2.Effects of 3 carbohydrates on CAP10 promoter activity of Cryptococcus neoformans capsule-associated gene
Zhiyu ZHAO ; Hai WEN ; Sunxiao CHEN
Academic Journal of Second Military Medical University 1982;0(02):-
Objective:To study the effects of glucose,mannose and galactose on CAP10 promoter activity of Cryptococcus neoformans capsule-associated gene.Methods: Yeast cells were transfected with plasmid containing a 951 bp length of 5′ upstream flanking sequence of CAP10 coding region and a reporter gene,chloramphenicol acetyl transferase(CAT);the transfectants were treated with different concentrations(10,20,40,60,80 mg/ml) of glucose,mannose and galactose.CAT activity was assessed by ELISA method and CAT activities of different groups were compared.Results: Different concentrations of glucose and mannose had no obvious influence on CAT activity;different concentrations of galactose had obvious influence on CAT activity and the influence was positively dependent with its dose within the experimental concentration range.Conclusion: Glucose and mannose have no obvious effect on the activity of CAP10 promoter;galactose has obvious inductive effect on activity of CAP10 promoter,suggesting that CAP10 gene might be related with galactose metabolism.
3.Relationship between viral gene activity and cytopathogenesis in human cytomegalovirus infection
Yang ZHAO ; Dunjin CHEN ; Liangzhen WEN
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To explore the relationship between human cytomegalovirus(HCMV) gene activity and cytopathogenesis in HCMV infection. METHODS: HCMV echelin-infected cell model was set up in vitro by coincubating passage cultured HEL and HCMV with different titers (group A: 10-5; group B: 10-3; and group C: 10-1). FQ-PCR was performed to evaluate the number of HCMV DNA copies. MCP mRNA was measured by RT-PCR. Meanwhile the development of cytopathologic effects (CPE) was observed under microscope and ultrastructural changes determined by TEM.RESULTS: Compared with group B and C in high HCMV titer, group A with low viral titer showed low HCMV DNA load (P
4.Analysis and Clinical Implication of Ha-ras, p53 and RB Gene Mutations and Infection of Bladder Cancer with Papillomavirus
Wenli ZHAO ; Qinzhong CHEN ; Peie WEN ; At ET ;
Chinese Journal of Cancer Biotherapy 1996;0(04):-
To clarify the clinical significance of alterations of Ha-ras, p53 and RB gene as well as infection with HPV16,18 in human bladder cancer, we examined the state of Ha-ras, p53 and RB genes, sequences of HPV16/18, and their association with clinicopathological parameters in 39 cases of bladder cancer and 7 cases of normal tissue, using nonisotopic PCR-SSCP and dot blot. The overall incidences of Ha-ras, p53 and RB gene mutation and HPV infection in tumor were 61.5% ,36% ,30.8% and 15.4% , respectively. The HPV positive rate was negatively correlated with clinical stage and pathological classification. Rather, the mutations of Ha-ras and p53 gene were positively correlated with the above clinical parameters. The incidence of Ha-ras gene mutation in recurring tumors was significantly higher than that in primary ones. A negative correlation between HPV infection and p53 mutation was also found. The results suggest that the above molecular events and their interaction play important roles in the development of bladder cancer, and that they wonld be of practical assistance in the prognosis and monitoring of bladder cancer.
5.Construction and evaluation of non-specific targeting cationic polymer lipid liposomes.
Tao CHEN ; Rutao WANG ; Zhao WANG ; Tingli LU ; Wen ZHAO
Acta Pharmaceutica Sinica 2010;45(3):359-64
A new class of dendrimer polylysine poly(ethylene glycol)-lipid was designed and synthesized. The cationic polymer liposomes were prepared by the lipid film-extrusion and post-insertion two methods with these dendrimer polylysine poly(ethylene glycol)-lipid and other lipids. The structural properties of obtained cationic polymer liposomes were studied by laser light scattering and fluorescence spectrometer. It was demonstrated that the nano sized liposomes with different density of surface cationic charges can be prepared by either lipid film-extrusion or post-insertion methods, but post-insertion process did not affect drug loading, did not influence drug loading capacity and did not induce liposomal morphology and particle size. The density of positive charge does not affect the size and distribution of different liposomes size and distribution. It was the better choice for manufacture because post-insertion method did not cause early release of drug and size changes. Cell binding experiments show that cationic polymer liposomes, especially dendrimer polymer liposomes had higher local charge density, and therefore have dramatic non specific cell targeting ability.
