Objective To explore the dynamic changes of tracheal stem cells during tracheal regeneration after injury induced by fluorouracil(5-FU) in rats. Methods Extracorporeal tracheal injury(Wistar rats) was induced by 5-FU.ABCG2 expression in tracheal epithelium during the process of regeneration was analyzed by indirect immunofluorescence and Western blotting. Results 1.After treatment with 5-FU for 12 hours,the tracheal epithelium shed and there were ABCG2 positive cells among residual cells in G-0.Three hours after the removal of 5-FU,the tracheal rings were covered with flattened epithelial cells. ABCG2 positive cells increased slightly.Six-nine hours after the removal of 5FU,the epithelial cells changed into cuboidal,correspondingly,the ABCG2 positive cells increased obviously;At 24 hours after the removal of 5-FU,most of the epithelial cells were cuboidal and merged into pieces,but the ABCG2 positive cells decreased obviously;Until 48 hours after the removal of 5-FU,only a few positive cells could be seen with the pseudostratified mucociliary epithelium restored similar to its original mode.There were no detectable ABCG2 positive cells in normal tracheal epithelium.2.Western blotting analysis showed that there were different ABCG2 levels at different times after the removal of 5-FU which in accordance with the change of immunofluorescene.ABCG2 was minimally detected after treatment with 5-FU for 12 hours,reaching a maximal level at 6 hours after the removal of 5-FU,and then decreased over time.Until 48 hours after the removal of 5-FU,very low ABCG2 level was detected.Conclusion The expression of ABCG2 is correlated to the number of tracheal stem cells,suggesting that ABCG2 may serve as a marker for isolating stem cells from tracheal epithelium.

Objective To evaluate the methods of measuring platelet-associated antibody PAIgG/ PAIgA/ PAIgM by flow cytometry(FCM) and enzyme linked immunosorbent assay(ELISA), and to investigate their diagnostic value for patients with idiopathic thrombocytopenic purpura(ITP).Methods With FCM and ELISA respectively, PAIg on the platelet membrane and in plasma were measured in 19 patients with ITP and 17 healthy volunteers, and were compared with each other in order to find out whether there were differences in these groups.Results FCM and ELISA measurement in patients with ITP were significantly higher than those in control group (P0.05). Compared with the results of ELISA, the positive percentage of PAIgG measured by FCM(84%) in ITP patients was slightly higher than that by ELISA(79%).Conclusion The platelet-associated antibodies of PAIgG/ PAIgA/ PAIgM, especially PAIgG,are important for diagnosing ITP. FCM, in combination with ELISA, may improve the reliability and the positive percentage of detection in ITP patients.

BACKGROUND: Special anatomical location makes eye lens expose to stressful situation in a long term, so it needs persistent protection to resist stress. Heat shock proteins (HSPs) exert self-protective defensive effect under the stress.OBJECTIVE: To observe the changes in mRNA expression of HSPs in the lens under the condition of exogenous stress.DESIGN: Repeated measurement, controlled observation trial.SETTING: Department of Ophthalmology, First Hospital Affiliated to Jiamusi University.PARTICIPANTS: Human lens endothelial cells B3 (LEC-B3) were obtained from the Department of Ophthalmology, First Hospital Affiliated to China Medical University. Reverse transcription-polymerase chain reaction (RT-PCR) kit was purchased from Bao Biotechnology Company, Japan.METHODS: This experiment was carried out in the Laboratory of Congenital Malformation, Ministry of Public Health,China Medical University between September 2003 and September 2004. LEC-B3 of the exponential growth was used for the experiment. Thermal or oxidative stress on LEC-B3 cells were produced by 30-minute exposure at 45 ℃ or incubated for 30 minutes in DMEM supplemented with 50 mmol/L H2O2. Some stressful cells were allowed to recuperate after exposure to stress in normal culture medium at 37 ℃ for 0, 2, 4, 6, 16 and 24 hours, separately. The expressions of HSP27 and HSP70 in the LEC-B3 were detected by RT-PCR.MAIN OUTCOME MEASURES: The expressions of HSP27 and HSP70 in LEC-B3.RESULTS: HSPs existed in both physiological and stressful situation. The levels of HSPmRNA were increased 2 hours after both stresses. But the expressions of HSP27 and HSP70 got to the summit at different intervals from 2 hours to 6 hours in each group. Subsequently, they were decreased gradually in each group, but they all could maintain a high level at 16 hours.CONCLUSION: Stressful environment induces the over-expressions of HSP27 mRNA and HSP70 mRNA in LEC-B3. This may play an important protective role in lens epithelial cells in responding to cellular stress.

