Knockdown of long non-coding RNA MIR4697 host gene inhibits adipogenic differentiation in bone marrow mesenchymal stem cells / 北京大学学报(医学版)

OBJECTIVE@#To preliminarily investigate the role of long non-coding RNA (lncRNA) MIR4697 host gene (MIR4697HG) in regulating the adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).@*METHODS@#For adipogenic differentiation, BMSCs were induced in adipogenic media for 10 days. The mRNA expression levels of lncRNA MIR4697HG and adipogenic marker genes including peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhanced binding protein α (CEBP/α) and adiponectin (ADIPQ) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) at different time points (0, 1, 2, 3, 5, 7, 10 days). The MIR4697HG stable knockdown-BMSC cell line was generated by infection of MIR4697HG shRNA-containing lentiviruses. To avoid off-target effect, two target sequences (shMIR4697HG-1, shMIR4697HG-2) were designed. And then cells were induced to differentiate in adipogenic medium. Oil red O staining, Western blot and qRT-PCR were used to detect the effect of MIR4697HG knockdown on adipogenic differentiation of BMSCs.@*RESULTS@#The mRNA expression level of MIR4697HG was significantly increased during adipogenic differentiation (P < 0.01), and adipogenic differentiation of BMSCs was evidenced by upregulated mRNA levels of specific adipogenesis-related genes including PPARγ, CEBP/α and ADIPQ. Observed by fluorescence microscopy, more than 90% transfected target cells expressed green fluorescent protein successfully after shMIR4697HG-1 group, shMIR4697HG-2 group and shNC group transfection for 72 h. And the transfection efficiency of MIR4697HG examined by qRT-PCR was above 60%. Then the BMSCs were treated with adipogenic media for 7 days and showed that the mRNA expression levels of adipogenesis-related genes including PPARγ, CEBP/α and ADIPQ were significantly decreased in the MIR4697HG knockdown group (P < 0.01), while the expression levels of PPARγ and CEBP/α proteins were decreased remarkably as well (P < 0.01). Consistently, MIR4697HG knockdown BMSCs formed less lipid droplets compared with the control BMSCs, which further demonstrated that MIR4697HG knockdown inhibited adipogenic differentiation of BMSCs.@*CONCLUSION@#lncRNA MIR4697HG played a crucial role in regulating the adipogenic differentiation of BMSCs, and MIR4697HG knockdown significantly inhibited the adipogenic differentiation of BMSCs. These data may suggest that lncRNA MIR4697HG could serve as a therapeutic potential target for the aberrant adipogenic differentiation-associated disorders including osteoporosis.

Discussion on relationship between autophagy and endometriosis based on pathogenesis of "dampness and stasis" / 中国中药杂志

Endometriosis(EMs) is a stubborn gynecological disease caused by persistent immune-inflammatory effects, and is known as "benign tumor" because of its similar characteristics to malignant tumors. National physician master Professor BAN Xiu-wen believes that the spread of damp-evil is the pathologic foundation for inflammatory response of ectopic endometrium; accumulation of blood stasis is the pathological product of continuous inflammatory attacks, and the combination of dampness and stasis is the main pathogenesis for refractory EMs. Modern researches have shown that immune-inflammatory effect is the key mechanism for development of EMs, and is closely related to cell autophagy, all of which have made it become the hot spots in research of the pathogenesis, diagnosis and treatment of EMs. Therefore, with immune-inflammatory effect as the breakthrough point in this research, and with reference to the related research of autophagy, the correlation between "combination of dampness and stasis" and abnormal autophagy-induced immune inflammatory response in ectopic endometrium was discussed, to provide guidance for the clinical application of traditional Chinese medicine and modern research.

Study on the Correlation between Caring Perception Status of Patients and Nurse-patient Relationship Trust Degree / 中国医学伦理学

Objective:To investigate the perception status of inpatients to nurses' caring behavior, and analyze the correlation between them and nurse-patient relationship trust degree. Methods:Adopting general information questionnaire, Caring Behaviors Inventory ( CBI) , nurse-patient relationship trust scale, a questionnaire survey was conducted among 226 inpatients in a third class A tertiary hospital in Tianjin. Results:The total score of ca-ring behavior perception was (96. 92 ± 15. 68), and among the four dimensions, the perception of patients to nur-ses' knowledge and skills was deepest, the perception to nurse' s contact with patients was worst; total score of nurse-patient trust scale was (124. 75 ± 19. 13). There was a positive correlation between the total score of pa-tients' caring perception and nurse-patient relationship trust degree (r=0. 554);multiple linear stepwise regres-sion analysis showed respect for patients, support and assurance were the main factors influencing the nurse-pa-tient relationship trust degree. Conclusion:The patients' perception on nurses' caring behavior and nurse-pa-tient relationship trust degree is closely related. Nurses should carry out targeted care to patients with different char-acteristics to improve patients' trust to nurses and construct a harmonious nurse-patient relationship.

