BACKGROUND:Although the methods of mechanical scrapping,tissue block transplantation,and enzyme digestion to culture endothelial cells have been quite mature,it has been constantly updated.OBJECTIVE:To explore the culture and identification of rabbit aortic endothelial cells.METHODS:Aorta was derived from one-week-old New Zealand rabbit.Following removing the adventitia,the endothelium was digested in the mixture of 2 g/L type Ⅰ collagen,2 9/L type Ⅱ collagen,2 g/L type Ⅳ collagen,and 2 g/L type Ⅴ collagen (1:1:1:1)for 20 minutes,and the digestionwas stopped with culturemedium(1:1).Cells in the endothelium were lightly scraped to make cell suspension,which was centrifuged to precipitate cells with DMEM culture medium containing 20%fetal bovine serum,1 μg/L VEGF,2 μg/L bFGF,and 6 U/L gentamicin.The cells were then blown into single cells and the culture liquid was changed every 48 hours.The cells were passaged according to the ratio of 1:2.Inverted phase contrast microscope was used to observe cell culture;immunohistochemistry and immunofluorescence were used to determine Ⅷ-related antigen;electron microscope was used to detect Weibel-Paladed body.RESULTS AND CONCLUSION:Ⅷ-related antigen and electron microscopy confirmed that the primary and 5-passage endothelial cells were successfully cultured,suggesting that rabbit aortic endothelial cells were cultured into cell line.Ⅷ-related antigen and electron microscopy determined that a combination with W-P body was a good method to determine endothelial cells.

Objective To explore the outpatients′experience of caring and provide references for improving their experience of medical care.Methods The phenomenology research method was adopted in this study,and twenty-one outpatients were subject to an in-depth interview to learn their outpatient experiences.Results Ten themes of experience with caring were extracted as follows:passionate and patient,careful and considerate,respect for others,offers to help,encouraging and comforting,apologies and courteously,humor,truthful considerations,conscientious and responsible,and a caring environment setup.Five themes of experience without caring were extracted as follows:impatient communication, careless seeing of outpatients,delayed service,passive reminders,and facilities without enough details. Conclusions The outpatients may have abundant experiences of caring at the outpatient clinic,also lack of caring sometimes as well.Outpatient managers should keep an eye on outpatients′needs and satisfaction of caring from time to time,to improve the quality of outpatient service.

Thin endometrium is an important factor influencing the in vitro fertilization and embryo transfer, and there is a lack of effective therapy in the treatment of thin endometrium.The aim of this study was to explore a stable animal model of effective thin endometrium, and to promote the research on thin endometrium pathogenesis and provide experimen-tal basis of treatment.To analyze a variety of establishments of endometrial damage animal model reported in the domestic and foreign literatures,it is concluded that perfusing 95% ethanol into uterine cavities of rats can establish a rat model of thin endometrium,and put forward some experience and methods for its improvement.

OBJECTIVE To investigate the delayed cytotoxicity effect of chlorpyrifos (CPF) with?drawal on primary hippocampal neurons. METHODS Hippocampal neurons were prepared from SD rat fetuses on the 17th day of gestation. Seven days after culture,neurons were exposed to CPF 10 and 30 μmol · L-1,respectively,for 72 h or for 48 h followed by CPF withdrawal for 24 h. CCK-8 kit and neuronal nuclei(NeuN), 5-bromodeoxyuridine(BrdU) and β Ⅲ tubulin immunofluorescence expression methods were used to evaluate the cell viability. RESULTS Compared with normal control, no significant cytotoxicity was found after CPF 72 h continuous exposure. However,CPF 48 h expo?sure followed by CPF withdrawal for 24 h induced evident cytotoxicity. The amount of BrdU positive and β Ⅲ tubulin positive hippocampal neurons were both decreased significantly(P<0.05),and cell survival and viability reduced after CPF withdrawal. CONCLUSION CPF exposure withdrawal can induce more seriously delayed cytotoxicity than continuous exposure in rat primary hippocampal neurons.

