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Objective The aim of this study was to investigate the expression of miR-455-5p in epithelial ovarian cancer and its effect on the development of epithelial ovarian cancer. Methods The miRNA expression data of normal ovarian epithelial tis-sues and epithelial ovarian cancer tissues GSE83693 were downloaded from the GEO database. Differential expression analysis was used to obtain differential expression data of miRNAs in epithelial ovarian cancer. The expression of miR-455 -5p was analyzed whether there is difference expression between normal ovarian epithelium and epithelial ovary cancer tissues; qRT-PCR was used to verify the differential expression prediction results; bio-informatics software was used to analyze the KEGG pathway enrichment and GO gene function annotation of miR-455-5p target genes,and to explore the disorders of dyregulated miR-455-5p in the devel-opment of epithelial ovarian cancer. Results A total of 101 cases of differentially expressed miRNAs were screened,34 cases were up-regulated and 67 cases were down-regulated. Among them,miR-455-5p was down-regulated significantly(P<0. 01),and the different fulds were -2. 9019. The results of qRT-PCR showed that the expression of miR-455-5p in epithelial ovarian cancer cells(SKOV-3,OVCAR-3 and A2780)was significantly lower than that in normal ovarian epithelial cells(IOSE-80),and the dif-ferential expression was statistically significant(P<0. 05). The results of KEGG pathway enrichment analysis showed that miR-455-5p regulated target genes mainly involved in five pathways,including TGF-β signaling pathway,Hippo signaling pathway,ECM-receptor interaction,transcriptional dysregulation pathway in cancer,and chronic granule cellular leukemia,which were associated with tumors. GO functional annotation analysis showed that the target genes regulated by miR-455-5p in the above pathway was mainly involved in protein phosphorylation,promoted cell proliferation and migration,inhibited apoptosis,promoted epithelial-mesenchymal transition,regulated transcription and regulated cell cycle,etc. ,which associated with tumorigenesis. Conclusion The expression of miR-455-5p is down-regulated in epithelial ovarian cancer. The miR-455-5p target genes are involved in the pathogenesis and function of epithelial ovarian cancer,and are associated with the development of epithelial ovarian cancer.
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Objective The aim of this study was to investigate the expression of microRNA-150(miR-150)in human epi-thelial ovarian cancer cells and its effect on proliferation,apoptosis,invasion and metastasis of human epithelial ovarian cancer cells. Methods The expression level of miR-150 in cells from each treatment group was detected by Real-Time PCR(qRT-PCR);effects of proliferation,apoptosis,invasion and metastasis of epithelial ovarian cancer cells was investigated by MTT,flow cytometry, and transwell assays. Results Compared with normal ovarian epithelial cells(T29),the expression of miR-150 was significantly de-creased in epithelial ovarian cancer cells(A2780 and OVCAR3)(P<0. 01); After transfection miR-150 mimic,the expression of miR-150 in A2780 and OVCAR3 cells was significantly increased(P<0. 01);After 3 d of transfection,the OD values of the miR-150mimicgroup(A2780:1.12±0.03;OVCAR3:1.91±0.03)werelowerthanthatintheblankgroup(A2780:2.35±0.09;OVCAR3:2.63 ±0.07)and the miR-150 NC group(A2780:2.18 ±0.07;OVCAR3:2.43 ±0.11)(P<0.01);The apoptotic rate in the miR-150 mimic group(A2780:16. 10 ± 0. 58% ;OVCAR3:15. 16 ± 1. 30% ) were significantly increased when compared to the blank group(A2780:10. 07 ± 0. 66%;OVCAR3:3. 81 ± 0. 24%) and the miR -150 NC group(A2780:10. 36 ± 1. 08%;OVCAR3:4.89 ±0.07%)(P<0.01);The number of transmembrane cells in the miR-150 mimic group(A2780:38.67 ±2.03;OVCAR3:28. 67 ± 2. 03)was higher than that in the blank group(A2780:76. 30 ± 7. 45;OVCAR3:55. 67 ± 3. 18)and the miR-150 NC group(A2780:74. 33 ± 5. 78;OVCAR3:56. 33 ± 3. 84)(P<0. 01). Conclusion The decreased expression of miR-150 in epi-thelial cancer cells may be one of the mechanisms of proliferation,invasion and metastasis of epithelial ovarian cancer. Up-regulation of miR-150 may inhibit the proliferation of epithelial ovarian cancer cells and promote apoptosis to reduce the abilities of invasion and metastasis in epithelial ovarian cancer cells.
