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Objective:To investigate the effect of miR-214 targeting sex determining region Y-box 4 (SOX4) on the proliferation and apoptosis of rheumatoid arthritis (RA) synovial fibroblasts.Methods:Synovial tissue samples from 45 patients with rheumatoid arthritis and 30 patients with joint trauma who underwent joint replacement surgery from 2017 to 2019 in Shangqiu First People's Hospital samples were collected. The expression levels of miR-214 and SOX4 in synovial tissues were detected by real time quantitative polymerase chain reaction (RT-qPCR); The expression levels of serum rheumatoid factor (RF) and C-reactive protein (CRP) were detected by enzyme-linked immunosorbent assay (ELISA) test. The correlation between miR-214 or SOX4 and RF, CRP, erythrocyte sedimentation rate (ESR) was tested by Pearson's analysis. Human fibroblast-like synoviocytes (HFLS)-RA cells were derived from the control group, miR-214 mimic group, mimic-NC group and miR-214 mimic+pcDNA3.1-SOX4 group and se-parated by Lipofectamine 3 000. The cell viability of each group was detected by CCK8, the apoptosis of each group was detected by flow cytometry, the target relationship was verified by dual luciferase experiment, Western-blot was used to detect the protein expression levels of related indicators. Independent sample t test was used for the comparison between the two groups, one-way nalysis of variance (ANOVA) was used for multi-group comparison, and LSD- t test was used for the comparison between the two groups. Results:Compared with the control group (4.6±0.7, 1.9±0.7) and HFLS cells (1.00±0.06, 1.00±0.09), miR-214 was expressed lower in RA tissues (2.6±0.9) and HFLS-RA (0.30±0.05) ( t=10.026, P<0.05; t=15.815, P<0.05). Compared with SOX4 was highly expressed in RA tissues (4.6±0.9) and HFLS-RA cells (3.89±0.41) ( t=14.772, P<0.05; t=12.020, P<0.05). Serum RF, ESR and CRP expression levels in RA group [(46±7) U/ml, (46.4±9.6) mm/1 h, (34.8±9.8) mg/ml] were signifi-cantly higherthan those in the control group [(16±4) U/ml, (9.2±2.0) mm/1 h, (2.1±0.7) mg/ml] ( t=22.906, P<0.05; t=25.338, P<0.05; t=22.314, P<0.05). miR-214 was negatively correlated with serum RF, CRP and ESR was negatively correlated with miRNA( r=-0.574, P<0.05; r=-0.448, P<0.05; r=-0.549, P<0.05), while was positively correlated with SOX4 ( r=0.492, P<0.05; r=0.369, P<0.05; r=0.325, P<0.05). Compared with the control group and the mimic-NC group, cell viability, Ki-67 and p-p65 protein expression decreased significantly after miR-214 mimic transfection for 24 h ( F24 h=16.980, P<0.05;, F48 h=42.735, P<0.05; F72 h=164.448, P<0.05; FKi-67=290.112, P<0.05; Fp-p65/p65=223.548, P<0.05), while the apoptosis rate and Bax protein expression increased significantly ( F=344.360, P<0.05; F=106.376, P<0.05). Overexpression of SOX4, reverses the effect of miR-214. Conclusion:miR-214 targets SOX4 to inhibit the proliferation of synovial fibroblasts and promote apoptosis in rheumatoid arthritis. This may relate to the inhibition of NF-κB signaling pathway by miR-214.
