AIM:To investigate the effect of the N-terminal 24-amino acid (N24) overexpression in p55γre-gulatory subunit of phosphatidylinositiol 3-kinase ( PI3K) on the cell adhesion of human gastric carcinoma cell MGC 803. METHODS:MGC803 cells, which stably expressed GFP-N24 fusion protein and GFP alone , were generated by transfec-tion with pEGFPN-24 plasmid and control plasmid pEGFP-C1, respectively.The morphological change of the cells was ob-served under inverted microscope .The expression of GFP-N24 fusion protein was detected by Western blot .The adhesion of the cells was determined by cell adhesion assay .The effects of GFP-N24 fusion protein on the expression of E-cadherin andβ-catenin were analyzed by Western blot .The expression and secretion of matrix metalloproteinase 9 (MMP9) and u-rokinase-type plasminogen activator ( uPA ) in the MGC803 cells were detected by gelatin zymography .RESULTS:MGC803/GFP-N24 cell line steadily expressed GFP-N24 fusion protein and MGC803/GFP cell line steadily expressing GFP were successfully established , but the expression of fusion protein GFP-N24 was lower than that of the control protein GFP.The morphological changes of the transfected cells from paving stone to fibroblast cell form after gene transfection , and the cytoplasm secretory granules were increased significantly .The cell adhesion to fibronectin and collagen decreased after GFP-N24 transfection .GFP-N24 fusion protein increased the expression of cell adhesion molecule E-cadherin and de-creased the wnt signal pathway molecule β-catenin in the MGC803 cells.However, it did not affect the expression and se-cretion of tumor metastasis-related proteins MMP9 and uPA.CONCLUSION:Overexpression of N24p55γinhibits cell ad-hesion by influencing the expression of E-cadherin and β-catenin in the MGC803 cells.

Objective To evaluate the effect of P-gp inhibitors of verapamil (VER) and GF120918 on 18F-FDG uptake in Bcap37 and Bcap37/multidrug resistancce (MDR)1 cell lines in vitro,and to explore the relationship between 18F-FDG uptake and P-gp expression at cellular level.Methods Bcap37 and Bcap37/MDR1 cells were seeded into 6-well plates at a density of 1 × 106 per well.Three days later,37 kBq/ml 18F-FDG,or 37 kBq/ml 18F-FDG + 100 μmoL/L VER,or 37 kBq/ml 18F-FDG + 50 μmol/L GF120918 were added into each well.Mter incubated for 10,30,60 and 120 min at 37 ℃ and in 5％ CO2,the medium was removed and the cells were washed three times with 1 ml ice-cold PBS immediately.The radioactivity of 18 F-FDG was measured using a gamma counter.The uptake of 18F-FDG was expressed as the ratio of 18F-FDG radioactivity in Bcap37 or Bcap37/MDR1 cells and the overall radioactivity added to the cells in each well.The t test was used for statistical analysis.Results 18F-FDG uptake was higher in Bcap37/MDR1 cells than that in Bcap37 cells after incubated for 10 min.The uptake rate was (1.88 ±0.19) ％ in Bcap37/MDR1 cells and (1.37 ± 0.18) ％ in Bcap37 cells (t =7.832,P ＜ 0.05).On the contrary,18 F-FDG uptake was significantly higher in Bcap37 cells than that in Bcap37/MDR1 cells after incubated for 60 and 120 min.The uptake rates were (2.29 ±0.23)％ and (2.34 ±0.15)％ in Bcap37 cells,(1.47 ±0.14)％ and (1.53 ±0.22)％ in Bcap37/MDR1 cells (t =8.437,8.283,both P ＜ 0.05).18 F-FDG uptake was significantly higher with VER or GF120918 in Bcap37/MDR1 cells than that without VER or GF120918 after the incubation of 60 and 120 min (t =9.032,9.243 and 8.765,8.803,all P ＜ 0.05).The uptake rates with VER or GF120918 were (2.45 ±0.21)％ and (2.46 ±0.25)％,(2.50 ±0.24)％ and (2.48 ±0.27)％.There was no significant difference of 18F-FDG uptake in Bcap37 cells with or without VER or GF120918.Conclusions 18F-FDG is a substrate of P-gp at cellular level.P-gp may act as an efflux pump to reduce 18F-FDG uptake in Bcap37/MDR1 cells.The uptake of 18F-FDG can be used to evaluate the function of P-gp in tumor cells.

