Objective To compare the differences of temperature results between mercury thermometer and infrared thermometer. Methods The temperature of three parts was recorded respectively by mercury thermometer in axilla and infrared thermometer in forehead and earlobe on the same patient, totally 98 patients were recorded in ICU. Results There was statistical difference of three parts' temperature in 98 cases. It could be assumed that axilla temperature ＞ earlobe temperature ＞ forehead temperature;There was no statistical difference of three parts' temperature in 17 cases with temperature 38.0～38.9 ℃ by mercury thermometer, but the difference had clinical significance. It could be assumed that axilla temperature ＞ earlobe temperature ＞ forehead temperature; There was statistical difference of three parts' temperature in 30 cases with temperature at 37.0～37.9 ℃ by mercury thermometer. It could be assumed that axilla temperature ＞ earlobe temperature ＞ forehead temperature. There was no statistical difference of three parts'temperature in 51 cases with temperature at 35.0～36.9 ℃ by mercury thermometer. Conclusions Temperature of the patients with normal axilla temperature could be monitored by infrared thermometer instead of mercury thermometer,but it is not applicable to the patients with fever.

AIM: To construct the mammalian cell expression vector for enhanced green fluorescent protein (EGFP) and HLA-A*0201 fusion protein and analyze its expression and subcellular localization in the transfected K562 cells. METHODS: The HLA-A*0201 cDNA was cloned by RT-PCR and the gene was inserted into pEGFP-N1 to construct a vector for the fusion protein. The expression of the fusion protein in K562 cells transfected with the vector was evaluated by flow cytometry and its subcellular localization was investigated by confocal microscopy. RESULTS: The full-length encoding region of HLA-A*0201 cDNA was cloned from two HLA-A2 positive donors and the expression vector for the HLA-A*0201-EGFP fusion protein was constructed by PCR using a primer pair to introduce a Kozak sequence before ATG and the stop codon was deleted. Five hours after K562 cells was transfected with the vector, the expression percentages of HLA-A*0201 and EGFP were 25.12?2.26 and 27.37?3.59, respectively and no significant increase was observed after 24 h. The fusion protein was predominantly located on the membrane with low level distribution within the cells. In contrast, no HLA-A*0201 but only EGFP was detected in the empty vector transfected K562 cells and the EGFP was dispersed within the cells. CONCLUSIONS: The expression vector for HLA-A*0201-EGFP fusion protein was constructed and the fusion protein expressed in K562 cells was primarily distributed on the membrane. The results suggest that the transfected K562 cells are potential antigen-presenting cells.

Objective To analyze the indoor radon level and distribution characteristic in different geological background by studying the indoor radon level in three typical areas in Zhuhai City.Methods The region of investigation includes three districts:granite area in Zhuhai District,granite area in Doumen District and the Quaternary sedimentary area in Doumen District.Activated charcoal adsorption method was used to measure the indoor radon concentrations.In some sampling sites,solid state nuclear track detectors were used at the same time for the indoor radon measurement.Results The average indoor radon level included 80 buildings was (66.0 ± 49.8) Bq/m3 using activated charcoal adsorption method and the maximum value was 1078.5 Bq/m3.The results of 23 sampling sites show that the average indoor radon level using solid state nuclear track detectors was (88.8 ± 49.1) Bq/m3,and (69.5 ± 37.7) Bq/m3 by activated charcoal adsorption method.The indoor radon concentration was (73.6 ± 61.0),(87.5 ± 58.3) and (48.6 ± 22.6) Bq/m3 in granite area in Zhuhai District,granite area in Doumen District and the Quaternary sedimentary area in Doumen District,respectively.Conclusions The surface lithology of an area has a certain impact on the indoor radon level.The indoor radon level in granite area in Zhuhai District and Doumen District is apparently higher than that in the Quaternary sedimentary area in Doumen District.The study of indoor radon level and distribution characteristic should be discussed in combination with geological background of area.