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Objective The purpose of this study is to explore the expression of 12 tumor Markerss in the serum of colon cancer patients by C 12 tumor marker protein chip ,screening most valuable Markers for diagnosis . Methods C12 tumor Marker of protein chips were used for 12 sorts of tumor marker detection among 60 CRC patients and 40 healthy controls ,address the most valuable tumor marker relative to colon cancer and contribution of combinations effect on diagnosis .Result CEA+CA199+CA242+Bete-HCG combination diagnosis has the highest effectivity and diagnosis sensitivity was greatly improved by combination detection .The optimized diagno-sis showed no significant tendency to be correlated with age ,sex,differentiation stage,pathology,Lymph node me-tastasis,clinical stage.Conclusion It is of importance to use sera CEA +CA199+CA242+Bete-HCG detec-tion as markers for colon cancer diagnosis ,which may improve diagnosis efficiency .
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Objective To investigate the association SLC25A12 and SCN2A2 gene single nucleotide polymorphisms(SNPs) and susceptibility to autism among 105 Japanese family trios consisting of fathers,mothers,and affected offsprings with autism.Methods Genomic DNA was isolated from the whole blood samples.The PCR-single stranded conformational polymorphism(SSCP) technique was used to test genotype of SNPs(rs3770448,rs3769955) at SLC25A12 and SCN2A2 genes.Results The distributions of genotypic and allelic frequencies of rs3770448 and rs3769955 were not deviated from the Hardy-Weinberg equilibrium.The results of transmission disequilibrium test(TDT) indicated that the allelic frequency transmitted from the heterozygote parents didn′t deviate 50%.Conclusion The polymorphism of rs3770448 in the SLC25A12 and rs3769955 in the SCN2A2 locus may not be associated with autism.But the association of the other SNPs at the SLC25A12 and SCN2A2 locus with the illness can not be ruled out.
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BACKGROUND: Skin transplantation is the most effective conventional method to cure large area full-thickness skin damage caused by empyrosis or some diseases, but present deficiency of skin donator is the largest barrier in front. The most ideal way to solve this problem is to use tissue-engineering skin reconstructed by self-skin cells as seed cells.OBJECTIVE: To investigate the effects of tissue engineered artificial animal skin in animal grafting experiment.DESIGN: A randomized controlled trial SETTING: Institute of frontier medical sciences and department of dermatology in a university.MATERIALS: Study was performed in the Cell-Engineering Institute of Jilin University from September 1998 to July 2001. Totally 20 newborn Wistar rats and 24 8-week old male nude mice were selected into our study.METHODS: Full-thickness wounds(diameter: 20 nn) were made on the backs of twenty-four nude mice to establish full-thickness skin defect animal model for grafting by the tissue-engineered reconstructive artificial skin made by chitosan(CH) as stromal scaffold. Twenty-four 8-week old nude mice were divided into artificial skin (AS) group, chitosan membrane(CH) group and control group (CG). All wounds were covered with AS, CH or petrolatun gauze correspondingly. The wounds of each groups were observed daily,techniques like histology and infrared-ray scan were used for a dynamical surveillance on the 3rd, 7th, 14th and 21st days.MAIN OUTCOME MEASURES: ① general observation; ② blood supply in recipient area under infrared-ray observation; ③ histological observation.RESULTS: Transplanted AS had a favorable fusion between tissue-engineered skin and self-skin on the 3rd day after grafting with a few of ingrowths of capillaries. The color of the AS was closed to self-skin. The capillaries gradually increased in the grafts over time. The new epidermis was clearly consisted of stratum basale, stratum spinosum, stratum granulosum, and stratum corneum. Keratinization enhanced with exfoliation. Cells in dermis increased and the scaffold gradually degraded. The secreted extracellular matrix increased as well. On the 14th day after grafting, the wounds almost recovered.The color of the grafted artificial skin was more similar to the nature skin with very little scaring, which indicated that a second grafting was unnecessary. The scabs did not completely fall off in CH group until the 14th day, and the wound was not healed. The color was darker than that of AS group. The scabs fell off in CG, and the wounds were big and deep with dark red color.CONCLUSION: The new reconstructive tissue-engineered artificial skin with CH as stromal scaffold has good histocompatibility, which can be applied in grafting for full-thickness wounds.
