1.Expression of cytokines in CD8~+ T lymphocytes of the peripheral blood of patients with condyloma acuminatum
Dong-Xian LIU ; Wen JIANG ; Xing-Ping CHEN ; Ying-Ling CHEN ; Dong-Liang YANG ;
Chinese Journal of Dermatology 2003;0(11):-
Objective To detect the levels of cytokine IL-2,IL-12,IFN-?and IL-4 secreted by peripheral CD8~+ T lymphocytes in patients with condyloma acuminatum (CA).Methods Flow cytometry was employed to study the expression of cytokines IL-2,-12,INF-?and IL-4 in CD8~+ T lymphocytes in the peripheral blood of 60 patients with CA and 20 healthy controls.Results The percentage of CD8~+ T lym- phocytes producing IL-2,IL-12 and IFN-?were significantly lower in CA patients than that in healthy con- trols (P
3.Inhibitory effect of neutralizing interleukin-17 antibody on corneal allograft rejection
Xue-dong, CHEN ; Shi-yong, ZHAO ; Xian-ling, TANG ; Hong-yan, GE ; Ping, LIU
Chinese Journal of Experimental Ophthalmology 2012;(10):897-901
Background Interleukin-17 (IL-17)is a potent pro-inflammatory cytokine and plays a pathogenic role in autoimmune disease.It was confirmed that IL-17 is implicated in allograft rejection of many transplanted organs.Recent studies have foensed on the effect of IL-17 antagonists on allograft rejection.Objective This study aimed to investigate the inhibitory effect of anti-mouse IL-17 monoclonal antibody (mAb) on corneal allograft rejection.Methods Twenty-five 8 to 10-week-old C57BL/6 mice and 50 BALB/c mice were collected.Donor cornea grafts with 2 mm diameter from 25 C57BL/6 mice was transplanted to 50 eye of BALB/c mice to establish a model of corneal transplantation.The recipients were randomized into 2 groups,and neutralizing mouse IL-17antibody or isotype control antibody was intraperitoneally injected immediately after transplantation for experimental treatment,respectively.Allografts were scored clinically at appropriate time points after treatment based on Plskova criteria,and ≥5 was confirmed as rejection.Infiltrating cells in corneal graft were detected qualitatively and quantitatively by immunohistochemistry and reverse transcription-PCR separately.The cytokine levels of T helper type 1 (Th1),Th2,and Th17 in recipients' spleen wer(c) analyzcd by ELISA.The use of the animals followed the Statement of ARVO.Results Compared with the isotype control antibody group,the survival of grafts was improved in the IL-17mAb group(P<0.05).The levels of neutrophile granulocyte mRNA,CD4+ and CD8+ T lymphotes mRNA were 2.22±0.10,1.64±0.04 and 1.32±0.10 in the IL-17 mAb group,showing a significant decline in comparison with those of the isotype control antibody group(3.61 ±0.08,2.69±0.06 and 2.17±0.04) (P=0.000,0.000,0.000).Interferon-γ(IFN-γ),IL-12 p40 and IL-17 concentrations in recipients ' splenocytes were (529.80 ± 13.83) ng/L,(539.58 ±10.74) ng/L and(173.70±8.11)ng/L in the IL-17 mAb group,and thosc in the isotype control antibody group were (741.48± 10.51) ng/L,(1156.90 ± 69.93) ng/L and (366.13± 7.93) ng/L,with significant differences between them (P=0.000,0.001,0.000).Conclusions Neutralization IL-17 bioactivity inhibits mouse corneal allograft rejection to a certain extent.
4.Effects of Long-term Treatment with Hydrochlorothiazide Combined Spironolactone or Captopril on Left Ventricular Hypertrophy in Hypertensive Patients
Ai-Jun XING ; Dong-Xian LI ; Xin DU ; Shou-Ling WU ; Hai-Yan ZHAO ; Li-Ming LING ; Dong-Qing LI ; Zheng-Xin CAO ; Gui-Ling WANG ; Qing YU ;
Chinese Journal of Hypertension 2007;0(05):-
0.05);2)After 12,24,36 months' treatment,BP was decreased significantly in each group (P0.05).Conclusion Both combined spirono- lactone/HCTZ and captopril/HCTZ significantly reduced BP and LVMI or LVMI and the maguitude of reduction was further enhanced after prolonged treatment.
5.Mangiferin promotes uric acid excretion and kidney function improvement and modulates related renal transporters in hyperuricemic mice.
