1.Genetic polymorphism and forensic application of five novel STR loci
Chinese Journal of Blood Transfusion 2001;0(06):-
0.85,and the exclusion probabilities ≥0.48.The independence test of 5 loci proved that there were no correlations between the two STRs in 5 pairs,which were LFG26 and LFG21,LFG20 and LFG24,LFG21 and LFG24,LFG24 and LFG26,LFG24 and LFG29.Therefore they could be used simultaneously.Conclusion The forensic study of 5 STRs on chromosome 21 revealed that LFG21,LFG24,LFG26 and LFG29 were good candidates as polymorphic markers for forensic DNA analysis.
3.The effects of CDK4 on proliferation of the cells in the epithelium with oral lichen planus
Zhou WEI ; Jin YAN ; Wu ZHIFEN
Journal of Practical Stomatology 1995;0(04):-
Objective: To investigate the cause and mechanism of oral lichen planus (OLP). Methods: The expression of cyclins dependent kinases (CDK4) in epithelial cells in the samples in 40 cases of OLP and in 10 cases of healthy controls was observed by immunohistochemical method. Results: The expression indexes of CDK4 in the epithelial basal layer and spinous layer in OLP were 0. 5637 ? 0. 0201 and 0. 5388? 0. 0120, those in the healthy control were 0. 4020? 0. 0155 and 0. 3480? 0. 0163, respectively(p
4.Development of a hydrogel microsphere delivery system for rhBMP2
Faming CHEN ; Zhifen WU ; Yan JIN
Journal of Practical Stomatology 1996;0(02):-
Objective: To design and synthesize a novel vector for rhBMP2 delivery system in tissue engineering. Methods:Dextran glycidol methacrylate(dex-GMA) was synthesized with dextran(dex) and glycidol methacrylate(GMA).Dex-GMA microspheres were prepared by suspension polymerization. The swelling behavior of the microspheres was evaluated by the swelling equilibrium parameter Q. The biodegradation properties of the dextran-based hydrogel microspheres were assessed by the surface morphology before and after biodegradation. Results:Microspheres of dex-GMA in the size(diameter) of 20 to 80 ?m with good configuration were produced. Q value of the microspheres with the diameter of 20-30 ?m was 10.9?3.3,that of those with 70-80 ?m 8.9?6.4.Stirring speed, span-80(emulsifer) quantity and the proportion of dex-GMA affected the size of the microspheres. rhBMP2 was enveloped into the microspheres. Complete degradation of the microspheres was observed during 20 to 40 days at 37 ℃ in normal saline. Conclusion: Dex-GMA hydrogel microspheres may be a release controlling system for rhBMP2 delivery.
5.Isolation and identification of advanced oxidation protein products
Yan SUN ; Xiongfei WU ; Xiyu JIN
Journal of Third Military Medical University 2003;0(10):-
Objective To isolate and identify advanced oxidation protein products from human serum albumin (AOPP-HSA), expecting to search for a method of preparing highly purified and bioactive AOPP-HSA. Methods AOPP-HSA crude products were prepared in vitro by exposing HSA to HOCl. AOPP-HSA was isolated by gel chromatography and ion-exchange HPLC. Its structural features and biological activities were characterized by UV and fluorescence spectrum, SDS-PAGE, and the experiment of TNF-? release from monocytes. Results The isolated protein was purified up to 99.4% and was dityrosine-containing protein cross-linking products with molecular weight of 700?10 3. It possessed the ability of triggering the considerable release of TNF-? from monocytes. Conclusion Highly purified and bioactive AOPP-HSA can be successfully prepared by above-mentioned two-step chromatography from AOPP-HSA crude products, which builds a basis for further study of AOPP.
