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Objective To explore the expression characteristic of T regulatory cell (Treg)/T help cell (Th)17 and the correlation to disease progression in chronic hepatitis B (CHB) patients undergoing antivirus treatment,and to explore their roles in pathogenesis of CHB.Methods A total of 53 patients with CHB (CHB group) and 21 healthy controls (healthy control group) were selected,CHB patients included mild 18 cases,moderate 16 cases and severe 19 cases.The expression of Treg and Th17 were detected by flow cytometry and compared.The levels of alanine aminotransferase (ALT),total bilirubin (TBIL),cholinesterase,albumin were measured by automatic biochemical machine and for correlation analysis.Results The levels ofTh17,Treg,Treg/Th17 were (2.13 ± 0.65)％,(2.99 ± 0.68)％ and (6.07 ± 1.18)％,(5.14 ± 0.96)％ and 2.86 ± 0.67,1.73 ± 0.45 in CHB group before and after treatment for 24 weeks and (3.59 ± 0.75)％,(4.02 ± 0.77)％,1.04 ± 0.34 in healthy control group.There were significant differences (F =14.78,10.12,17.19; P ＜ 0.01).The level of Th17 in mild,moderate and severe CHB was gradually decreased,but there was no significant difference (F =1.10,P =0.337).The level of Treg in mild,moderate and severe CHB patients was gradually increased,but there was no significant difference (F =0.54,P =0.585).The level of Treg/Th17 was gradually increased with aggravation (2.58 ± 0.59,2.76 ± 0.63,3.21 ± 0.71),and there was significant difference (F =3.15,P ＜ 0.01),the level of Treg/Th17 in severe CHB patients was significantly higher than that in mild,moderate CHB patients (P ＜0.05).The level of Treg/Th17 had significantly positive correlation with ALT,TBIL (r =0.272,P=0.000; r =0.226,P=0.000).Conclusion Treg/Th17 balance not only relates with the pathogenesis of CHB,but also with the related immune inflammatory of liver tissue.
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Objective To investigate the optimized parameters of dye (SYBR Green Ⅰ) realtime fluorescent polymerase chain reaction (RF-PCR) for detecting αβT lymphocyte clones in the peripheral blood and its application in monitoring specific T cell clone in the peripheral blood mononuclear cells (PBMC) from patients with chronic hepatitis B (CHB). Methods The total RNA was extracted from the PBMC of six healthy donors, and was reversely transcripted into cDNA. Then the cDNA was amplified using RF-PCR with the primers specific for T cell receptor β viable region (TCRBV) gene families as upstream primers and the primer for T cell receptor (TCR) β constant region (TCRBC) as downstream primer. The annealing temperature,concentration of primers and the total number of cycles were comparatively analyzed. The optimized PCR was performed to investigate the 24 TCRBV gene families from 12 patients with CHB, and the PCR products were monitored by melting curve analysis, and the clone expansion of peripheral blood T cell was detected by peak-motif of melting curve analysis. Results The optimized annealing temperature, final premier concentration,the number of cycles were 60.6 ℃, 0.5 μmol/L and 40 cycles, respectively. The begin temperature for melting curve analysis was better as 80 ℃ compared to 75 ℃. There was mono-peak on melting peak chart for TCRBV gene families in PBMC from patients with CHB, and PCR products of the single peak were determined as monoclonal T cell by sequencing. Conclusions The optimized reaction parameters of RF-PCR for monitoring 24 TCRBV gene families are determined. The melting peak chart could be used to monitor the clone expansion of the peripheral lymphocytes and to detect the clone-specific T cells in the peripheral blood from patients with CHB.
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Objective To investigate the role of Fas antigen,Fas ligand(FasL)and soluble Fas(sFas)in the pathogenesis of vitiligo.Methods The expression of Fas and FasL on peripheral blood lym-phocyte(PBLC)was detected by flow cytometry.Serum sFas was quantitated using a dual antibody sandwich enzyme-linked immunosorbant assay(ELISA).Results The expression of Fas and FasL was significantly de-creased in patients with vitiligo vulgaris(43.45%and58.40%)than those in normal controls(58.30%and64.65%)(P
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Objective To investigate the relationship between platelet-derived growth factor(PDGF)and granulocyte colony-stimulating factor(G-CSF)and psoriasis vulgaris.Methods Sera were taken from60patients with psoriasis vulgaris and42healthy controls.The serum levels of PDGF and G-CSF were measured by a dual antibody sandwich enzyme-linked immunosorbent assay(ELISA).Clinical severity of psoriasis vul-garis was assessed by psoriasis area and severity index(PASI)score.Results The serum PDGF levels were significantly higher in patients with psoriasis vulgaris than those in normal controls(P
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Objective To investigate the expression of costimulatory molecules CD80and CD86on peripheral blood lymphocytes(PBLC)in patients with systemic lupus erythematosus(SLE)and its significance.Methods The expression of CD80and CD86on PBLC was measured by flow cytometric assay in30patients with SLE and25normal controls.Results Expression of CD86on PBLC in patients with SLE was significantly lower than that of normal controls(P