Objective To investigate the effects of different modes of mechanical ventilation on lung function during open heart surgery in children with pulmonary hypertension (PH). Methods Twenty-four patients with moderate PH aged 3 mon-12 yr 2mon (group M) and twenty-four patients with severe PH aged 3 mon-7yr 9mon (group S) were further divided randomly into 3 subgroups based on the modes of mechanical ventilation used: pressure-controlled ventilation(PCV) only and alveolar pressure (Palveo) was maintained at 0 cm H2O during CPB (group M1/S1 n = 8); PCV + PEEP (2 cm H2O) and Palveo was maintained at 2 cm H2O during CPB (group M2/S2 n = 8); and PCV+ PEEP (4 cm H2O) and Palveo was maintained at 4 cm H2O during CPB (group M3/S3 n=8). The patients were premedicated with atropine 0.01 mg ?kg -1 and midazolam 0. 1 mg?kg-1 . Anesthesia was induced with fentanyl 20?g?kg-1 and vecuronium 0. 1 mg?kg-1 and maintained with vecuronium infusion(70?g?kg-1?h-1 ) and inhalation of N2O-O2 and isoflurane. The children were mechanically ventilated after intubation and the ventilatory parameters were VT = 10-12 ml ?kg-1, F = 20 bpm and I : E = 1: 2. Lungs were lavaged and 2-3 ml of aspirate were taken for determination of surfactant. Total protein (TP), total phospholipid -(TPL), saturated phosphatidylcholine (Sat PC) and respiratory system compliance(Crs) was measured before skin incision( T0 ), before CPB (T,), at the end of CPB ( T2) and at the end of surgery(T3 ) . Results (1) Crs decreased significantly at T2 and T3 as compared with that at T0 and T1 in group M1/S1 and M2/S2 but in group M3/S3 there was no significant difference in Crs between T0, T1 and T2-3. (2) Sat PC/TPL and Sat PC/TP decreased significantly after CPB (T2, T3 ) in group M,/S, and M2/S2 but in group M3/S3 there was no significant difference in Sat PC/TP and Sat PC/TPL between T0-1 and T2-3. Conclusion PEEP 4 cm H2O can effectively prevent postoperative lung function from worsening in children with pulmonary hypertension undergoing open heart surgery with CPB.

OBJECTIVE:To establish the quality standard of Yanyan pills.METHODS: The constituents such as Lysimachia christinae,Radix et Rhizoma Glycyrrhizae and Radix Astragali were identified qualitatively by TLC,and the content of caffeotannic acid in L.christinae was determined by HPLC.RESULTS: The TLC spots were distinctive and well-isolated.The linear range of caffeotannic acid was 0.036～0.18 ?g(r=0.999 9)with an average recovery of 100.8%(RSD=0.67%,n=6).CONCLUSION: The established standard is applicable for the quality control of Yanyan pills.

Objective To understand biochemistry study situation in clinical medicine undergraduates and to improve biochemistry teaching quality.Methods Totally 120 clinical medicine undergraduates were enrolled randomly to receive investigation on problems related to biochemistry.Corresponding statistic analysis was made using softwares of Excel and SPSS 16.0 in view of the main problems.Results One hundred and twenty copies of survey questionnaires were finished.There were 114 effective copies and the effective rate was 95％ after checking on these questionnaires.61.4％ of the students neither liked nor disliked biochemistry course; 56.1％ of the students thought that learning biochemistry was difficult; students thought that chapter 4 sugar metabolism,chapter 5 lipid metabolism and chapter 6 biological oxidation to be very difficult.Only 11％ of the students did preview,further analysis found positive relation between preview and understanding (rj =0.288,P ＜ 0.01 ).Conclusion Clinical medicine undergraduates like biochemistry insufficiently with few doing preview.It suggests that teachers should pay attention to the cultivation and stimulation of students' interest in biochemistry course,improve teaching method for the difficult sections,strengthen experiment teaching and try combination and separation teaching mode.

