1.Analysis of GDNF(△N39)-R9 Fusion Protein Delivery Across The Cellular Membrane,Blood Brain Barrier and Biological Function
Progress in Biochemistry and Biophysics 2006;0(08):-
In order to study the application of glial cell line-derived neurotrophic factor(GDNF)in clinic,gene mutation,fusion protein expression in E.coli and purification methods have been used to obtain the fragments of GDNF,GDNF(△N39),GDNF(△N39)-R9.Using primary cultured dopaminergic neurons and PC12 cells with transfected with GFR?1 and Ret to observe their biological function and cytotoxicity.Using B-Endo3 cells and Transwell method to analyze their delivery across the cellular membrane and blood brain barrier.The results show that GDNF(△N39)-R9 has the same neurotrophic function with wild GDNF and nearly no cytotoxicity to dopaminergic neurons and PC12-GFR?1-Ret cells and can get through effectually the cellular membrane and simulacrum of blood brain barrier with matrigel and B-Endo3.
2.Enhancement of antitumor immunity of dendritic cells pulsed with hyperthermia lymphoma cells
Limei AI ; Jing PAN ; Yuanyuan SUN
Chinese Journal of Immunology 2010;26(3):232-235
Objective:To investigate the immuno-effects triggered by dendritic cells (DCs) pulsed with Heat-treated lymphoma cells.Methods:Mononuclear cells from healthy human peripheral blood(PBMC) were isolated and DCs were generated in vitro with normal methods.Lymphoma cells were heated (42℃,2 h) and then loaded onto DCs.Immunotypes of DCs and lymphocytes were detected with flow-cytometric analysis.IFN-γ releasing was detected by ELISPOT,lymphocyte cytotoxicity was analyzed by MTT method after mixing with DC reaction.Results:After pulsing DCs with hyperthermia lymphoma cells,CD80~+,CD86~+ and HLA-DR existed higher than those of the control group,IFN-γ releasing and cytotoxicities were also strengthened,but there was no difference between them,cytotoxitic lymphocytes (CTLs) were produced more as CD8~+,CD25~+ and CD56~+ cells by flow-cytometric analysis.Conclusion:Anti-lymphoma immuno-effects could be strengthened by loading DC with hyperthermia lymphoma cells.
3.Preparation and quality evaluation of paclitaxel loaded solid lipid nanoparticles
Jing HAN ; Zhi LI ; Qin CEN ; Yuefeng PAN ; Limei YIN
Chinese Traditional and Herbal Drugs 1994;0(07):-
Objective The paclitaxel loaded solid lipid nanoparticles (PTX-SLNs) were prepared by an ultrasonic-dispersion emulsification technique. The stability of PTX-SLNs was investigated in this study. Methods The technology was preferenced by stability, Zeta potential, particle diameter, and entrapment efficiency as indexes. The doses of lipid materials and coemulsifier, the ultrasonic time, and the ultrasonic power were investigated in detail. Results The optimum prescriptions were definited by one-factor and orthogonal test. The adjuvant: glyceryl monostearate (100/150 mg), Fabaceous Lecithin (100 mg), coemulsifier Pluronic F68∶Tween 80 (2∶1). The samples were sonicated with an energy output of 300 W in 20 min after emulsified at (75?5) ℃. Conclusion The PTX-SLNs are successfully prepared and PTX-SLNs with high stability are fairly dispersed in colloidal solution. This technology has a nice prospect with safety and reliability.
4.Improvement of Hydroscopicity of Traditional Chinese Drug Extractum by Spray Drying
Zhi LI ; Jing HAN ; Qin CEN ; Yuefeng PAN ; Limei YIN
China Pharmacy 2007;0(27):-
OBJECTIVE:To improve hydroscopicity and fluidity of traditional Chinese drug extractum by spray drying and to improve affixes to the wall of the drier during drying for providing reasonable relative humidity of production.METHODS:The compounding of adjuvant and the technology were optimized by orthogonal experiment,and the hydroscopicity and fluidity of powdered extract were investigated.RESULTS:The hydroscopicity of traditional Chinese drug extractum can be greatly decreased by adding 3% gum arabic and 7% ?-CD.The optimum technological conditions were as follows:extractum density of 1.10g? mL-1,temperature of intake airflow of 170℃,atomization pressure of 0.5Mpa,and feed material speed of 400mL? h-1 by orthogonal experiment.The critical relative humidity of the optimum formula was 64%.CONCLUTION:The problems of time consuming and moisture absorption of traditional Chinese drug extractum existed in the traditional old technology can be improved in this new technology.
