Objective:To observe the effects of electroacupuncture(EA)on gut microbiota and serum inflammatory factors interleukin(IL)-1β and tumor necrosis factor(TNF)-α in Crohn disease(CD)model rats. Methods:Thirty-six Sprague-Dawley rats were randomly divided into a normal control(NC)group with 10 rats and a modeling group with 26 rats.In the modeling group,the CD rat model was prepared with 2,4,6-trinitrobenzene sulfonic acid(TNBS)enema.After successful modeling,the rats were randomly divided into a CD model(CD)group,an EA group,and a Western medicine(WM)group.The NC and CD groups received no treatment;the EA group was treated with EA for 20 min each time,with 7 consecutive days'intervention;the WM group received mesalazine enteric-coated tablet solution by gavage once a day for 7 d.The changes in body mass and disease activity index(DAI)were observed.Serum IL-1β and TNF-α were determined by enzyme-linked immunosorbent assay.Hematoxylin-eosin staining was used to observe the pathological changes of colon tissues,and 16S rDNA sequencing was used to analyze the structural changes of gut microbiota. Results:Compared with the NC group,the body mass of rats in the CD group decreased(P<0.01),and the DAI score increased(P<0.01);the colon tissue structure was disordered,and many inflammatory cells were present;also,IL-1β and TNF-α increased significantly(P<0.01).As a result,the diversity of gut microbiota decreased,and the abundance of some conditional pathogenic bacteria(such as Prevotella)increased,while the abundance of beneficial bacteria(such as Lactobacillus,Rochella,and Spirillum)decreased.After the intervention,compared with the CD group,the body mass of rats in the EA group and WM group increased(P<0.01);the DAI score decreased(P<0.01),the colon tissue structure improved,and the IL-1β and TNF-α levels decreased(P<0.01);the diversity of gut microbiota increased(P<0.05),and the abundance of some conditional pathogenic bacteria decreased while the abundance of beneficial bacteria increased in the EA group;whereas the diversity of gut microbiota in the WM group was not statistically different(P>0.05). Conclusion:EA can reduce the damage of colon mucosa,regulate the imbalance of gut microbiota,and inhibit the serum inflammatory factor IL-1β and TNF-α expression in CD rats.

Objective:To investigate the risk factors for positive margins after endoscopic submucosal dissection (ESD) for early gastric cancer and precancerous lesions, and to follow up the recurrence.Methods:The endoscopic, clinical and pathological data of 489 patients with early gastric cancer or precancerous lesions treated by ESD in Fujian Provincial Hospital from January 2015 to December 2020 were retrospectively collected. They were categorized into a negative group (371 cases), a low-grade intraepithelial neoplasia (LGIN)-positive group (79 cases), and a high-grade intraepithelial neoplasia (HGIN) or cancer-positive group (39 cases) according to the different margins. Logistic regression was used to analyze the risk factors for positive margins, the Kaplan-Meier method and log-rank test to compare the risk of recurrence in different margin groups, and the Cox proportional risk regression model to explore the associated factors that caused recurrence in those with positive margins.Results:In the 489 patients, the positive resection margin rate was 24.1% (118/489), of which HGIN or cancer accounted for 33.1% (39/118). LGIN-positive margin was more likely to occur for lesions larger than 10 cm 2 ( OR=1.58, 95% CI: 1.13-2.08, P=0.033), in the presence of ulcers ( OR=2.92, 95% CI: 1.37-4.54, P=0.012) and for 1-2 years of ESD experience [ OR=1.69 (1-2 years VS 5-6 years), 95% CI: 1.51-1.94, P=0.026]. Those located in the upper 1/3 of the stomach [ OR=3.64 (upper 1/3 VS lower 1/3), 95% CI: 1.27-5.50 P=0.010] and submucosal infiltration (SM1 VS M1+M2: OR=2.37, 95% CI: 1.04-5.72, P=0.028; SM2 VS M1+M2: OR=6.08, 95% CI: 1.31-12.75, P=0.002) were high risk factors for HGIN/cancer-positive margin. Postoperative follow-up was completed in 337 patients, with a median follow-up time of 26.0 (22) months. The overall cumulative recurrence was 5.3% (18/337), 2.1% (5/239) in the negative margin group, 8.3% (6/72) in the LGIN-positive margin group, and 26.9% (7/26) in the HGIN/cancer-positive group, with statistically significant differences among the 3 groups ( P<0.05). Risk factors for recurrence in the positive margin group included positive basal margins ( HR=5.17, 95% CI: 1.47-14.09, P=0.011) and SM1 invasion ( HR=4.82, 95% CI: 1.38-14.77, P=0.013). Conclusion:Positive margins after ESD for early gastric cancer and precancerous lesions are related to lesion location, size, presence of ulceration, depth of infiltration, and endoscopists' experience. The overall risk of recurrence is higher in those with positive margins than in those with negative margins. Additional treatments need to be considered comprehensively for those with submucosal invasion and positive basal margins.

