Objective To culture and expand primary rat aortic vascular stem cells in vitro and evaluate their characteristics as mesenchymal stem cells.Methods The thoracic and abdominal aortas isolated from 2-to 3-week-old Sprague-Dawley rats were cut into vascular segments 2.0 mm in length and cultured in culture flasks till adhesion and solidification of the outer membranes.The primary cells were further cultured to 80%-90%confluence before passaging.The morphology and growth characteristics of the cells were observed under a microscope,and the expressions of surface marker CD molecules on the cells was analyzed using flow cytometry.Adipogenic and osteogenic differentiation assays were performed to assess the capacity of the cells for multilineage differentiation.Results After 3 days of culture,a small number of spindle,star-shaped or polygonal cells migrated out from the peripheral of the vascular segments.At 5-6 days,island-like cell clusters occurred and the cells began to proliferate rapidly.The cell clusters expanded radially and showed signs of cell cloning.At 7-8 days,the cells fused into sheets and displayed a vortex-like distribution.The cells in the third passage presented with a uniform morphology,showing a typical fibroblast-like arrangement.Flow cytometry showed that the cells expressed predominantly CD44(80.3%),CD73(62.2%)and CD90(46.8%)with low expressions of CD34(1.1%),CD45(0.2%)and CD11b/c(0.2%).Adipogenic and osteogenic differentiation experiments demonstrated that the cells were capable of lipogenic and osteogenic differentiation in vitro.Conclusion Rat aortic vascular stem cells with mesenchymal stem cell characteristics can be successfully isolated and cultured by adherent culture of the segmented outer membrane.

Objective To culture and expand primary rat aortic vascular stem cells in vitro and evaluate their characteristics as mesenchymal stem cells.Methods The thoracic and abdominal aortas isolated from 2-to 3-week-old Sprague-Dawley rats were cut into vascular segments 2.0 mm in length and cultured in culture flasks till adhesion and solidification of the outer membranes.The primary cells were further cultured to 80%-90%confluence before passaging.The morphology and growth characteristics of the cells were observed under a microscope,and the expressions of surface marker CD molecules on the cells was analyzed using flow cytometry.Adipogenic and osteogenic differentiation assays were performed to assess the capacity of the cells for multilineage differentiation.Results After 3 days of culture,a small number of spindle,star-shaped or polygonal cells migrated out from the peripheral of the vascular segments.At 5-6 days,island-like cell clusters occurred and the cells began to proliferate rapidly.The cell clusters expanded radially and showed signs of cell cloning.At 7-8 days,the cells fused into sheets and displayed a vortex-like distribution.The cells in the third passage presented with a uniform morphology,showing a typical fibroblast-like arrangement.Flow cytometry showed that the cells expressed predominantly CD44(80.3%),CD73(62.2%)and CD90(46.8%)with low expressions of CD34(1.1%),CD45(0.2%)and CD11b/c(0.2%).Adipogenic and osteogenic differentiation experiments demonstrated that the cells were capable of lipogenic and osteogenic differentiation in vitro.Conclusion Rat aortic vascular stem cells with mesenchymal stem cell characteristics can be successfully isolated and cultured by adherent culture of the segmented outer membrane.

Objective    To investigate the feasibility and safety of hybrid coronary revascularization (HCR) in patients with multivessel coronary artery disease (MVCAD). Methods    A total of 50 patients with MVCAD who underwent HCR technique in our heart center from May 2016 to April 2019 were included in this study (a HCR group), including 38 patients who underwent two-stage HCR and 12 patients one-stop HCR. There were 39 males and 11 females, with an average age of 62.4 (46-82) years. Another 482 patients who underwent conventional median incision under off-pump coronary artery bypass grafting (OPCAB) at the same period were selected as control (an OPCAB group), including 392 males and 90 females, with an average age of 64.2 (48-84) years. The safety and feasibility of HCR were evaluated and compared with conventional OPCAB technique. Results    There was no perioperative death in both groups. Compared with the OPCAB, HCR was associated with shorter operation time, less chest tube drainage, lower requirement of blood transfusion, shorter mechanical ventilation time and shorter postoperative intensive care unit (ICU) stay (P<0.05). There was no statistical difference in the incidence of major adverse cardiac or cerebrovascular events during the follow-up of 6 to 36 months between the two groups. Conclusion    HCR provides favorable short and mid-term outcomes for selected patients with MVCAD compared with conventional OPCAB.

