Objective:To investigate the role of recombinant E.coli LLO/OVA on regulating the function of murine CD4+ CD25+ Treg cells.Methods:After E.coli LLO/OVA or E.coli OVA vaccination,the murine spleen CD4+ CD25+ Treg,CD4+ CD25- T and CD11c cells were collected respectively by magnetic beads sorting.The concentration of IL-10 in the supernatant of mix cocultured CD4+ CD25+ Treg and CD11c cells,and the suppression role of CD4+ CD25+ Treg cells on the proliferation of CD4+ CD25- T cells were determined.The percentage of OVA specific CD8+ T cells in mouse spleen was analyzed by flow cytometry.The number of metastatic tumor nodules in lungs of the mice transplanted with B16-OVA subcutaneouly was compared before and after dilition of CD4+ CD25+ Treg cells in mice.Results:Compared to E.coli OVA,E.coli LLO/OVA significantly downregulated IL-10 secretion of CD4+CD25+Treg cells and attenuated the suppressive effect of CD4+ CD25+ Treg on the proliferation of CD4+ CD25- T cells(P

OBJECTIVE:To improve the level of pharmaceutical experiment teaching for pharmacy major. METHODS:The ex-periment teaching of pharmacochemistry for pharmacy major in our school was taken as a pilot,and the teaching contents,teach-ing methods and appraisal system were explored. RESULTS:The teaching contents of“designing experiment”of pharmacochemis-try for pharmacy major in our school were added,multimedia teaching mode was introduced and scientific and comprehensive as-sessment mode was established. CONCLUSIONS:The reform of experiment teaching has cultivated the independent thinking and creative ability of students,improved enthusiasm and initiative,achieved good teaching quality and promoted the improvement of professional quality of teaching staff.

Objective To investigate the intelligence development status of the low birth weight infants and its influence factors. Methods The intelligence developments of 135 low birth weight infants aged 6-30 months were investigated using Bayley Scales of Infant Development, and the self-development influential factors were analyzed with questionnaire.Results The average of MDI was 86.76±18.95 in 135 infants,of which the detection rate of MDI<80 points accounted for 17.8%and the detection rate of PDI<80 points accounted for 29.6%;birth weight of infant,age,the degree of parents,culture and feeding way had significant effects on MDI in infants;age and birth weight had significant effects on PDI in infants. Conclusion The development level of infants is influenced by multi-factors. More attention should be paid to reduce premature and low birth weight infants,improve the degree of parents' culture, advocate breastfeeding and conduct early intervention in order to assure the intelligence development of infants.

Nature is abundant in protein scaffolds.By selecting suitable protein scaffold,display and screening methods,the rational and constrained random peptide library(RPL)can be constructed.Compared with the non-constrained RPL,it offered more opportunities for obtaining novel protein structures and more higher affinity ligands against the target molecules.At present,the protein scaffold constrained RPLs have been shown great potential in applications such as target selection,basic research,clinical diagnosis,medical therapy and so on.It is systematically introduced the structure bases,classification and construction of constrained RPL based on scaffolds,as well the recent great advances of application in selection against target molecules with S-S constrained scaffolds,antibodies,Zinc finger protein,Z domain,FN3 domain as important examples.

Objective To design and formulate a electronic preoperative preparation checklist for applying in the preoperative preparation,to reduce missing rate in preoperative preparation and transfer,to improve patient satisfaction,to avoid operation delay and medical accident caused by inappropriate preparation.Methods A total of 145 patients with surgery from March 2013 to February 2013 were as experimental group,and 158 patients with surgery from March 2012 to February 2012 were as control group.The experimental group was used electronic preoperative preparation checklist for preoperative preparation and transition,and the control group was used conventional methods.The incidence of mistake for preoperative preparation and transfer and both surgeon's and patient's satisfactory were compared between two groups.Results After applying the electronic preoperative preparation checklist,the incidence of mistake for preoperative preparation and transfer in experimental group reduced significandy to 1.37％(2/145) and 4.83％ (7/145),compared with the incidence of control group 6.33％(10/158),11.39％(18/158),and the differences between two groups were statistically different (x2=4.870,4.305,P ＜ 0.05).Both surgeon's and patient's satisfactory were improved dramatically,the satisfactory in experimental group improved to 100.00％(50/50)and 97.93％(142/145),compared with the satisfactory of control group 90.00％(45/50) and 90.51％(143/158),and the differences between two groups were statistically different(x2=5.263,7.459,P ＜ 0.05).Conclusions Implementing the electronic preoperative preparation checklist can reduce the incidence of mistake before operation and ensure patient operation schedule.Therefore,it could improve nursing care quality and efficiency.