Objective:To investigate the longitudinal changes and influencing factors of out-of-hospital functional exercise compliance in patients with lumbar disc herniation treated conservatively based on patient-based compliance curve (PBCC) .Methods:From April to September 2020, the convenient sampling method was used to select 88 patients with lumbar disc herniation treated conservatively in Hangzhou Hospital of Traditional Chinese Medicine as the research objects. The score of out-of-hospital functional exercise compliance questionnaire of patients with lumbar disc herniation was measured from the time of discharge, once every 2 weeks, and the PBCC of functional exercise was drawn after continuous measurement to the 24th week , taking time as independent variable ( x) and functional exercise compliance index as dependent variable ( y) . Univariate and multiple linear regression analyses were used to explore the influencing factors of 24-week compliance index. A total of 88 questionnaires were distributed, among which 3 patients withdrew due to surgery and 11 patients were lost to follow-up. A total of 74 valid questionnaires were finally recovered, with an effective recovery rate of 84.09% (74/88) . Results:Within 24 weeks of discharge, the PBCC formula of the total functional exercise score of patients was y=0.005 x3-0.196 x2+1.436 x+59.197 ( R2=0.898, F=26.517, P<0.01) . The PBCC formula of physical exercise was y=0.008 x3-0.337 x2+3.616 x+56.153 ( R2=0.878, F=21.570, P<0.01) . The formula of exercise effect supervision PBCC was y=-0.006 x3+0.309 x2-5.315 x+76.371 ( R2=0.910, F=30.458, P<0.01) . The PBCC formula of actively seeking advice was y=0.012 x3-0.460 x2+3.931 x+44.071 ( R2=0.865, F=19.217, P<0.01) . Except for the PBCC of effect supervision, the changes of other PBCC showed a trend of first increasing and then decreasing. The average compliance index in 24 weeks was (57.73±8.20) , the high peak appeared in the 4th week and the index was (64.08±8.91) . The low peak appeared in the 18th week and the index was (52.42±7.81) . The results of univariate analysis showed that there were statistically significant differences in the compliance level of rehabilitation exercise in patients with lumbar protrusion in different age, education level, course of lumbar protrusion, pain degree, accompanying symptoms, functional exercise methods and disease knowledge ( P<0.05) . The results of multiple linear regression analysis showed that the main influencing factors of functional exercise compliance of patients were disease knowledge, pain degree, course of lumbar protrusion and functional exercise method ( P<0.05) . Conclusions:The functional exercise compliance of lumbar disc herniation patients shows a horizontal "S" -like dynamic change. During the period of decreased compliance, regular rehabilitation guidance and health education, pain management and prevention of recurrence are necessary measures to improve functional exercise compliance of patients.