Objective:To evaluate the results of butterfly cartilage myringoplasty for anterior quadrant tympanic perforation under endoscope.Methods:Thirty-eight patients with anterior quadrant tympanic perforations who were subjected to endoscopic butterfly cartilage myringoplasty from April 2016 to October 2018 were included in this study, including 16 males and 22 females, with an average age of (34.5±14.2) years. The patients were reviewed retrospectively, and the pre-and post-operative pure tone audiometry (PTA) thresholds, pre-and post-operative air-bone gaps (ABG), post-operative graft success rates and complications were evaluated. SPSS 23.0 was used to analyze data.Results:Mean post-operative follow-up duration was (9.4±3.1) months (range 6-18 months). The graft survival rate was 94.7% (36/38) . The preoperative and postoperative mean PTA was (30.9±8.9) dB HL and (21.4±7.7) dB HL respectively. Preoperative and postoperative mean ABG was (18.4±6.3) dB and (10.8±6.0) dB respectively. There was significant difference between pre-and postoperative PTA and ABG ( t=5.353 and 4.162, P<0.05 for both). A postoperative ABG reduction of (8.3±1.5) dB was reached. Two (4.7%) patients had postoperative myringitis, two (4.7%) had recurrent perforation, and one (2.4%) had lateral healing of transplanted tympanic membrane in the postoperative follow-ups. No intratympanic cholesteatoma was observed. Conclusions:Endoscopic butterfly inlay myringoplasty is a reliable, minimally invasive alternative method to repair anterior tympanic membrane perforations, with high closure rate and low risk of complications.

Objective To identify the predictors of postoperative acute kidney injury in patients undergoing surgery for Stanford type A acute aortic dissection. Methods A total of 220 patients who underwent surgery for type A acute aortic dissection in Qingdao Municipal Hospital from September 2010 to September 2017 were divided into two groups including a group A and a group B based on whether acute kidney injury occurred or not after surgery. There were 40 patients with 29 males and 11 females with the mean age of 54.6±9.2 years in the group A, 180 patients with 133 males and 47 females with the mean age of 48.5±7.9 years in the group B. Univariate and multivariate analyses (logistic regression) were used to identify the predictive risk factors. Results Overall in-hospital mortality was 5.5%. In univariate analysis, there were statistically significant differences with respect to the age, preoperative creatinine, preoperative white blood cell, the European system for cardiac operative risk evaluation (EuroSCORE), total cardiopulmonary bypass (CPB) time, deep hypothermic circulatory arrest (DHCA) time, arch replacement, red blood cell transfusion intraoperative and in 24 hours postoperatively, postoperative mechanical ventilation time, ICU stay duration, hospital stay duration and in hospital mortality. Multivariate logistic analysis showed that preoperative creatinine, preoperative white blood cell, CPB time, and red blood cell transfusion intraoperative and in 24 hours postoperatively were the independent predictors for postoperative acute kidney injury. Conclusion The incidence of acute kidney injury is high after surgery for acute Stanford type A aortic dissection. It can be predicted based on above factors, for patients with these risk factors, more perioperative care strategies are needed in order to induce the incidence of acute kidney injury.

Otorhinolaryngology is a high-risk department of a hospital, where there are many emergency critical diseases, common multiple diseases and major malignant diseases. Therefore, it is easy to cause many medical ethical problems. This paper analyzed the clinical status and characteristics of various otolaryngology diseases systematically, and expounded the related medical ethical issues in the diagnosis and treatment of otorhinolaryngology, including doctor-patient trust, safety and informed consent. Finally, the paper put forward a number of measures to do well the psychological evaluation and nursing care of patients, improve the professional skills of medical and nursing care, formulate the treatment plan of diseases, enhance the supervision and management of the network, and promote the social support of patients. The aim was to alleviate the "doctor-patient conflict" and create a harmonious medical environment.

