OBJECTIVETo investigate the role and significance of T help cells 17(Th17) in pathogenesis of idiopathic thrombocytopenic purpura (ITP).

METHODSPeripheral blood samples from ITP patients and normal controls were examined for Th17 cell proportion by flow cytometry (FCM). Expression of IL-17, IL-23, IL-6 and TGF-β1 in hematoplasma was detected by ELISA. The mRNA expression level of IL-17 and RORγt in peripheral blood mononuclear cells (PBMNC) from patients with ITP and normal controls were measured by RT-PCR technique, and expression levels of pSTAT3 and RORγt proteins were analyzed by Western-blot.

RESULTSTh17 cells in peripheral blood from patients with ITP was greatly increased when compared with normal control group (P<0.05). Expressions of IL-17, IL-23, IL-6 and TGF-β1 in hematoplasma of ITP patients were all significantly higher than those in normal control group (all P<0.01). mRNA expression levels of IL-17 and RORγt in PBMNC from patients with ITP were much higher than those in normal controls (P<0.05). Protein expressions of pSTAT3 and RORγt in PBMNC of ITP patients were greatly increased as compared with those in control (P<0.05).

CONCLUSIONTh17 cell subgroup may play a role in incidence and development of ITP, which may participate in the pathogenesis of ITP by increasing Th17 cell proportion and altering the expression level of Th17-related cytokines as well as regulatory and transcriptional factors.

Objective To determine the effects of noncoding repressor of NFAT (NRON) overexpression or silencing on human umbilical vein endothelial cells (HUVECs)functions.Methods Stable HUVECs cell lines with NRON overexpression and short hairpin RNA (shRNA) interference were obtained.HUVECs,the empty vector pBABE-cell line and the empty vector pSuper-cell line served as controls.Cell proliferations of these cell lines were tested using MTS method,tube formation capacity and migration function were also examined.Results MTS experiments evidenced dose-dependent cells proliferations in all cell lines after 48 h culture with fetal bovine serum (HUVECs,r =0.91 ;pBABE empty vectors cell-line,r =0.88 ; NRON overexpression cell-line,r =0.89 ; pSuper empty vectors cell-line,r =0.95 ; shRNA infererence cell-line,r =0.97).Proliferation capacity was lower in NRON overexpressed HUVECs and was higher in NRON silencing HUVECs compared with pBABE empty vectors treated and normal HUVECs (all P ＜ 0.05).Tube formation and migration functions were also reduced in NRON overexpressed HUVECs [(8.33 ±0.12) roots,(1857 ±65) cells] and increased in shRNA infererence of NRON treated HUVECs [(36.00 ±0.51) roots,(6987 ±50) cells] compared with pBABE empty vectors treated HUVECs [(19.67 ± 1.42) roots,(4411 ± 117) cells],pSuper empty vectors treated HUVECs [(17.33 ± 2.93) roots,(3883 ± 109) cells] and normal HUVECs [(23.33 ± 3.01) roots,(5145 ± 72)cells,all P ＜ 0.05].Conclusion NRON overexpression could reduce and NRON silencing could increase proliferation,tube formation and migration capacities of HUVECs.

OBJECTIVESTo investigate the relationship between HBV (hepatitis B virus) polymerase gene 180 and 204 sites mutation and lamivudine resistance.

METHODSOne hundred forty-one patients with lamivudine resistance after lamivudine treatment and 60 chronic hepatitis B patients without lamivudine treatment were enrolled in this study. The serum HBV DNA mutation was analyzed by sequence detection via polymerase chain reaction (PCR). The sequences of the same patient were analyzed before and after lamivudine treatment.

RESULTSOne hundred and nine lamivudine resistance patients had HBV YMDD (tyrosine-methionine-aspartate-aspartate) mutation. Among them, 45 patients had rtL180M/M204V mutation (41.28%), 28 patients had rtL180M/M204I mutation (25.70%) and 36 patients had rtM204I mutation (33.02%). There were 6 patients with rtL180M mutation in 32 lamivudine resistance patients. Sixty chronic hepatitis patients without lamivudine treatment had no mutations.

CONCLUSIONSHBV mutations, which play an important role in lamivudine resistance usually locate at polymerase gene 204 site; 180 site mutation was also observed in these patients. Evaluation of the anti-virus therapy by surveillance of the two sites mutations is of importance.

China ; epidemiology ; DNA Mutational Analysis ; methods ; DNA, Viral ; genetics ; Drug Resistance, Viral ; Gene Products, pol ; genetics ; Genetic Predisposition to Disease ; epidemiology ; Genetic Testing ; methods ; Hepatitis B ; drug therapy ; genetics ; Hepatitis B virus ; drug effects ; enzymology ; genetics ; Humans ; Incidence ; Lamivudine ; therapeutic use ; Polymorphism, Genetic ; Risk Assessment ; methods ; Risk Factors

Objectives: To investigate the relationship between HBV (hepatitis B virus) polymerase gene 180 and 204 sites mutation and lamivudine resistance. Methods: One hundred forty-one patients with lamivudine resistance after lamivudine treatment and 60 chronic hepatitis B patients without lamivudine treatment were enrolled in this study. The serum HBV DNA mutation was analyzed by sequence detection via polymerase chain reaction (PCR). The sequences of the same patient were analyzed before and after lamivudine treatment. Results: One hundred and nine lamivudine resistance patients had HBV YMDD (tyrosine-methionine-aspartate-aspartate) mutation. Among them, 45 patients had rtL 180M/M204V mutation (41.28%), 28patients had rtL180M/M204I mutation (25.70%) and 36 patients had rtM204I mutation (33.02%). There were 6 patients with rtL180M mutation in 32 lamivudine resistance patients. Sixty chronic hepatitis patients without lamivudine treatment had no mutations. Conclusions: HBV mutations, which play an important role in lamivudine resistance usually locate at polymerase gene 204 site; 180 site mutation was also observed in these patients. Evaluation of the anti-virus therapy by surveillance of the two sites mutations is of importance.