2.The Influence of ?-Synuclein Overexpression on Mitochondrial Membrane Structure with Atomic Force Microscopy
Chun-Li ZHAO ; Yuan-Gang ZHU ; Chun-Li DUAN ; Ling-Ling LU ; Ling ZHANG ; Hui YANG ;
China Biotechnology 2006;0(11):-
Objective:To identify the effect of ?-synuclein overexpression on mitochondrial membrane structure with atomic force microscopy. Methods:?-syn expression was mediated by AAV (adeno-associated viral vector) and Recombinant AAV/?-syn and AAV/LacZ viral particles were stereotaxically injected in the left side of rat substantia nigra (SN) for rat model of ?-synuclein overexpression. Mitochondria were isolated from rats SN of Brain. Mitochondria were analysis with JC-1 staining,atomic force microscopy and Western blot. Results:By 16 weeks post-infection of AAV-?-syn,the level of ?-syn increased about 2 times in mitochondrial fraction with Western blot and mitochondrial membrane potential (??) decreased with JC-1 staining. Furthermore,mitochondria swelling and porous like structure formed on the mitochondrial membrane with atomic force microscopy. Conclusion:The data suggested that ?-syn could accumulate in mitochondria,might form mitochondrial membrane pores and lead to ?? decreases. ?-syn might lead to mitochondrial dysfunction in Parkinson's disease.
3.Study on the Role of ?-synuclein in Mitochondria Dysfunction Caused by Small Dosage Rotenone
Ling-Ling LU ; Yuan LIANG ; Chun-Li DUAN ; Chun-Li ZHAO ; Huan-Ying ZHAO ; Hui YANG ;
China Biotechnology 2006;0(12):-
Mitochondrial dysfunction has been implicated in the aetiology of sporadic Parkinson's disease but its role in the disease mechanism remains unclear.To investigate the effect of synuclein on mitochondrial dysfunction induced by rotenone.The human dopaminergic SH-SY5Y cells were used as a cell model.The cells over-expressed the wild-type ?-synuclein were treated with complex I inhibitor rotenone.The cell viability,complex I activity,Mitochondrial swelling and O2-content were tested at different time point-1w,2w,4w after rotenone treated.CCK-8 test results showed that the cell viability of overexpressed ?-synuclein(SH-SY5Y-Syn)was much lower than the control group(SH-SY5Y-Ctr).After administrating with rotenone about 1w or 2w the cell viability of SH-SY5Y-Syn became higher than that of SH-SY5Y-Ctr.On the 4th week the results were contrary to the first 2 weeks.Similar results were got when test the mitochondrial function.In the first 2 weeks after roteoone administrating,the mitochondrial function of SH-SY5Y-Syn was better than that of SH-SY5Y-Ctr.This suggest that the ?-synuclein could protect the mitochondrial against the injury induced by rotenone in the early stage-1w,2w,while this effect disappeared in the final stage-4w.
4. Preparation and performance test of vegetable enteric hollow capsules by one-line molding process
Chinese Pharmaceutical Journal 2015;50(23):2057-2063
OBJECTIVE: To develop vegetable enteric hollow capsules with sodium alginate (Alg-Na), starch and excipients by one-line molding process and investigate their properties. METHODS: The influence of the ratio of raw materials on capsule shape index was analyzed by multiple factors trials. The light transmittance and mechanical properties were evaluated, and DSC, FT-IR, and SEM analysis were conducted. RESULTS: The optimal formula composed of medium viscosity Alg-Na 0.6%, ultralow viscosity Alg-Na 2.5%, starch 10%, gellan gum 0.6%, and glycerin 2% could produce vegetable enteric hollow capsules with good performance. The main property indexes were as follows: liquid viscosity 434 mPa·s, capsule thickness 0.11-0.12 mm, water content 10.09%, disintegration time 8-15 min in simulated intestinal fluid (not disintegrated within 2 h in simulated gastric fluid), light transmittance of capsule film 85.5%, tensile strength 27.97 MPa, and elongation at break 2.0%. CONCLUSION: This study lays theoretical foundation for replacing gelatin capsules with vegetable enteric hollow capsules.
5.The Influence of Body Positions on Blood Pressure in Diabetic Patients
Xiao-Li YE ; Ling-Chun LU ; Shan LI ; Tie-Min WEI ; Chun-Lai ZENG ;
Chinese Journal of Hypertension 2007;0(03):-
Objective To investigate the difference between the blood pressure readings between sitting and supine position,and to study the factors that associated with the sitting-supine blood pressure difference in patient with diabetes.Methods We measured the sitting blood pressure first then followed by the supine pressure in 356 diabetic patients,using a standard mercury sphygmomanometer.Patient's body weight,height and blood glucose levels were also measured.Results SBP and DBP were significantly higher in the supine position than in sitting position in diabetic patients(by 3.5?7.6/1.5?4.9 mm Hg,P
6.Optimized alloxan-induced diabetic rabbit model
Wen-Tao, SUN ; Chun-Ling, LEI ; Si-Hai, ZHAO ; Chun-Chao, BI ; Lu, ZHANG
International Eye Science 2010;10(10):1848-1850
AIM: To explore the frequency of drug injection of alloxan diabetes on the established model of rabbit.METHODS: Thirty-six healthy rabbits, weighing 2-2.5kg, were randomly divided into one time drug injection group (group A, n=12), two times drug injection group (group B, n=12) and three times drug injection group (group C, n=12). Each rabbit was injected with a total amount of 150mg/kg of alloxan. Fasting blood glucose was measured. The success rate and death rate of each group were also calculated.RESULTS: The success rate of diabetic rabbit model in group B was higher than that in group A (P<0.01) and its death rate was lower than that of group A (P<0.01); the success rate of diabetic rabbit model in group C was highest and the death rate was the lowest in three groups(P<0.01). CONCLUSION: Multiple administration of alloxan can improve success rate in establishing diabetic rabbit model with decreased death rate and increased stability.
