ABSTRACT
Objective:To investigate the expression level of circular RNA circ_0020123 in ovarian cancer (OC) , and then to evaluate the diagnostic value of circ_0020123 level in ovarian cancer.Methods:Fifty-five ovarian cancer patients who were diagnosed and treated in Hangzhou Linping District Maternal and Child Health Hospital from Oct. 2016 to Mar. 2017 were selected, and serum was collected for high-throughput gene expression sequencing analysis. The circ_0020123 level in serum of ovarian cancer patients and healthy controls was determined by real-time quantitative PCR, and the diagnostic efficacy of circ_0020123 level on ovarian cancer was evaluated by drawing receiver operating characteristic curve (ROC) . The correlation between the expression of circ_0020123 and the clinical characteristics of ovarian cancer patients was investigated by t-test. The survival curve was drawn to analyze the relationship between the serum circ_0020123 level and the prognosis and survival of ovarian cancer patients. Western blot was used to detect the expression levels of autophagy-related proteins LC3-II/LC3-I and p62.Results:Compared with the serum of healthy people (1±0.25) , the expression of circ_0020123 was significantly up-regulated in the serum of ovarian cancer patients (1.24±0.23) ( t=5.23, P<0.001) . The ROC curve showed that circ_0020123 had high diagnostic performance for the detection of ovarian cancer specimens (AUC=0.7579, P<0.001) . Comprehensive analysis of t test and logistic analysis showed that the high expression of circ_0020123 was significantly correlated with FIGO stage ( t=5.46, P<0.001) and tumor size ( t=6.37, P<0.001) . The measurement of survival data showed that the 5-year overall survival of the circ_0020123 high expression group was significantly shorter than that of the circ_0020123 low expression group ( P=0.019) . Knockdown of circ_0020123 in cells down-regulated the expression of autophagy-related proteins LC3-II/LC3-I (0.45±0.05) ( t=9.31, P=0.001) , and up-regulated the expression of p62 (1.94±0.11) ( t=12.62, P<0.001) . Conclusion:The results of this study indicate that circ_0020123 is an important ovarian cancer-related circular RNA, which provides a potential diagnostic, prognostic biomarker and therapeutic target for ovarian cancer patients.
ABSTRACT
OBJECTIVE: To investigate the effects of manganese(Mn) and high fat diet(HFD) co-exposure on the neurological behavior and gut microbiota in mice, and to observe the correlation between them. METHODS: Specific pathogen free adult male C57 BL/6 mice were randomly divided into 4 groups. Mice in control group and Mn exposure group were fed with normal diet, while the HFD group and co-exposure group were fed with HFD. Both the Mn exposure group and the co-exposure group were exposed to 10 mg/(kg·d) manganese chloride by intraperitoneal injection, while the control group and HFD group were treated with 0.9% sodium chloride solution of the same volume, once per day for 60 consecutive days. At the end of exposure, the mice were subjected to experiments of neurological behaviors. Then, the mice were sacrificed and intestinal feces were collected. The relative abundance of gut microbiota(relative abundance>1.000%) was detected by high-throughput sequencing. RESULTS: After exposure, the body weight of the HFD group and the co-exposure group increased significantly(P<0.05), while that of the Mn exposure group decreased(P<0.05), compared with the control group. The latency, time in central, crossing, total distance and open arm time(OT%) of mice in the Mn exposure group were lower than that of the control group(P<0.05), and close arm time(CT%) prolonged(P<0.05). Compared with the control group and the HFD group, the latency, rearing, time in central, crossing, total distance, OT% and open arm entry(OE%) of mice in the co-exposure group decreased(P<0.05), and CT% increased(P<0.05). The total distance of mice in the co-exposure group was lower than that of the Mn exposure group(P<0.05). The relative abundance of Firmicutes increased(P<0.05), those of Bacteroidetes and Actinobacteria decreased in mice in the HFD group at the phylum level(P<0.05) compared with mice in the control group. The relative abundance of Firmicutes, Proteobacteria and Cyanobacteria increased(P<0.05), and Bacteroidetes and Actinobacteria decreased(P<0.05) in mice in the Mn exposure group. The relative abundance of Oscillospira, Bacteroides and Prevotella of mice in the HFD group reduced at the genus level(P<0.05) compared with the control group. The relative abundance of Lactobacillus increased in Mn exposure group(P<0.05), and Oscillospira, Bacteroides and Prevotella decreased(P<0.05). The relative abundance of Firmicutes, Cyanobacteria and Lactobacillus of mice in the co-exposure group increased(P<0.05), and those of the remaining 6 bacteria were lower(P<0.05) compared with mice in the other 3 groups. Among the mice of co-exposure group, the latency was positively correlated with Bacteroidetes(P<0.05). The rearing was positively correlated with Firmicutes(P<0.05) and negatively correlated with Actinobacteria(P<0.01). The OE% was negatively correlated with Firmicutes(P<0.05) and positively correlated with Actinobacteria(P<0.05). The crossing was positively correlated with Prevotella(P<0.05). CONCLUSION: Manganese combined with HFD had a synergistic effect on the abnormality of neurological behavior of mice. There are some correlation between the abnormality of neurological behavior and the homeostatic imbalance of intestinal flora in mice.
