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.Monitoring density of mosquitoes and resistance of major vector mosquitoes to insecticides in Zigong, Sichuan
LI Chun-yan ; TIAN Feng-yuan ; LING Yuan-nong ; DU Yu
China Tropical Medicine 2022;22(09):866-
Abstract: Objective To investigate the population densities composition and seasonal change of mosquitoes, and resistance of Culex pipiens quinquefasciatus and Aedes albopictus several commonly used insecticides in Zigong, so as to provide a theoretical basis for guiding the scientific rational selection and use of chemical insecticides, as well as the prevention and control of mosquito-borne diseases. Methods The surveillance data on mosquitoes by traps were collected from 2018 to 2020 in Zigong. The wild Culex pipiens quinquefasciatus and Aedes albopictus were collected throughout the city and reared to F1 generation in lab. The resistance of adult Culex pipiens quinquefasciatus and Aedes albopictus to these commonly insecticides was detected by contact tube method recommended. The statistical analysis on the monitoring results was used by Excel 2010. Results The annual average mosquito densities of monitoring sites were 1.11, 10.71, and 4.81 mosquitoes/(lamp·h), respectively from 2018 to 2020. The seasonal density of mosquitoes showed a single peak curve distribution throughout the year from 2018 to 2020 in Zigong, with the highest in July. There were obvious differences in mosquito density between different habitats, with the highest density of mosquitoes in barn. The 24 h mortality of Aedes albopictus and Culex pipiens quinquefasciatus to beta-cypermethrin, lambda-cyhalothrin, deltamethrin, permethrin, malathion, fenitrothion, chlorpyrifos, propoxur, and bendiocarb were 70.71% and 3.85%, 26.53% and 1.68%, 19.79% and 1.94%, 40.80% and 0.81%, 97.98% and 3.85%, 100.00% and 7.52%, 100.00% and 66.34%, 96.15% and 33.60%, 100.00% and 71.43%, respectively. The adult Aedes albopictus larvae in Zigong was sensitive to fenitrothion, chlorpyrifos, bendiocarb, showed resistance to beta-cypermethrin, lambda-cyhalothrin, deltamethrin, permethrin, and showed resistance dubiously to malathion and propoxur. The adult Culex pipiens quinquefasciatus larvae in Zigong have developed certain resistance to these commonly insecticides. Conclusion Both Culex pipiens quinquefasciatus and Aedes albopictus had high resistance to pyrethroid insecticides, which suggested that we should strengthen the patriotic hygiene campaigns, reduce pyrethroid insecticides, use alternately different types of insecticides for delay the development of resistance and better mosquito prevention and control.
5.Circulated extraction of flavonoids from Glechoma longituba by ultrasonic wave.
China Journal of Chinese Materia Medica 2007;32(5):385-387
OBJECTIVETo investigate the optimum condition of extraction for the flavonoids in Glechoma longituba by ultrasonic wave.
METHODUsing orthogonal test, the effects of ultrasonic power, ultrasonic time, extraction temperature and solvent concentration were considered, the comprehensive evaluation was guided by the content of the flavonoids determined by ultraviolet spectrophotometer.
RESULTThe optimum condition was as follow: ultrasonic power; 800 W, ultrasonic time 90 min, extraction temperature 40 degrees C, solvent concentration 65%. The flavonoids concent is 5.228%.
CONCLUSIONUsing ethanol as solvent, circulated extraction of the flavonoids from G. longituba with ultrasonic wave is feasible with the optimum conditions in low temperature, short time and high production yield.
Flavonoids ; isolation & purification ; Lamiaceae ; chemistry ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Technology, Pharmaceutical ; instrumentation ; methods ; Temperature ; Time Factors ; Ultrasonics
6.Preventive Effect on Nosocomial Infection with Herbal Medicine Atractylodes chinensis Disinfectant for Air Sterilization in Cardiothoracic Surgery
Hong-Yan HE ; Li SHENG ; Xue-Qin HUANG ; Fu-Ling LV ; Yuan-Chun HUANG ;
Chinese Journal of Nosocomiology 1994;0(04):-
OBJECTIVE To observe the preventive effect on nosocomial infection by the herbal medicine Atractylodes chinensis disinfectant for air sterilization in cardiothoracic surgery.METHODS A.chinensis disinfectant and ultraviolet irradiation were used to disinfect the patients′ rooms of our cardiothoracic surgery department.The total number of germ and fungus in the indoor air before and after using the two methods were collected.The nosocomial infection rates of the same period were also studied.RESULTS A.chinensis disinfectant had strong effect on sterilization of indoor germs and fungi,and the sterilization rate was 92.7%.The comparison between A.chinensis disinfectant and ultraviolet irradiation showed significant difference in their disinfection effects,while no significant difference in their nosocomial infection rates.CONCLUSIONS The herbal medicine A.chinensis disinfectant can achieve good sterilization effect and prevent nosocomial infection.