5.Total ginsenosides improve cardiac function via promoting angiogenesis in a rat model of acute myocardial infarction
Ying WEI ; Limei YU ; Yuying WANG ; Guanping YAO ; Li PAN
Chinese Pharmacological Bulletin 2016;32(4):559-563,564
Aim To investigate the effects of total gin-senosides ( TG) on microvascular regeneration and car-diac function in rat after acute myocardial infarction ( AMI ) . Methods The acute myocardial infarction model was created with left coronary artery ligated in male Sprague Dawley rats. The model rats were ran-domly divided into sham, model, TG low and high dose groups. TG groups were injected into abdominal cavity with TG(20 mg·kg-1·d-1, 40 mg·kg-1· d-1 ) . Sham and model groups were injected with e-qual-volume normal saline. On the 35th day post-opera-tion, heart function was examined by color doppler ul-trasoundination. HE, Masson and immunohistochemis-trical staining were used to observe the histopathologi-cal changes of myocardium and micro vessel density. The level of vascular endothelial growth factor( VEGF) and basic fibroblast growth factor( bFGF) mRNA were detected by real-time PCR. Results Compared with the model group, high and low dose TG obviously de-creased the left ventricular end diastolic dimension, the left ventricular end-systolic dimension, the left ventric-ular end-diastolic volume and the left ventricular end-systolic volume(P<0. 05 and P<0. 01), and signifi-cantly increased the ejection fraction and the fractional shortening ( P <0. 01 ) . The histopathological changes of myocardium on myocardial infarction and fibrosis were dramatically reduced by TG. But ventricular wall was thicker. Two dose TG remarkably increased the expressions of VEGF and bFGF mRNA and micro ves-sel density of compositive CD31 + cells in ischemic myocardial tissue and around of infarct area(P<0. 01, P <0 . 0 5 ) . Conclusions TG could improve the car-diac function of acute myocardial infarction rat. The mechanism may be related to the upregulation of VEGF and bFGF gene expression, the promotion angiogene-sis, then the improvement of blood supply in myocardi-al infarction area.
6.Liponeurocytoma in petrous bone and supratentorial region of cerebellum: report of a case.
Limei QU ; Zhenyu PAN ; Hongxi MA ; Yinping WANG
Chinese Journal of Pathology 2015;44(11):809-810
Bone Neoplasms
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pathology
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Cerebellum
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pathology
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Humans
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Neurocytoma
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pathology
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Petrous Bone
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pathology
7.Efficacy of targeted monitoring and intervention on multidrug-resistant organisms in intensive care units
Yanan CHEN ; Jing LIU ; Limei TIAN ; Shanshan LIU ; Liye PAN ; Ting LIU ; Huiyi WU
Chinese Journal of Infection Control 2017;16(1):58-61,65
Objective To understand the status of infection with multidrug-resistant organisms (MDROs) in intensive care units(ICUs),and evaluate the intervention efficacy of targeted monitoring.Methods Prospective study was adopted,patients who were admitted to ICUs in 2014-2015 were selected (January-December 2014 was as preintervention stage,January-December 2015 was as intervention stage),trend of MDRO infection before and after intervention were compared and analyzed.Results Before and after intervention,297 and 217 strains of MDROs were isolated respectively,except carbapenem-resistant Pseudomonasaeruginosa (CRPA),the isolated strains of carbapenem-resistant Acinetobacterbaunannii (CRAB),carbapenem-resistant Enterobacteriaceae (CRE),methicillin-resistant Staphylococcus aureus(MRSA),and vancomycin-resistant Enterococcus (VRE) declined after intervention.MDRO infection rate declined from 7.17 % before intervention to 3.88% after intervention,infection rate of CRAB and CRE after intervention were both lower than before intervention (both P<0.05);MDRO infection rates in general ICU and internal medicine ICU increased from 8.75% and 7.84‰ before intervention to 4.39‰ and 2.28% after intervention,respectively (both P<0.05).After taking comprehensive intervention measures,compliance to prevention and control measures,such as ordering rate of doctor's advice on contact isolation for 24 hours,hand hygiene,health care workers' awareness all enhanced significantly(all P<0.05).Conclusion Targeted monitoring and intervention measures can reduce isolation rate of MDROs in ICUs.