Objective: To investigate the effect of human antigen R on lysosomal acidification during autophagy in mouse cardiomyocytes cultured in vitro. Methods: The hearts of 20 C57BL/6 mice aged 1-2 days no matter male or female were isolated to culture primary cardiomyocytes which were used in the following experiments. (1) The cells were divided into 5 groups according to the random number table (the same grouping method below), i. e., normal control group and sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups. The cells in normal control group were routinely cultured for 54.0 h with Dulbecco′s modified Eagle medium/nutrient mixture F12 (DMEM/F12) medium (the same regular culture condition below), and the cells in sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups were firstly regularly cultured for 53.5, 53.0, 51.0, 48.0 h and then cultured with replaced sugar-free serum-free medium for 0.5, 1.0, 3.0, and 6.0 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 Ⅱ (LC3Ⅱ), autophagy-related protein 5, and adenosine triphosphatase V1 region E1 subunit (ATP6V1E1) were detected by Western blotting. (2) The cells were divided into normal control group and sugar-free serum-free 3.0 h group. The cells in corresponding groups were treated the same as those in experiment (1), and the cell lysosomal acidification level was observed and detected under a laser scanning confocal microscope. (3) Two batches of cells were grouped and treated the same as those in experiment (1). The protein expression of human antigen R in the whole protein of cells of one batch and its protein expression in the cytoplasm and nucleus protein of cells of the other batch were detected by Western blotting. (4) The cells were divided into normal control group, simple control small interfering RNA (siRNA) group, simple human antigen R-siRNA1 (HuR-siRNA1) group, simple HuR-siRNA2 group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group. After 48 hours of regular culture, the cells in simple control siRNA group and sugar-free serum-free+ control siRNA group were transfected with negative control siRNA for 6 h, the cells in simple HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA1 group were transfected with HuR-siRNA1 for 6 h, and the cells in simple HuR-siRNA2 group and sugar-free serum-free+ HuR-siRNA2 group were transfected with HuR-siRNA2 for 6 h. Hereafter, the cells in these 8 groups were continuously cultured for 48 h with regular conditon, and then the cells in normal control group and each simple siRNA-treated group were replaced with DMEM/F12 medium, the cells in the other groups were replaced with sugar-free serum-free medium, and they were cultured for 3 h. The protein expression of human antigen R in the whole protein of cells was detected by Western blotting. (5) Two batches of cells were divided into sugar-free serum-free+ control siRNA group and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The distribution and expression of human antigen R in the cells of one batch were observed and detected by immunofluorescence method, and the lysosomal acidification level in the cells of the other batch was observed and detected under a laser scanning confocal microscope. (6) Three batches of cells were divided into sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group, and the cells in corresponding groups were treated the same as those in experiment (4). The protein expressions of cathepsin D in the whole protein of cells of one batch, human antigen R in the cytoplasm protein of cells of one batch, and ATP6V1E1 in the whole protein of cells of the other batch were detected by Western blotting. (7) The cells were divided into normal control group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The mRNA expression of ATP6V1E1 in cells was detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. The sample number of each experiment was 3. Data were processed with independent data t test, one-way analysis of variance, least significant difference t test, and Bonferroni correction. Results: (1) Compared with those of normal control group, the protein expressions of LC3Ⅱ and ATP6V1E1 in the whole protein of cells of sugar-free serum-free 1.0, 3.0, and 6.0 h groups were significantly increased (t=12.16, 4.05, 4.82, 11.64, 3.29, 8.37, P<0.05 or P<0.01). Compared with that of normal control group, the protein expression of autophagy-related protein 5 in the whole protein of cells of sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups was significantly increased (t=6.88, 10.56, 5.76, 9.91, P<0.05 or P<0.01). (2) Compared with 1.03±0.08 of normal control group, the lysosomal acidification level in the cells of sugar-free serum-free 3.0 group (2.92±0.30) was significantly increased (t=6.01, P<0.01). (3) There was no statistically significant difference in the overall comparison of protein expression of human antigen R in the whole protein of cells among the 5 groups (F=1.09, P>0.05). Compared with that of normal control group, the protein expression of human antigen R in the cytoplasm protein of cells was significantly increased in sugar-free serum-free 1.0, 3.0, and 6.0 h groups (t=43.05, 11.07, 5.39, P<0.05 or P<0.01), while the protein expression of human antigen R in the nucleus protein of cells was significantly decreased in sugar-free serum-free 3.0 and 6.0 h groups (t=11.18, 12.71, P<0.01). (4) Compared with that of simple control siRNA group, the protein expression of human antigen R in the whole protein of cells of simple HuR-siRNA1 group and simple HuR-siRNA2 group was significantly decreased (t=4.82, 4.44, P<0.05). Compared with that of sugar-free serum-free+ control siRNA group, the protein expression of human antigen R in the whole protein of cells of sugar-free serum-free+ HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA2 group was significantly decreased (t=4.39, 6.27, P<0.05). (5) Compared with those of sugar-free serum-free+ control siRNA group, the distribution of human antigen R in the cytoplasm of cells and its expression level were significantly decreased in sugar-free serum-free+ HuR-siRNA1 group (t=10.13, P<0.01). Compared with 1.00±0.06 of sugar-free serum-free+ control siRNA group, the lysosomal acidification level (0.73±0.06) in the cells of sugar-free serum-free+ HuR-siRNA1 group was significantly decreased (t=3.28, P<0.01). (6) Compared with those of sugar-free serum-free+ control siRNA group, the protein expressions of cathepsin D in the whole protein of cells, human antigen R in the cytoplasm protein of cells, and ATP6V1E1 in the whole protein of cells were significantly decreased in sugar-free serum-free+ HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA2 group (t=4.16, 3.99, 4.81, 5.07, 11.68, 12.97, P<0.05 or P<0.01). (7) Compared with that of normal control group, the mRNA expression of ATP6V1E1 in the cells of sugar-free serum-free 3.0 h group was significantly increased (t=5.51, P<0.05). Compared with that of sugar-free serum-free+ control siRNA group, the mRNA expression of ATP6V1E1 in the cells of sugar-free serum-free+ HuR-siRNA1 group was significantly decreased (t=5.97, P<0.05). Conclusions After sugar-free serum-free treatment in vitro, the autophagy in mouse primary cardiomyocytes is activated, the lysosomal acidification is enhanced, and the expression of human antigen R in cytoplasm is increased. Human antigen R function is activated and involved in maintaining lysosomal acidification during autophagy in mouse cardiomyocytes.

Objective: To explore the effects of decline of pH value on cardiomyocyte viability of rats, and to analyze the possible mechanism. Methods: Hearts of five newborn Sprague-Dawley rats were isolated, and then primary cardiomyocytes were cultured and used in the following experiments. (1) The primary cardiomyocytes were divided into pH 7.4+ 6 h, pH 7.0+ 6 h, pH 6.5+ 6 h, pH 6.0+ 6 h, pH 6.5+ 1 h, and pH 6.5+ 3 h groups according to the random number table, with 4 wells in each group. After being routinely cultured for 48 h (similarly hereinafter), cells in pH 7.4+ 6 h, pH 7.0+ 6 h, pH 6.5+ 6 h, and pH 6.0+ 6 h groups were cultured with pH 7.4, pH 7.0, pH 6.5, and pH 6.0 DMEM-F12 medium (similarly hereinafter), respectively, and then they were cultured for 6 h. Cells in pH 6.5+ 1 h and pH 6.5+ 3 h groups were cultured with pH 6.5 medium, and then they were cultured for 1 h and 3 h, respectively. Viability of cells was detected by methyl-thiazolyl-tetrazolium (MTT) method. (2) The primary cardiomyocytes were divided into pH 7.4, pH 6.5, and pH 6.5+ taxol groups according to the random number table, with 2 wells in each group. Cells in pH 7.4 group were cultured with pH 7.4 medium, while cells in pH 6.5 and pH 6.5+ taxol groups were cultured with pH 6.5 medium. Cells in pH 6.5+ taxol group were added with taxol of a final molarity of 0.2 μmol/L in addition, and then they were cultured for 6 h. Morphology and density of microtubule of cells was detected by immunofluorescence assay. (3) The primary cardiomyocytes were grouped and treated as in