[Objective]To establish a novel noninvasive fluorescent animal model for endometriosis in vitro and in vivo.[Methods]Adenovirus encoding enhancing green fluorescent protein(Ad-eGFP)was used to transfect endometrial glandular cells and stromal cells(cells transfection and injection,Method No.1),and fragments(tissues transfection and injection,Method No.2).Transfection efficiencies were compared between the two methods in vitro.Then GFP transfected glandular cells and stromal cells suspension were injected into nude mice subcutaneously(Method No.1),taking Method No.2 as a comparison.In vivo observation last for 25 days,and positive rates and duration times of fluorescent lesions were calculated.Histological examination was used to confirmed lesion formation.[Results]On the fifth day after injection,lesion positive rate of Method No.1 was 88.9%,which was statistically significantly higher than that of Method No.2(22.2%),P=0.015＜0.05.The fluorescent positive duration of Method No.1 and No.2 were 12 ± 8 days and 7±4 days.The structures of lesions were all identified as human original endometrium by histological examination,including HE staining and immunofluoresceney.[Conclusion]Noninvasive animal model of endometriosis can be built up by subcutaneously injection of Ad-EGFP transfected endometrial glandular cells and stromal cells suspension with higher positive rate and longer observation time

Background Subretinal transplantation of retinal pigment epithelium (RPE) cells for the treatment of age-related macular degeneration (AMD) have accelerated the drive to develop xeno-free cultivation system that support the rapid differentiation of human embryonic stem cells (hESCs) into ES-RPE cells.Objective This study was to report a modified xeno-free culture system and method for accelerating derivation of hESCs to differentiate into RPE cells.Methods This study was approved by Ethic Committee of Zhongshan Ophthalmic Center.HESC H1 line was cloned and cuhured in Vitronectin XFTM-coated 6-well dish with xenogenetic-free medium.Cells were cultured in 50 ng/ml noggin,10 ng/ml DKK-1 and 10 ng/ml insulin like growth factor-1 (IGF-1) medium for 2 days,and then the concentration of noggin was decreased to 10 ng/ml and 5 ng/ml basic fibroblast growth factor (bFGF) and cultured for the following 2 days.Sequentially,noggin and bFGF were removed and cultured for 2 days.Finally,1 μmol/L CHIR99021 was added in medium for 6 days.Morphological changes in the progress of ESCs differentiation into RPE were observed by Living Cell Imaging System.The expression of Mitf and RPE65,RPE cellsspecific markers,in the cells were detected by immunofluorescence technique,and the relative expression levels of RPE cells-specific marker mRNA were assayed using real time fluorescent quantitation PCR.Results Polygonalshape monolayer cells which contained pigments were initially observed at day 14 after cultured with the cobblestonelike arrangement.Mitf and RPE65 were strongly expressed in the hES-derived RPE cells 35 days after induced,showing red fluorescence,and the cells presented hexagonal shape at cultured day 60 with numerous pigment granules in cytoplasm.Compared with before differentiation,the expression levels of Mitf mRNA in hES-RPE cells increased by (3.43±2.77) folds and (8.91 ± 2.83) folds,and the expression levels of RPE65 mRNA increased by (14.60 ± 3.94) folds and (87.16 ±9.32) folds at day 7 and day 14 after differentiation,respectively (all at P＜0.05).Conclusions A defined xeno-free culture system is successfully established by adding niacinamide,DKK-l,noggin,IGF-1 and CHIR99021 in xeno-free medium,and this system can accelerate the derivation and differentiation of hESCs into RPE-like cells.

Objective To investigate whether catalpol affects senile plaque formation and spatial learning and memory ability in the amyloid-β protein precursor/presenilin 1 (APP/PSI) double transgenic mice.Methods Three month-old APP/PS1 double transgenic mice were randomly divided into catalpoltreated and saline-treated groups (n =10),with C57 mice of the same age and genetic background as normal control group (n =10).The catalpol (in a dose of 5 mg · kg-1 · d-1) and the same amount of saline were peritoneally injected into Alzheimer' s disease (AD) model mice for 3 weeks.Immunohistochemical staining was performed to examine senile plaques in the brain of AD model mice,and Morris water maze was used to assess the spatial learning and memory abilities of the mice.Results Compared with the saline-treated AD model mice (6.0 ±0.6),the number of senile plaques of catalpol treated AD mice significantly decreased (2.3± 0.7; t =3.500,P =0.025); Mice in each groups had similar latency and path length to reach platform in visible platform test; In hidden platform test,catalpol-treated mice had a significant lesser latency and path length compared with saline-treated mice,furthermore,catalpol-treated mice had much more platform-crossing times (6.4 ± 0.8) than saline-treated mice (2.9 ± 0.4 ; t =5.592,P =0.001).Conclusion Catalpol can significantly decrease the senile plaque formation and improve the spatial learning and memory abilities of APP/PS1 double transgenic mice.