Expression of GDNF in rat Sertoli cells cultured in vitro at different temperatures / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the mechanism of hyperthermia inducing infertility by observing the expression of glial cell line-derived neurotrophic factor (GDNF) in rat Sertoli cells cultured in vitro at different temperatures.</p><p><b>METHODS</b>Using combination enzyme digestion and selective adhesion, we isolated Sertoli cells from male Wistar rats and cultured them in vitro at different temperatures, followed by observation of the changes in their adhesion and morphology and identification by FasL immunohistochemical staining. We divided the Sertoli cells into a control group (35 degrees C) and four experimental groups (36 degrees C, 37 degrees C, 38 degrees C, and 39 degrees C), measured their proliferation by CCK-8, observed their morphology and structure by HE staining, and determined the expression of GDNF by RT-PCR, immunofluorescence and Western blot.</p><p><b>RESULTS</b>Sertoli cells were successfully isolated and in vitro-cultured, with a purity of (95.30 +/- 2.15)% (n = 10). The CCK-8 assay showed that the proliferation of the Sertoli cells was the highest at 36 degrees C, gradually decreasing with the temperature above 36 degrees C, and significantly inhibited at 39 degrees C (P < 0.01). Immunofluorescence revealed the expression of GDNF in the cytoplasm, with the highest fluorescence intensity at 36 degrees C. RT-PCR and Western blot exhibited a decreasing trend of the GDNF expression with the increasing temperature above 36 degrees C. There were statistically significant differences in the expression of GDNF between the control group and the four experimental groups (P < 0.01).</p><p><b>CONCLUSION</b>The proliferation and GDNF expression of in vitro-cultured Sertoli cells differ significantly at different temperatures. At > 36 degrees C, the higher the temperature is, the lower the Sertoli cell proliferation and GDNF expression are. Our findings suggest that high temperature above 36 degrees C suppresses the function of Sertoli cells and may also damage spermatogenesis.</p>

Effect of H2O2,extracellular matrix and out segment of photoreceptor on expression of focal adhesion kinase in RPE cell / 中华实验眼科杂志

Background The underlying mechanism of choroidal neovascularization(CNV) is multifactorial and complex.Focal adhesion kinase(FAK) plays a crucial role in controlling essential cellular processes and influencing distinct steps of the angiogenic response.But to our knowledge,seldom study on the effect of FAK on CNV formation has been reported previously.Objective In this study,the effect of several CNV risk factors on the expression of FAK in cultured retinal pigment epithelium(RPE) cells was investigated to illuminate effect of FAK on CNV.Methods Human RPE cells were isolated from donor eyes and exposed to H2O2,swallow of outer segment of photoreceptors(POS) and extracellular matrix(ECM) separately with the treating as follows:RPE cells were co-cultured with 10,20,50 and 100μmol/L H2O2 for 20 days;POS(1×106/ml) were co-cultivated with RPE cells for 20 days(setting control group,POS group,hypoxia group with 200μmol/L CoCl2,and POS+hyoxia group);RPE cells were cultured on the plates coated with 100mg/L fibronectin(FN),laminin(LN) or collagen typeⅠfor 30minutes or 1 hour.The expression of FAK and pFAK in RPE cells were examined by Western blot analysis.Results FAK was highly expressed in the 20μmol/L and 50μmol/L H2O2 groups compared with control group(P＜0.01);while he expression level of pFAK was reduced after treated with H2O2 in comparison with the control group(P＜0.01).After cultured with POS for 20 days,the undigested lysosome could be observed in RPE cells.The expressions of FAK and pFAK in RPE cells were not significantly changed between control group and POS groups(P＞0.05),but those in hypoxia group were significantly up-regulated in comparison with control group(P＜0.01).Compared with the hypoxia group,the expression amount of pFAK was elevated in POS+hyoxia group(P＜0.01).In comparison with control group,the increased pFAK expression was seen in FN,LN and collagen typeⅠtreating for 1-hour groups(P＜0.05,P＜0.01).Conclusion FAK pathway participates in several CNV-initiated signaling,such as H2O2,POS and ECM,in cultured RPE cells.It is reasonable to believe that FAK potentially plays an important role in CNV-dependent disorder.