Objective To understand the needs to reform curriculum for the graduate students in health management, and to provide evidence for the reform of the curriculum system. Methods From Jan-uary 2015 to May from China's colleges and universities carrying out health management professional post-graduate education, graduate students and supervisors from 3 universities were selected, and self-designed questionnaires were used to investigate their views on training objectives, abilities and curriculum settings, as well as the needs and suggestions for curriculum reform. The questionnaire survey data were collected and analyzed by SPSS 17.0. Results 120 questionnaires were issued and 100 were recovered in the study, with a recovery rate of 83.3％. 84.0％ of the graduate students did not manage relevant work experience before entering the school. The graduate students thought that we should focus on the research and teaching ability training related to the management of health services, and then the cultivation of management ability. There were 60 questionnaires for instructors, and 51 were recovered, with a recovery rate of 85.0％. With regard to the demand for ability training , teachers and graduate students tended to agree on the goal and focus of the training. 100.0％instructors believed that learning was an important course, only 11.2％ of the students thought that the course of study was not important, but to the current status of health manage-ment professional postgraduate courses, 79.3％ instructors believed that the current curriculum was reasonable, and only 53.5％of the students thought it reasonable. Conclusion The emphasis should be given to optimizing the course setting, adding practical and innovative courses, and strengthening the training of scientific research, etc., which is the only way to improve the quality of health management postgraduate education and meet the needs of health development.

Objective To establish a mouse embryonic stem cell test (mEST) model and human embryonic stem cell test (hEST) model, to evaluate the embryotoxicity of di(2-ethylhexyl) phthalate (DEHP). Methods We developed mEST and hEST models according to the European Centre for the Validation of Alternative Methods (ECVAM). We used penicillin G (PN-G) as the standard negative reference and 5-fluorouracil (5-FU) as the standard positive reference, respectively, to verify validity of the models. Based on model validity, mouse embryonic stem cells D3 (mESC-D3), mouse Balb/c-3T3 (3T3), and human embryonic stem cells H9 (hESC-H9) were administered different concentrations of DEHP (15.6, 31.2, 62.5, 125.0, 250.0, 500.0, and 1 000.0 μg/ml) for 7 days. A cell counting Kit-8 was used to detect the 50%inhibitory proliferation concentration (IC50) of mESC-D3 cells, 3T3 cells, and hESC-H9 with DEHP. mESC-D3 and hESC-H9 were treated with DEHP (15.6, 31.2, 62.5, 125.0, 250.0μg/ml, and 500.0μg/ml) for 10 days based on the cytotoxicity results. At day 10, the expression of cardiomyocyte differentiation gene alpha-myosin heavy chain (α-MHC) was detected by real-time PCR and the 50% inhibition of cardiomyocycte differentiation (ID50) determined. Based on the values of IC50 and ID50, functionsⅠ,ⅡandⅡcould be calculated by three linear discriminant functions in the EST model and the embryotoxicity of DEHP described by comparing the three functions. Results Nontrophoblast lineage both ES cells were cultured under optimal conditions and highly expressed hESC markers OCT4 , SSEA4, and TRA-1-60. The embryoid bodies formed were uniform in size and shape, and these results were highly repeatable. The PN-G and 5-FU results coincided with the prediction by ECVAM. Validation of our EST models was satisfactory. Results of the three endpoints of DEHP in mEST were 197.3 μg/ml (IC50 3T3), 210.0 μg/ml (IC50 D3) and 246.8μg/ml (ID50 D3). DEHP was evaluated to be a nonembryotoxic compound based on values of functionⅠ(7.78), functionⅡ(7.58) and functionⅢ(-7.79). The three endpoints of DEHP in hEST were 195.4μg/ml (IC50 3T3), 184.8 μg/ml (IC50 D3), and 84.3 μg/ml (ID50). By comparing the values of function Ⅰ (3.21), function Ⅱ (5.77), and function Ⅲ (-6.46), DEHP was evaluated to be weakly embryotoxic. Conclusion DEHP was determined to be a nonembryotoxic compound by mEST and weakly embryotoxic by hEST. Therefore, hEST is a more sensible model for the evaluation of DEHP embryotoxicity.