Objective To explore the effects of under-expression of large tumor suppressor 1 (LATS1) on activation of Hippo signaling pathway and differentiation, proliferation, migration of bone marrow mesenchymal stem cells (mMSCs) of micein vitro.Methods mMSCs of C57BL/6 mice were divided into normal control (MSC) group, empty vector control (MSC-GFP) group, LATS1-over-expressing (MSC-LATS1) group, empty vector without LATS1 shRNA control (MSC-shControl) group and LATS1-under-expressing (MSC-shLATS1) group. Lentiviral vectors with activated,inactivated LATS1 (the key molecule of Hippo signaling pathway) modifications and empty vectors were constructed and were used to infect mMSCsin vitro. The transduction efficiencies mediated by the lentiviral vectors were evaluated by fluorescence microscopy and flow cytometry. The mRNA expression of LATS1 was quantified by quantitative real-time polymerase chain reaction (qRT-PCR), and the protein expressions of LATS1, YAP (p-YAP), 14-3-3 were quantified by Western Blot to evaluate the activation of Hippo signaling pathway. Osteogenic and adipogenic differentiation of mMSCs were evaluated through measurement of Runx2, OSX and C/EBPα, PPAR-γ mRNA by qRT-PCR, as well as Alizarin Red S and Oil red O staining. Proliferation of mMSCs was evaluated using methy thiazdyl tetrazolium (MTT) assay. The scratch test and Transwell chamber test were used to analyze the horizontal and vertical migration ability of mMSCs.Results The transduction efficiencies mediated by the lentiviral vectors were 94.74%-96.10%. Compared with MSC-GFP group, the activation of Hippo signaling pathway was promoted in MSC-LATS1 group [LATS1 mRNA (2-ΔΔCT): 4.37±0.21 vs. 1.20±0.04, LATS1 protein (gray value): 2.21±0.06 vs. 1.09±0.10, p-YAP/YAP protein (gray value): 1.51±0.13 vs. 0.98±0.05, 14-3-3 protein (gray value): 1.92±0.18 vs. 1.10±0.09, allP < 0.05], osteogenic and adipogenic differentiation of mMSCs were decreased in MSC-LATS1 group [mineralization (A value):0.13±0.02 vs. 0.40±0.03, Runx2 mRNA (2-ΔΔCT): 0.51±0.02 vs. 0.98±0.09, OSX mRNA (2-ΔΔCT): 0.41±0.04 vs. 1.04±0.09, lipid accumulation (A value): 0.10±0.02 vs. 0.25±0.03, C/EBPα mRNA (2-ΔΔCT): 0.33±0.03 vs. 1.11±0.09, PPAR-γ mRNA (2-ΔΔCT): 0.29±0.02 vs. 1.04±0.10, allP < 0.05], the proliferation rate of mMSCs at 4-7 days was decreased in MSC-LATS1 group and so were the horizontal and vertical migration of mMSCs [wound healing rate: (18.65±3.53)% vs. (40.29±1.87)%, migrated cells (cells/MP): 35.99±6.18 vs. 103.67±17.77, bothP <0.05]. Compared with MSC-shControl group, the activation of Hippo signaling pathway was inhibited in MSC-shLATS1 group [LATS1 mRNA (2-ΔΔCT): 0.16±0.01 vs. 0.98±0.03, LATS1 protein (gray value): 0.38±0.03 vs. 1.04±0.07, p-YAP/YAP protein (gray value): 0.58±0.04 vs. 1.05±0.06, 14-3-3 protein (gray value): 0.14±0.02 vs. 1.02±0.09, allP < 0.05], osteogenic and adipogenic differentiation of mMSCs were increased in MSC-shLATS1 group [mineralization (A value): 0.93±0.13 vs. 0.44±0.05, Runx2 mRNA (2-ΔΔCT): 1.44±0.12 vs. 0.95±0.04, OSX mRNA (2-ΔΔCT):1.67±0.06 vs. 1.10±0.11, lipid accumulation (A value): 0.47±0.06 vs. 0.28±0.04, C/EBPα mRNA (2-ΔΔCT):3.98±0.61 vs. 0.99±0.10, PPAR-γ mRNA (2-ΔΔCT): 3.05±0.36 vs. 0.98±0.14, allP < 0.05], the proliferation rate of mMSCs at 3-7 days was increased in MSC-shLATS1 group and so were the horizontal and vertical migration of mMSCs [wound healing rate: (80.18±6.98)% vs. (46.18±1.01)%, migrated cells (cells/MP): 212.69±41.21 vs. 115.87±35.15, bothP < 0.05].Conclusions Under-expression of LATS1 promotes the differentiation, proliferation, migration of mMSCs by inhibition of Hippo signaling pathwayin vitro.