Objective To evaluate the regulatory effect of OX40 co-stimulatory signal on the expression of Foxp3 in inductive regulatory T cells (iTreg) in vitro.Method CD4+ CD25+ naive T cells were isolated from C57BL/6 mouse lymphocyte suspension by MASC CD4+ CD25+ regulatory T cell isolation kit.Inductive Tregs were generated by stimulation of naive T cells in the presence of transforming growth factor beta (TGFβ1),anti-CD3,anti-CD28 and IL-2.The regulatory effect on iTregs was shown by use of OX40 stimulation monoclonal antibody (OX86) or control antibody.Using flow cytometric analysis (FACS),we examined the antibody-based identification of Tregs surface markers CD4 and CD25,along with the intracellular activation marker FoxP3.Results The ratio of CD4+ CD25+ nTregs isolated from mouse lymphatic node was (5.0 ± 0.4)％ vs.(71.8 ± 13.4)％ of TGFβ1-driven iTregs.The ratio of CD4+ CD25+ Tregs was (80.0 ± 1.6) ％ in OX40 stimulation McAb group vs.(86.0 ± 1.4)％ in control antibody group.Furthermore,the expression of Foxp3 was (59.2 ± 0.7) ％ in OX40 stimulation McAb group vs.(70.0 ± 0.8) ％ in control antibody group (P＜0.05).Conclusion TGFβ1-dependent protocol may induce the conversion of naive CD4+ T cells into CD25+ Foxp3+ iTregs.OX40 stimulation can down-regulate the expression of Foxp3 in CD4+ CD25 + iTreg significantly.Thus OX40 molecular may become an attractive target in Tregs-induced transplant tolerance.Further study should be performed to increase the suppressive activity of iTregs through blockade of OX40 signal.

Objective To compare the growth of three different cancer cell lines on chick chorioallantoic membrane (CAM), to select the best transplanted cancer cell line for establishing a transplanted tumor model and to observe the biological characteristics.Methods The human lung cancer cell line A549, human tongue cancer cell line TCA8113 and human liver cancer cell line QGY7703 were respectively inoculated into CAM at the 7th day of age.The chick embryo survival rate, tumor survival rate, tumor formation rate and induced angiogenesis were detected and the growth characteristics of the transplanted tumor model were observed.Results Compared with the groups inoculated with A549 cells and QGY7703 cells, the tumor formation rate of TCA8113 cells was the highest (P < 0.05), to be the best cancer cell line for transplanted tumor.The optimal inoculated number of cells was 8.0×106/chick embryo, the optimal growth period of the tumor was 4~8 d, and the best experiment time was 7 d after inoculation.Conclusion The TCA-CAM transplanted tumor model of tongue squamous cell cancer is successfully established for further study of the biological characteristics and mechanisms of tumor growth, angiogenesis, invasion and metastasis, and provide a good experimental animal model for anti-tumor drug screening.

Objective To study the effects caused by novel antipsychotics quetiapine and aripiprazole on concentrations of platelet 5-HT in patients with schizophrenia,and to explore the relationships among the change of platelet 5-HT concentrations, psychiatric symptoms and the sensitivity of therapy. Methods Sixty-eight patients with schizophrenia meeting International Classification of Diseases-10 (ICD-10) were enrolled in the study. They were divided into quetiapine group(n=34) and aripiprazole group(n=34) according to the sequence of admis-sion. The platelet 5-HT concentrations were measured with high performance liquid chromatography method(HPLC-ECD). The positive and negative syndrome scale (PANSS) was used to evaluate the psychopathology. Treatment emergent symptom scale (TESS) was used to evaluate the side effects. PANSS,TESS were used at pre-treatment,the end of 4th and 8th week of treatment. Results The platelet 5-HT concentrations in two groups were higher af-ter 8 weeks treatment,but had no statistically significant difference (P>0.05). In quetiapine group, the platelet 5-HT concentration was (458.89±233.36) ng/10~9 at the pre-treatment, and (554.31±313.22) ng/10~9 at the end of treatment (t=1.709, P=0.099). In aripiprazole group, the platelet 5-HT concentration was (409.83±149.32)ng/10~9 at the pretreatment, and (421.27±245.96)ng/10~9 at the end of treatment (t=0.321, P=0.819). There was no difference between two groups(P>0.05). Stepwise regression analysis showed the changing rate of platelet 5-HT concentrations was positively correlated with the dosage of quetiapine (r=0.385, P=0.039).It was positively correlated with the decreasing rate of PANSS in aripiprazole group(r=0.391, P=0.040). Con-dusion The changes of psychiatric symptoms and the dosages of antipsychotics maybe have intimate relationships with platelet 5-HT concentrations in the course of medication.