Objective:To explore the DNA methylation patterns and methylation differential genes of patients with coal-burning-borne endemic fluorosis, and to provide a basis for study of the pathogenesis of fluoride-induced body injury.Methods:A case-control study was conducted in Shuicheng County, Liupanshui, Guizhou Province, ten patients with severe fluorosis were selected as the fluorosis group in Douqing Township, where people burning high fluorine coal in open range all year round; and ten people without fluorosis phenotype were selected as the control group in Huaga Township, where firewood was the main fuel. Peripheral blood samples were collected from the two groups of people. Reduced representation bisulfite sequencing (RRBS) technique was used to detect the whole genome DNA methylation pattern ( n = 4) and DNA differentially methylated region (DMR), the DMR differential degree (log 2Ratio) and KEGG pathway enrichment analysis were used to screen the methylation differential genes, and real-time PCR was used to verify the mRNA expression levels of the candidate methylation differential genes( n = 10). Results:The methylation pattern analysis results showed that the methylation levels of all C bases in the genome DNA of the fluorosis group and the control group were (61.53 ± 0.59)% and (62.48 ± 1.53)%, respectively; among them, the methylated levels at CG sites were (63.75 ± 0.65)%, (64.36 ± 1.01)%, at CHG sites were (13.79 ± 0.72)%, (16.69 ± 4.06)%, and at CHH sites were (25.12 ± 1.72)%, (29.77 ± 3.97)%. Compared with the control group, patients in the fluorosis group had 1 000 DMR distributed on different autosomes; and the chromosome 19 was the most with 104 segments. There were 978 DMR-related genes, including 265 hypermethylation genes and 713 hypomethylation genes; KEGG pathway enrichment analysis showed that methylation differential genes were mainly involved in cell metabolism, cancers, and phosphatidylinositol 3-kinase-protein kinase B (PI3K-AKT) and other signaling pathways; combined with the differential degree of DMR, the hypermethylated succinate dehydrogenase complex flavoprotein subunit A pseudogene 3 (SDHAP3, log 2Ratio = 3.487) and hypomethylated nuclear factor κB inhibitor kinase regulatory subunit γ (IKBKG, log 2Ratio =-4.436) were selected as the candidate genes. There were statistically significant differences in the mRNA expression levels of SDHAP3 (0.54 ± 0.08, 1.00 ± 0.00) and IKBKG (1.32 ± 0.39, 1.00 ± 0.00) between fluorosis group and control group ( F = 22.94, 15.09, P < 0.01 or < 0.05). Conclusion:There are a large number of methylation differential genes in the genomes of patients with coal-burning-borne endemic fluorosis and controls, the hypermethylated SDHAP3 and hypomethylated IKBKG may be involved in fluoride induced body injury.

Objective To compare the long-term efficacy between two radiochemotherapy regimens for locally advanced nasopharyngeal carcinoma (NPC):intensity-modulated radiotherapy with concurrent chemotherapy (CCRT) versus neoadjuvant chemotherapy (NACT) followed by CCRT.Methods A retrospective analysis was performed on the clinical data of 278 patients with locally advanced NPC who were admitted to our hospital from 2001 to 2008.Of the 278 patients,133 received CCRT,and 145 received NACT followed by CCRT (NACT + CCRT).Results The follow-up rate was 96.6％.The 5-year overall survival (OS),distant metastasis-free survival (DMFS),recurrence-free survival (RFS),and progression-free survival (PFS) were 78.1％,78.0％,90.6％,and 72.0％,respectively.There were no significant differences between the CCRT group and NACT + CCRT group in 5-year OS (79.9％ vs.76.4％,P =0.443),DMFS (77.1％ vs.78.9％,P=0.972),RFS (91.6％ vs.89.8％,P=0.475),and PFS (71.6％ vs.72.2％,P=0.731).Subgroup analysis showed that compared with CCRT,NACT + CCRT did not significantly improve 5-year RFS in T3-4N0-1 patients (90.7％ vs.86.9％,P=0.376) and did not significantly improve 5-year DMFS in patients with advanced N-stage disease (57.6％ vs.69.7％,P =0.275).There were significantly higher numbers of individuals with neutropenia,decrease in hemoglobin,and upper gastrointestinal reactions in patients treated with NACT + CCRT than in those treated with CCRT (100 vs.52,P=0.000;64 vs.35,P=0.010;90 vs.63,P=0.044).Conclusions Compared with CCRT,NACT + CCRT does not significantly improve the prognosis in patients with locally advanced NPC and leads to significant increases in grade ≥ 3 toxicities (neutropenia,decrease in hemoglobin,and upper gastrointestinal reactions).The role of NACT in the treatment of locally advanced NPC needs further study