OBJECTIVE: To investigate the characteristics of adductor spasmodic dysphonia phonatory break in mandarin Chinese and select the stimuli phrases. METHOD: Thirty-eight patients with adductor spasmodic dysphonia were involved in this study. Standard phrase " fù mŭ xīn" and a speech corpus in mandarin Chinese with 229 syllables covering all vowel and constant of mandarin Chinese were selected. Every patient read the phrases above twice in normal speed and comfortable voice. Two auditory perpetual speech pathologists marked phonatory break syllables respectively. The frequency of phonatory break syllables and their located phrases were calculated, rated and described. The phrases including the most phonatory break syllables were selected as stimuli phrases, the phonatory break frequency of which was also higher than that of standard phrase "fù mŭ xīn". RESULT: Phonatory break happened in the reading of all patients. The average number of phonatory break syllables was 14 (3-33). Phonatroy break occurred when saying 177 (77.3%) syllables in the speech corpus. The syllables "guŏ, rén, zāng, diàn, chē, gè, guăn, a, bā, ne, de" broke in 23.1%-41.0% patients. These syllables belonged to the phrases "pĭng guŏ, huŏ chē, shì de, nĭ shì gè hăo rén, wŏ mén shì yŏu zŏng shì bă qĭn shì nong dé hĕn zāng, wŏ mén nà biān yŏu wăng qiú yùn dong chăng, cān gŭan, jiŭ bā hé yī gè miàn bāo dìan, tā shì duō me kāng kăi a,wŏ yīng gāi zài xìn lĭ xiĕ yī xiē shén mē ne?". Thirty-seven patients (97.3%) had phonatory break in above mentioned words. Ratios of these words phonatory break also were more than "fù mŭ xīn". CONCLUSION Adductor spasmodic dysphonic patients exhibited different degrees of phonatory break in mandarine Chinese. The phrases" shì de, pĭng guŏ, huŏ chē, nĭ shì gè hăo rén, wŏ mén nà biān yŏu wăng qiú yùn dong chăng, cān gŭan, jiŭ bā hé yī gè miàn bāo dìan, tā shì duō me kāng kăi a" were recommended as stimuli phrases for adductor spasmodic dysphonia evaluation.
Dysphonia ; diagnosis ; physiopathology ; Female ; Humans ; Language ; Male ; Phonation ; Spasm ; diagnosis ; physiopathology ; Voice

Objective To investigate the safety and feasibility of dyclonine combined with propofol in the application of painless gastroscopy. Methods A total of 90 patients received the painless gastroscopy in our hospi-tal were enrolled from September to December 2016. They were divided into 3 groups according to the random number table(n=30):Dyclonine+propofol(DP)group,Fentanyl+propofol(FP)group,Propofol(P)group. The hemodynamic changes,adverse reaction,propofol dosage,time of gastroscopy examination and time of conscious recovery were observed and recorded. Results Compared with P group,the incidence of hypertension,tachycardia, choking cough,body movement and the dosage of propofol in DP group and FP group were significantly decreased (P<0.05,respectively). Compared with DP group,the incidence of respiratory depression,the time of gastroscopy examination and the time of Conscious recovery in FP group and P group were significantly increased (P < 0.01 , respectively). Compared with FP group,the incidence of nausea and vomiting in DP and P group were significantly decreased (P < 0.05 ,respectively). Conclusions Dyclonine combined with propofol reduced the incidence of cardiovascular response,choking cough,body movement,respiratory depression,and nausea and vomiting,with the reduced dosage of propofol ,the shorten gastroscopy examination time and the recovery time. Therefore ,dyclo-nine combined with propofol is a safe and feasible anaesthesia management for the painless gastroscopy.

Diagnosis, Differential ; Humans ; Liver Diseases, Alcoholic ; diagnosis ; Non-alcoholic Fatty Liver Disease ; diagnosis

Teaching evaluation is an important part of the whole teaching process. In view of the shortcomings of the traditional final assessment, formative assessment has received wide attention from the education community with its characteristics of diverse assessing subjects, diverse assessment contents and diverse assessment tools. This paper introduces formative assessment into flipped classroom, and tries to construct formative assessment index system, which may provide the theoretical basis for the implementation of formative assessment in flipped classroom.

