Objective　To investigate the potentiality of adhesion of S. pn to type Ⅱ pneumocyte (A549) in inducing tyrosine phosphorylation of A549 cell proteins and the function subcomponents involved in. Methods　Labelled S. pn with FITC for the observation of kinetic characters of adhesion in vitro; The tyrosine phosphorylation of A549 cell proteins induced by S. pn was examined by immunohistochemistry and ELISA. Results　The adhesion of S. pn to A549 cells was in a time-and dose-dependent fashion suggesting the specific interaction; Tyrosine phosphorylation induced by adhesion was dose-and time-dependent and was mediated by the surface proteins of S. pn. Conclusion　S. pn adheres to A549 cells and triggers the signal cascade in which the surface proteins of S. pn play a vital role.

As required to the standards of the model curricula and with the years of continuous teaching research and reforming,the clinical biochemistry curriculum has kept developing from the teaching staff,content of course,teaching method and means and the teaching condition etc.,thus forming model curricula with special features and notable results.

For the reform and development of clinical laboratory education, based on the previous course reform, Laboratory Medicine College of Chongqing Medical University has put the leading teachers'responsibility system of laboratory medicine course into practice. In the recent 3 years, the course is better organized.The students are more interested in the course and they communicate more with teachers than ever before. The effect of the course is obvious. The leading teachers' responsibility system in laboratory medicine course should be promoted.

Objective To evaluate the protective effects elicited by ClpP fusion protein in animal protection tests. Methods Pneumococcal antigens were purified from recombinant Escherichia coli express-ing CIpP cloned gene. 6- to 8-week-old female BALB/c mice were immunized intraperitoneally with either ClpP or PBS plus alum. Every mouse received three doses of 20 μg antigen or PBS in 100 mg of alum adju-vant at 14 d intervals. Sera were collected from mice and analyzed by ELISA 1 week after the third immuni-zation, Intraperitoneal-challenge experiments with 12 different serotypes of Streptococcus pneumoniae were carried out 2 weeks after the third immunization, and we compared their median survival times and survival rates respectively by Mann Whitney U test and Fisher exact test. Results ELISA analysis demonstrated high titer specific antibody responses to ClpP. The median survival times for mice immunized with ClpP pro-tein antigens in adjuvant were significantly longer than those for mice that received the adjuvants alone. Con-clusion A highly expressed recombinant ClpP protein has been successfully obtained and proved to exert the protection against invasive pneumococcal infection without relation of serotype, suggesting ClpP can be a promising candidate vaccine.

Objective To determine the vitamin D status in children with sepsis/severe sepsis in pediatric intensive care unit (PICU) in order to explor the association between vitamin D status and clinical outcomes,in turn to provide evidence for optimizing nutrition support.Methods It was a prospective,observational,multi-center study,carried out in patients with sepsis/severe sepsis from March 1,2013,to March 30,2014,in the PICUs of three tertiary-care children's hospitals.Total serum 25-hydroxy vitamin D [25 (OH) D] was measured by using an enzyme-linked immunosorbent assay at admission.The association of vitamin D status at admission with length of PICU length of stay,total hospital stay,in-hospital mortality,28-days mortality and costs were analyzed.Results A total of 194 patients includng 117 boys (60.3％)and 77 girls (39.7％) were enrolled.There were 96 patients with sepsis and 98 with severe sepsis.The mortality on discharge and 28 days were 6.7％ and 24.2％ respectively.The median vitamin D level was 9.79 ng/mL (5.32,18.46) at admission.Of them 77.8％ (151/194) had vitamin D deficiency and 50.5％ (98/194) had severe vitamin D deficiency.Patients with severe vitamin D deficiency,had higher mortality on discharge (P =0.011).Vitamin D status had no significant correlations with 28 days mortality,length of PICU stay,total hospital stay and costs.Conclusions More than three-quarters (77.8％) of children with sepsis/severe sepsis in PICUs had Vitamin D deficiency.Patients with severe vitamin D deficiency at admission had higher risk of mortality at discharge.

