1.Study on animal models for food allergy
International Journal of Pediatrics 2012;39(1):68-70
Animal models play an important role in study on food allergy.This paper reviews the potential benefits and pitfalls of different animal models,including animal species and strains,test proteins,routes of exposure,the incorpration of adjuvants and evaluting indictor of allergic models,which can provide theoretical basis for evaluting the potential allergenicity of novel proteins and preventing food allergic diseases.
2.Suppressing effects of MMPs inhibitor GM6001, MMP-2/9 inhibitor Ⅰ ,MMP-2/9 inhibitor Ⅱ on migration of human lens epithelial cells
Chinese Journal of Experimental Ophthalmology 2015;33(9):811-815
Background The primary pathologic mechanism of posterior capsular opacification (PCO) is proliferation,migration and epithelial-mesenchymal transformation of residuary lens epithelial cells (LECs) following cataract surgery.Matrix metalloproteinases (MMPs) play a role during the migration of LECs.Researches showed that GM6001,a broad inhibitor of MMPs,can arrest the migration of LECs,but as specific inhibitors of MMPs,the efficacy and safety of MMP-2/9 inhibitor Ⅰ and Ⅱ on LECs migration remain unclear.Objective This study was to determine and compare the inhibitory efficacy among GM6001,MMP-2/9 inhibitor Ⅰ and Ⅱ on human LECs and search the clinical medication to prevent PCO.Methods Human LECs were cultured and passaged in vitro,and the cells of 3-4 generation were incubated in 6-well plates.Then the cells of 70% confluent monolayer were cultured in DMEM without fetal bovine serum for 12 hours.GM6001,MMP-2/9 inhibitor Ⅰ and Ⅱ at different concentrations (0.25,0.50,1.00,2.00,4.00,8.00,16.00,32.00,64.00,128.00 μmol/L) were added into the culture medium for 24 hours separately,and regularly cultured cells served as the control group.A bare area was made by a 200 μl sterile spear on the cell layer,and the migrated distance and inhibitory rate were calculated.The second or third generation of cells were incubated in 96-well plates at a density of 5×105/ml (200 μl/well).GM6001 (128.00 μmol/L),MMP-2/9 inhibitor Ⅰ (64.00 μmol/L) and Ⅱ (32.00 μmol/L) were added into the culture medium for 24 hours,and the cell viability was assayed by using MTT assay.Results Cultured cells grew well with irregular arrangement and presented the polygon in shape.The migrated distance was gradually reduced as the increase of concentrations of GM6001,MMP-2/9 inhibitor Ⅰ and Ⅱ,showing significant differences among the various concentration groups (GM6001:F=248.647,P<0.05;MMP-2/9 inhibitor Ⅰ:F=357.125,P<0.05;MP2/9 inhibitor Ⅱ:F=396.374,P< 0.05).The cell migrated distance in the control group was set to 1,the relative migrated distances were 0.478 ± 0.091,0.294±0.088 and 0.191 ±0.081 in the GM6001 group,MMP-2/9 inhibitor Ⅰ group and MMP-2/9 inhibitor Ⅱ group at the concentrations of 32.00 μmol/L,respectively,showing a significant difference among the groups (F =116.031,P<0.01),and cell migrated distance was obviously shorter in the MMP-2/9 inhibitor Ⅱ group than that in the GM6001 group or MMP-2/9 inhibitor Ⅰ group (all at P<0.01).The A values were 0.607±0.016,0.567±0.015,0.583±0.010 and 0.595 ±0.0138 in the control group,GM6001 group (128.00 μmol/L),MMP-2/9 inhibitor Ⅰ group (64.00 μmol/L) and MMP-2/9 inhibitor Ⅱ group (32.00 μmol/L),respectively,without significant difference among the groups (F=1.403,P>0.05).Conclusions GM6001,MMP-2/9 inhibitor Ⅰ and Ⅱ reduce the mobility of human LECs effectively but do not affect the viability of the cells in vitro.MMP-2/9 inhibitor Ⅱ appears to be most dominant in inhibiting migration of human LECs.
3.Expressions of apoptosis associated factors and adhesion molecules in acute rejection of rat cardiac allografts
Feng LIU ; Yulan LIU ; Junhong SUN
Chinese Journal of Organ Transplantation 2003;0(06):-
Objective To study the effects of apoptosis associated factors Fas/Fas, Bcl-2/Bax, adhesion molecules including ICAM-1, VCAM-1, PECAM-1, P-selectin, L-selectin and E-selectin in rat heart allograft immune response at different time points. Methods Rat acute cardiac allograft rejection model (PVG heart to DA rats) were established. Heart samples were collected on the day 1, 3, 5, 7 postoperatively ( n =3 in each group). Apoptosis was monitored by TUNEL. Fas, FasL, ICAM-1 and Selectin was examined by means of immunohistochemistry. The expression of Bcl/Bax mRNA, VCAM-1 mRNA, PECAM-1 mRNA was detected by means of situ-hybridization. Results In acute cardiac allograft rejection group, the expression of apoptosis, Fas/FasL, Bax, ICAM-1, VCAM-1 and PECAM-1 was increased. Apoptosis, Fas/FasL and Bax were mostly detected in cardiomycotes and ICAM-1, VCAM-1 and PECAM-1 were mostly detected in vascular endothelium and infiltrated cells. There was no expression of apoptosis associated factors and adhesion molecules in normal rat heart.Conclusion The high expression of apoptosis, Fas/FasL, Bax, ICAM-1,VCAM-1, PECAM-1 may be related with cardiac allograft rejection.
