Objective To investigate whether the Notch-1 signaling pathway is involved in the acquisition of the epithelial-mesenchymal transition (EMT) phenotype of gefitinib-acquired resistant lung cancer cells.Methods The PC9 cell line (harboring EGFR exon 19 deletion) and PC9/AB2 cells (gefitinibacquired resistant PC9 cells) were used.siRNA targeting Notch-1 eukaryotic expression vector (siNotch-1) was constructed and PC9/AB2 cells were transfected with siNotch-1.The protein expression of EGFR,Akt,Erk,Notch receptors and ligands,TGF-β receptors,E-cadherin,Vimentin,and Snail were detected by Western blot assay.DNA sequencing and fluorescence in situ hybridization (FISH) analysis were processed to detect mutation of EGFR exon 20 and MET amplification,respectively.For cytotoxicity assay,cell viability was assessed by Cell Counting Kit 8.Results Gefitinib resistant cell line PC9/AB2 had no evidence of MET amplification or EGFR T790M mutation.The expression of Notch-1 was upregulated in gefitinib resistant PC9/AB2 cells compared with that in gefitinib-sensitive PC9 cells.There were no significant protein expression differences of other Notch receptors,Notch ligands or TGF-β receptors between both paired cell lines.Western blot results showed that protein expression of E-cadherin was greatly reduced in PC9/AB2 cells,while elevated levels of Vimentin and Snail were observed.A significant reduction of the expression of Snail and Vimentin in Notch-1 siRNA transfected PC9/AB2 cells with increased E-cadherin expression was found by Western blot assay.PC9/AB2 cells displayed a round like cell morphology after Notch-1 siRNA transfection.Silence of Notch-1 decreased colony-formation ability but enhanced sensitivity of gefitinib on PC9/AB2 cells.Conclusion Notch-1 might play a novel role in acquired resistance to gefitinib,which could be reversed by inhibiting Notch-l.

Objective To explore the current status of humanistic care among medical students in clinical practice in obstetrics and gynecology department and its related influencing factors and to propose the corresponding countermeasures.Methods Scale of humanistic care quality was used in the survey for clinical practice students in obstetrics and gynecology department.ResultsThe total score rate of human care quality of medical students before clinical teaching were (84.12 ± 9.24)with a scoring rate of 72％.Significant differences were observed in the medical students with different medical professional(P ＜0.05).The total score of humanistic care quality of medical students after clinical teaching were (96.41 ± 1 1.53 ),significant higher than that of before training ( P ＜ 0.05 ).ConclusionHumanistic care quality in medical students needs to be strengthen.Clinical training in obstetrics and gynecology department combined with humanities education has important practical significance to improve the quality of medical students.

Objective To determine the diagnostic and assessment value of soluble urokinase plasminogen activator receptor (suPAR) level in septic patients.Methods Totally 82 septic patients in the Department of Intensive Care Unit of The First Affiliated Hospital,Sun Yat-Sen University were prospectively analyzed from June 2013 to March 2014.Another 29 patients with systemic inflammatory response syndrome (SIRS) and 15 healthy subjects served as controls.Septic patients were divided into sepsis group (n =27),severe sepsis group (n =27) and septic shock group (n =28) according to the severity,and there was no significant difference in age and sex among these groups.Measurement of plasma suPAR,serum procalcitonin (PCT) and C-reactive protein (CRP) levels,and calculation of acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) and sequential organ failure assessment (SOFA) score were performed.Comparison of group differences for continuous variables was done by oneway ANOVA or nonparametric Kruskal-Wallis test.Spearman rank correlation analysis was applied to establish the relation between variables.Receiver operating characteristics (ROC) curve was created and area under curve (AUC) was calculated to determine the diagnostic value of these variables in sepsis.Results The levels of plasma suPAR in SIRS group,sepsis group,severe sepsis group,septic shock group,and healthy control group were (8.22±0.61),(11.45±1.12),(12.99±1.28),(15.75± 1.23) and (4.65 ±0.30) ng/mL,respectively.Plasma suPAR levels in SIRS group and sepsis group were higher than that in healthy control group (P ＜ 0.01),and elevated plasma suPAR was accompanied by increased severity of sepsis (P ＜ 0.05).PCT levels of sepsis group (17.66 ± 8.42) ng/mL,severe sepsis group (9.67 ±3.56) ng/mL and septic shock group (29.19 ± 10.78) ng/mL were greater than that in SIRS group (1.10 ± 0.78) ng/mL,P ＜ 0.01.CRP levels elevated in all groups,but there were no significant differences among them.When suPAR and CRP were applied to distinguishing sepsis from SIRS,the AUC values from suPAR and combination of suPAR and PCT were 0.817 (P ＜ 0.01,95 ％ CI:0.714-0.921) and 0.927 (P＜0.01,95％ CI:0.870-0.985),respectively.Using9.52 ng/mL suPAR as the best cut-off to distinguish sepsis from SIRS,there were 71.93％ sensitivity and 95.46％ specificity.The levels of plasma suPAR positively correlated with PCT levels (r =0.326),APACHE Ⅱ score (r =0.492) and SOFA score (r =0.386),P ＜ 0.01.Conclusions Plasma suPAR levels significantly elevated in septic patients and correlated with the severity of sepsis.Sepsis and SIRS may be discerned by plasma suPAR levels.Joint use of suPAR and PCT could greatly increase the specificity of diagnosis of sepsis.