experiment (2), with 2 wells in each group. The expressions of polymerized microtubulin and free microtubulin were determined with Western blotting. (4) The primary cardiomyocytes were grouped and treated as in experiment (2), with 4 wells in each group. Viability of cells after treated with taxol was detected by MTT method. Data were processed with one-way analysis of variance and LSD-t test. Results: (1) The viability of cells in pH 7.4+ 6 h, pH 7.0+ 6 h, pH 6.5+ 6 h, pH 6.0+ 6 h, pH 6.5+ 1 h, and pH 6.5+ 3 h groups were 1.00±0.08, 0.90±0.08, 0.85±0.06, 0.83±0.04, 0.91±0.10, and 0.89±0.10, respectively. Compared with that in pH 7.4+ 6 h group, viability of cells in pH 7.0+ 6 h, pH 6.5+ 6 h, pH 6.0+ 6 h, pH 6.5+ 1 h, and pH 6.5+ 3 h groups were all decreased in different degrees (t=2.476, 4.002, 4.996, 2.168, 2.400, P<0.05). (2) Microtubules of cells in pH 7.4 group were radially distributed around the nucleus with clear tubular structure. Compared with that in pH 7.4 group, the skeleton of microtubules of cells in pH 6.5 group was obviously damaged, with broken structure of microtubule and reduced density. Compared with that in pH 6.5 group, the damage degree of microtubules of cells in pH 6.5+ taxol group was obviously alleviated, and the structure of microtubules basically returned to normal. (3) Compared with that in pH 7.4 group, the expression of free microtubulin of cells in pH 6.5 group was significantly increased (t=3.030, P<0.05), while the expression of polymerized microtubulin of cells was significantly decreased (t=8.604, P<0.05). Compared with that in pH 6.5 group, the expression of free microtubulin of cells in pH 6.5+ taxol group was significantly decreased (t=4.559, P<0.05), while the expression of polymerized microtubulin of cells was significantly increased (t=5.472, P<0.05). (4) Viability of cells in pH 7.4, pH 6.5, and pH 6.5+ taxol groups were 1.00±0.10, 0.83±0.04, and 0.93±0.10, respectively. Compared with that in pH 7.4 group, the viability of cells in pH 6.5 group was obviously declined (t=4.412, P<0.05). Compared with that in pH 6.5 group, the viability of cells in pH 6.5+ taxol group was obviously increased (t=2.461, P<0.05). Conclusions The decline of pH value reduces the viability of cardiomyocytes of rats through destroying the skeleton of microtubule. Stabilizing microtubule skeleton can significantly reduce acidic treatment-induced damage and ameliorate cardiomyocyte viability.

Objective To investigate current status of teachers' informational teaching ability in medical vocational college and provide reference for the improvement of teachers' informational teaching ability. Methods Convenience sampling method was adopted in survey. From September to December 2016,a total of 204 teachers from medical vocational college were investigated with the general information questionnaire and informational teaching ability questionnaire.Resluts The score of medical vocational college teachers' informational teaching ability was (3.50±0.66).Age and educational degree were the main factors influencing teachers' informational teaching ability (P<0.05).Conclusions Teachers' informational teaching ability in medical vocational was at medium level.In the future, we should pay attention to the two factors of age and education degree, and carry out the hierarchical and targeted training of teachers' informational teaching ability in medical vocational college.

Objective To investigate influence of nicotinic acid adenine dinucleotide phosphate (NAADP) on autophagy in hypoxic cardiomyocytes of rats and its mechanism.Methods Five neonatal Sprague-Dawley rats were collected and sacrificed to harvest the hearts,and primary cardiomyocytes were separated for the following experiments.(1) Primary cardiomyocytes were collected and divided into normoxia group,hypoxia 9 h group,and hypoxia 9 h + NAADP group according to random number table,with 5 wells in each group.Cells in normoxia group were cultured routinely in the constant temperature incubator at 37 ℃ for 9 hours.Cells in hypoxia 9 h group and hypoxia 9 h + NAADP group were cultured in hypoxic incubator with volume fraction 94％ nitrogen,5％ carbon dioxide,and 1％ oxygen for 9 hours.Before hypoxia,cells in hypoxia 9 h + NAADP group were dealt with final amount-of-substance concentration 10 μmol/L NAADP.Cell counting kit 8 was used to measure cell viability.