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Objective:To investigate the regulatory effect of miRNA-21-5p on peripheral blood B lymphocyte proliferation and apoptosis in systemic lupus erythematosus (SLE) patients by targeting program-med cell death protein 4 (PDCD4) .Methods:Thirty patients with SLE diagnosed clinically in our hospital were enrolled. Peripheral blood lymphocyte (PBL) was extracted by gradient centrifugation, and B cells were separated by magnetic beads. The proportion of B lymphocyte in peripheral blood was detected by flow cytometry. The cells were divided into five groups by electrotransfection: blank control group, miRNA-21-5p negative control (NC) group, miRNA-21-5p group and miRNA-21-5p inhibitor group, PDCD4 negative control group and PDCD4 siRNA group. Cells were collected 48 hours after transfection. The expression levels of miRNA-21 and PDCD4 were detected by real-time polymerase chain reaction (RT-PCR). Western blotting assay was used to detect the expression of PDCD4 in cells of each group. The targeting relationship between miRNA-21-5p and PDCD4 was verified by double luciferase target experiment. Flow cytometry was used to detect the apoptosis of cells in each group, and CCK-8 method was used to detect the proliferation of cells in each group. Western blotting and RT-PCR were used to detect the expression levels of Fas, FasL, CD40 and CD40L, respectively. Independent sample t test was used to compare the data between the two groups; single factor analysis of variance was used to analyze the results of multiple samples; chi square test was used to compare the positive rate of anti dsDNA antibody. Results:The levels of serum complement [C3 (0.85±0.11) g/L and C4 (0.54±0.09) g/L] in patients with SLE were lower ( t=7.524, P<0.05; t=38.471, P<0.05) than [C3 (1.16±0.17) g/L and C4 (1.57±0.09) g/L] in healthy controls. The levels of anti-dsDNA antibodies (47%), IgG(15.46±0.75) g/L, and IgA (2.68±0.20) g/L were increased than the levels of anti-dsDNA antibodies (17%), IgG (11.95±0.80) g/L, and IgA (2.16±0.11) g/L in healthy controls ( χ2=4.427, P<0.05; t=15.218, P<0.05; t=10.125, P<0.05). The proportion of B lymphocyte [(6.78±0.29)%] and the expression levels of miRNA21-5p (7.52±0.59) in peripheral blood of SLE patients was significantly higher than the proportion of B lymphocyte [(2.03±0.24)%] and the expression levels of miRNA21-5p (3.60±0.62) in healthy controls ( t=59.064, P<0.05; t=19.317, P<0.05), while the expression levels of PDCD4 gene (1.54±0.35) in peripheral blood of SLE patients was significantly lower than that (4.42±0.42) in healthy controls ( t=19.126, P<0.05). Compared with the blank control group and the miRNA-21-5p NC group, cell proliferation in the miRNA-21-5p Inhibitor group was inhibited, and the proportion of apoptotic cells increased ( F=5.244, P<0.05; F=37.903, P<0.05). Compared with the blank control group and PDCD4 NC group, cell proliferation in PDCD4 siRNA group was significantly enhanced, and apoptotic rate decreased ( F=5.956, P<0.05; F=25.431, P<0.05). The results of double luciferase reporter gene assay showed that PDCD4 is the target gene of miRNA-21-5p. Conclusion:miRNA-21-5p may promote the proliferation of peripheral blood B lymphocyte in SLE patients by inhibiting the expression of PDCD4, leading to abnormal lymphocyte apoptosis. miRNA-21-5p can be used as a new target gene for the treatment of systemic lupus erythematosus.
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Objective To investigate the characteristic of top-down attentional control of acute stress disorder children.Methods According to SCID-IV-TR,23 acute stress disorder children were chosen as the experinent group and 23 normal children were chosen as the control group.They were asked to perform a visual search task.Results (1) The reaction time of acute stress disorder children((1 253±158)ms) was significantly longer than normal children's((1 194± 146) ms) (P＜0.05).(2) The reaction time of valid condition((1 172± 144) ms)was significantly shorter than neutral condition ((1 229± 156) ms) and invalid condition ((1 269± 157) ms) (P＜0.05),there was no difference of reaction time between neutral condition((1 229± 156)ms) and invalid condition ((1 269±157)ms) (P＞0.05).(3) The reaction time of simple display condition((1 182±127)ms) was significantlv shorter than complex display condition ((1 264± 177)ms) (P＜0.01).Conclusion The performance of acute stress disorder children on top-down attentional control is less than normal children,the reason is that inhibiting capacity of acute stress disorder children is lower than normal children.It indicates that trauma event have negative influence on children's inhibiting capacity.