AIM: To investigate the amount and patterns of expressing CD69 , IL-4 and IFN-? on TCRV?24 +NKT cells, and compare with that of CD3 +T cells from human peripheral blood in response to in vitro stimulation. METHODS: The whole blood was stained with three-color immunofluorescence directly or after cultured with PDB+ionomycin(Ion) for 6 h, then the mononuclear cells were separated by lysing red blood cells. The expression rates of CD69, IL-4 and IFN-? on TCRV?24 +NKT cells and CD3 +T cells were estimated by flow cytometer. RESULTS: As a proportion of mature T cells, the ratio of TCRV?24 + NKT cells to CD3 +T cells was about(1.34?0.42)%. The expression rates of CD69 on TCRV?24 + NKT cells and CD3 +T cells in response to PDB + Ion for 6 h were (96.71?1.33)% and (98.60?0.47)%, respectively, while the ratio were (11.47?2.86)% and (1.07?0.45)% in the unstimulated group, and there were significant difference between them. The expression rates of IL-4 and IFN-? on TCRV?24 +NKT cells stimulated with PDB+Ion for 6 h were (48.62?2.44)% and (46.65?8.91)%, respectively ,which were significantly higher than that of unstimulated group [(31.57?3.31)%, (13.45?6.29)%] and that of stimulated CD3 +T cells, though the expression rates on stimulated CD3 +T cells were significantly higher than that of unstimulated CD3 +T cells. CONCLUSIONS: There is small amount of NKT cells in adult human peripheral blood. The expression rates of IFN-? and IL-4 on these lymphocytes are higher than CD3 +T cells, suggesting that NKT cells are important immunomodulatory cells in special microvironments.

AIM: To investigate the effect of berberine (Ber) on the activation and proliferation of T lymphocytes and its mechanism of action. METHODS: Whole peripheral blood from normal subjects was stimulated with phytohemagglutinin (PHA) or phorbol ester (PDB) plus ionomycin (Ion) and the expression levels of CD69 and CD25 were evaluated with flow cytometry after the staining with appropriate fluorescent monoclonal antibody. The distribution of cell cycles was analyzed by propidium iodide staining and dead cells by 7-aminoactinomycin live staining. RESULTS: 100 ?mol/L and 50 ?mol/L of Ber had significant inhibition of the expression of CD69 on T cells stimulated with PDB plus Ion or PHA, while effect of 25 ?mol/L Ber was not significant. And as time of action extended, the extent of inhibition decreased. For the expression of CD25, Ber at the concentrations as above all exerted significant inhibitory effect in a dose-dependent manner. Moreover, Ber could block lymphocytes cell cycle progression from G_0/G_1 phase to S and G_2/M phase without phase specificity. Besides, live staining analysis revealed that Ber did not have significant cytotoxicity on lymphocytes. CONCLUSIONS: Ber significantly inhibits the expression of early and mid activation antigens of T cells and also blocks the progression of lymphocytes cell cycles. These results suggest that Ber exerts immunosuppression effect through inhibiting the activation and proliferation of T cells.

AIM: To investigate the details of age-related changes of T lymphocytes in order to seek for sensitive biomarker for immunosenesence. METHODS: Heparin anticoagulated venous blood was collected freshly from young (20-35 years) and elderly (50-75 years) volunteers and three or four color immunofluorescence staining was performed. The nucleated cells were acquired and the phenotypes of T lymphocyte subpopulations were analyzed with flow cytometry. RESULTS: There was no significant difference in percentages of pan-T (CD3 +), helper T (CD4 +) and cytotoxic (CD8 +) T subsets between young and elderly, whereas the density of CD3 molecule (MFI) on T cells in elderly group decreased significantly. It was also found that the rates of CD44 + and CD62L + T cell subsets in young group did not have statistical difference from elderly. However,the rates of CD95 + pan-T, helper T and cytotoxic T subsets of elderly group were all markedly higher than that in young group. CONCLUSIONS: The relative rates of T cell and its subsets displayed no age-related changes while the density of CD3 was down-regulated during aging in these groups investigated. Moreover, the expression percentage of CD95 (Fas) on T cells increased as aging, suggesting that it is a potential biomarker for evaluating immunosenescence.