Aim To investigate the ameliorative effects pseudoberberine(Y53), a berberine(BBR) analogue, on diabetic nephropathy( DN) in streptozotocin( STZ)-induced diabetic mice. Methods Diabetes mellitus ( DM) of the C57BL/6J mice was induced by intraper-itoneal injection of STZ at 120 mg·kg-1 . The diabetic animals were divided into 4 groups, which were orally treated with saline, 50 mg · kg-1 of BBR, 50 mg · kg-1 of Y53 or 5 mg · kg-1 of rosiglitazone ( ROSI ) , respectively. During and after the experiment, the u-rine, blood, serum and kidney of the animals were harvested for determination of relevant parameters by kits. Kidney tissues of the mice were subjected to pathological examination by hematoxylin & eosin( HE) staining;mRNA and protein expression levels of target genes in the kidney were determined by quantitative re-al-time reverse transcriptase-polymerase chain reaction ( qRT-PCR) and Western blot, respectively. Results Y53 greatly reduced the fasting blood glucose ( FBG ) and glycosylated hemoglobin( GHb) , improved diabet-ic symptoms such as polyphagia and polyuria in the di-abetic mice( P<0. 01 vs DM control group) . Y53 po-tently reduced the blood urea nitrogen ( BUN ) , serum creatinine( Scr) , 24 h urinary protein, kidney index, serum and kidney advanced glycation end-products ( AGEs) and nitric oxide( NO) , as well as kidney cho-lesterol( CHO ) and triglyceride ( TG ) contents ( P <0. 05 or P<0. 01 vs DM control group) . In the patho-logical examination, Y53 greatly restored kidney mor-phology and suppressed glomerular sclerosis( P<0. 001 vs DM control group). In addition, Y53 significantly reduced the renal expression of fibrosis-related genes, such as the transforming growth factor-β1 ( TGF-β1 ) and smad2(P<0. 01 vs DM control group). The reno-protective efficacies of Y53 were superior to those of BBR and ROSI in our study ( P<0. 05 or P<0. 01 ) . Conclusions The BBR analogue Y53 has potent ac-tivities in ameliorating renal injury and restoring renal function in STZ-induced diabetic mice. Y53 may be developed as a novel kind of agent for the treatment of DN in the future.

Objective To discuss the anticancer role of arsenic trioxide (ATO) with cisplatin on human oral carcinoma HSQ-89 cells. Methods The human oral epidermoid HSQ-89 cells were chosen as the subjects. Different concentrations of ATO were added into Cisplatin(DDP)-treated cells. The inhibition rate of tumor cells was detected by MTT assay. Results Different concentrations of ATO (0,2.5,5,7.5,10,12.5 μmol/mL) were added into oral cancer HSQ-89 cells which have been treated with DDP (15 μg/mL). The inhibition rate of tumor cells were 26.9%, 67.5%, 73.0%, 88.5%, 90.4%, 98.7%respectively; The combined application of ATO with cisplatin could improve the inhibition rate of HSQ-89 cells in a dose-dependent relation. Conclusion The combined application of ATO and DDP can produce a synergistic action of inhibition on oral cancer cell.