Objective To establish a mouse embryonic stem cell test (mEST) model and human embryonic stem cell test (hEST) model, to evaluate the embryotoxicity of di(2-ethylhexyl) phthalate (DEHP). Methods We developed mEST and hEST models according to the European Centre for the Validation of Alternative Methods (ECVAM). We used penicillin G (PN-G) as the standard negative reference and 5-fluorouracil (5-FU) as the standard positive reference, respectively, to verify validity of the models. Based on model validity, mouse embryonic stem cells D3 (mESC-D3), mouse Balb/c-3T3 (3T3), and human embryonic stem cells H9 (hESC-H9) were administered different concentrations of DEHP (15.6, 31.2, 62.5, 125.0, 250.0, 500.0, and 1 000.0 μg/ml) for 7 days. A cell counting Kit-8 was used to detect the 50%inhibitory proliferation concentration (IC50) of mESC-D3 cells, 3T3 cells, and hESC-H9 with DEHP. mESC-D3 and hESC-H9 were treated with DEHP (15.6, 31.2, 62.5, 125.0, 250.0μg/ml, and 500.0μg/ml) for 10 days based on the cytotoxicity results. At day 10, the expression of cardiomyocyte differentiation gene alpha-myosin heavy chain (α-MHC) was detected by real-time PCR and the 50% inhibition of cardiomyocycte differentiation (ID50) determined. Based on the values of IC50 and ID50, functionsⅠ,ⅡandⅡcould be calculated by three linear discriminant functions in the EST model and the embryotoxicity of DEHP described by comparing the three functions. Results Nontrophoblast lineage both ES cells were cultured under optimal conditions and highly expressed hESC markers OCT4 , SSEA4, and TRA-1-60. The embryoid bodies formed were uniform in size and shape, and these results were highly repeatable. The PN-G and 5-FU results coincided with the prediction by ECVAM. Validation of our EST models was satisfactory. Results of the three endpoints of DEHP in mEST were 197.3 μg/ml (IC50 3T3), 210.0 μg/ml (IC50 D3) and 246.8μg/ml (ID50 D3). DEHP was evaluated to be a nonembryotoxic compound based on values of functionⅠ(7.78), functionⅡ(7.58) and functionⅢ(-7.79). The three endpoints of DEHP in hEST were 195.4μg/ml (IC50 3T3), 184.8 μg/ml (IC50 D3), and 84.3 μg/ml (ID50). By comparing the values of function Ⅰ (3.21), function Ⅱ (5.77), and function Ⅲ (-6.46), DEHP was evaluated to be weakly embryotoxic. Conclusion DEHP was determined to be a nonembryotoxic compound by mEST and weakly embryotoxic by hEST. Therefore, hEST is a more sensible model for the evaluation of DEHP embryotoxicity.

Objective To explore the application effect of problem-based health education mode in patients with upper urinary tract calculi .Methods Effect of two different health education modes for urolithiasis patients in Peking University Third Hospital were researched .The patients were divided into observation group with 104 cases and control group with 103 cases, according to their order of admission of odd and even numbers . Patients in the observation group were assessed by a self-designed questionnaire concerning urolithiasis knowledge (with full score 20 points).Health education was conducted according to the patients′scores on key parts of the disease .Health education of the patients was recorded , according to which problem-based health education was conducted .Routine comprehensive health education mode was conducted to patients in the control group.When leaving hospital , routine discharge education and continuing nursing care were conducted to both groups.4 weeks after the operation , satisfaction of patients were evaluated , and disease-related knowledge of the patients was evaluated again .Results Patients′knowledge and attitude to health education were not statistically different (P>0.05).The patients′scores of knowledge 4 weeks after the operation were (15.49 ±2.85) in the observation group, higher than those in the control group (10.22 ±3.41) (t =12.09, P <0.001).The patients′scores of satisfaction were (9.97 ±0.17) in the observation group, higher than those in the control group (9.89 ±0.31) (t=2.249, P=0.026).Conclusions Assessment of the patients′health knowledge by questionnaires when admitted to the hospital helps to determine their need of knowledge concerning the disease . Establishment of health education record helps to give prominence to the key points in the education .Consistent in content, fine in effect, health education has won high satisfaction among patients .