AIM : To construct and analyze eukaryotic expression plasmid inserted by Cx50 with V64G mutation through bioinformatics software.METHODS: The full coding domain sequence of Cx50 with V64G mutation was acquired from the blood of patients with cataract and was cloned into pcDNA3.1 /Amp (+).The constructed plasmid was identified with PCR , enzyme digestion and sequencing. The analysis of Cx50 with V64G mutation was performed with bioinformatics software.RESULTS : Cx50 with V64G mutation was successfully amplified and its eukaryotic expression plasmid was constructed. Valine-64 is well conserved in the first extracellular loop of connexin 50 in different species and also in different human α -type gap junctional proteins.CONCLUSION : The successive reconstruction and verification of eukaryotic expression plasmid containing Cx50 with V64G mutation established the foundation for further studying the mechanism of cataract.

Objective To investigate the effects of ischemic preconditioning on pneumocyte apoptosis and the expression of HSFT0 after lung isehemia-reperfusion(I/R) in rats and discuss its possible mechanism of extenu-ating ischemia-repedusion injury. Method Thirtysix male Sprague-Dawley rats were randomly divided into three groups [ sham operation(SO ) group, ischemia-teperfusion(L/R) group, and ischemic preconditioning(IP) group],twelve in each group. Lung croas-clamping was used to build the L/R model. In IP group, three cycles of 5-minute-ischemia + 5-minute-reperfusion were given to the pulmonary artery before the procedure. Sham operation rats had a thoracotomy only. Two hours(or five hours) reperfusion was given to both L/R and IP group. Tenninal-deoxynucleotidyl Transferase Mediated d-UTP Nick End Labeiing(TUNEL) was used to evaluate apoptosis. Expression of HSP/0 in lung was observed by immunohistochemical stain and image analysis. Index of quantitative assessment of histologic lung injury(IQA), wet to dry weight ratio(W/D) were measured. The pathological change of lung tissue was observed under both hght and electron microscopy. Statistical analysis was carried out by One-way Anova. Scheffe test was used for intragroup comparison. Results The apoptosis index and expression of HSP70、W/D,IQA of hng tissue in I/R group were higher than those in the sham operation group (P<0.01). Compared with the L/R group, the apoptosis index and expression of HSP70, W/D, IQA of lung tissue significantly decreased (P<0.01), the levels of expression of HSPTO increased significantly in IP group ( P<0.01 ). The pathological and ultrastructure change of lung tissue was better in IP group than those in I/R group. Condusions Ischemic preconditioning can extenuate lung I/R injury by the possible mechanism of increasing the expression of HSPT0 which inhibits the apoptosis during lung I/R injury.

Objective To knockout the murine Txnip gene using microinjection of transcription activator-like effector nuclease ( TALEN) mRNAs.Methods TALEN knockout site recognizing Txnip was designed by tools on line, then constructed the vectors and assayed its cleavage activity at cellular level.TALEN mRNA was transcribed in vitro and microinjected into C57BL/6J mouse zygotes.F0 mice were verified at DNA level with BamHI and TXNIP-knockout mice were obtained.Results We designed and constructed TALENs which recognized and cut the first exon of Txnip, and got four TXNIP knockout mice, among which two were frameshift mutation, demonstrating that the TXNIP-knockout mice were generated by TALEN technique.Conclusions Microinjection of in vitro transcribed TALEN mRNAs into murine zygotes is a highly effective and convenient way to develop TXNIP-knockout mouse model.

OBJECTIVE: To establish Caco-2 (a human colon adenocarcinoma cell line) cell monolayer model and the standard operation procedure for studying and assessing intestinal absorption of chemical components of traditional Chinese medicine. METHODS: Caco-2 cell monolayer model was established and evaluated by morphology feature using scanning electron microscope, inverted microscope and transepithelial electrical resistance (TEER) assay. Additionally, the model was further tested for the activity of alkaline phosphatase and the apparent permeability (Papp) of standard compounds, i.e. propranolol and atenolol, which were the control substances for high and poor transcellular transport marker, respectively. RESULTS: The integrality of cell monolayer, cell differentiation (reflected by expression of alkaline phosphatase and cell monolayer morphology), and the Papp value of standard compounds in the established Caco-2 cell model were satisfactory. All parameters tested were in good agreement with those reported in the literature. CONCLUSION: The established Caco-2 cell model can be used to study the intestinal absorption of orally administrated chemical components of traditional Chinese medicine and their absorption mechanism.