3.WT1 gene and glomerular diseases.
Jing-jing WANG ; Li-yan YE ; Zi-hua YU
Chinese Journal of Pediatrics 2009;47(3):233-237
Humans
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Kidney Diseases
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genetics
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Mutation
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WT1 Proteins
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genetics
5.Identification of the SjARG Novel Gene and Study on Its Protective Potential as a Vaccine
Zi LI ; Xinbing YU ; Zhongdao WU ; Xuchu HU
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(02):-
Objective To recognize and identify the arginase(ARG)gene of Schistosoma japonicum(Sj),and to study its protection potential as a vaccine.Methods The 5'-end of the ARG gene from the Sj cercariae cDNA library was amplified by nested-PCR and the sequence was identified by bioinformatics.The complete coding sequence(CDS)was cloned into pET30a(+)vector,and a recombinant SjARG protein(rSjARG)was expressed,purified and used to raise antibodies.ARG's activity as an enzyme was tested by ornithine-ninhydrin reaction.Western blotting was used to compare the immunologic characteristics of rSjARG with that of the native one in Sj adult worm.Indirect immunofluorescence assay was used to immunolocalize it.For evaluating the protection potential of rSjARG,mice were immunized by the recombinant protein and challenged by cercariae of S.japonicum.Results The CDS length of the SjARG novel gene was identified as 1095bp.rSjARG showed enzyme activity and the same immunologic characteristics with the native arginase in adult worm.SjARG located in the genital organ and gut of both sexes.The worm reduction rate and egg reduction rate in rSjARG group were 55.8% and 48.8% respectively,higher than that of the rSj26GST group(28.6% and 6.89% respectively).Conclusion SjARG gene was identified,which shows a higher protection than the Sj26GST.
7.Effect of Modified Hangqi Chifeng Decoction Containing Serum on the Expression of Col IV, MMP-2, and TIMP-2 in Glomerular Mesangial Cells Induced by LPS.
Hong-xia LIU ; Yu ZHANG ; Peng LI ; Yan-hong GAO ; Shuang LI ; Zi-kai YU
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(5):592-596
OBJECTIVETo explore the effect of Modified Hangqi Chifeng Decoction (MHCD) on levels of collagen type IV (Col IV), matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-2 (TIMP-2) in extracellular matrix (ECM) of glomerular mesangial cells (GMCs) in LPS induced mice.
METHODSNormal serum and telmisartan, high, medium, low dose MHCD containing serums were prepared by using serum pharmacology method. GMCs were cultured in vitro. The proliferation of mesangial cells were induced using LPS as stimulating factor. GMCs were divided into six groups, i.e., the normal group, the model group, the telmisartan group, high, medium and low dose MHCD groups. Col IV content in the supernatant of mesangial cells was detected using ELISA. Protein expressions of MMP-2 and TIMP-2 were detected using Western blot.
RESULTSCompared with the normal group, Col IV content obviously increased in the model group after 72-h LPS stimulation; protein expressions of MMP-2 and TIMP-2 were obviously up-regulated, and MMP-2/TIMP-2 ratio was down-regulated in the model group (P < 0.01). Compared with the model group, Col IV content obviously decreased in high and medium dose MHCD groups and the telmisartan group (P < 0.01); protein expressions of MMP-2 were obviously down-regulated in medium and low dose MHCD groups (P < 0.01, P < 0.05); the protein expression of TIMP-2 was obviously down-regulated in high, medium, low dose MHCD groups and the telmisartan group (P < 0.01). The pro- tein expression of TIMP-2 was obviously lower in the high dose MHCD group than in the low dose MHCD group (P < 0.01). MMP-2/TIMP-2 ratio was obviously up-regulated in the telmisartan group, high and medium dose MHCD groups (P < 0.01).
CONCLUSIONMHCD could regulate disordered MMP-2/TIMP-2 ratio in LPS induced ECM, inhibit excessive production of Col IV in ECM, promote the degradation of ECM, reduce the accumulation of ECM, thereby, delaying the process of glomerular sclerosis.