4.Expression of Fas system and apoptosis in rat allografted liver
Yulan LIU ; Feng LIU ; Junhong SUN
Chinese Journal of General Surgery 1993;0(02):-
Objective To investigate the effects of apoptosis, Fas/FasL and Bax/Bcl-2 in rat liver allograft immune response. MethodsDifferent rat allograft models were established:(1)Acute liver allograft rejection group: PVG liver to Lewis rats; (2) Liver allograft tolerance group: PVG liver→DA rats; (3) F1(P/D)→D group: F1(PVG-DA) liver to DA rats; (4)HLTx(P/2→D)group: PVG/2 liver→DA rats. Liver samples were collected on days 1, 3, 5, 7 postoperatively (each n=3). Apoptosis was monitored by TUNEL(terminal deoxynucleotidyl transferase(TdT) mediated dUTP nick-end-labeling). The expressions of Fas/FasL and Bcl-2/Bax mRNA were examined by means of immunohistochemistry and in situ-hybridization respectively. Results Compared with liver allograft tolerance group, in acute liver allograft rejection group, the apoptosis cell numbers increased in hepatocytes, FasL expression was high in infiltrated cells in portal tract areas and low in hepatocytes, and the expression of Fas and Bax mRNA increased. The expression of molecules detected in F1(P/D)→D group was similar with those in acute liver allograft rejection. The expression of molecules detected in HLTx(P/2→D)group was similar with those in liver allograft tolerance group. On the fifth day postoperatively, the apoptosis index in portal tract areas had a positive correlation with the expression of FasL in hepatocyte and a negative correlation with the expression of Fas and Bax. Conclusions The high expression of FasL in infiltrated cells of portal tract areas, Fas and Bax in hepatocytes may be involved in rejection response. The high expression of apoptosis in infiltrated cells of portal areas, FasL in hepatocytes may relate with the establishment of tolerance.
5.Preliminary inquiry on relationship between blood pressure variability and target-organ damage
Junhong ZHOU ; Yanfeng FENG ; Sumei LIU
Chinese Journal of Interventional Cardiology 1996;0(04):-
Ambulatory blood pressure monitor (ABPM) was done on 103 hypertensive patients according to clinical stages and left ventricular measure (LVM) was randomly done on a portion of the patients. The results showed that the variability of stage-Ⅱ-group's 24 hour systolic pressure and stage-Ⅲ-group's systolic and diastolic pressures was significantly higher than that of stage-Ⅰ-group (P
6.The Factor Relateted to the Prognosis in HFRS Patients with ARF Oliguresis stay.
Yanhong LIU ; Junhong WANG ; Tao CHEN
Journal of Medical Research 2006;0(09):-
Objective To investigate the factor relateted to the prognosis in HFRS patients with ARF oliguresis stage and the applied chance of hemodiaglysis in HFRS patients. Methods 46 cases of HFRS patients with ARF, with hemodiaglysis applied hospitalized in the department of infectious disease of Luohe centre hospital during 1996 to 2004 were retrospectively investigated and the factor related to the prognosis were analyzed,divided into three groups:successful group,delayed group, died group, and 14 sorts of clinical factors and common lab test items compared. Results Between live groups, the following factors are statistics value,such as the age, platelet,serum albution before HD,frequency of HD and the days of oliguresis stage, the difference of the other factors such as chance of HD,laboratory indicators and complication occuring rate of are no notable. The age of died group is in one′s prime, died in 3 hospitalized days, given HD only 1~2 times,so,chance of HD and days of oliguresis stage were without comparing value,but,the rate of acute complication is 100 percent,the difference is notable.Conclusions The influence of HD time is no notable,and, to prevent acute complication before HD is the key factor relateted to the prognosis.
7.Linkage Disequilibrium and Mutation Rate Analysis of Sixteen X-STR Loci
Li LI ; Junhong LIU ; Ruxin ZHU ; Yuan LIN
Journal of Forensic Medicine 2014;(6):437-440
Objective To assess the patterns oflinkage disequilibrium (LD ) of16 STR loci on X chrom o-som e and investigate the genetic stability. Methods G enom ic DNA samples extracted from blood stains from 500 unrelated individuals and 885 lineage m em bers from E astern C hinese H an population were genotyped through m ultiplex am plification using ID typerX-16 kit by our independent research followed by capillary electrophoresis. LD was assessed by PowerM arker v3.25 software and m utation rate of every locus was analyzed. Results LD were not found at the 16 X-STR loci. A llele m utations were observed at 10 loci. A m ong them , m utation rates of DXS6809 and DXS7132 were both up to 0.004 8. Conclusion W hen the 16 X-ST R loci included in ID typerX-16 kit were used for parentage testing, product princi-ples can be applied to calculate the likelihood, but m utation should be taken into consideration in the case that the genotypes do not m eet the genetic law(especially at DXS6809 and DXS7132).