6.Effect of Nourishing Qi,Activating Blood Circulation and Dispersing Phlegm Recipe on the Pathology of Chronic Obstructive Pulmonary Disease Rat Models
Cuiling FENG ; Yan JIN ; Hongli WU
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(12):-
Objective To establish chronic obstructive pulmonary disease(COPD)rat models,and to study the intervention of the formula of nourishing Qi,activating blood circulation and dispersing phlegm recipe on the morphology.Method To establish rat COPD models by intratracheal instillation of lipopolysaccharide and exposure to cigarette smoke.To instill intervention drug daily either the formula of nourishing Qi,activating blood circulation and dispersing phlegm recipe or the roxithromythin,starting on the 20th,30th and 40th day of the experiment respectively(the groups was named h 20,h 30,h 40,r 20, r 30 and r 40 for short),and to observe the effect on the morphology by means of collagen staining and image analyzer.Results The pathological changes and lung function in the model group were accorded with the human COPD.In drug intervention groups,airway inflammation and epithelial proliferation were alleviated to different degree compared to the model group.In model group,the collagen deposition was increased predominant type I collagen compared to the health comtrol group,and the deposition in the drug intervention groups were decreased compared to the model group,according to the Sirius redpolarizing microscopy morphometry method.The thickness of the airway wall in the model group was significantly increased compared to the health control group(P
7.The effect of human Slitrk1 gene on proliferation and differentiation of PC12 cells
Yanbin JIN ; Yan WU ; Xiaowen WANG
Medical Journal of Chinese People's Liberation Army 2001;0(11):-
Objective To study the effect of human Slitrk1 gene on proliferation and differentiation of PC12 cells. Methods The CDS sequence of Slitrk1 was amplified by PCR, and then cloned into pcDNA4 vector. The recombinant plasmid pcDNA4/Slitrk1 were transfected into rat PC12 cells by lipofectamine. The stable expression cell clones were screened by RT-PCR. MTT method was used to detect the proliferation rate of PC12 cells. The morphologic changes in PC12 cells were observed microscopically. Results The stable cell lines expressing pcDNA4 (pcDNA4/PC12) and pcDNA4/Slitrk1 (ST1/PC12) were established. Compared to pcDNA4/PC12 cells, the growth rate of ST1/PC12 cells was decreased. In addition, pcDNA4/PC12 cells tend to grow as well as the normal PC12 cells. However, most of the ST1/PC12 cells adhered to the plate with one or two neurites. Conclusion Over-expression of human Slitrk1 gene inhibited the proliferation of PC12 cells and promoted the outgrowth of neurites. It is suggested that human Slitrk1 gene may be involved in differentiation of PC12 cells.
8.Preparation and Evaluation of BMP_2 Carrying Gel Microspheres by Dextran-glycidyl Methacrylate
Faming CHEN ; Zhifen WU ; Guofang WANG ; Yan DU ; Yan JIN
China Pharmacy 2001;0(12):-
OBJRCTIVE:To study the feasibility and the preparation of bone morphogenetic protein-2(BMP 2 )carrying gel microspheres by dextran-glycidyl methacrylate(DEX-GMA)and to make an initial study on the drug-loading and drug releasing function of gel.METHODS:The orthogonal test was conducted with the reaction temperature,the addition of DEX-GMA and Span-80,the stirring speed and so on as investigation factors,the best preparation technics of BMP 2 -DEX-GMA gel microspheres was optimized and the finished products of which were given an initial determination.RE-SULTS:The BMP 2 -DEX-GMA gel microspheres preparation could be made by suspension polymerization,the optimized preparation technics including the reaction temperature was30℃,the addition of DEX-GMA and Span-80were0.6%and0.06%respectively,the stirring speed was300r/min,the BMP 2 -DEX-GMA gel microspheres from the above formulation take good shapes with the particle diameter at20?m~50?m,the envelopment ratio was(78.52?4.34)%,the quantity of drug-loading was(10.68?1.34)%,and the swelling ratio was85.9%,which has a good stability and redispersibility.CON-CLUSION:The preparation technics of gel microspheres drug-loading system is simple and the loading dosage is high,which can be used as a bioactive drugs carrier.