Objective: To investigate the association between the polymorphism of HLA-A, HLA-B genes and pre-eclampsia. Methods: HLA-A, HLA-B genotyping was performed by polymerase chain reaction sequence-specific primer (PCR-SSP) in 119 preeclampsia patients, 117 normal pregnant women and their neonates. Results: The study showed that 16 HLA-A and 39 HLA-B alleles were obtained in pre-eclamptic patients and normal pregnant women. 15 HLA-A and 37 HLA-B alleles were obtained in their neonates. No significant difference was found in maternal or neonatal HLA-A, HLA-B alleles be-tween pre-eclampsia group and control group (Pc>0. 05). The frequencies of HLA-A11, HLA-A24,HLA-B13, HLA-B14, HLA-B15, HLA-B52 maternal/fetus genetic assoications were significantly different between pre-eclampsia group and control group (P<0. 05). Conclusion: Some HLA-A, HLA-B maternal/fetus special bindings may be associated with the susceptibility or protective of pre-eclampsia.

Objective To study the killing effect of cytokine-induced killer cells (CIK cells) on human lung adenocarcinoma cell line (A549) and the lung adenocarcinoma' s radiation resistant cell line (A549RR).Methods Peripheral blood mononuclear cells (PBMC) of healthy volunteers were stimulated by different cytokines,and were induced into killer activity CIK cells.The phenotype of CIK cells were analyzed by flow cytometer.A549 and A549RR cell lines were cultured separately with the CIK cells.The absorbance value (A) of the cells was measured by CCK8,and the killing rates of all cells which were cultured for 24 and 48 hours with the CIK were calculated.Results The rate of CD3+ CD56+ cell was 45.8 ％ after culture for 14 d.The killing rates of CIK cells to lung adenocarcinoma A549 cells and its radiation resistant cells A549RR were increased with the rise of the ratio of effective cells to target (5∶1-40∶1) and the increasing of culturing time (all P ＜ 0.001).The killing effect of CIK to A549 and A549RR cells had no obvious difference in the same culturing time and the same ratio of effective cells to target(all P ＞ 0.05).Conclusion CIK cells have strong anti-tumor effect against lung adenocarcinoma and its radiation resistant cells with high clinical application value.

〔Abstract〕 The paper introduces the application of association rules in the library .With circulation data of freshmen in different ma-jors, in terms of different majors and grades of the college , it extracts partial records suitable for data mining , analyzes phases of data preparation and data mining by use of association rules , and provides technical support for in -depth conduction of reader services .

BACKGROUND: Special anatomical location makes eye lens expose to stressful situation in a long term. Whether the environmental stress can up-regulate the expression of heat shock proteins in human lens epithelial cells? Whether the synthesis increase occurs in the level of trenscdption or translation, remains unclear.OBJECTIVE: To observe the expression and location of heat shock protein 27 (HSP27) in human lens epithelial cells under the conditions of high temperature and oxidative stress, and to investigate the pathogenesis of the cataract.METHODS: Human lens epithelial cells cultured in vitro were exposed to heat (45 ℃) and oxidative stress (50 mmol/L H_2O_2) for 30 minutes, respectively, then allowed to recover normal conditions. At different intervals (0, 2, 4, 6, 16, 24 hours),immunocytochemistry and reverse transcription polymerase chain reaction were used to determine the expression and localization of HSP27.RESULTS AND CONCLUSION: HSP27 was shown to express in both physiological and stressful conditions. The expressions of HSP27 mRNA and protein ware remarkably increased at 2 hours following heat and oxidative stress, and reached the peak at 6 hours. HSP27 could maintain a high level for 16 hours. The stress-induced HSP27 protein positive particles transferred from the cytoplasm to the nucleus, and gradually shift back to the cytoplasm along time. It is proved that HSP27 exists in lens epithelial cells and can be increased after stress. The data suggested it may play an important protective role in lens epithelial cells in respond to cellular stress.