ABSTRACT
BACKGROUND:There is a very close relationship between osteoporosis and periodontal disease in postmenopausal women, but the mechanism remains unclear. OBJECTIVE:To investigate the expression of nuclear factor-κB in the alveolar bone andinterleukin-17 in the serum and gingiva in the mouse model of osteoporosis caused by ovariectomy. METHODS:Female mice aged 3 months were randomly divided into ovariectomy and sham operation groups. At 6 months after surgery, the mouse models were evaluated histologically on the submandibular bone and thigh bone stained with hematoxylin and eosin. In the submandibular bone, the expression levels of OCN and Runx2 were detected by RT-PCR, and the expression level of nuclear factor-κB was detected by immunohistochemical staining and western blot assay. Besides, the expression level of interleukin-17 in the serum and gingival homogenate was evaluated using Cytometric Beads Array. RESULTS AND CONCLUSION:The thigh bone in the ovariectomy group revealed the thin cortical bone, enlarged marrow cavity, and increased resorption lacunae, as well as fewer, thinner trabeculae with lower density and irregular structure. Compared with the sham operation group, the expression levels of OCN and Runx2 in the alveolar bone were decreased in the ovariectomy group. The activation of nuclear factor-κB (P65)appeared with P65 positive expression in the submandibular bone in the ovariectomy group, and the relative expression level was higher than that in the sham operation group. The serum level of interleukin-17 in the ovariectomy group was higher than that in the sham operation group, but the level in the gingival tissue showed no significant difference between the two groups. These results indicate that estrogen deficiency after ovariectomy can activate nuclear factor-κB signal pathway to play a role in periodontal osteolysis. However interleukin-17 in the local periodontal tissue may not be a key cytokine to damage the periodontal tissue.
ABSTRACT
Objective:To explore the effects of exosomes derived from bone-marrow endothelial progenitor cells(EPCs-Exos)on the angiogenesis and collagen deposition in vitro,and to illustrate the possible mechanism for EPCs-Exos to accelerate the cutaneous deficiency repair.Methods:The endothelial progenitor cells (EPCs) were isolated, cultivated and identified;at the same time, the EPCs-Exos were also isolated and identified.The EPCs-Exos uptake in EPCs was observed and analyzed.16 rats were randomly divided into 4 groups, and then the models of skin defect were established, respectively.The equal volume of PBS and 50, 100, 150 mg·L-1 EPCs-Exos were injected into the area around skin defect of the rats in 4 groups.The wound closed sizes on the 0, 3rd, 7th and 14th days were measured, and Masson''s trichrome staining and CD31 immunofluorescence staining were performed on the 14th day for evaluating the tissue healing efficacy after EPCs-Exos treatment.In vitro,the mediums containing PBS and 50, 100, 150 mg·L-1 EPCs-Exos were used to culture the human umbilical vein endothelial cells (HUVECs), respectively.Scratch test and tubule formation assay were used to detect the migration and capillary network formation of HUVECs.At the same time, Western blotting was used for analyzing the expression level of angiogenesis related gene vascular endothelial growth factor A (VEGFA) in HUVECs.Results: The primary EPCs were isolated and identified successfully, and EPCs-Exos were purified and characterized.The CD31 immunofluorescence staining and double staining of DiL-ac-LDL and FITC-UEA-I of EPCs were positive.The electron microscope results showed that EPCs-Exos were nearly spheroidal, with the diameter about 40-100 nm.For the models of rat skin injury treated by EPCs-Exos, with the increasing of injection doses, the sizes of skin defect scar were gradually reduced, the degrees of scar healings were gradually increased,and the differences between various groups were statistically significant (P< 0.05).EPCs-Exos promoted the collagen maturity of healing skin in a dose-dependent manner;on the 14th day, the effect in 150 mg·L-1 EPCs-Exos group was the most significant.In vitro, EPCs-Exos promoted the migration and capillary network formation of HUVECs and increased the expression level of VEGFA;the migration rate,the net number of branches and the expression level of VEGFA in 150 mg·L-1 EPCs-Exos group were significantly higher than those in 50 mg·L-1 EPCs-Exos group and PBS group (P< 0.05).Conclusion: EPCs-Exos can promote the repair of traumatic skin defect of the rats by positively regulating the vascular endothelial cell function.