7.Effects of tidal volume with different amount of gas after recruitment maneuver ventilation on lung vascular endothelial diastole function in rats with acute lung injury
Jianqiang WANG ; Chun PAN ; Ling LIU ; Liang JIN ; Yi YANG ; Guomin LI ; Dong YUAN ; Haibo QIU
Chinese Journal of Emergency Medicine 2010;19(9):931-937
Objective To observe the effects of recruitment maneuver (RM) and tidal volume with different amount of gas after RM ventilation on lung diastole function in rats with acute lung injury (ALI). Method ALI rat models were induced by intravenous infusion of lipopolysaccharide (LPS) in dose of 6 mg/kg. Twenty-five rats were randomly(random number) divided into control group ( n = 5), ALI group ( n = 5), low tidal volume group (LV group,VT= 6 mL/kg, n = 5), sustained inflation (SI) with low tidal volume (SI+ LV group, VT=6 mL/kg, n = 5), and SI with moderate tidal volume group (SI+ MV group, VT= 12 mL/kg, n = 5). The RM carried out by using SI with airway pressure 30 cmH-2O for 30 seconds, and the positive end-expiratory pressure (PEEP)was set at 5 cmH2O. Lung tissue was taken after mechanical ventilation for 5 hours. The mean arterial pressure (MAP) was monitored throughout the entire course of experiment. Endothelin-1 ( ET-1 ), endothelial nitricoxide synthase (eNOS), and acetylcholine-(Ach-) induced endothelium-dependent relaxation response of isolated pulmonary artery rings were investigated after mechanical ventilation for 5 hours. Results The LPS increased the ET-1 level in lung tissue, decreased the level of eNOS in lung tissue, and impaired the Ach-induced endothelium-dependent relaxation response in pulmonary vassals, without obvious influence on systemic hemodynamics. SI + LV significantly reduced LPS-induced elevation of ET-1 level, and increased the level of eNOS, and significantly lessened endothelial dysfunction and ameliorated dysfunction od endothelium-dependent relaxation in pulmonary vas sals. Conclusions RM with high tidal volume or lowtidal volume ventilation could improve the lung vascular endothelial function of rats with acute lung injury, and RM with low tidal volume ventilation could lessen more the injury of lung vascular endothelial diastole function in rats with acute lung injury.
8.An Intracerebral Hemorrhage Model Established by Autologous Blood Injection Three Times in Rat
Hong-ling LI ; Rui-chun LIU ; Yuan-chong GOU ; Chunyan LI
Chinese Journal of Rehabilitation Theory and Practice 2006;12(8):651-653
ObjectiveTo establish an animal model of intracerebral hemorrhage (ICH) in rat and to observe the changes in behavior, brain edema and tissue structure during absorption of the hematoma.MethodsAn animal model of ICH was established by stereotactically infusing 100 μl (the first time 20 μl, the second time 40 μl, the third time 40 μl) of autologous caudal artery blood into caudate nucleus in the rats. The behavior of rats with ICH was assessed by Bederson score, beam-walking test, bilateral forepaws grasp and measurement of forelimb placing. The brain edema and the change in histology were observed.ResultsThe successful ratio of this model was 75%. There were significant behavioral changes and brain edema at 6 h after the ICH in rats of the experimental group compared to those of the control group. The worse behavioral changes and brain edema was at 48~72 h after hemorrhage. In the earlier period, the nerve cells were swelling. A lot of glial cells appeared around the hematoma in the late period.ConclusionThis ICH model is reproducible, and the obvious behavioral change and histological changes are similar to clinical patient with ICH.
9.Sequence Analysis of ORF3 and Partial ORF1 Region from Two Patients Infected with New Genotype of Hepatitis E Virus(HEV)
You-chun, WANG ; Hua-yuan, ZHANG ; Wen-jie, GU ; Ling, ROGER ; He-min, LI ; Harrison J TIM
Virologica Sinica 2001;16(1):28-33
ORF3 and partial ORF1 regions were amplified with RT-PCR f rom two patients (T1 and T11)infected with new genotype of hepatitis E Virus. Th e PCR products were cloned and sequenced. The results showed that G-C rich regi on in ORF3 was deleted when amplified with normal PCR reaction. However, PCR rea ction containing G-C melt solution can overcome this problem. The sequence anal ysis showed that T1 and T11 belong to a new genotype of HEV which differs from g enotype I,II and III reported.T1 and T11 have 79%~82%, 80%~81% and 83%~85% id entical to genotype I,II and III respectively.
10.Utilization of ?-Red Recombination System and Balanced Lethal System to Reconstruct the Plasmid with Antibiotic Resistant Gene Using in Live Vaccine
Sheng-Ling YUAN ; Peng WANG ; Xiang-Xin LIU ; Yan-Chun WANG ; De-Wen ZHAN ; Zhao-Shan ZHANG ;
Microbiology 1992;0(02):-
Recombination plasmid pMM085 possessed both immunogens heat-labile enterotoxin(LT) and fimbriae antigen K88 of enterotoxigenic Escherichia coli (ETEC). Althouth vaccine strain MM-3 carrying pMM085 had good effect to protect piglets against diarrhea due to ETEC infections,it was not ideal live vaccine for pMM085 bringing chloramphenicol resistance gene (cat). To solve the problem,the host-plasmid balanced lethal system was introduced which including the replacement of cat gene by asd gene and transformation the new plasmid to the strain X6097 which asd gene was knocked out in its chromosome. Considering pMM085 was a big plasmid (23kb) and traditional genetic manipulations was not easy to carry on,?-Red recombination system was adopt in this work to realize the replacement of cat gene by asd gene. The results indicated that ?-Red recombination system was convenient and efficient to reconstruct big plasmid.