(2) Primary cardiomyocytes were collected and divided into normoxia group,hypoxia 9 h group,hypoxia 9 h + NAADP group,hypoxia 9 h + tran-Ned-19 group,and hypoxia 9 h + trans-Ned-19 + NAADP group according to the random number table,with 2 wells in each group.Cells in normoxia group were cultured routinely in the constant temperature incubator at 37 ℃ for 9 hours.And cells in the other 4 groups were cultured in hypoxic incubator as that in experiment (1) Before hypoxia,cells in hypoxia 9 h + NAADP group were dealt with amount-of-substance concentration 10 μmol/L NAADP,cells in hypoxia 9 h + tran-Ned-19 group were dealt with amount-of-substance concentration 1 μmol/L trans-Ned-19,and cells in hypoxia 9 h + trans-Ned-19 + NAADP group were dealt with amount-of-substance concentration 10 μmol/L NAADP and 1 μmol/L trans-Ned-19.Protein expressions of microtubule associated protein 1 light chain 3-Ⅱ and P62 were detected by Western blotting.(3) Primary cardiomyocytes were collected and grouped as those in experiment (1).The lysosomal acidity was determined by immunofluorescence method.Data were processed with one-way analysis of variance and least-significant difference test.Results (1) The cell viability in normoxia group was 1.114 ± 0.024,which was significantly higher than 0.685 ± 0.079 of cells in hypoxia 9 h group (P ＜ 0.01).The cell viability of hypoxia 9 h + NAADP group was 0.886 ± 0.061,which was obviously higher than that of cells in hypoxia 9 h group (P ＜0.05).(2) Expressions of microtubule-associated protein 1 light chain 3-Ⅱ and P62 of cells in hypoxia 9 h group were significantly higher than those of cells in normoxia group (P ＜ 0.0l).Compared with those in hypoxia 9 h group,expression of P62 in hypoxia 9 h + NAADP group was significantly decreased (P ＜ 0.01),while expression of microtubule-associated protein 1 light chain 3-Ⅱ did not change significantly (P ＞ 0.05).There were no significantly statistical difference in expressions of microtubule-associated protein 1 light chain 3-Ⅱ and P62 between hypoxia 9 h group and hypoxia 9 h + trans-Ned-19 group (P ＞ 0.05).Compared with those of cells in hypoxia 9 h + NAADP group,expression of P62 of cells in hypoxia 9 h + trans-Ned-19 + NAADP group was obviously increased (P ＜ 0.01),while expression of microtubule-associated protein 1 light chain 3-Ⅱ did not change significantly (P ＞ 0.05).(3) The intensity of green fluorescence of cells in normoxia group was strong and co-localized well with red fluorescence,and internal environment of lysosome was with stronger acidity.The intensity of green fluorescence in cells of hypoxia 9 h group was significantly lower than that of cells in normoxia group,and acidity of internal environment of lysosome was weakened.The intensity of green fluorescence and acidity of internal environment of lysosome in hypoxia 9 h + NAADP were significantly stronger than those of cells in hypoxia 9 h group,but significantly lower than those of cells in normoxia group.Conclusions NAADP can improve myocardial cell viability through acidifying internal environment of lysosome of cardiomyocyte after hypoxia,promoting degradation of autophagosomes,reducing autophagic lysosomal accumulation,and repairing damaged autophagie flow.

Objective To explore the application of mobile app-based blended teaching inNursing Fundamentals.Methods Totally 101 nursing students from two classes of the School of Clinical Medicine and Nursing, Suzhou Vocational Health College were selected in September 2016. The two classes were divided into the treatment group (n=50) and the control group (n=51) respectively by drawing lots. The control group received conventional teaching, while the treatment group received mobile app-based blended teaching. The theoretical and practical performance were compared between the two groups at the end of course. Nursing students in the treatment group were evaluated with the self-designed Teaching Model Evaluation Questionnaire. Nursing students in the two groups were evaluated using the Autonomous Learning Competency Scale for Nursing Undergraduates before and after intervention.Results The theoretical and practical performance of the treatment group was (82.76±9.05) and (84.94±5.93) post intervention, both higher than that of the control group (t=-3.32, -7.99;P<0.05). The nursing students in the treatment group scored (94.82±7.31) post intervention, higher than that of the control group (t=-2.31,P<0.05). All nursing students in the treatment group believed the mobile app-based blended teaching was better than conventional teaching. Conclusions The mobile app-based blended teaching can improve the learning outcome of nursing students.