BACKGROUND: To predict the functional outcome basing on hospitalization data as earlier as possible has important significance for evaluating the prognosis and discharged recovery. In recent years, researches on the balance function of stroke patients has drew more and more attentions, but less related reports about the relationship between balance and discharge functional outcome are available.OBJECTIVE: To predict the discharg functional outcomes of stoke patients basing on hospitalization data, as well as the relationship between it and scores for Fugl-Meyer balance(FMB) and Berg balance scale(BBS).DESIGN: Multiple factors and multi-variable study based on patient' s clinical presentation.SETTING: Rehabilitative department in a college hospital.PARTICIPANTS: Between August 2002 and May 2003, 40 patients with stoke for the first time were hospitalized in Rehabilitative and Neurological Department of Tongji Hospital, who accorded with the enrolling standards.METHODS: The available hospitalization data and discharge functional outcomes were subjected to single factor and multiple factor analysis, and collected data includes the history of disease, physical examination, scores for FMB and BBS when hospitalization, scores for daily life ability(ADL)and functional independence measure(FIM) . Functional outcome was presented by the discharge FIM.MAIN OUTCOME MEASURES: Simple correlative analysis of BBS, FMB and FIM, regressive analysis of discharge FIM influencing factors.RESULTS: Simple liner correlative analysis indicted that scores for BBS and FMB was strongly correlated with the scores for FIM at hospitalization and discharge( P ＜ 0. 001 or P ＜ 0.05), regressive analysis revealed that scores for FIM, ADL and BBS at hospitalization could be used for predicting the discharge functional outcomes of stroke patients.CONCLUSION: Predictors for discharge functional outcomes includes scores for FIM, ADL and BBS when hospitalization, which are closely correlated with BBS.

Objective To observe the effects of hyperbaric oxygen (HBO) exposure on the expression of aquaporin-1 (AQP1) and aquaporin-5 (AQP5) in rats with simulated high-altitude pulmonary edema (HAPE).Methods Fifty-six rats were randomly divided into five groups:control (normal),HAPE (high altitude pulmonary edema model),1 HBOT (HAPE model and HBO therapy for 1 time),2 HBOT (HAPE model and HBO therapy twice) and NOT (normal pressure oxygen therapy) groups,and was intervened accordingly.Western blotting and real-time PCR techniques were used to analyze the expression of AQP1 and AQP5 in their lungs.The wet-todry (W/D) weight ratio and morphology of the lungs was also examined.Results The protein and gene expression of AQP1 and AQP5 in the HAPE group decreased significantly compared with the control group.There were obvious differences in the protein and mRNA expression of AQP1 and AQP5 between the 2 HBOT group and the HAPE group and between the 2 HBOT group and the 1 HBOT group.Compared with the control group and the 1 HBOT group,marked lung injury could be seen in the HAPE group.Compared with the NOT group and the 1 HBOT group,lung injury in the 2 HBOT group was relieved significantly.Conclusions HAPE in rats is associated with down-regulation of the expression of AQP1 and AQP5 in the lungs.This down-regulation can be attenuated and lung injury can be alleviated by HBOT.Two sessions of HBOT could be more helpful than one for promoting this improvement.

Objective To discuss the effect of the teaching mode of technological pedagogical and content knowledge in biochemistry theory teaching. Methods 400 students in 8 classes of clinical medicine undergraduate in Grade 2013 were selected and randomly divided into 2 groups. The traditional teaching mode was applied in the control group of 198 students in 1-4 classes, while the technological pedagogical and content knowledge teaching mode was applied in the experimental group of 202 students in 5-8 classes. 385 students in 8 classes of clinical medical undergraduates in Grade 2014 were selected and randomly divided into 2 groups. The control group of 189 students in 1-4 classes adopted the traditional teaching mode, while the experimental group of 196 students in 5-8 classes adopted the technological pedagogical and content knowledge teaching mode, which used the micro-lesson and network platform as learning resource superior to the flipped classroom. Flipped classroom was divided into two major learning links:extracurricular self-study and class digestion. Through the network platform the micro-lesson was presented to the learners. Learners could make self-study according to their own specific circumstances and in the classroom many activities were increased such as the mutual cooperation between the students, the students' PPT teaching, the students' questions and the discussion, and the interaction between teachers and students, etc. The results of the examinations of the two terms students of the same profession and the questionnaire were analyzed. The related data were processed by SPSS 15.0, and the data between groups were compared by t test . Results The test scores analysis showed that the individual test scores in experimental group of Grade 2013 were [(17.94±2.02) vs. (12.28±4.17)], and the individual test scores in experimental group of Grade 2014 were [(18.21 ±1.78) vs. (12.45 ±5.13)], which were obviously higher than the control group, and there was statistical significance. The final exam scores in experimental group of Grade 2013 were [(78.28±11.18) vs. (68.65±12.51)], and the final exam scores in experimental group of Grade 2014 were [(81.73 ±9.12) vs. (74.41 ±11.87)], which were obviously higher than the control group, and there was statistical significance. The results of survey showed that the students thought the teaching mode aroused their study interests while 393 (93.7%), thought the teaching mode developed their self-study ability while 357 (89.7%), thought the teaching mode beneficial to cultivating their ability of solving the problems. Conclusion The teaching mode of technological pedagogical and content knowledge is of certain signifi-cance to break the plight of the traditional teaching, inspire the students interest in learning, improve the teaching quality of biochemistry, and make for the teachers' professional development.

