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Objective To investigate the role of natural killer cells (NK cells) in patients with chronic hepatitis B (CHB). Methods Blood samples and liver biopsies from 108 CHB patients including 43 mild, 28 moderate and 37 severe CHB patients were collected. The levels of interferon (IFN) gamma in liver tissues and peripheral blood were detected by intracellular cytokine staining.The data were analyzed by t test. Results The levels of IFN gamma produced by NK cells in liver tissues and peripheral blood of patients with severe CHB were (8. 12±4. 26) % and (4. 59±2. 95) % ,respectively, which were significantly higher than those in patients with mild CHB [(3. 53±1. 56)%and (2. 26±0.61)%, respectively; t=3. 80 and t = 4. 30, respectively; both P0. 05). The levels of IFN gamma produced by NK cells in liver tissues and blood samples in severe phase of severe CHB were (10. 02 ± 4. 15)% and (6. 78 + 2. 91)%, respectively, those in recovery phase were (6. 13±2. 01)% and (3. 13± 1. 52)%, respectively (t = 1. 89 and t=1. 74, respectively; both P<0. 05]. Conclusion NK cells may play a pivotal role in the pathogenesis of severe CHB.
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Objective To establish the methodology of flow cytometry for detecting human cells in human/goat chimerisra.Methods Human hemopoietic stem/progenitor cells (CD+34 cells) or MIG-tranadueed-GFP CD+34 cells were transplanted into the peritoneal cavity of fetal goats in utero to obtain human/goat chimera modeL The peripheral blood cells from the chimeras were labeled with multiple mouse anti-human antibodies and the monoelonal antibodies that were specific for human but had not or only minimal cross-reaction with goat were screened as the primary antibodies for routine analysis in flow cytometry.Human cord blood was proportionally (25% ,50% ,75%,100%) added into the blood of the untransplanted goats and the cells were labeled with CD+34 monoclonal antibody.The region and size of the "gate" were chosen based on to the distribution of CD+34 cells or human cord blood.One human/goat chimera marked with GFP (MIG goat) was sacrificed and the substantial liver cells from its perfused liver were analyzed for the GFP+cells percentage and DNA contents by flow cytometry.Results CD7,CD15,CD38,CD45CD20CD34CD14and GPA monoclonal antibodies were chosen as the primary antibodies in rou tine detection by flow cytometry.The size and area of the "gate" were also defined.29.1% (29100/100 000 ) of the substantial liver cells from the MIG goat expressed GFP.DNA content analysis showed that the GFP+ cells obtained from the liver of MIG goat mainly manifested two peaks that were correspond to those of human.Conclusions Flow cytometry is rapid,simple and effective for the investigation of differentiation,homing and biological characteristics of stem cells in vivo.The selections of suitable surface antibodies and the "gate" are very important for detecting human cells accurately in the human/goat chimerism.
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Objective: To investigate the effects of the exogenous testosterone propionate on apoptosis of rat germ cells and the mechanisms thereof. Methods: Thirty 35-day-old male SD rats were randomly divided into experimental group and the control group. The rats in experimental group were injected (i.m.) testosterone propionate and the control group with an equal volume of saline. By using terminal deoxynueleotidy transferase nediated dUTP nick-end labeling (TUNEL), flow cytometry (FCM), radioimmunoassay (RIA) and electron microscopy, the quantity and quality of apoptosis of germ cells were evaluated. Results:(1) Compared with the control, the apoptotic number of rat germ cells was increased in the experimental group, especially the primary spermatocyte. The apoptotie rate was 11.3% detected by FCM in experimental group,while 3.6% in the control group (P ＜ 0.01). (2) The percentages of 1C were 21.8% in experimental group and 33.8% in control group (P ＜ 0.01).The percentages of 2C were 52.6% in experimental group and 37.1% in control group (P ＜ 0.01). (3) The serum levels of testosterone were (3 486.8±333.3) ng/L in experimental group and (846.9±167.5) ng/L in control group (P ＜ 0.01). The serum levels of follicle-stimulating hormone (FSH) were (2.5±0.8) IU/L in experimental group and (5.2±1.7) IU/L in control group (P ＜0.01). Conclusion: The exogenous testosterone propionate might induce apoptosis of germ cells by retroinhibition of the hypothalamie-pituitary-gonadal axis, thus having contraceptive effects.
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Scm in volume(42/60),multiple site involvement(44/60),blood type"O"(31/41),in comparison with those of survival group,and the difference was statistically significant.C-erbB-2,p16,p53,P-gp,CD_(44) and CD_(25)expression were not significantly different in these two groups. Conclusion The clinical stage, lymph node metastasis,lymphatic tumor emboli and/or neural involvement,infiltration depth,histological dif- ferentiation,tumor volume,involvement extension are important prognostic factors in patients with gastric can- cer,while the significance of cancer-related gene expression in gastric carcinomas needs to be studied further.