9.Predominant antigenic epitopes on Hap adhesin of nontypeable Haemophilus influenzae and their immunogenicity
Zhongxiu WU ; Rongshan WANG ; Yanying HUANG ; Hongxing JIN ; Jie YAN
Chinese Journal of Microbiology and Immunology 2017;37(8):594-600
Objective To investigate the distribution and sequence conservation of Hap adhensin encoding gene (hap) in clinical isolates of nontypeable Haemophilus influenzae (NTHi), to screen out and identify the predominant T-and B-cell (T-B) combined antigenic epitopes on Hap protein and to analyze their immunogenicity.Methods Sequence conservation of hap genes in NTHi strains and T-B combined antigenic epitopes were predicted using bioinformatic softwares.PCR was used to amplify the 156 bp segment at 5′-end and the 855 bp segment at 3′-end of hap gene (hap-5′-156 and hap-3′-855) and the amplified products were sequenced.Phage display systems of seven T-B combined antigenic epitopes located on the 55 aa segment at N-terminal and the 285 aa segment at C-terminal of Hap protein (Hap-N52 and Hap-C285) were constructed.Western blot assay and ELISA were performed to detect the antigenicity and immunoreactivity of different T-B combined epitopes displayed by recombinant phage PⅢ protein (rPⅢ).Results Hap protein encoded by the hap gene in NTHi was located on membrane surface.Sequences of the 156 bp segment at 5′-end and the 855 bp segment at 3′-end of hap genes extracted from different NTHi strains were relatively conservative, but many mutations were found in sequences at the middle regions of these hap genes.All of the 56 NTHi strains carried hap-5′-156 and hap-3′-855 segments and shared 92.3%-100% identities in nucleotide and amino acid sequences of these segements.Hap-N5-24 in the Hap-N52 segment as well as Hap-C4-27, Hap-C28-47, Hap-C114-129, Hap-C150-173, Hap-C200-227 and Hap-C241-267 in the Hap-C285 segment was predicted as the T-B combined antigenic epitope with a higher score and less mutations.Results of Western blot assay and ELISA confirmed that the rPⅢ-displayed Hap-C4-27 and Hap-C150-173 epitopes presented clear hybridization bands with NTHi antisera, and 96.9% (63/65) and 92.3% (60/65) of serum samples from children with NTHi infection were positive for antibodies against Hap-C4-27 and Hap-C150-173 epitopes, respectively.Conclusion The gene of hap is widely distributed in clinical isolates of NTHi.Moreover, sequences of the 156 pb segment at 5′-end and the 855 bp segment at 3′-end of hap gene are conservative.Hap-C4-27 and Hap-C150-173 are the predominant T-B combined antigenic epitopes on Hap protein, suggesting that they can be used as epitope candidates for developing multiple antigenic peptide vaccines against NTHi.
10.Selection of two types of rat strains for easily diabetic retinopathy
Wei, ZHOU ; Chunjie, JIN ; Zhaolian, MENG ; Jianguo, WU ; Hua, YAN
Chinese Journal of Experimental Ophthalmology 2014;32(1):32-35
Background Streptozotocin (STZ)-induced type 1 diabetic model is an acceptable model and is often used to the study of diabetic retinopathy (DR).However,the model is often established using retinal digest preparation in vitro in albino rats,and therefore it is difficult to evaluate the change of retinal vessels by fundus fluorescein angiography (FFA) in vivo.Recently,pigmented rats are being used as the DR model.But the study on the comparison between albino rat model and pigmented rat model is seldom.Objective This study was to observe and compare the manifestations of FFA and retinal digest preparation in early diabetic pigmented diabetic rat and albino diabetic rat,and to select the right model of DR using FFA.Methods Type 1 diabetic models were induced in 15 six-week-old health male BN rats and 15 six-week-old health male SD rats by the injection of STZ (55 mg/kg) via tail vein.The models were thought to be successful in the rats with the blood glucose level ≥ 16.7 mmol/L.The right eyes of the rats were extracted 6 weeks after injection of STZ.Lens were examined by slit lamp microscope.Retinal vascular changes were examined by fundus photography,FFA and periodic acid Schiff staining of retinal digest preparation.The manifestations of FFA and retinal digest preparation were contrasted between BN rats and SD rats.The number of eyes with lens opacification was compared by Chi-square test and the ratio of vascular endothelial cells and perithelial cells (E/P) was compared between BN rats and SD rats.The use and care of experimental animals complied the Statement of Ethic Committee of Tianjin Medicine University.Results Six weeks after injection of STZ,11 BN rats and 10SD rats were included in this study.The blood levels were (24.73±2.98) mmol/L and (22.36±3.65) mmol/L in BN rats and SD rats,respectively,without significant difference between the 2 types of rats (t =7.873,P>0.05).Lens opacification occurred in 6 BN rats and in 5 SD rats (P=0.717).FFA showed the clear retinal vascular under the brown background.Evident vessel disorder and fluorescence leakage like background stage of DR were seen in 9 eyes.However,in the SD rats,retinal vessel abnormality could not exhibited owing to the interference of choroid fluorescent light from choroidal vessels and vortex vein.Retinal digest preparation exhibited that unevenness of vessel diameter,decrease of perithelial cells and increase of endothelial cells in both types of rats.The E/P values were 11.50±3.68in the BN rats and 12.86±3.94 in the SD rats,without significant difference between them (t=9.785,P>0.05).Conclusions The abnormality of fundus vascular morphology can be better displayed in pigmented diabetic rats than albino rats by FFA in vivo.