BACKGROUND: Thin endometrial diseases are a challenge in clinical treatment at present. Scholars have found that bone marrow mesenchymal stem cells (BMSCs) transplantation has its unique curative effect and advantages, but few studies have been conducted on pathway or gene control. OBJECTIVE: To observe the effect of BMSCs transplantation in rats with thin endometrium based on the HOXA10 regulatory network. METHODS: Twenty-one adult female Sprague-Dawley rats of SPF grade (provided by the Animal Experimental Center, Guangzhou University of Chinese Medicine in China) were randomly divided into three groups (n=7/group): control group, model group, and BMSCs group. In the latter two groups, a thin endometrium model was prepared in each rat by filling the uterine cavity with 95％ ethanol. In the control group, normal saline was injected to fill the uterine cavity of rats. After extraction of ethanol or normal saline, the rats in the BMSCs group were injected intrauterinely with 1 mL of BMSCs suspension (1×1010 cells/L) , and those in the control and model groups were given the same volume of normal saline. After two estrous cycles, the uterus of each rat was removed. Hematoxylin-eosin staining was used to measure the thickness of the endometrium. Immunohistochemistry was used to detect the expression of vimentin, keratin, vascular endothelial growth factor, leukemia inhibitory factor and integrin αvβ3. qRT-PCR was used to detect the relative transcription of HOXA10 and miR-196 b. RESULTS AND CONCLUSION: (1) Compared with the control group, the endometrial thickness of the rats were significantly thinner in the model and BMSCs groups (P < 0.05) , while the endometrial thickness in the BMSCs group was thicker than that in the model group (P < 0.05). (2) The mean absorbance values of endometrial vimentin, keratin, vascular endothelial growth factor, leukemia inhibitory factor and integrin αvβ3 were highest in the control group, higher in the BMSCs group and lowest in the model group, and there were significant differences between groups (P < 0.05). (3) The relative transcript level of HOXA10 gene in the model and BMSCs group was significantly lower than that in the control group, while the relative transcript level of HOXA10 gene in the BMSCs group was significantly higher than that in the model group (P < 0.05). The relative transcript level of miR-196 b in the model and BMSCs groups was significantly higher than that in the control group (P < 0.05) , while the relative transcript level of miR-196 b in the BMSCs group was lower than that in the model group (P < 0.05). (4) HOXA10 was negatively correlated with miR-196 b gene, HOXA10 was positively correlated with the protein expression to different extents, and miR-196 b gene was negatively correlated with the protein expression to different extents. These findings suggest that BMSCs transplantation can improve the endometrial thickness and relevant protein levels of thin endometrium rats to some extent, which may be attributed to the negative regulation of HOXA10 gene by miR-196 b, and HOXA10 gene further promotes the expression of vascular endothelial growth factor, leukemia inhibitory factor and integrin αvβ3 proteins.

Objective To explore the methods of the standardized management used in humanistic nursing care and to assess its effects.Methods Measures of the standardized management began to be used in such care at the hospital since January 2013.Questionnaires of patient satisfaction with nursing care for inpatients and those discharged were used to assess the effects of the standardized management measures periodically and synthetically.Results The patient satisfaction with humanistic nursing care keeps increasing year by year from 2013 to 201 5.The differences among three sets of data are statistically significant(P < 0.05 ).Conclusions Humanistic nursing care calls for promotion of standardized management.It is high time to build such a standardized management system at the hospitals.

Objective:To present the surgical technique and to evaluate the results of the modified ar-throscopic Latarjet procedure.Methods: Arthroscopic Latarjet procedure has proven to be a reliable method of treatment for difficult anteroinferior instability of the shoulder joint.However, there is no ante-rior capsule reattachment and too much subscapularis damage for the classic procedure.From February 2013 , we modified the classic procedure with reattachment of anterior joint capsule and muscle-tendon junction splitting of subscapularis.Coracoid graft position was evaluated using CT scanning.Results:From March 2012 to August 2014, 51 modified Latarjet procedures were successfully performed arthro-scopically for patients with anterior shoulder instability.According to the CT scanning at the final follow up, the graft was flush with the glenoid in 94.1%, and medially placed in 5.9%.Vertical positioning was perfect in 96.0%(2 to 5 o’clock), too high in 2.0%, and too low in 2.0%.There were no cases of recurrent dislocation or subluxation.Conclusion: The modified arthroscopic Latarjet procedure has shown satisfactory results with good graft positioning.It is a minimal invasive and accurate approach, which combines the advantages of the open procedure.