7.Experimental autoimmune myositis in rat
Yao XIE ; Xin LU ; Guo-Chun WANG ; Tai-Ling WANG ; Hong LI ; Jion GUO ;
Chinese Journal of Rheumatology 2003;0(11):-
Objective The aim of our study is to establish and characterize the animal model for au- toimmune myositis.Methods Fifty male SD rats were randomly divided into 2 groups:model group(n=40) and control group(n=10).The model group rats were immunized with muscle homogenate every week for 5 weeks and received an injection of 2?g pertussis toxin at the first and second week.As controls,10 SD rats were injected with an equal volume of normal saline.Tissue specimens from limb skeletal muscles were ob- tained at 1,2,3,4,5 weeks after injection.At the same time,the blood samples were collected,and the level of CPK was measured.Results The model group had significantly elevated serum CPK levels.There were multiple inflammatory lesions in the skeletal muscles.Local degeneration and necrosis of muscle fibers with disappeared transverse striation,mononuelear cell infiltration in the interstitial could be observed.The patho- logic grade was mainly 2a.The infiltrating mononuclear cells were predominantly CD8~+T cells that mainly lo- cated in the endnmysium.MHC classⅠantigen expression on muscle fiber membranes in the model group was upregulated.Conclusion The experimental autoimmune myositis induced by syngeneic skeletal muscle ho- mogenate in SD rat is pathologically similar to human myositis.It can be used as a good model for human myositis and provides the basis for the etiopathology and therapeutical studies.
8.Medium Optimization for Lipopeptide Produced by Bacillus natto TK-1 Using Response Surface Methodology
Xiao-Hong CAO ; Ping CAI ; Fan LI ; Chun-Ling WANG ; Mei-Fang LU ;
China Biotechnology 2006;0(04):-
Response Surface Methodology was applied to optimize the culture components for lipopeptide production by Bacillus natto TK-1. In the first step, two level factorial design of Plackett-Burman was used to evaluate the influence of six related factors. It showed that three factors playing the important roles in the medium, including peptone, yeast extract powder and CaCl_2. The path of steepest ascent was used to approach the optimal region of the fermentation conditions subsequently. In the third step, the concentrations of those three main factors were further optimized by using Box-Behnken and Response Surface Analysis. By solving the quadratic regression model equation, the optimal concentrations of the variables were determined as: peptone 1.73%, yeast extract powder 0.063 %, CaCl_2 1.385?10-4mol/L. Under the optimal culture conditions, the diameter of haemolysis zone increased 29.3 % than before. HPLC analysis showed the precise production of lipopeptide was 30.2% higher than preliminary culture. Furthermore, at three batches cultivation, the experiment values under the optimal conditions agreed with the predictive values. It showed that Response Surface Methodology was proper and a good choice for optimization.
9.Studies of Temperature Shift Fermentation for Eicosapentaenoic Acid Production by Nitzschia laevis
Xiao-Hong CAO ; Yu-Hua ZHAO ; Mei-Fang LU ; Jing LEI ; Chun-Ling WANG ;
China Biotechnology 2006;0(12):-
Fermentation for Eicosapentaenoic Acid(EPA) production by Nitzschia laevis at various temperature between 10℃ and 30℃ was investigated and the dynamics characteristics during fermentation process were also analyzed.Based on the results,a varying temperature nursing method of two stage control strategy is proposed:During the first stage,which comprises the delay phase and the initial index phase,the temperature is maintained at 25℃;then the temperature is shifted to 20℃ and kept up till the end of the fermentation process.By this method,a EPA content of 6.0% and a yield of 291.60 mg/L have been gained.These are 24.07% and 18.81% higher than that of fixed temperature(25℃) fermentation,respectively.
10.The Structural Identification and Antitumor Activity on MCF-7 Cells of Surfactin from Bacillus subtilis TK-1
Xiao-Hong CAO ; Run-Zhi JIAO ; Chun-Ling WANG ; Le YAN ; Mei-Fang LU ;
China Biotechnology 2006;0(02):-
This thesis aimed at the Bacillus natto TK-1 screened out from Natto.The lipopeptide was purified using Thin-Layer Chromatography(TLC),and investigated its anti-tumor activity.After acid precipitation and methanol extraction,the lipopeptide was separated on TLC.Then the authors get the monomer surfactin which molecular weight is 1036Da through the High performance liquid chromatography(HPLC),electro spray ionization-mass spectrometry(ESI-MS) and infrared(IR).MTT method was implied to testify the anti-tumor activity of the purified sample from TLC.The results indicated a concentration and time-dependent relationships.After 48h,their IC50 were 40 mg/L.The detection with inverted microscope fluorescence microscope displays that the surfactin will cause a series of Morphological changes to the cells.In TUNEL experiment,the authors noticed that surfactin has the ability to induce apoptosis,besides this inhibition shows an obvious time-dependent relationship.