Qing-hua HU ; Xian ZHANG ; Yu WANG ; Ling-dong KONG
Acta Pharmaceutica Sinica 2010;45(10):1239-1246
The effects of mangiferin on uric acid excretion, kidney function and related renal transporters were investigated in hyperuricemic mice induced by potassium oxonate. Mice were divided into normal control group, and 5 hyperuricemic groups with model control, 50, 100, and 200 mg x kg(-1) mangiferin, and 5 mg x kg(-1) allopurinol. Mice were administered by gavage once daily with 250 mg x kg(-1) potassium oxonate for seven consecutive days to create the model. And 3 doses of mangiferin were orally initiated on the day 1 h after potassium oxonate was given, separately. Serum uric acid, creatinine and urea nitrogon levels, as well as urinary uric acid creatinine levels were measured. Mouse uromodulin (mUMOD) levels in serum, urine and kidney were determined by ELISA method. The mRNA and protein levels of related renal transporters were assayed by RT-PCR and Western blotting methods, respectively. Compared to model group, mangiferin significantly reduced serum uric acid, creatinine and urea nitrogon levels, increased 24 h uric acid and creatinine excretion, and fractional excretion of uric acid in hyperuricemic mice, exhibiting uric acid excretion enhancement and kidney function improvement. Mangiferin was found to down-regulate mRNA and protein levels of urate transporter 1 (mURAT1) and glucose transporter 9 (mGLUT9), as well as up-regulate organic anion transporter 1 (mOAT1) in the kidney of hyperuricemic mice. These findings suggested that mangiferin might enhance uric acid excretion and in turn reduce serum uric acid level through the decrease of uric acid reabsorption and the increase of uric acid secretion in hyperuricemic mice. Moreover, mangiferin remarkably up-regulated expression levels of renal organic cation and carnitine transporters (mOCT1, mOCT2, mOCTN1 and mOCTN2), increased urine mUMOD levels, as well as decreased serum and kidney mUMOD levels in hyperuricemic mice, which might be involved in mangiferin-mediated renal protective action.
Animals
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Blood Urea Nitrogen
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Carrier Proteins
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genetics
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metabolism
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Creatinine
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blood
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Glucose Transport Proteins, Facilitative
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genetics
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metabolism
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Hyperuricemia
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blood
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chemically induced
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physiopathology
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urine
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Kidney
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metabolism
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physiopathology
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Male
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Membrane Proteins
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genetics
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metabolism
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Mice
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Octamer Transcription Factor-1
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genetics
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metabolism
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Organic Anion Transport Protein 1
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genetics
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metabolism
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Organic Anion Transporters
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genetics
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metabolism
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Organic Cation Transport Proteins
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genetics
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metabolism
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Organic Cation Transporter 2
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Oxonic Acid
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Protective Agents
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pharmacology
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RNA, Messenger
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metabolism
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Random Allocation
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Solute Carrier Family 22 Member 5
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Uric Acid
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blood
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urine
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Uromodulin
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blood
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urine
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Xanthones
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pharmacology
6.Activation of JAK/STAT1 signal transduction pathway in different organs of MRL/lpr lupus mice
Jing DONG ; Qi-Xin WANG ; Shao-Chun WANG ; Xian-Fu MA ; Xiu-Ling JIA ; Qing-Rui YANG ; Yuan-Chao ZHANG ;
Chinese Journal of Rheumatology 2003;0(10):-
Objective To explore the activation and function of Janus protein-tyrosine kinase (JAK)/ signal transducer and activator of transcription (STAT1) signal transduction pathway in kidney,lung and brain of MRL/lpr mice.Methods MRL/lpr mice with systemic lupus erythematosus (SLE) were studied at the age of 12 weeks up.Non-SLE MRL/lpr mice were used as controls.We used phosphospecific antibodies to detect STAT1 activation in kidney,lung and brain by immunohistochemistry and Western blots.Gene expression of the STAT induced feedback inhibitors of cytokine signaling 1 (SOCS-1) was investigated by SYBR green I real-time reverse transcriptase polymerase chain reaction (PCR).Results Phosphorylation of STAT1 protein was markedly activated in these three organs,although renal and pulmonary STAT1 activation were much more evidently activated.SOCS-1 gene expression increased in all three organs,while renal SOCS-1 gene expres- sion increased less than lung and brain.Conclusion The activation of JAK/STATI signal transduction path- way may be pathogenic in the organ involvement and progression of SLE.The pathogenesis of lupus nephritis may also be associated with the down-regulation of SOCS-1 feedback inhibition.
