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 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.

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.

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 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 .

Objective To investigate and compare the predictive value of 2 prediction scoring systems for diagnosis of coronary heart disease (CHD) in patients with suspected symptom, and provide information for diagnosis and therapy. Methods By prospectively studying a database of 272 patients with suspected CHD,the total score was calculated by prediction scoring system including PROCAM (The Prospective Cardiovascular Munster Study) and SCP(Suspected CHD Prediction Scoring System) with the data of clinic parameters and risk factors. All patients received coronary angiography and they were categorized into the CHD group (n =94) and non CHD group ( =178) according to the angiography result. The relationship between total scores and the SYNTAX score was evaluated by Spearman analysis and the value of the prediction scoring system was evaluated by the ROC (receiver operating characteristic) system. Results The score of PROCAM was from 6. 00 -77. 00(41. 76 ± 19. 91), and the score was significantly correlated with the extent and severity of coronary artery atherosclerosis (rs = 0. 420,P = 0. 023). The score of SCP was from 1. 00 - 13. 00(8. 64 ± 3. 42), and it was significantly correlated with the SYNTAX score (rs = 0. 482,P = 0. 016). The areas under ROC was 0. 770 (P = 0. 007) in PROCAM and that was 0. 733 (P = 0. 012) in SCP. Conclusions The nature and extent of coronary artery atherosclerosis could be evaluated by the scoring system effectively,which had a good correlation with CAG result.

Objective To express virus-like particles(VLP) of enterovirus 71 (EV71) in Han-senula polymorpha.Methods The coding sequences of P1 and 3CD genes of EV71 were optimized accord-ing to codon usage bias of Hansenula polymorpha for achieving high expression , and then cloned into the ex-pression vector PMV of Hansenula polymorpha .The recombinant expression vector PMV-P1-3CD was trans-formed into Hansenula polymorpha AU 0501 .The transformants were stably cultured in selective medium (Yeast Nitrogen Base) and screened for strains with positive P1 and 3CD genes by PCR.Then an induced cultivation on the recombinant strains were performed in a medium supplemented with methanol to a final concentration of 1.0%and the expressed products were analyzed by SDS-PAGE and Western blot assays to select high expression strains .The high expression strains were cultured in 30 L fermentor and its fermenta-tion products were analyzed by electronic microscope after purification .Results EV71 recombinant expres-sion strains were successfully constructed .The results of SDS-PAGE showed that the expressed products had obvious VP3, VP1, VP0 protein bands with molecular weights of 26×103, 33×103 and 35×103, respective-ly, which were consistent with the expected molecular weight of the fusion proteins .Western blot demonstra-ted that the expressed products could be specifically recognized by the polyclonal antibody against EV 71-VP1 at 33 ×10 3 , indicating its high immunoreactivity .ELISA confirmed that the expression level of EV 71 fermen-tation products was reached to 200 mg/L.Electronic microscope analysis showed that the VLP of recombi-nant EV71 were 24-30 nm in diameter with normal structure .Conclusion The virus-like particles of human enterovirus 71 are successfully expressed in Hansenula polymorpha , which provides a foundation for the fur-ther development of EV 71 VLP vaccine .

BACKGROUND:Previous tudies have shown that the anti-aging effects of stem cel s with lycopene are more significant, and can also significantly improve the aging body immune function.
OBJECTIVE:To investigate the anti-aging effects of human umbilical cord mesenchymal stem cel s combined with lycopene on the aging beagles.
METHODS:Sixteen aging beagles (6-7 years old) were randomly divided into two groups:aging control group and aging treatment group;young beagles (3-4 years old) were chosen as young control group. In the aging treatment group, human umbilical cord mesenchymal stem cel s combined with lycopene was given;while in the other two group, an equal amount of DMEM/F12 cel culture medium and sunflower oil was given. Each dog's general conditions were observed regularly during the whole progress. The changes of superoxide dismutase,
malondialdehyde, glutathione peroxidase in the serum were detected at regular time of the whole process, and the structure changes of each organ were observed at 24 weeks of treatment.
RESULTS AND CONCLUSION:(1) Before treatment, the levels of superoxide dismutase and glutathione peroxidase in the aging control and treatment groups were lower than those in the young control group (P<0.05), while the malondialdehyde content was higher than that in the young control group (P<0.05). However, there was no significant statistical difference between aging control and treatment groups (P>0.05). (2) For the aging treatment group at 24 weeks of treatment:the beagle fur became clearer and smoother, motility was strengthened, appetite became better;and the activity of superoxide dismutase in serum at 8 to 24 weeks of treatment increased significantly compared with before treatment (P<0.05), the activity of glutathione peroxidase significantly increased from 6 weeks (compared with treatment before, P<0.05), the malondialdehyde content decreased significantly from 4 weeks of treatment to the completion of the experiment (compared with before treatment, P<0.05). (3) After the experiment, the microscopic observation showed that compared with the aging control group, the tissues and organ structures of the aging treatment group were al clear, had no inflammatory infiltrates, no obvious necrosis and fibrosis lesions. These results were mainly consistent with the observations of young control group. The above results show that umbilical cord mesenchymal stem cel s combined with lycopene therapy on the natural aging beagles may enhance the activities of antioxidant enzymes, reduce malondialdehyde content, and their combination also can repair tissue structures and promote the functions, which has obvious anti-aging effects.