence protein (GFP), and to explore the cell membrane penetrating activity of Tat-GFP in MCF-7 cells. Methods The plasmid pET-24a-Tat-GFP was transformed into Escherichia coli BL21 cells. Different concentrations (0.5 and 1.0 mmol · L-1 ) of isopropyl-β-D-thiogalactopyranoside (IPTG ) and cell culture temperatures (22℃ and 37℃)were used to optimize the protein expression.The Tat-GFP proteins in supernatant were purified using Ni-IDA resins. Western blotting analysis was used to identify the Tat-GFP protein, and confocal laser scanning microscope (CLSM ) was used to examine the cell penetration of Tat-GFP protein. Results There was no significant difference in the Tat-GFP protein production induced by 0.5 and 1.0 mmol·L-1 IPTG;however,the low temperature (22℃)-induced BL21 cells expressed more Tat-GFP proteins than that at 37℃ induction.The Western blotting analysis results showed that GFP antibody could specifically recognize the proteins in PVDF membranes in dose-dependent manner;the CLSM results indicated the distribution of green fluorescence in cytoplasm and nucleus of MCF-7 cells.Conclusion The Tat-GFP protein highly expresses in the supenatant of Escherichia coli i BL2 1 cells at low temperature;the obtained Tat-GFP protein with green fluorescence preserves the cell penetrating activity.
2.Observations on the Efficacy of Nape-eight-needle Acupuncture plus a New-type Moxibustion Box in Treating Cervical Spondylotic Radiculopathy
Shuohuan JIN ; Xiaoping YU ; Ansheng YU ; Hanqin JIANG ; Xin GUAN
Shanghai Journal of Acupuncture and Moxibustion 2015;(11):1091-1093,1094
Objective To investigate the clinical efficacy of a new-type moxibustion box in treating cervical spondylotic radiculopathy.Methods Sixty patients with cervical spondylotic radiculopathy were randomly allocated to treatment and control groups, 30 cases each. The treatment group received nape-eight-needle acupuncture plus new-type moxibustion box therapy and the control group, nape-eight-needle acupuncture plus conventional moxibustion box therapy. After 10 treatments, the NPQ score, the symptom and sign score and the VAS score were recorded in the two groups and the clinical therapeutic effects were compared between the two groups.Results The total effective rate was 100.0% in the treatment group versus 83.3% in the control group, and the difference was statistically significant (P<0.05). There were statistically significant pre-/post-treatment differences in the NPQ score, the symptom and sign score and the VAS score in the two groups (P<0.01,P<0.05). There were statistically significant post-treatment differences in the NPQ score, the symptom and sign score and the VAS score between the treatment and control groups (P<0.05).ConclusionNape-eight-needle acupuncture plus a new-type moxibustion box is an effective way to treat cervical spondylotic radiculopathy.
3.Stability Predictions of 5% Glutaraldehyde Solution Using Classic Constant Temperature Accelerated Test Method
Yujun CHEN ; Shaowen ZHANG ; Hongzhen GUAN ; Yu FENG ; Xin LIU
China Pharmacy 2007;0(32):-
OBJECTIVE: To investigate the stability of 0.5% glutaraldehyde solution. METHODS: Classic constant temperature accelerated test method and sample observation method were applied with the content of glutaraldehyde as index. The content of glutaraldehyde was determined by titration which was determined by titration. RESULTS: The content change of 0.5% glutaraldehyde solution was in line with the first order kinetics, and the results of both methods were similar. The validity duration of 0.5% glutaraldehyde solution was 85.7 days at room temperature (25 ℃). CONCLUSION: 0.5% glutaraldehyde solution should be used up as soon as possible and the perfect time is less than 2 week.
4.Structure Design of Exoskeleton Device for Rehabilitation of Hands
Jicai LI ; Long GUAN ; Xin HU ; Hongliu YU
Chinese Journal of Rehabilitation Theory and Practice 2013;19(5):412-415
Objective To design an exoskeleton mechanical device for hand dysfunction rehabilitation. Methods Based on the bionic principles of the motion and the structure of finger joints, a micro exoskeleton mechanical structure with multi-joint fingers and multi-DOF motion was established, which included wearable finger exoskeleton and micro-motor drive mechanism. With the motion analysis software,the motion simulation analysis of the design was carried out on the basis of the three-dimensional model of the mechanical device. Results and Conclusion The structure of training device is compact and light, and the exoskeleton finger mechanism can move normally in the range of 0~67° without interference. Simulation data show that the mechanical device can complete planning exercises, and meet the design requirements of kinematics and mechanical properties.