Objective To investigate the anti-inflammatory/immune modulatory effects of high-expressing membrane bound M-CSF-hematopoietic malignant cells on macrophages. Methods After coculturing RAW264.7 and murine macrophage cell line with Namalwa-M, a cell line stably expressing mM-CSF, and companing with Nainalwa-V, a cell line stably transfected with the empty vector as the control, flow cytometry was used to detect the expression of the marker of alternatively activated macrophages, CD206, and intracellular expression of IL-10, IL-12, IL-6 and TNF-α to study the immunophenotype of RAW264.7; phagocytic assays to investigate their functional activity in vitro. Results RAW264.7 cocultured with Namalwa-M consistently showed high-level expression of CD206, which indicated activities of these macrophage cells were increased. Furthermore, these RAW264.7 expressing high levels of IL-10, TNF-a and low levels of IL-12, IL-6, as determined by intracellular staining were suggested that the phagocytic activity was increased. Functionally, RAW264.7 cocultured with Namalwa-M showed a higher level of phagocytic activity. Conclusion Macrophage generated in vitro after cocultured with mM-CSF-expressing hematopoietic malignant cell line could be transformed into abnormal macrophage with an immunophenotype defined as IL-10-high, IL-12-low, IL-6-low and TNF-α-high.

Objective To investigate the relationship between suspected ankylosing spondylitis and HLA‐B27 antigen by detec‐ting the positive frequency of HLA‐B27antigen in 872 suspected AS patients ,and evaluate its clinical significance .Methods The positive frequency of HLA‐B27 on the T lymphocyte membrane were detected by flow eytometer in 872 suspected AS patients .Re‐sults Among the 872 suspected AS patients the ratio between male and female was 1 .8∶1 ,the positive rate of antigen HLA‐B27 was 27 .29% ,and the male and female patients′positive rates of HLA‐B27 antigen were 32 .50% and 17 .95% ,respectively (P<0 .05) .The male and female patients′ expression percentage of B27+ /B7- monoclonal antibody were 39 .16 ± 42 .79 and 20 .96 ± 33 .86 ,respectively(P<0 .05) .The male and female patients′mean fluorescence intension of B27+ /B7- monoclonal antibody were 5 .35 ± 5 .44 and 3 .35 ± 3 .87 ,respectively(P<0 .05) .Conclusion The patients with AS are strongly associated with HLA‐B27 an‐tigen .Detection of HLA‐B27 antigen expression intensity in suspected AS patients with FCM is helpful to diagnosis and differential diagnosis of AS .

Objective To investigate the effects of hypoxia on the expression of Stat3 and p-Stat3 ,and assessed the apoptosis ability of JAR cells in vitro .Methods JAR cells were cultured under hypoxic conditions .Western blot were used to determine the protein expression of Stat3 and p-Stat3 .Cellular apoptosis was monitored by flow cytometry analysis .Results Abnormal morpholo-gy changes in trophoblast cells under low oxygen conditions .After 48 h hypoxic treatment ,the protein of Stat3 and p-Stat3 were significantly decreased(P< 0 .05) ;however ,the level of apoptosis was significantly increased (P < 0 .05) .Conclusion Stat3 and p-Stat3 protein levels were decreased under hypoxia circumstance ,while the cell apoptosis ability was increased in JAR cells .