Objective To detect PAH gene mutations in classical PKU patients by HRM analysis. MethodsMutation scanning of PAH gene were performed in 17 classical PKU patients by HRM analysis ( LightScanner), covering the 13 exons and exon-intron boundaries. The HRM results were further confirmed by DNA sequencing, and the sensitivity and specificity of HRM method in PKU diagnosis were also evaluated. In addition, prenatal diagnosis was performed in two fetuses at risk for classical PKU. Results In the 17 patients, two mutations were identified in 16 patients, three mutations were identified in 1 patient.In this subject, a total of 22 different pathogenic mutations : 194V( c. 280A ＞ G), IVS4nt-1 G ＞ A( c. 442-1G ＞ A), R158Q( c. 4736 ＞ A), Q160X( c. 478C ＞ T), W187X( c. 561G ＞ A), E6nt-96A ＞ G( c. 611A ＞G), G239D( c. 716G ＞ A), R241 C( c. 721C ＞ T), R243Q( c. 728G ＞ A), G247R (c. 739G ＞ C), G247V (c. 740G＞T), R261X(c. 781C ＞T), PR261Q(c. 782G ＞ A), H264R (c. 791A ＞ G), F302fsX39 (c. 904delT), E305K( c. 913G ＞ A), G312V( c. 935G ＞ T), Y356X( c. 1068C ＞ A ), V399V ( c. 1197A ＞T), R408Q(c. 1223G ＞ A), T418P(c. 1252A ＞ C) , A434D(c. 1301C ＞ A), 3 silent mutations Q232Q (c. 696G ＞ A), V245V(c. 735G ＞ A), L385L(c. 1155C ＞ G), and one single nucleotide polymorphism rs2280615 ( c. 402A ＞ C) were identified, of which 194V ( c. 280A ＞ G), Q160X ( c. 478C ＞ T), H264R (c. 791A ＞ G), G312V( c. 935G ＞ T) and E305K ( c. 913G ＞ A) were novel mutations identified in PAH gene. The prenatal diagnosis results of the two fetuses : one was diagnosed as normal, the other was diagnosed as a carrier. In this study, the sensitivity and specificity for mutation detection by HRM were 100％, and the HRM results were consistent with DNA sequencing results. Conclusions HRM analysis is a simple,accurate, rapid, high-throughput and low-cost genetic analysis approach. It could be applied to mutation scanning of classical PKU of PAH gene and rapid prenatal diagnosis in parents with known mutations.

BACKGROUND:Bone marrow is the main source of mesenchymal stem cells(MSCs)at present,but its application has been limited,because of some reasons such as inconvenience of isolation,and the quantity of cells decreases with human increased age.Umbilical cord as a new source of MSCs has been widespread concerned recently.OBJECTIVE:To explore the approach of isolating and culturing MSCs from Wharton's jelly of human umbilical cord,and the methods of identifying the surface antigens and the differentiation potential.METHODS:MSCs were isolated and amplified via tissue-cultivation,and cultured by FasGrow medium.Morphology of MSCs from Wharton's jelly of human umbilical cord was observed under the optical microscope.Its immunophenotypes were detected using immunohistochemistry.The differentiation of MSCs into the osteoblasts was determined utilizing Gomori calcium-cobalt alkaline phosphatase staining,von Kossa calcium node staining,and tetracyclinefluorescence labeling.The differentiation of MSCs into the adipocytes was detected using oil red O staining.RESULTS AND CONCLUSION:MSCs were easily obtained from Wharton's jelly of human umbilical cord via the proposed approach.The primary cells grew up to 70%-80% confluence after 12-16 days of culture,and meanwhile the undifferentiated state was maintained and proliferation was stabilized after passage.The cell cycle of double increase was about 2 days,and proliferation in vitro reached twenty generation above.Surface antigen analysis showed that CD44,CD105,CD133,MHC-I were positively expressed,while CD34,CD45 were negatively expressed.Experiments of differentiation in vitro indicated that the obtained cells were capable of differentiating into fat,osteoblast and nerve-like cells.

Objective To investigate the expression difference of vascular endothelial growth factor (VEGF)between lung cancer and lung benign disease,and its relation to clinical characteristics and prognosis of non-small cell lung cancer(NSCLC),and to research the possible role of VEGF in the growth of lung can- cer.Methods The expression of VEGF and microvascular density(MVD)were detected in 60 lung cancer tissues and 30 lung benign disease tissues after operation by immunohistochemical method and put up statisti- cal analysis.Results The positive rate of VEGF expression(63%)and MVD(45.13?10.27)in lung cancer tissues were both higher than those in lung benign disease tissues(27%,33?6.49)(P