Animals ; Cells, Cultured ; Collagen Type IV ; metabolism ; Extracellular Matrix ; metabolism ; Kidney Glomerulus ; cytology ; Matrix Metalloproteinase 2 ; metabolism ; Mesangial Cells ; drug effects ; Mice ; RNA, Messenger ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism
8.Studies on drug resistance genes among 49 strains of extended-spectrum beta-lactamase-producing bacteria
Zi-Yong SUN ; Li-Ming CHEN ; Xu-Hui ZHU ; Li LI ; Bei ZHANG ; Yun-Song YU ;
Chinese Journal of Laboratory Medicine 2003;0(09):-
Objective To investigate the drug resistance genes of extended-spectrum beta- lactamase-producing bacteria in 49 strains.Methods Extended-spectrum ?-lactamase -producing strains were detected by the disc diffusion test.The techniques of polymerase chain reaction,sequence analysis, pulsed-field gel electrophoresis were used to analyze the genotype and homology of extended-spectrum ?- lactamase-producing strains.Results The incidence of ESBL-producing strains from E.coli,K pneumoniae,K oxytoca,was 20% in 2000,and 40% in 2003.Among the 49 ESBLs producers the most common genotype was CTX-M-14( n=33).The others were CTX-M-3,CTX-M-9,CTX-M-12,CTX-M-15, CTX-M-24 and SHVSa.Both two CTX-M subtypes,CTX-M-3 and CTX-M-14,were detected in one strain. However,4 ESBL-producing strains confirmed by phenotype remained untyped.The results showed that the ESBLs producers were not closely related,except for two strains of E.coli and two strains of K.pneumoniae which were homgenic respectively.Concolusions The incidence of ESBL-producing strains increases with years.The most common genotype of ESBLs is CTX-M.There is no evidence for epidemiologic spread of ESBL-producers by pulsed-field gel electrophoresis.
9.Diagnostic value of ovarian morphology by ultrasonography in pubertal polycystic ovary syndrome
Ya-Xiao CHEN ; Dong-Zi YANG ; Lin LI ; Xiao-Li CHEN ; Yu LI ;
Chinese Journal of Obstetrics and Gynecology 2000;0(09):-
0.05).Setting the threshold of MOV at 6.4 cm~3 offered the best compromise between sensitivity (84.8%)and specificity(87.5%),and setting the threshold of MaxOV at 8.6 cm~3 offered the best compromise between sensitivity(75.8%)and specificity(95.2%)and setting the threshold of MFN at 8 offered the best compromise between sensitivity(86.7%)and specificity(78.3%).Conclusions Ovarian morphology by ultrasonography yields satisfactory diagnostic accuracy for adolescent PCOS.Taking MOV≥ 6.4 cm~3 or MaxOV≥8.6 cm~3 or MFN≥8 as an ultraphonic criterion for pubertal PCOS offer the best compromise between sensitivity and specificity.
10.Effect of compound Chinese traditional medicine PC-SPES II in inhibiting proliferation of human prostate cancer cell LNCaP and on expressions of AR and PSA.
Bi-yan ZHANG ; Yu-feng LI ; Yun LAI ; Yun-sen LI ; Zi-jun CHEN
China Journal of Chinese Materia Medica 2015;40(5):950-956
To investigate the effect of compound Chinese traditional medicine PC-SPES II I in inhibiting proliferation of human prostate cancer cell LNCaP based on the androgen receptor (AR) signaling pathway. The effect of PC-SPES II on LNCaP cell proliferation was detected by MTT assay. According to the findings, at the mass concentration of 180-1 440 mg x L(-1), PC-SPES II significantly inhibited the proliferation of LNCaP cells; the IC50 of PC-SPES II at 24 h and 48 h were 311.48, 199.01 mg x L(-1), respectively. The flow Cytometry detection showed 240 mg x L(-1) PC-SPES II arrested cells in G2/M phase, and an obvious apoptotic peak appeared before G0/G1 peak and rose over time. Meanwhile, Hoechst 33258 staining revealed apoptotic cellular morphology. Annexin V-FITC/PI staining manifested an increase in apoptotic cell ratio at the PC-SPES II concentration of 480 mg x L(-1) in a dose dependent manner. The prostate specific antigen (PSA) secretion of LNCaP cells was tested by PSA ELISA kit. Besides, compared with 25 mg x L(-1) Bic, 480 mg x L(-1) PC-SPES II significantly reduced the cell secretion of PSA. The AR and PSA mRNA and protein expressions were detected by qRT-PCR and Western blot. According to the results, after the induction of LNCaP cells with synthetic androgen 25 μg x L(-1) R1881, 240-480 mg x L(-1) PC-SPES II notably down-regulated the AR and PSA mRNA and protein expressions and inhibited the translocation of AR from cytoplasm to nucleus. In summary, PC-SPES II significantly can inhibit the in vitro proliferation of LNCaP cells and arrest cell cycle arrest in G2/M phase. Its mechanism may be associated with the down-regulation of the AR and PSA expressions and the inhibition of AR nuclear translocation.
Antineoplastic Agents, Phytogenic
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pharmacology
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Cell Line, Tumor
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Cell Proliferation
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drug effects
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Drugs, Chinese Herbal
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pharmacology
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Humans
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Male
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Prostate-Specific Antigen
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genetics
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metabolism
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Prostatic Neoplasms
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drug therapy
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genetics
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metabolism
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physiopathology
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Receptors, Androgen
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genetics
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metabolism
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Signal Transduction
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drug effects