Objective To investigate the clinical and pathological characteristics between types 1 and 2 endometrial uterine cancers.Methods The clinical materials of 9 437 patients with uterine cancer were collected with retrospective analysis from 62 hospitals during 2000 to 2010.Results The mean age of type 1 endometrial cancers was less than type 2.There were more young patients in type 1 endometrial cancers.The mean menopause age of type 2 endometrial cancers was greater than type 1.The mean age of menarche,obesity,diabetes,hypertension,infertility,and nulliparous were not significant differences between types 1 and 2 endometrial cancers.There were more patients with advanced tumor,deep myometrium invasion,estrogen receptor (ERs) negative,progesterone receptor (PR) negative,P53 positive,lymph vascular space involvement,cervical stromal invasion,adnexal metastasis,and lymph node metastasis in type 2 endometrial cancers.Conclusions Type 2 endometrial uterine cancers occurred in elder people with more pathological risk factors and more malignant biological activities.

Objective To investigate the effects of repeated intraperitoneal dexmedetomidine on the cognitive function in rats with chronic cerebral ischemia.Methods Forty-eight male Sprague-Dawley rats,aged 3-4months,weighing 250-300 g,were randomly divided into 4 groups (n =12 each) ∶ sham operation group (group S),chronic cerebral ischemia group (group IS),dexmedetomidine treatment 1 group (group DXM1) and dexmedetomidine treatment 2 group (group DXM2).Dexmedetomidine 5 μg/kg was injected intraperitoneally at 30 min before occlusion of bilateral common carotid arteries and 3,12,24 and 48 h after occlusion in group DXM1,and at 3,12,24 and 48 h after occlusion in group DXM2.The cognitive function was assessed by Morris water maze 2 weeks after occlusion.The apoptosis was examined by TUNEL.The expression of Bcl-2 protein in hippocampus was detected by Western blot.Results Compared with group S,the escape latency was significantly prolonged from 2nd day to 5th day after the place navigation test in group IS and on 2nd day after Morris water maze test in groups DXM1 and DXM2,and the time of staying in 1 st quadrant was significantly shortened,the apoptotic rate was increased,and the expression of Bcl-2 was up-regulated in groups IS,DXM1 and DXM2 (P ＜ 0.05).Compared with group IS,the escape latency was significantly shortened from 3rd day to 5th day after the place navigation

Objective:To investigate the protective effects of Ghrelin on LPS-induced apoptosis of human alveolar epithelial A549 cells,along with the possible molecular mechanisms.Methods: CCK-8 assay was used to examine the cell viability of A549 treated by LPS.Apoptosis of A549 cells was measured by TUNEL.NO(Nitric oxide)production was detected by NO-specific fluorescent probe 3-Amino,4-aminomethyl-2′,7′-difluoresceindiacetate(DAF-FM DA).Western blot was also performed to examine the expressions of iNOS(inducible nitric oxide synthase),AKT,ERK,p-AKT,p-ERK and apoptotic proteins,such as Bcl-2,Bax,and cleaved caspase-3.Results: LPS exposure impaired cell viability and increased apoptosis of A549 cells significantly in concentration-and time-dependent manners accompanied with increased Bax and cleaved caspase-3 production,coupled with decreased Bcl-2 levels.Meanwhile,LPS promoted iNOS expression and the production of NO.Ghrelin pretreatment ameliorated LPS-caused alterations in the ratio of Bax/Bcl-2 and cleaved caspase-3 expression.TUNEL analysis showed that Ghrelin could decrease the apoptosis induced by LPS in A549(P<0.05).Simultaneously,LPS remarkably decreased the expression of p-AKT and p-ERK in A549 cells,which was abrogated by Ghrelin pretreatment.However,Ghrelin had no significant effect on NO production induced by LPS.Conclusion: Ghrelin reduces LPS-induced apoptosis of human alveolar epithelial cells partly through activating the AKT and ERK pathway,but the level of iNOS derived NO could not be reduced.