8.Effects of basic fibroblast growth factor on expressions of collagen and fibronectin in normal skin and hypertrophic scar fibroblasts
Rui SONG ; Huining BIAN ; Wen LAI ; Dehua CHEN ; Kesen ZHAO
Chinese Journal of Tissue Engineering Research 2010;14(41):7784-7790
BACKGROUND:Basic fibroblast growth factor(bFGF)can promote production of collagen,fibronectin and matrix enzyme in healing wounds.However,dysregulation of this process,such as the abnormal coordination of cell proliferation,extracellular.matrix and neovasculadzation formation,or remodeling of the wound matrix will lead to excess accumulation of scar tissues.OBJECTIVE:To investigate effects of bFGF on normal skin wound healing and hypertrophic scar formation.METHODS:Normal and hypertrophic scar fibroblasts from tissue biopsies from 5 patients who underwent plastic surgery for repairing hypertrophic scars were isolated and cultured.The expressions of collagen,fibronectin and protein synthesis were detected by RT-PCR and ELISA.The mitochonddal membrane potential changes were measured using JC-1 staining and flow cytometry.Simultaneously,adenosine tdphosphate(ATP)levels were determined by chemiluminescence method.The effects of bFGF on these indexes of normal and hypertrophic scar fibroblasts were observed.RESULTS AND CONCLUSION:Hypertrophic scar fibroblasts become slower after being exposed to bFGF,which selectively inhibited type Ⅰ collagen production in hypertrophic scar fibroblasts(P<0.05).Although bFGF inhibited type]collagen production,it had no effect on type Ⅲ collagen expression in both normal and hypertrophic scar fibroblasts.However,fibronectin expression in the normal fibroblasts was up-reguleted after bFGF treatment(P<0.05).In addition,the mitochonddal membrane potential tended to depolarization,although no statistical difference,in hypertrophic scar fibroblasts treated with bFGF(10 or 100 μg/L).bFGF treatment increased the cellular ATP levels in the normal fibroblasts,while there were no significant alterations in the hypertrophic scar fibroblasts over a treatment of bFGF(10 or 100 μg/L,P<0.05).The results suggest that there are differential effects and mechanisms on the skin fibroblasts with bFGF treatment in normal wound healing and hypertrophic scar formation.
9.Purification, identification and characterization of the major allergen from Linnaeus
Qihua ZHAO ; Xizhong WANG ; Lijin CHEN ; Wen LI ; Yongping LIU
Chinese Journal of Immunology 1985;0(03):-
Objective:To purify and characterize the major antigenic components of sand swimming crab,and provide theoretical evidences for the research of standardization of allergenic vaccine.Methods:The crude extract of Linnaeus was prepared using classical method, and electrophoresis of SDS-PAGE was used to separate the proteins of each group. The allergen proteins of 26 crab were identified using Western blot. To distinguish between the primary allergen proteins and secondary allergen proteins, FPLC(Gel Chromatography and ion exchange chromatography) was used to filtrate and identify the allergen protein.Results:SDS-PAGE analysis revealed that sand swimming crab proteins were composed of at lease 9 discrete protein bands, which molecular weight ranging from 13 000 to 90 000. Major bands were of 20 900, 24 200, 27 100, 29 200, 33 700, 38 900, 48 700, 74 700, 89 100 respectively. Western blot assay indicated that the crude extract reacted with sera obtained from 26 crab allergenic subjects and contained 5 allergen bands altogether, and the bands of 74 400 and 48 700 were the major allergenic components. The positive rates of the two major allergen proteins were both 100%, indicating the allergenic components maintained the immunocompetence after yielding from chromatography.Conclusion:The 74 400 and 48 700 bands are the major allergens of sand swimming crab. The major allergen proteins can be purified by chromatography.
10.Treatment of complex renal stones in solitary kidneys under percutaneous nephrolithotomy combined with retrograde intrarenal surgery
Guohua ZENG ; Zhijian ZHAO ; Wen ZHONG ; Wenzhong CHEN ; Wenqi WU
Chinese Journal of Urology 2013;(4):268-271
Objective To evaluate the efficacy and advantages of the technique by combined PCNL and retrograde intrarenal surgery (RIRS) in a second stage to treat the complex renal stones in solitary kidney cases.Methods PCNL most with a single 18-24 F tract was performed first and RIRS was carried out at a second stage in solitary kidneys of 21 patients,of which congenital in 14.3% (3 cases),contralateral nephrectomy in 42.8% (9 cases),and functional solitry kidneys in 42.8% (9 cases).Of the 21 patients,the average age was 45 years with 15 males and 6 females.The stones were 8 multiple,6 partial staghorn,and 7 complete staghorn with a mean size of 4.6 (3.8-6.8) cm.Results Of the 21 PCNL cases,a single tract,double,triple tracts were established in 18 (85.7%),2 (9.5%),1 (4.8%) cases,respectively.Mean operation time of PCNL was 95 (45-175) min.After 1 day of PCNL,all case had residual stones with a mean size of 1.9 (1.0-3.5) cm.Two case occurred fever after PCNL and one case presented bleeding resolved by selective renal artery embolization.The mean operation time of RIRS was 72 (35-95) min.Stone-free rate after RIRS was 85.7% (18/21).The final stone free rate increased to 95.2% (20/21) after one case received a second-look PCNL and two cases accepted ESWL.Two cases occurred fever and steinstrasses after RIRS resolved by rigid ureteroscopy.At the 3 months follow-up,renal function became stable,improved and worse in 71.4% (n=15),23.8% (n=5),and 4.8% (n=l) of patients.Conclusions PCNL combined with RIRS could be an effective and safe option for complex stones in solitary kidneys with less bleeding,reduced tracts,minor complications and good renal function preservation.