Objective To analyze the relevant factors influencing 18F-FDG uptake in the primary lesion of breast invasive ductal carcinoma (BIDC).Methods A total of 160 female patients underwent 18 F-FDG PET/CT examination from 2010 to 2015 and breast lesions were revealed.Lesions were divided into benign group (n =118) and malignant group (n =49,BIDC) according to pathological results.KruskalWallis H test and Mann-Whitney u test were performed to compare SUVmax of the two groups,and to investigate the relationship between the SUVmax of breast malignant lesion and patients' age as well as clinical pathological parameters including T stage,lymphatic vessel invasion,nuclear grade,route of metastasis,ER,PR,HER2 and Ki-67 expression,and subtype of breast cancer.The diagnostic efficiency of 18F-FDG PET/ CT in differentiating benign and malignant breast lesions was analyzed using ROC curve analysis.Results The SUVmax of BIDC was 6.09(3.88,9.26),higher than that of breast benign lesion (1.35 (0.95,2.35);u=341.0,P＜0.05).The SUVmax of BIDC showed statistically significant difference between groups with different T stage,with or without lymphatic vessel invasion,with different nuclear grade,different routes of metastasis and different Ki-67 expression (u:117.5-209.5,H=7.70,P＜0.01 or 0.05).For all breast lesions,lesions with the maximum diameter ≤ 2.0 cm and lesions with the maximum diameter ＞2.0 cm,the optimum cutoff values of SUVmax were ＞2.60,＞ 1.71 and ＞3.97,respectively.When the optimum cutoff values of SUVmax for breast lesions with the maximum diameter ≤2.0 cm were selected as ＞ 1.71 and ＞2.60,the Youden indexes were 0.66 and 0.61(z=0.566,P＞0.05).When the optimum cutoff values of SUVmax for breast lesions with the maximum diamter ＞2.0 cm were selected as ＞3.97 and ＞2.60,the Youden indexes were 0.89 and 0.81(z=0.748,P＞0.05).Conclusions T stage,lymphatic vessel invasion,nuclear grade,route of metastasis and Ki-67 expression of BIDC influence the uptake of 18F-FDG by tumor tissues.The SUVmax of the primary lesion of BIDC is related to the size of lesion,and thus the diagnostic threshold of SUVmax should be decreased appropriately for small lesions.

The wild type cells of human stomach glandular carcinoma, cell line BGC 823, were treated firstly by a chemical carcinogen (MNNG) for the induction of mutagenesis, then the cells were selected in medium with gradually increasing amount of 8-AG (8-azaguanine) from 1-20?g/ml. It was found that the mutant cells could grow vigorously in the medium containing 20?g/ml of 8-AG but not in HAT medium. The HGPRT assay showed an obvious quantitative difference between the wild type and HGPRT mutant cells of BGC 823.