Objective To explore the effect of speech therapy on articulation disorder of indifferent kinds of children with cerebral palsy(CP).Methods 49 CP children with articulation disorder were divided into the common group(n=21)and family rehabilitation group(n=28).All children in the two groups were treated with systemic speech therapy,but those in the family rehabilitation group were added with family rehabilitation.The changes of articulation disorder of children in two groups before and after treatment were observed.Results After treatment,all children got improvement,but the effect of children with spastic type was superior to those with other CP types.The efficiency rate of the family rehabilitation group was 39.3%,that of the common group was 14.3%,there was a significant difference between two groups(P<0.05).Conclusion The children with different CP type have different therapeutic effects for articulation disorder,the family rehabilitation can improve the therapeutic effect.

Objective:To investigate effects of tumor specific TCR gene Vα12.2-Vβ7.1 modification on recognition of tumor antigen and activation of anti-tumor reactivity of T cells.Methods: T cells were transduced using recombinant Ad 5F35-TRAV-TRBV adenovirus ,and multiplicity of infection was optimized.Specific lysis of T cells was evaluated by calcein release assay.The frequency of apoptotic cells in target cells was detected by Annexin V /PI double-labeled FACS.The expression of FasL on T cells was analyzed by FACS.The secretion of cytokine IFN-γand IL-2 of T cells was determined by ELISA assays.Results: The highest tranduce efficiency was obtained at MOI 100 by recombinant Ad5F35-TRAV-TRBV adenovirus.The frequency of TCRVα12+Vβ7+cells reached above 25%3 days after transduction.TCR gene modification enhanced the ability of T cells to lyse HLA-A2+AFP+target cells(P<0.001), the ability of T cells to induce HepG-2 apoptosis(P<0.001),and expression of FasL on T cells(P<0.001).TCR gene modification also enhanced T cells to secret IFN-γafter coculture with antigen positive tumor cells ( P<0.001 ).Conclusion: Specific TCR gene modification by recombinant adeno virus effectively promotes T cells to recognize antigen positive tumor cell and exert anti -tumor reac-tivity.

In order to demonstrate the taxonomic position of paramyxovirus Tianjin strain and explore its mechanism of pathogenesis. Bioinformatics methods were used to analyze the deduced amino acid sequences of NP, P, M, and L protein of Tianjin strain. Phylogenetic analysis based on NP, P, M, and L protein sequences demonstrated that Tianjin strain belonged to the genus Respirovirus, in the subfamily Paramyxovirinae and most likely a new genotype of Sendai virus. Sequence similarities comparisons indicated that Tianjin strain P protein was poorly conserved, sharing only 78.7%~91.9% amino acid identity with 6 known Sendai viruses, while L protein was the most conserved, having 96.0%~98.0% amino acid identity with other Sendai viruses. Multiple-sequence alignments of Tianjin strain NP, P, M, and L protein with those of 6 known Sendai viruses showed that Tianjin strain possessed a lot of unique amino acid substitutions in protein sequences, 15 in NP, 29 in P, 6 in M, and 29 in L. The presence of these unique amino acid substitutions suggests that Tianjin strain maybe has a significant difference in host or pathological characteristics from the known Sendai viruses.

OBJECTIVETo evaluate the detective efficacy of Chromogenic Coliform and Escherichia Coli Agar (CCEA).

METHODSA new chromogenic medium CCEA prepared by Huankai laboratory was used to compare with a classical medium of violet red bile agar (VRBA), and other two Chromogenic media Agar I and Agar II by detecting separately 11 reference strains, thirteen sterile samples with Coliform or E.coli and other four samples, and the accordant rates of detection were observed.

RESULTSCCEA had the good selectivity. To seven kinds of quality strains in the resultant analysis, CCEA with VRBA and Agar I had not shown salience difference (P > 0.05), and CCEA with Agar II had significant difference (P < 0.05). CCEA showed more advantages than the Agar II. To thirteen sterile samples with Coliform or E.coli in resultant analysis, CCEA with Agar I and Agar II had shown no significant difference (P > 0.05), while CCEA with VRBA had significant difference (P < 0.05). CCEA might be more advantageous than the VRBA. In analysis of the four actual samples of Coliform, CCEA with VRBA, Agar I and Agar II showed no significant difference (P > 0.05). The accordant rates were 90%, 71.88%, 86.25% and 81.25% respectively, showing CCEA > Agar I > Agar II > VRBA. To two actual samples of E.coli in the resultant analysis, the CCEA with Agar I and Agar II had not shown significant difference (P > 0.05). The accordant rates were 100% respectively.

CONCLUSIONSThe CCEA might be more advantageous than the VRBA, having the same efficacy as with Agar I and Agar II.