8.The effect of shenfu injection on immune function in severe trauma patients
Limei LEI ; Xiang LAN ; Yan KAO ; Hao ZHENG ; Jie PAN ; Dingding HUANG
Chinese Journal of Primary Medicine and Pharmacy 2015;(19):2928-2930,2931
Objective To investigate the effect of shenfu injection on immune function in severe trauma patients.Methods 60 severe trauma patients were divided into the control group (n =30)and shefu group (n =30) by random number table.Other 30 cases were chosen as the health control group at the same phase.All patients were received conventional treatments,however,patients of the shenfu group were additionally received the shenfu injection treatment in the early stage.The CD +3 ,CD +4 ,CD +8 cell,human leukocyte antigen (HLA -DR),interleukin -1(IL -1),interleukin -6(IL -6)were detected on 3rd and 7th day by double -antibody sandwich enzyme -linked immu-nosorbent assay (ELISA).Results Compared to the health control group,the IL -1,IL -6 in the control and shenfu group were significantly higher than the health control group(f=7.128,q =9.212,10.112,all P <0.05).The IL -1and IL -6 in the control and shenfu group were significantly increased on 3rd day (t =11.126,10.013,all P <0.05)and decreased on 7th day(t =17.121,14.213,all P <0.05).The IL -1 and IL -6 in shenfu group were sig-nificantly lower than that of the control group(χ2 =4.113,10.117,all P <0.05).The CD +3 ,CD +4 ,CD +8 ,HLA -DR and CD +3 /CD +8 rate in the control and shenfu group were significantly lower than the health control group(f=11.071, q =10.229,12.032,all P <0.05).On 3rd day,the CD +3 ,CD +4 ,CD +8 ,HLA -DR and CD +3 /CD +8 rate in shenfu group were significantly increased(t =10.013,P <0.05).On 7th day,CD +3 ,CD +4 ,CD +8 ,HLA -DR and CD +3 /CD +8 rate in the control and shenfu group were both increased(t =11.126,15.932,all P <0.05).And the CD +3 ,CD +4 ,CD +8 ,HLA -DR and CD +3 /CD +8 rate in shenfu group were significantly higher than the control group(χ2 =3.771,P <0.05).Conclusion Shenfu injection can regulate immune function in severe trauma and improve clinical treatment.
9.Extraction of total RNA and cloning of sgDHAR gene from Siraitia grosvenorii.
Rongchang WEI ; Huan ZHAO ; Xiaojun MA ; Ke MI ; Changming MO ; Limei PAN ; Longhua BAI ; Qi TANG
Acta Pharmaceutica Sinica 2014;49(1):115-23
Abstract: Total RNA was isolated from Siraitia grosvenorii fruit by the method of modified Trizol, according to S. grosvenorii fruit characteristics of rich phenols, polysaccharide, oil and proteins. The OD260/280, OD260/230, RNA integrity (RIN) and yield of the total RNA with this method were 2.01, 2.02, 9.50 and 260 mirog.g-1, respectively. The open reading frame (ORF) of dehydroascorbate reductase (DHAR), named as SgDHAR, was cloned by rapid amplification of cDNA ends (RACE) and RT-PCR method from S. grosvenorii. The GenBank accession number for this gene is KC907731. The SgDHAR gene contains a full-length cDNA of 1,252 bp including ORF of 819 bp and encodes a predicted protein of 272 amino acids. The molecular mass is 30.217 7 kD and the isoelectric point is 8.76. Homology comparison showed that it shared 87% nucleotide sequence homology with Cucumis sativus. Expression patterns using qRT-PCR analysis showed that SgDHAR was mainly expressed in fruit and stem, followed by flower, and was lowest in root, while the expression level was 6.83 times in triploid. T than that in diploid. Therefore, SgDHAR gene may be involved in abortion of triploid seedless S. grosvenorii.
10.Isolation, Screening, and Identification of Actinomycetes with Antifungaland Enzyme Activity Assays against Colletotrichum dematium ofSarcandra glabra
Lisha SONG ; Ni JIANG ; Shugen WEI ; Zuzai LAN ; Limei PAN
Mycobiology 2020;48(1):37-43
A serious leaf disease caused by Colletotrichum dematium was found during the cultivationof Sarcandra glabra in Jingxi, Rong’an, and Donglan Counties in Guangxi Province, whichinflicted huge losses to plant productivity. Biological control gradually became an effectivecontrol method for plant pathogens. Many studies showed that the application of actinomycetesin biological control has been effective. Therefore, it may be of great significance tostudy the application of actinomycetes on controlling the diseases caused by S. glabra.Strains of antifungal actinomycetes capable of inhibiting C. dematium were identified, isolatedand screened from healthy plants tissues and the rhizospheres in soils containing S.glabra. In this study, 15 actinomycetes strains were isolated and among these, strains JT-2F,DT-3F, and JJ-3F, appeared to show antagonistic effects against anthracnose of S. glabra.The strains JT-2F and DT-3F were isolated from soil, while JJ-3F was isolated from plantstems. The antagonism rate of strain JT-2F was 86.75%, which was the highest value amongthe three strains. Additionally, the JT-2F strain also had the strongest antagonistic activitywhen the antagonistic activities were tested against seven plant pathogens. Strain JT-2F isable to produce proteases and cellulase to degrade the protein and cellulose componentsof cell walls of C. dematium, respectively. This results in mycelia damage which leads toinhibition of the growth of C. dematium. Strain JT-2F was identified as Streptomyces tsukiyonensisbased on morphological traits and 16S rDNA sequence analysis.