Objective To investigate the pediatric intensive care unit (PICU) in critically ill children with pediatric critical score (PCIS) and pediatric death risk score (PRISM),comparing its value.Methods A total of 100 cases of matching children with acute respiratory distress syndrome (ARDS) to stay in our hospital during 2007.07 ～2012.07 was retrospectively analyzed.Check PICU most serious disease in this group of children,living PICU most severely ill when diagnosed with acute respiratory distress syndrome (ARDS),when it was suffering from ARDS most serious line PCIS with PRISM score,and then using the Logistic multiple regression to analyze the scores of the two scoring ability as predicting the ARDS death in children with risk factors,and using two-factor analysis of variance of the two scoring methods by judging linear correlation existence.Results (1)PCIS with PRISM score showed a negative correlation (r=-0.6031,P ＜0.01),the linear regression equation y =-0.2389x +74.816 (P＜0.01).(2) There was no statistical difference between the ARDS death group and survival group (P ＞ 0.05) ; By PCIS PRISM score LOGISTIC regression analysis of the risk of death in children,there was no statistical difference (P ＞ 0.05).Conclusions PCIS and PRISM score cannot be the standards to forecast the death of the ARDS patients,but can provide an objective referring standard of ARDS's treatment.

Teachers introduced the means of the scientific thinking and scientific research prac-tice in the curriculum of proteomics in order to train the postgraduates to joint the theory and the practical research work in the curriculum course. For example, to cultivate students to truly understand the 2-DE protein separation technique, teachers used the various proteomic profile obtained from our research to points out its problems and limitations based on the classical 2-DE, then compared the improved 2-DE protein separation chromatography with the classical 2-DE graphic and analyzed what is the problem and how to solve them in the practical work. To improve the ability of scientific thinking for postgraduates , teachers took the tumor as the teaching cases and asked the postgraduates to search the references of can-cer associated with their professional , to put forward the scientific problems based on carefully reading these references, to design their research project and to seek the answer to their issue. These new teaching methods were found to be beneficial in training scientific talents of medical postgraduates.

Objective To investigate whether S. pn can provoke filamentous actin (F-actin) rearrangements in vitro , which will further lead to S. pn invasion of A549 and the relationship between PLC signaling molecule and. the invasion events. Methods Labelled F-actin with FITC-phalloidin, we observed F-actin rearrangements by S. pn adhesion of type Ⅱ pneumocytes ( A549). S. pn invasion of A549 cells was determined by pretreating A549 cells with Cytochalasin D . To investigate whether F-actin rearrangements can be blocked by PLC inhibitor, A549 cells were pretreated with PLC inhibitors U73122. Results Intact S. pn can promote F-actin rearrangements. Cytochalasin D is able to prevent S. pn invasion of A549 cells. Inhibitors of PLC signal transduction molecules block F-actin rearrangements dose dependently. Conclusion S. pn can provoke F-actin rearrangements through PLC signaling pathways, which will further lead to S. pn invasion of A549 cells.

Objective To investigate the inhibitory effect of Berberine hydrochloride alone or combined with antibiotics on the formation of Staphylococcus epidermidis biofilm and the expressions of relative genes in order to explore a new way to treat the infection induced by the bacteria.Methods Biofilm of Staphylococcus epidermidis was constructed in vitro first,and then Berberine hydrochloride alone or combined with norvancomycin,ciprofloxacin or erythromycin was used to affect the biofilm formation.Then the inhibition was detected by CRA,and the expressions of icaA and agr by the way of fluorescent quantitation RT-PCR.Results The effectiveness of Berberine hydrochloride on the expressions of icaA and agr in Staphylococcus epidermidis was not as better as the other antibiotics.When different-dosed berberine hydrochloride was combined with norvancomycin,ciprofloxacin or erythromycin to affect the biofilm,the inhibition on the expressions of icaA and agr showed a negatively dose-dependent fashion.Conclusion Berberine hydrochloride can inhibit the formation of the biofilm of Staphylococcus epidermidisthrough suppressing the relative genes,though not as good as the antibiotics,such as norvancomycin,ciprofloxacin and erythromycin.It should not be combined with these antibiotics because the effectiveness is antagonism.

Guided by the demands of society,we propose a new professional training objectives of medical laboratory technology,which is to cultivate comprehensive talents covering the entire industrial chain of medical laboratory technology with profound foundation,broad caliber,high quality and outstanding ability.According to the training objective,we build up 4321 talents training system and try to carry out preliminary practice and exploration on talent cultivation model from the following aspects,such as the construction of curriculum system,the reform of the teaching contents and methods,training of students' professional skills and entrepreneurial innovation spirit.