AIM: Peripheral blood mononuclear cells (PBMC) were cultured in vitro to study the effect of gossypol, a polyphenolic antifertility agent, on the activation of normal human T cells. METHODS: Double fluorescent staining together with flow cytometry was adopted to analyze the influence of gossypol on expression of the early activation antigen CD69 on T-lymphocytes under stimulation of mitogen or phorbol ester. RESULTS: Analysis of T cell activation in vitro revealed that preincubation of PBMC with 100 μmol/L gossypol could completely inhibit the expression of early activation marker CD69 on CD3+ T cells in response to 10　mg/L PHA, and block T cell activation by 10-7 mol/L PDB as well. The suppression of CD69 expression was dose-dependent and IC50 of gossypol on PDB and PHA were (35.7±2.9) μmol/L and (32.8±1.5) μmol/L(眘), respectively. Besides, gossypol had similar inhibitory effect on CD69 expression of CD3- lymphocytes. However, it did not have any significant effect on T cell surface molecule CD3 down-regulation. CONCLUSION: Gossypol could inhibit T cell activation in vitro in response to polyclonal activators, both PHA and PDB, suggesting that its action site may be at PKC or its downstream and that gossypol possessed potential immuno-regulatory effect.

AIM: To study the effect of trichosanthin (TCS), a type I ribosome-inactivating protein, on the induction of apoptosis in human leukemic cell line HL-60 cells and the influence of cycloheximide (CHX) on TCS-induced apoptosis. METHODS: Flow cytometry together with fluorescent microscopy were adopted to investigate the apoptotic cell death in HL-60 cells treated with TCS. RESULTS: Flow cytometric analysis indicated that TCS was able to induce significant apoptosis in HL-60 cells. The rates of apoptotic cells in HL-60 cells treated with TCS (20 mg/L) for 48 h was 48.7%±2.3%(±s), which was significantly higher than that of control (6.3%±1.0%)(P＜0.05). Under the same condition, the rate of apoptosis caused by CHX (5 mg/L) was 65.3%±3.9%. TCS-induced apoptosis was further confirmed by fluorescent microscopy observation and DNA gel electrophoresis, in which typical nuclear morphological changes such as chromatin condensation, nuclear fragmentation, were observed in many of the cells treated with TCS， and DNA extracted from these cells displayed typical ladder pattern. Furthermore, the effect of TCS was significantly enhanced with the pretreatment of CHX (0.2 mg/L) which did not induce any significant apoptosis when used at 0.2 mg/L seperately. TCS-induced apoptosis was time- and dose-dependent. CONCLUSION: TCS was able to induce apoptosis in HL-60 cells, which was enhanced by CHX. It was suggested that TCS-induced apoptosis was independent of new protein synthesis.

AIM: Peripheral blood mononuclear cells (PBMC) were cultured in vitro to study the effect of gossypol, a polyphenolic antifertility agent, on the activation of normal human T cells. METHODS: Double fluorescent staining together with flow cytometry was adopted to analyze the influence of gossypol on expression of the early activation antigen CD69 on T-lymphocytes under stimulation of mitogen or phorbol ester. RESULTS: Analysis of T cell activation in vitro revealed that preincubation of PBMC with 100 ?mol/L gossypol could completely inhibit the expression of early activation marker CD69 on CD3 + T cells in response to 10 mg/L PHA, and block T cell activation by 10 -7 mol/L PDB as well. The suppression of CD69 expression was dose-dependent and IC 50 of gossypol on PDB and PHA were (35 7?2 9) ?mol/L and (32 8?1.5) ?mol/L( ?s ), respectively. Besides, gossypol had similar inhibitory effect on CD69 expression of CD3 - lymphocytes. However, it did not have any significant effect on T cell surface molecule CD3 down-regulation. CONCLUSION: Gossypol could inhibit T cell activation in vitro in response to polyclonal activators, both PHA and PDB, suggesting that its action site may be at PKC or its downstream and that gossypol possessed potential immuno-regulatory effect. [

Objective To investigate the status of cellular immunity from Th cell polarization in pa-tients with different courses of condyloma acuminatum(CA).Methods The isolated PBMC were polarized by PHA and self-plasma for72hours,then followed by two-color immunofluorescent staining with anti-CD4-PE and anti-CCR5-FITC,or with anti-CD4-FITC and anti-CCR3-PE.Finally the stained cells were analyzed by flow-cytometry.Results The percentages of Th1/Th2cells of the short-course group and long-course group were(25.82?2.22)%/(14.80?1.14)%and(12.20?1.37)%/(13.74?0.99)%,respectively;the differ-ences between normal control and two CA groups were significant(P