6.Effects of cerebroprotein hydrolysate for injection (II) on neuritogenesis and its underlying mechanisms
WEI Dasha ; GUAN Xin ; ZHANG Shengbin ; YU Fang ; WANG Cunfang ; ZHOU Yu ; PANG Tao
Journal of China Pharmaceutical University 2021;52(2):219-226
In most mammalian central nervous system diseases, axons are damaged.Due to the limited ability of damaged neurons to promote axonal regeneration, the formation of glial scar and the release of inhibitory nutrients, it is difficult to regenerate axons of damaged neurons. The purpose of this study was to investigate the effect of cerebroprotein hydrolysate for injection (II) (CBL) on neuritogenesis and its underlying mechanism. Immunofluorescence staining was used to detect the axon length of mouse neuroma cells (Neuro-2a) and mouse primary cortical neuronal cells. Western blotting was used to detect the expression of phosphorylated TrkB protein in Neuro-2a cells and mouse primary cortical neuronal cells. The results showed that CBL could increase the axon length of Neuro-2a cells or mouse primary cortical neuronal cells, and that the phosphorylation level of TrkB in neuronal cells was significantly increased when 5 μg/mL CBL was applied to neuronal cells for 1 h. In conclusion, CBL can promote neuritogenesis, and increase the expression of phosphorylated TrkB, which may be related to the activation of TrkB signaling pathway.
7.The study of the relationship between influenza virus infection and the dysfunction of vascular smooth muscle cells
Xiuru GUAN ; Xinyuan JIA ; Xiaochen YU ; Na ZHANG ; Yuling JIANG ; Xiaomin XIN
Chinese Journal of Microbiology and Immunology 2012;(10):841-844
Objective To research the influenza virus infection on rat vascular smooth cells number,proliferation,apoptosis,the amount of IL-6,sFas,platelet-derived growth factor(PDGF) and the mechanism of atherosclerosis.Methods Flow cytometry,enzyme-linked immunosorbent assay and cell count experiments were used to detect these indicators at 0 h,6 h,12 h,24 h,48 h.Results After influenza virus infected at 0 h,proliferation,apoptosis condition were 10.39%,0.44%,respectively; at 6 h,proliferation,apoptosis respectively increased to 12.68%,0.73% ; proliferation reached the peak at 12 h (18.01%),instead apoptosis decreased to 0.14% ; at 24 h,proliferation decreased to 12.89% and apoptosis markedly increased to 1.09% ; at 48 h,proliferation further reduced to 7.07% and apoptosis reached the peak(4.61%).The number of cells and the cytokine secretion were statistically significant to control group (P<0.05).Conclusion Influenza virus infection might lead to change of cell proliferation and apoptosis and involve the atherosclerosis form and development,and cytokines played an important role in them.
8.The expression and clinical significance of heparanase and fascin in human breast carcinoma
Ying-Chao LI ; Guan-Yu ZHANG ; Li-Li XU ; Xin DU ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(09):-
0.05),but there was significant correlation between the expression between the size of tumor,lymph nodes metastasis,reoccurrence or prognosis and clinical grade(P
9.Study on the High Yield Strain of Gentamicin Producer by Protoplast Fused and Fermentor Test
Yu-Xia GUAN ; Ji-Xian LAN ; Guang-Xin YAN ; Jing-Shi ZHENG ;
China Biotechnology 2006;0(08):-
The protoplasts of Micromonospora purpurea,the high yield strains of gentammicin producer were mutagenized by diethyl sulfate(DES)and ultraviolet radiation(UV)respectively,then fused,screened by gentamicin resistance and regenerated.The average fermentation unit 2200?U/ml could be achieved by shake flask for 10 batches.The average fermentation unit 1900?U/ml could be obtained by 5L fermentor for 7 batches.The quality of the end product conformed to CP2000,BP2000 and USP26 pharmacopoeia.
10.Practice of Constructing a Platform of Excavating and Protecting Characteristic Technology of Diagnosis and Treatment in Research-based Hospital of Traditional Chinese Medicine
Lanying HUANG ; Xiaoping YU ; Jing AI ; Peng GONG ; Xin GUAN ; Xianying WANG ; Shuiyin JIANG ; Kangmei INSTRUCTORZHU
Chinese Journal of Information on Traditional Chinese Medicine 2014;(10):4-6
A plan of constructing a platform of excavating and protecting characteristic technology of diagnosis and treatment in research-based hospital of TCM was put forward by Shuguang Hospital Affiliated with Shanghai University of Traditional Chinese Medicine and Research Institution of Characteristic Technology of Diagnosis and Treatment Affiliated with Shanghai Academy of Traditional Chinese Medicine. The platform has begun to take shape on the basis of previous theoretical thought and preliminary practice. It has its own management system and working standards and contents with a clear and definite developing direction. Its working contents include excavating and protecting clinical experience of famous and aged practitioner of Chinese medicine, folk proved recipe and technology of diagnosis and treatment of TCM, intangible cultural heritage of TCM. Some problems appeared during the work, such as the evaluation criterion of the folk proved recipe and technology of diagnosis and treatment, the management of clinical proving projects, the issue of intellectual property protection and so on, which must be further solved.