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Objective To explore the effect and mechanism of miR-581 on the autophagy of ovarian cancer SKOV3 cells. Methods miR-581 mimics and miR-581 NC were transfected into SKOV3 cells, and the transfection efficiency was detected by qRT-PCR. After successful transfection, Western blot was used to detect autophagy-related proteins expression in SKOV3 cells. TargetScanHuman database predicted miR-581 target genes, and Western blot verified the role of miR-581 and target genes. Results Overexpression of miR-581 significantly inhibited the expression of autophagy-related proteins LC3 Ⅱ and Beclin1 (P < 0.01); miR-581 negatively regulated FOXO1 expression (P < 0.01) and FOXO1 promoted autophagy of SKOV3 cells (P < 0.01). Conclusion miR-581 affects the autophagy of ovarian cancer SKOV3 cells by regulating the expression of FOXO1.
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Objective To obtain the Cyclin D1,the Cyclin D1 gene was cloned and expressed in Pichia pastoris.Methods The total RNA was extracted from liver cancer tissue.The Cyclin D1 cDNA was obtained by RT-PCR.The Cyclin D1 cDNA was sequenced.And sub-clonedto the Ppic9k,the Pichia pastoris expressed was used to obtain the Cyclin D1.Results The 483 bp Cyclin D1 cDNA was obtained.And the sequence of Cyclin D1 was corrected.The 36KD Cyclin D1 was obtained by Pichia pastoris expression.Conclusion The Cyclin D1 cDNA was cloned and Cyclin D1 was expressed in Pichia pastoris.
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Objective To establish the rheumatoid arthritis (RA) mouse model induced by glucose-6-phosphate isomerase (GPI),and explore the mechanism of GPI in RA.Methods Totally 36 DBA/1 mice were randomly divided into three groups:test group (injection GPI),positive group (injection of bovine collagen Ⅱ),and negative group (saline).The rates and changes of weight were observed.The score of the arthritis,the ankle histopathological changes and serum GPI content were detected.Results Toes swollen slightly,joint swelling,deformity and accompanied by block were appeared at 35th day in the test group.Compared to the control group,the rates and changes of weight in test group showed a significant difference (P ＜ 0.05).The score of arthritis was showed by x ± s.Compared to the negative group,the test group and positive group were showed significant difference (P ＜ 0.05).A lot of lower synovial lining exudate macrophages,fibroblasts,and other inflammatory cells were increased in the test group.The GPI content in the test group [(0.39 ±0.11)μg/ml] was significantly higher than the negative group [(0.10± 0.06) μg/ml,P ＜ 0.05].Conclusions GPI could induce rheumatoid arthritis in mice.It provides the experimental basis to diagnose RA.
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Objective To obtain the Cyclin D1 through cloning and prokaryotic expression of Cyclin D 1 gene.Methods The total RNA was extracted from liver cancer tissue .The Cyclin D1 cDNA was obtained by reverse transcriptase polymerase chain reaction (RT-PCR).The Cyclin D1 cDNA was sequenced, and sub-cloned to the PET32a+.The prokaryotic expressed was used to obtain the Cyclin D1.Results The 483 bp Cyclin D1 cDNA was obtained.The sequence of Cyclin D1 was corrected.The 36 KD CyclinD1 was obtained by prokaryotic expression .Conclusions The Cyclin D1 cDNA was obtained.Cyclin D1 was expressed in BL21.
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Objective To investigate the clinical application of fecal calprotectin in inflammatory bowel disease (IBD).Methods Colonoscopy took 79 patients with IBD that were diagnosed with pathology,including 47 cases of ulcerative colitis (UC) patients,32 cases of Crohn's disease (CD).Moreover,42 cases of IBD patients without abdominal pain,diarrhea and other intestinal inflammation were used as disease control group,and 34 cases of healthy people were used as healthy control group.The level of fecal calprotectin in each group was detected by enzyme-linked immunosorbent assay (ELISA).Results The positive rate of fecal Calprotectin in IBD group,disease control group and the healthy control group was 57.0％,19.0％,and 0,respectively; each positive rate in IBD group was significantly higher than the other two groups (P ＜ 0.05).The serum concentration of fecal calprotectin in IBD group [(493.86 ±204.18) μg/g] was significantly higher than the disease control group [(71.46 ± 60.51) μg/g] and the healthy control group [(36.19 ± 13.46) μg/g] (P ＜ 0.05) ; IBD active calprotection [(1015.23 ± 324.96) μg/g] was significantly higher than resting [(52.69 ±34.71) μg/g] (P ＜0.01).Conclusions Fecal calprotectin test benefits early diagnosis of IBD,and may be taken as the diagnostic index of IBD activity.It has extensively clinical value.
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Objective To investigate the value of urinary trehalase (T) in patients with renal proximal tubular damage.Methods 134 patients with kidney disease (male:66 female:68 age:18-59 ; 31cases with acute renal failure,30 cases with chronic renal failure,20 cases with drug-induced renal impairment,21 cases with renal transplantation and 32 cases with nephritic syndrome) and 101 healthy controls (58 males and 43 females) were selected.Urinary T,N-acetyl-D-glucosaminidase (NAG),β2-MG,gamma-glutamyl transferase (GGT) were detected.Data were analyzed by SPSS 11.5,including nonparametric rank test,ROC analysis.Results The level of urinary trehalase in control group was normally distributed (7.1 ± 4.1) μmol/h · g Cr (0-25 μmol/h · g Cr).There was no significant difference between men and women (t =0.63,P =0.53).Urinary T levels were significant higher in all kidney disease groups than in control group (Z =6.80,5.90,5.23,6.00,8.04,P ＜0.01).According to ROC curve,the area of urinary T under the ROC curve (AUC) in 134 patients was 0.9,significantly different with NAG,β2-MG,GGT area (P ＜ 0.01),the AUCs of T were 0.94,0.85,0.90,0.90,0.91 in acute and chronic renal failure group,drug-induced renal impairment group,renal transplantation group and nephritic syndrome group,respectively; Youden index were 0.85,0.65,0.77,0.66,0.72 respectively.Corresponding to the Youden index,sensitivity and specificity were 90.3％ and 95.1％,73.3％ and 92.1％,85.0％ and 92.1％,81.0％ and 85.2％,87.5％ and 84.2％ respectively.Conclusions The Urinary trehalase is better than other markers in the diagnosis of the proximal renal tubular damage.It was better to evaluate the proximal tubular function early in time.The diagnostic value of urinary trehalase played a key role in diagnosis,treatment and prognosis of kidney diseases.