7.Insulinllike growth factor 2 imprinting status and promoter usage in the placenta of macrosomia
Jin-Cui YAO ; Ya-Li HU ; Zhi-Qun WANG ; Yi-Min DAI ; Jing-Xian LING ; Xiao-Dong YE ;
Chinese Journal of Obstetrics and Gynecology 2001;0(05):-
Objective To study the mechanism of marcosomia by investigating insulin-like growth factor 2(IGF_2)imprinting status,expression level and the promoter usage in the placenta of macrosomia. Methods We selected heterozygous cases for Apa Ⅰ polymorphism in exon 9 of IGF_2 gene and then analyzed its imprinting status in 168 placentas of macrosomia and normal pregnancies.IGF_2 transcription levels and promoter usages in macrosomic and normal placenta were evaluated by using semi-quantitative RT- PCR assay.Results Thirty specimens of macrosomic placenta and 30 of normal placenta were identified as heterozygous for IGF_2.All of the heterozygous specimens showed maintenance of imprinting.The expression of placental IGF_2 mRNA(2.2?1.2)was significantly higher in macrosomia than that of normal weight group (1.6?0.6,P 0.05).Conclusion It is possible that over expression of IGF_2 in placenta contributes to macrosomia while the promoter usage and imprinting status are not associated with macrosomia.
8.Not Available.
Hong wu SHEN ; Hua guo CHEN ; Qing bo GOU ; Xian ling FAN ; Guo yan ZHANG ; Dong ZHAO
Journal of Forensic Medicine 2022;38(4):560-562
9.Solid-phase synthesis and biological characterization of S12A-HNTX-IV and R29A-HNTX-IV: two mutants of hainantoxin-IV.
Xia XU ; Xia XIONG ; Dong-Ling LI ; Yu-Cheng XIAO ; Xian-Chun WANG ; Song-Ping LIANG
Chinese Journal of Biotechnology 2005;21(1):92-96
Hainantoxin-IV (HNTX-IV) purified from the venom of the spider Selenocosmia hainana is a potent antagonist that acts on tetrodotoxin-sensitive (TrX-S) sodium channels. It is a 35-residue polypeptide and includes three disulfide bridges. In order to investigate the structure-function relationship of HNTX-IV, two mutants (S12A-HNTX-IV and R29A-HNTX-IV) of HNTX-TV in which Ser12 and Arg29 were replaced by Ala respectively, were synthesized by solid-phase Fmoc chemistry, followed by oxidative refolding of purified peptides under the optimal conditions. The synthetic mutants were analyzed by MALDI-TOF mass spectrometry, nuclear magnetic resonance spectroscopy (NMR) and electrophysiological experiments for molecular weight, conformation and physiological activity, respectively. The results show that the mutants and native HNTX-IV (nHNTX-IV) have almost identical three-dimensional structures. The bioactivity level of S12A-HNTX-IV is also about the same as that of nHNTX-IV, suggesting that Ser12 does not play any important role for the bioactivity of this toxin. The bioactivity of R29A-HNTX-IV is reduced by at last 155 times, indicating that Arg29 is a key residue relative to the bioactivity of HNTX-IV. It is presumed that the decrease in activity of R29A-HNTX-IV is due to the changes of the property in the binding site rather than the change in the basic conformation of the molecule.
Amino Acid Substitution
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Animals
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Mutation
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Sodium Channel Blockers
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Sodium Channels
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drug effects
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physiology
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Spider Venoms
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chemical synthesis
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genetics
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Structure-Activity Relationship
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Tetrodotoxin
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pharmacology
10.In vitro expression of hemophilia B gene mediated by lentivirus.
Dong-Mei YAN ; Kai-Lin XU ; Bing DU ; Ling-Yu ZENG ; Qun-Xian LU ; Xiu-Ying PAN
Chinese Journal of Hematology 2008;29(9):583-586
OBJECTIVETo construct a three plasmids lentiviral vector containing canine coagulation factor IX (cFIX) gene with ubiquinone promoter (PUB) and observe the expression of cFIX gene.
METHODSLentivirus was generated by transient three-plasmid transfection, namely, the VSV-G envelope expression cassette, the delta NRF packaging plasmid and the PTK 164 plasmid. Viral particles were used to infect the target cell, third passage mesenchymal stem cells (MSCs) and 293T cell respectively at MOI 3: 1. The cFIX activity was detected in cultured cells with one-stage clotting assay.
RESULTSThe MSCs were obtained in vitro. The lentivirus infected MSCs and 293T cells all expressed the active factor IX with the activity of (26.30 +/- 2.10)% and (19.70 +/- 1.53)%, respectively, which are significantly higher than that of control (1.00 +/- 0.05)%.
CONCLUSIONSThe lentiviral vector of three plasmids with ubiquinone promoter (PUB) was constructed and can transfect the MSCs and 293T cells.
Animals ; Bone Marrow Cells ; metabolism ; Cells, Cultured ; Dogs ; Factor IX ; genetics ; metabolism ; Genetic Vectors ; Hemophilia B ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; Plasmids ; genetics ; Transfection