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AIM: To observe the difference of cerebral injury following ischemia/reperfusion and mRNA expression of TGF-β1 between diabetic and non-diabetic rats.METHODS: At first, Wistar rats were divided into two groups,non-diabetes and diabetes, and then two groups followed by sham, middle cerebral artery occlusion(MCAO) 2h and reperfusion 24 h after MCAO 2 h respectively. TGF-β1 mRNA expression was measured by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR); Cerebral damage was evaluated by histopathology.RESULTS: In the same condition of ischemia or ischemia/reperfusion, injuried area enlarged in DM groups; The expression level of TGF-β1 mRNA increased at the time of 2 h after MCAO in non-diabetic group and diabetic group, especialy significantly in non-diabetic group with MCAO 2 h, and decreased at the time of reperfusion 24 h after MCAO 2 h, but still higher than that in the sham group.CONCLUSION: Diabetes mellitus exacerbated brain lesion following ischemia/repefusion, increased TGF-β1 mRNA expresion after MCAO may be an anti-injury reaction,and anti-injury ability is decreased under diabetic condition
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AIM: To study apoptotic injury induced by aeactive oxygen species - hydrogen peroxide (H2O2) on cardiac myocytes. METHODS: Cultured rat neonatal cardiac myocytes were treated with H2O2 of various concentra- tion to observe apoptoitic injury of cardiomyocytes by agarose gel electrophoresis, Giemsa - stained smears of cell, and flow cytometry, meanwhile lactate dehydrogenas (LDH) and malondialdehyde (MDA) were determined to assess the effect of H2O2 on lipid peroxidation and permeability of the plasma membrane. RESULTS: 5 mmol/L H2O2 caused culted cardiomyocytes apoptotic morophological characteristics, including nucleosomal DNA fragmentation in my- ocytes by agarose gell electrophoresis (DNA ladder), cell shrinkage, nuclear condensation, and chromatin margin by Giemsa-stained cell smears, and aneuploid peak (AP) - apoptotic bodies occurence by flow cytometry. CONCLU- SONS: H2O2 - induced apotosis in myocytes was a the - and concentration - dependent process, Treatment with low concentration of H2O2(10 mmol/L) rapidly induced a necrotic form of death characterized by smeared patterns of DNA digestion on agarose gel electrophoresis and lethal membrane disuption (as measured by LDH release). Exposure of 5 - 10 mmol/ L H2O2 induced cardiomyocytes apoptosis concurrently with biochemical changes of LDH and MDA increase in the medium.
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AIM: To study forms of cell death following cerebral ischemia/reperfusion in diabetic rats. METHODS: Based on the modles of diabetes and middle cerebral artery occlusion(MCAO), characteristics of cell death after ischemia/reperfusion were evaluated synthetically by the pathological, flow cytometry(FCM), TUNEL and the DNA agarose electrophoresis.RESULTS: The occurrence of cerebral injury after ischemia/reperfusion were accompanied by cell necrosis and cell apoptosis. And cell apoptosis was mainly located in ischeamic penumbra(IP) zone around the densely ischemic focus. Ischemic centre(IC)was characterized by cell necrosis. At the same time, the results showed that the process of ischemic cerebral injury worsen by diabetes mellitus was related to inducing cell apoptosis in IP and Mid zone.CONCLUSION: Neuronal damage following focal cerebral ischemia/reperfusion included cell necrosis and apoptosis, IC zone was mainly characterized by the former, however IP zone by the latter, and there had close internal relationship between them. Brain damage following cerebral ischemia/reperfusion was worsen instinctly under diabetic condition.
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AIM: To observe the difference of cerebral injury following ischemia/reperfusion and mRNA expression of TGF-? 1 between diabetic and non-diabetic rats.METHODS: At first, Wistar rats were divided into two groups,non-diabetes and diabetes, and then two groups followed by sham, middle cerebral artery occlusion(MCAO) 2h and reperfusion 24 h after MCAO 2 h respectively. TGF-? 1 mRNA expression was measured by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR); Cerebral damage was evaluated by histopathology.RESULTS: In the same condition of ischemia or ischemia/reperfusion, injuried area enlarged in DM groups; The expression level of TGF-? 1 mRNA increased at the time of 2 h after MCAO in non-diabetic group and diabetic group, especialy significantly in non-diabetic group with MCAO 2 h, and decreased at the time of reperfusion 24 h after MCAO 2 h, but still higher than that in the sham group.CONCLUSION: Diabetes mellitus exacerbated brain lesion following ischemia/repefusion, increased TGF-? 1 mRNA expresion after MCAO may be an anti-injury reaction,and anti-injury ability is decreased under diabetic condition.
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In the present study, Mongolian gerbil was used to make delayed neuronal damage (IND) animal model. In 6 different brain regions, 5 kinds of biochemical indexes in the aspect of free radical metabolism were measured at 7 time phases during ischemia and reperfusion. The results showed that, after transient cerebral ischemia and during reperfusion, some unique changes appeared tn hippocampal CA1 sector, which were characterized by the persistent increase of free radical content and persistent decrease of the Mn-SOD activity. The fluctuating changes could be seen in Cu, Zn-SOD activity and lipid peroxide content. There were no significant changes in the brain regions other than CA1 sector. It is concluded that the disturbance of free radical metabolism play a key role in the occur. fence of DND. It was also observed that Aniracetam, Helicid and Nieardipine had some control effects to a certatn extent on the free radical disturbance above in hippocampal CA1 sector of DND animals.