Objective To evaluate the relationship between 18F-FDG uptake and P-gp expression in Bcap37 or Bcap37/MDR1 tumor-bearing BALB/c nude mice.Methods Bcap37 or Bcap37/MDR1 cells were injected into BALB/c nude mice (1× 107cells/ml,0.2 ml/mouse) to construct mice models.Bcap37 (n=5) or Bcap37/MDR1 (n=5) tumor-bearing mice fasted for 6 h before imaging.After anesthesia,the mice were injected with 7.4 MBq of 18F-FDG via tail vein.The dynamic microPET scans were carried out for 90 min.On the microPET images,the ROI was drawn and the TAC was obtained.The next day,those 10 mice underwent dynamic microPET scans after injected with elacridar (GF120918) and 18F-FDG.Another 10 mice,5 with Bcap37 tumors and 5 with Bcap37/MDR1 tumors,were used.After 7.4 MBq 18F-FDG with or without 2.0 mg/kg GF120918 was administered via tail vein,microPET images were acquired at 60 min.ROI was drawn over the tumors and SUV was obtained.Two-sample t test was used to analyze the data.Results GF120918 did not significantly alter the 18F-FDG accumulation curve in Bcap37 tumors,but significantly enhanced the 18F-FDG accumulation in Bcap37/MDR1 tumors.GF120918 did not influence 18F-FDG uptake (SUV) in Bcap37 tumors (1.052±0.028,1.028±0.045,t =1.792,P＞0.05),but significantly increased the SUV in Bcap37/MDR1 tumors (1.015±0.043,0.712±0.031,t=3.365,P＜0.05);The SUV of 18 F-FDG in Bcap37 tumors was significantly higher than that in Bcap37/MDR1 tumors without injection of GF120918 (t =3.952,P＜0.05).The SUV was not significantly different when GF120118 was injected (t=1.835,P＞0.05).Conclusions 18F-FDG is a substrate of P-gp.18F-FDG imaging combined with GF120918 injection may be an effective noninvasive method for the detection of tumor's MDR.

Objective:To investigate the expression and clinical significance of effector T cell (Teff), regulatory T cell (Treg) and their cytokines in patients with vitiligo.Methods:84 patients with vitiligo (38 cases in stable stage, 46 cases in advanced stage) who were admitted to the Dermatology Department of the Dezhou People's Hospital between June 2018 and June 2019 and 30 healthy people (control group) were enrolled in the study. The levels of Teff subsets and Treg cells in peripheral blood were detected by flow cytometry. The levels of Teff subsets and Treg cells-associated cytokines were detected by enzyme-linked immunosorbent assay (ELISA). The relationship between Teff subsets and Treg cells levels in peripheral blood, relationship between Teff subgroups, Treg cells-associated cytokines and course of vitiligo, skin lesion area were analyzed.Results:The levels of T helper cell (Th)17 and Th22 in peripheral blood of vitiligo group were significantly higher than those of control group, while levels of Treg cells were significantly lower than control group ( P<0.05). Compared with the stable vitiligo group, the levels of Th17 and Th22 in peripheral blood were higher and levels of Treg cells were lower in the advanced vitiligo group ( P<0.05). Treg cell level was negatively correlated with Th17 cell level ( r=-0.303, P=0.001), and negatively correlated with Th22 cell level ( r=-0.200, P=0.033). The levels of serum interleukin (IL)-17 and IL-22 in vitiligo group were significantly higher than those in control group, while the level of transforming growth factor-β (TGF-β) was significantly lower than that in control group ( P<0.05). Compared with the stable vitiligo group, levels of serum IL-17 and IL-22 were higher and level of TGF-β was lower in the advanced vitiligo group ( P<0.05). Correlation analysis showed that levels of IL-17 and IL-22 were positively correlated with skin lesion area ( r=0.361, 0.288, P<0.05), and level of TGF-β was negatively correlated with skin lesion area ( r=-0.312, P<0.05). Conclusions:Teff/Treg cells are unbalanced in patients with vitiligo, showing obvious shift to Teff subsets. The progression of vitiligo may be related to Teff subsets and Treg cells-associated cytokines.

Objective To investigate the risk factors of pancreatic cancer in Wenzhou area.Methods A case control study was conducted on 220 cases with pancreatic cancer and 220 matched controls using conditional logistic regression analysis.Results Univariate analysis showed BMI ＞ 30 kg/m2,smoking,reproductive history,diabetes,cholecystitis,chronic pancreatitis,history of appendectomy,history of partial gastrectomy,history of cancer and familial history of cancer were associated with pancreatic cancer,while multivariate conditional logistic regression analysis showed that the risk factors of pancreatic cancer included smoking (OR =3.624,95％ CI i.474 ～ 8.907),BMI ＞ 30 kg/m2 (OR =1.789,95％ CI 1.030 ～ 3.108),diabetes (OR =3.191,95％ CI 1.094 ～ 9.309),chronic pancreatitis (OR =4.972,95％ CI 1.593 ～ 14.898),and cholecystitis (OR =2.289,95 ％ CI 1.024 ～5.116).Conclusions BMI ＞ 30 kg/m2,smoking and diabetes are risk factors for pancreatic cancer in Wenzhou area.