Objective To evaluate the clinical application of HLA-peptides tetramer staining flow cytometry for determining HBV specific CD8+ cells.Methods HBV specific CD8+ cells in whole blood samples of chronic hepatitis B patients were stained with tetramer complex of HLA-A2 and HBV core 18-27 peptide and counted by flow cytometry. Results The medians of percentages of HBV specific CD8+ cells of total CD8+ cells were 0.20%(0.02%～2.04%) in 11 acute hepatitis B patients and 0.05%(

Objective To analyze CD127 expression on the memory CD8+ lymphocytes from hepatitis B e antigen (HBeAg) positive chronic hepatitis B (CHB) patients treated with peginterferon α-2a (Pegasys). Methods Thirty HBeAg positive CHB patients were treated with peginterferon α-2a 180 μg once a week for 48 weeks and followed up for 24 weeks. The memory CD8+ lymphocytes were characterized by expressing CD45RA and CD27 markers. CD127 expression on cell surface was measured by four-colour flow cytometry. The difference of mean values between groups was evaluated by Mann-Whitney test. Results The CD127 expression on CD8+ T lymphocytes was significantly lower in HBeAg positive CHB patients compared to healthy controls (Z=2.889, P＜0.05), which was negatively correlated with serum hepatitis B virus (HBV) DNA level and HBeAg titers. The CD127 expression increased along with the decrease of HBV DNA and HBeAg after 24-week, 48-week and 72-week treatment in patients showing good response to peginterferon α-2a, while CD127 expression didn't change markedly in non responders (Z24w = 1.954, Z48w = 2.789, Z72w = 2. 989; all P＜0. 05). Conclusion CD127 expression on memory CD8+ lymphocytes increases along with effective anti-HBV treatment in CHB patients, which can be used as a marker for evaluating the effectiveness of anti-viral treatment.

Objective:To explore the protective effect of zonisamide (ZNS) on oxygen-glucose deprivation (OGD) cell model of traumatic brain injury (TBI), and its underlying mechanism.Methods:Human neuroblastoma cells (SH-SY5Y) were cultured in vitro and divided into the control group, OGD group, and drug administration group (OGD+ZNS group) according to the random number table method. The OGD method was used to establish a TBI cell model. After modeling, the cell activity, the release of lactate dehydrogenase (LDH), and β-galactosidase staining were detected to evaluate cell function and senescence. Additionally, mitochondrial morphology and potential membrane changes were observed using Mito Tracker Red and JC-1 mitochondrial membrane potential staining. ATP concentration was measured, and protein was extracted from SH-SY5Y cells and then subjected to Western blot analysis to detect endoplasmic reticulum stress-related markers, including glucose-regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), protein disulfide isomerase (PDI), and β-actin.Results:The OGD group had a significantly lower cell survival rate compared to the control group ( P<0.01), while the OGD+ZNS group had a significant higher cell survival rate than the OGD group ( P<0.01). The LDH release rate was significantly higher in the OGD group than in the control group ( P<0.01), while the OGD+ZNS group had a significant lower LDH release rate compared to the OGD group ( P<0.01). Moreover, the cell staining results indicated that compared to the control and OGD+ZNS groups, the cells in the OGD group exhibited significant damage and senescence with darker staining while the mitochondrial staining results demonstrated a significant reduction in mitochondrial linear junctions and decreased mitochondrial activity in the OGD group compared to the control and OGD+ZNS groups. Compared to the control and OGD+ZNS groups, the OGD group exhibited a significant reduction in mitochondrial staining red fluorescence, a significant increase in green fluorescence, and a significant decrease in mitochondrial membrane potential. The OGD group demonstrated a significant decrease in ATP concentration compared to the control group ( P<0.01), whereas the OGD+ZNS group exhibited a significant higher ATP concentration compared to the OGD group ( P<0.01). Western blot analysis revealed significant upregulation of GRP78, CHOP, and PDI in the OGD group compared to the control group (all P<0.05), while in the OGD+ZNS group, the expression levels of these proteins were significantly downregulated compared to the OGD group (all P<0.05). Conclusions:Zonisamide can protect OGD TBI cell model by preserving mitochondrial activity and inhibiting endoplasmic reticulum stress.