8.Population Difference Analysis of Allele Frequencies of 24 Y-STR Loci
Ruxin ZHU ; Junhong LIU ; Qi ZHAO ; Yuan LIN ; Li LI
Journal of Forensic Medicine 2016;32(3):189-192
Objective To investigate the population genetic polymorphisms of 24 Y-STR loci in unrelat-ed individuals in Eastern Chinese Han population, and to compare the difference of Han group between Eastern China and Guangdong.Methods The population genetics of 24 Y-STR loci in 268 unrelated Han individuals from Eastern China were analyzed by GFS 24 Y-STR amplification kit. The allele fre-quencies in Eastern Chinese Han population were compared with the data in Guangdong Han population, and the difference analysis between two groups was performed.Results Among the 24 Y-STR loci of 268 unrelated Han individuals from Eastern China, 235 alleles and 267 haplotypes were observed. GD value ranged from 0.5649 to 0.9668. The difference between 12 loci(DYS622,DYS552,DYS443etal.) of Han population in Eastern China and in Guangdong was statistically significance.Conclusion GFS 24Y STR amplification system shows favorable polymorphisms, which can be used in patrilineal genetic relationship identification.
9.Preparation and in vitro Transdermal Penetration of Capsaicin Liposomes
Junhong XU ; Liuyi LIU ; Zhuoyan DENG ; Qingguo LI
China Pharmacist 2014;(10):1622-1626
Objective:To prepare capsaicin liposomes and study the feasibility by in vitro percutaneous penetration test. Meth-ods:Capsaicin liposomes were prepared by a film-ultrasonic method. The best formula was screened by orthogonal test based on single factor studies with the entrapment efficiency as the index. The improved Franz diffusion cells were used to study the transdermal pene-tration of capsaicin suspensions, capsaicin liposomes and capsaicin ointments, and the cumulative penetration amount through the isola-ted rat skin was compared. Results:The optimal formula of capsaicin liposomes were as follows:the ratio of capsaicin to lipids was 1∶5;the amount of Tween-80 was 100 mg;the amount of vitamin E was 50 mg;10 ml dichloromethane was used as the solvent. The op-timal pH value of the external phase was 6. 5 with the volume of 10 ml. The ultrasonic time was 8 min. The order of 12-hour cumulative penetration amount was capsaicin liposomes>capsaicin-PBS suspensions>capsaicin ointments. Capsaicin liposomes had the highest 12-hour cumulative penetration amount and showed obvious sustained-release property. Conclusion:Capsaicin liposomes have high en-trapment efficiency, good percutaneous penetration and sustained-release property, and the preparation technology is simple.
10.RNAi-silenced Survivin and hTERT gene inhibits proliferation and induces apoptosis of human colorectal carcinoma cell line SW 480
Lin XIAO ; Ping WANG ; Junhong DONG ; Shouxun WANG ; Shunmei LIU
Basic & Clinical Medicine 2015;(1):38-43
Objective To investigate the Influence of Survivin and hTERT gene on cell proliferation and apoptosis in human colorectal carcinoma cell line SW 480 and to find experiment evidence for gene therapy of colorectal carci-noma .Methods Plasmids carrying shRNAs targeting survivin and hTERT were designed , constructed and trans-fected into SW480 cells.SW480 cells were then divided into blank group , blank Plasmid control group , survivin RNAi group , hTERT RNAi group and Survivin-hTERT RNAi group .The telomerase activity was examined by TRAP-PCR-ELISA analysis 48h after hTERT-shRNA transfection.Survivin and hTERT mRNA and protein expres-sion was analyzed by RT-PCR and Western blot .Cell apoptosis , proliferation were measured by flow cytometry , CCK-8 assay.Results Telomerase activity of SW480 cells in Survivin-hTERT RNAi groups were significantly decreased compared with the blank group ( P<0.01 ) .The expression of survivin and hTERT mRNA, proteins in the Survivin-hTERT RNAi group was reduced by 82.8%and 73.6%( P<0.01 ) ,79.2%and 66.7%( P<0.01 ) respectively .The inhibitory rate of cell proliferation of Survivin-hTERT RNAi group was 43.6% ±0.1%( P <0.01 ) .The apoptosis rate was 39.2%±2.3%( P<0.01 ) in the Survivin-hTERT RNAi group .Conclusions The Survivin-hTERT RNAi group could significantly reduces the protein expression of survivin and hTERT mRNA, in-hibit cell proliferation and induces cell apoptosis in human colorectal carcinoma cell line SW 480 .