Objective To investigate the differences and similarities between drug-induced liver injury (DILI) and autoimmune hepatitis (AIH) in serum biochemical parameters and liver pathology,and to provide some thoughts for clinical diagnosis and differentiation of these two diseases.Methods A retrospective analysis was performed for the biochemical,immunological,autoantibody,and liver pathological data of 106 DILI patients and 63 AIH patients who were hospitalized,diagnosed,and treated in our hospital from January 2012 to October 2014.The patients' general data,biochemical parameters,immunological data,Ishak score,and qualitative changes in liver tissue were analyzed and compared.The Kruskal-Wallis test was used for comparison of nonparametric data between multiple groups,the Nemenyi test was used for comparison of nonparametric data between any two groups,the Wilcoxon rank sum test was used for comparison of Ishak scores,and the chi-square test was used for comparison of constituent ratio of categorical data.Results There were significant differences between AIH group and DILI hepatocyte injury group/mixed-type DILI group in the following serum biochemical parameters:alanine aminotransferase (187.2 U/L vs 1 326.5 U/L and 455.6,P ＜ 0.05),aspartate aminotransferase (172.2 U/L vs 759.5 U/L and 349.5 U/L,P ＜0.05),alkaline phosphatase (209.3 U/L vs 157.3 U/L and 169.4 U/L,P ＜ 0.05),gamma-glutamyl transferase (254.8 U/L vs 176.5 U/L and 170.5 U/L,P ＜ 0.05),total bilirubin (37.2 μmol/L vs 95.8 μmol/L and 52.6 μmol/L,P ＜ 0.05),serum iron (18.9 μmol/L vs 36.2 μmol/L and 23.9 μmol/L,P ＜ 0.05),serum ferritin (122.5 μmol/L vs 410.4 μmol/L and 186.5 μmol/L,P ＜ 0.05),immunoglobulin G (18.4 g/L vs 12.6 g/L and 12.3 g/L,P ＜ 0.05),and immunoglobulin M (1.8 g/L vs 1.3 g/L and 1.1 g/L,P ＜ 0.05).There were also significant differences between AIH group and DILI hepatocyte injury group/mixed-type DILI group in the Ishak score for interface inflammation (2.2±0.8 vs 1.3±0.7 and 1.3±0.6,P ＜ 0.05),Ishak score for portal inflammation (2.3±0.9 vs 1.5±0.7 and 1.4±0.8,P ＜ 0.05),and fibrosis score (2.8±1.1 vs 1.5±0.7 and 1.3±0.7,P ＜ 0.05).There were significant differences between AIH group and DILI hepatocyte injury group/mixed-type DILI group in the proportion of wax-like deposition (0 vs 29.2％ and 34.5％,P ＜0.05) and proportion of iron deposition (11.1％vs 52.1％ and 25.9％,P ＜ 0.05).Conclusion There are differences in biochemistry,immunology,and liver histology between DILI and AIH patients.AIH patients have more serious interface inflammation and portal inflammation and a higher fibrosis degree compared with DILI patients,while DILI patients have greater proportions of wax-like deposition and iron deposition compared with AIH patients.

Objective? To explore the effects of graded psychological care on patients receiving radical surgery for stomach cancer. Methods? Totally 150 stomach cancer patients admitted between May 2015 and January 2017 were selected by convenient sampling and randomized into the observation group (n=75) and the control group (n=75). Patients in the control group received conventional psychological care, while patients in the observation group received graded psychological care on this basis. The Symptom Checklist 90 (SCL-90), Pittsburgh Sleep Quality Index (PSQI), Visual Analogue Scale (VAS) and satisfaction questionnaire were used to compare the effects of intervention between the two groups. Results? The total SCL-90 score of the observation group averaged (1.45±0.28) after intervention, lower than that of the control group (t=4.479,P＜ 0.01). The PSQI score of the observation group after intervention was (7.10±3.08), while that of the control group was [(8.51±3.17); (t=2.763,P＜0.01)]. The postoperative pain score of the observation group post intervention was (2.75±0.34), lower than that of the control group;The nursing satisfaction score of the observation group was(89.15±4.23), higher than that of the control group, and the differences were statistically significant (t=7.859, 11.707; P＜ 0.01). Conclusions? Graded psychological care, when used in patients receiving radical surgery for stomach cancer, can improve the patients' psychological symptom and their satisfaction.