Objective To understand the new standards for nursing grading,different level of care on the feelings of patients for nursing service,in order to improve the nursing service mode and accelerate hospitals to promote sustained and healthy development of the high quality nursing service and provide the beneficial reference. Methods Using stratified sampling,self-designed questionnaire was utilized to cross-sectional investigate the multi-dimensional situation of 286 patients in three class Ⅲ grade A hospitals in Guangdong province from June 2015 to October 2015. Results Different levels of care for patients with nursing services in six aspects of the feelings were different,and the ward management,interpersonal support and comprehensive feelings compared that the difference was statistically significant(P < 0.05). Three level care patients in these three aspects of the feelings were poor,but special care of patients with different dimensions were better. Conclusions Three-level care patients has a relatively poor experience in nursing services,so it is recommended to enhance the concept of rehabilitation through ward management to interpersonal support,so as to meet their individual needs. Simultaneously,in order to implement the new nursing grading standards well, we can try to refine and quantify the classification of care and combined with the patient's feelings to determine the classification of care,so it more accurately reflect the actual situation of patients, and help improve the quality of care services.

Objective To detect and explore the expression and clinical significance of dual specificity tyrosine phosphorylation regulated kinase1b (Dyrk1b) in the specimens and cells of cervical lesions. Methods (1)All the data were collected from 75 patients with cervical cancer and 52 cases with squamous intraepithelial lesion(SIL)admitted in the First Affiliated Hospital of Dalian Medical College during Jan. 2011 to Dec. 2013 and confirmed by pathological examination, included 60 cases of stageⅠand 15 cases of stageⅡ, 12 cases with low-grade squamous intraepithelial lesion(LSIL)and 40 cases with high-grade squamous intraepithelial lesion(HSIL). While, 28 cases with chronic cervicitis were chosen as the control group. The protein expression of Dyrk1b was detected by immunohistochemistry among the four groups.(2)The expression of Dyrk1b in HeLa and SiHa cells were detected by western blot method and the expression of Dyrk1b protein were also detected after treatment of AZ191 (5, 10 μmol/L) for 48 hours in HeLa and SiHa cells.(3)The cellular survival and proliferation of HeLa and SiHa cells treated by different concentrations of AZ191(2.5, 5, 10, 25, 50, 100 μmol/L)for 48 hours were detected by methyl thiazolyl tetrazolium (MTT) assay.(4)The rate of apoptosis of HeLa and SiHa cells was detected by flowcytometry after treatment of AZ191 (5, 10μmol/L) for 48 hours. Results (1)The positive rates of Dyrk1b protein in chronic cervicitis, LSIL, HSIL and cervical squamous cancer by immunohistochemistry were 11%(3/28), 1/12, 42%(17/40)and 71%(53/75), respectively. The expression of Dyrk1b in cervical squamous cancer and HISL were higher than those in LSIL and chronic cervicitis (P<0.01), there were significant difference between cervical squamous cancer and HSIL, or between HSIL and LSIL(all P<0.05), while there were not significant difference between LSIL and chronic cervicitis(P>0.05). Expression of Dyrk1b was correlated with stromal invasion depth of cervical cancer (P<0.05), but not with age, clinical stage, lymph node metastasis, and serum squamous cell carcinom antigen(SCC-Ag)levels (all P>0.05). (2) Dyrk1b protein was expressed in different levels in HeLa and SiHa cells, and the expression of Dyrk1b was decreased gradually as the increased of the concentration of AZ191 in both HeLa and SiHa cells by treatment of AZ191 for 48 hours. (3) Different concentration of AZ191 treated on cervical cancer cells could inhibit the cellular proliferation and induce cell apoptosis in a concentration-dependent manner(P<0.01), concomitant to the decreased cell survival rate. The apoptosis rate of HeLa and SiHa were increased significantly after 10μmol/L AZ191-treatment for 48 hours, but no any difference induced by 5 μmol/L AZ191-treatment compared to control group. Also,there was no any difference between Hela and SiHa cells in either inhibitory effect or apoptosis rate induced by AZ191. Conclusions Dyrk1b is over-expressed in either specimens or cells of cervical cancer. The expression of Dyrk1b protein in cervical lesions is increased as the progression of disease. Dyrk1b inhibitor AZ191 could inhibit cellular proliferation and induce apoptosis in a concentration-dependent manner in cervical cancer cells.