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Objective To investigate the value of positron emisson tomography-computed tomography ( PET-CT) in monitoring the residual lesions in lipiodol sedimentary region and extrahepatic metastastic lesions of hepatocellular carcinoma ( HCC) with alpha fetoproteins ( AFP) positive after interventional therapy. Methods The data of 20 cases with primary HCC confirmed by histopathology were retrospectively analyzed. Their AFP levels decreased to normal range after interventional treatments, but rose to abnormal high level during following-up. After the abdominal routine imaging examinations, the definite diagnosis of the residual lesions in lipiodol sedimentary region or extrahepatic lesions can't be made confidently. All cases were scanned by PET-CT, and according to their PET-CT results, the further treatments were given and the therapeutic results were monitored with radiology and AFP tests. Results In all 20 cases, 10 of them were detected to have the extrahepatic metastastic lesions by PET-CT, including 4 with abdominal wall metastasis upon the liver, 3 with solitary pulmonary metastasis with diameter less than 1 cm, 2 with mesenteric mestastasis, 1 with mestastasis of operative incisions, but these lesions were neglected by abdominal routine imaging examinations. Eight cases showed the uneven lipiodol sedimentary region in the primary lesion by CT or MRI examination, but can't be diagnosed whether it was residual lesion with other examinations including DSA. A definite diagnosis was obtained by PET-CT. In 2 cases, lymph nodes less than 1. 5 cm were found in the hepatic portal area(PHA) and retroperitoneum on CT images,which was determined to be metastatic by PET-CT. All the detected lesions were given further treatments of surgery or interventional therapy. Most patients showed decreased AFP levels except the 2 patients with lymph node metastasis. The imaging examinations also indicated that the treatments had a good effect on lesions. Conclusion In the patients with HCC and abnormal high levels of AFP, in addition to abdominal routine imaging examinations, PET-CT is beneficial for those who is suspected of residual lesions in lipiodol sedimentary region or extrahepatic metastasis.
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Objective To compare the diagnostic values of US-guided optical imaging system (OPTIMUS) and full-field digital mammography (FFDM) in breast neoplasms. Methods Sixty-four patients suspected of breast neoplasms underwent OPTIMUS and FFDM before surgery. All patients were proved histopathologically. The accuracy of diagnosis was compared with chi-square test. Results Sixty-five masses were delineated by OPTIMUS and FFDM. The diagnostic accuracy, sensitivity and specificity of OPTIMUS were 86. 2% ( 56/65 ), 87.0% ( 20/23 ) and 85.7% ( 36/42 ), respectively. The positive likelihood ratio was 6. 09 and the negative likelihood ratio was 0. 175. The diagnostic accuracy, sensitivity and specificity of FFDM were 73.8% (48/65), 82. 6% ( 19/23 ) and 71.4% (30/42), respectively. The positive likelihood ratio was 2. 892 and the negative likelihood ratio was 0. 243. There were no significant differences between OPTIMUS and FFDM ( x2 = 3. 077 ,P ＞ 0. 05). Conclusion OPTIMUS is similar and supplementary to FFDM for the diagnosis of breast neoplasms.
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Objective To explore the feasibility of detecting atherosclerosis with 7.0T MR and Micro-PET. Methods Ten 46-week-old ApoE-/- mice with high lipid diet for 6 months were selected to establish atherosclerosis models. Among them, 5 mice underwent MRI before and 12 h, 24 h, 36 h after injection of SPIO, respectively, and the other 5 mice were injected with ~(18)F-fluorodeoxyglucose (~(18)F-FDG) through tail vein and observed with Micro-PET after 1 h, 2 h and 3 h. The specimens of abdominal aorta were taken for pathologic examination. Results Atherosclerotic plaques were observed in all animals with 7.0T MRI after 6 months high lipid diet. Thirty-six hours after the injection of SPIO, the high signal rings were thinner and the lumen of blood vessels were wider than those before injection on T2WI. Radioactive concentration was observed in abdominal aorta and both sides of iliac artery 3 h after the injection of ~(18)F-fluorodeoxyglucose (~(18)F-FDG). Pathological examination showed the formation of atherosclerotic plaques and the aggregation of the macrophages. Conclusion 7.0T MRI and Micro-PET can be used to observe the macrophage-rich plaque and to judge the vulnerability of plaque, thus provide theoretical basis for early detection, diagnosis and treatment of atherosclerosis.
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Objective To investigate the tissue characteristics within vascular stent and transjugular intrahepatic portosystemic shunt(TIPS)on swine and to provide more information for the understanding and prevention of vascular stent and TIPS restenosis.Methods Animal models for TIPS were built in 6 swine and vascular stents were implanted in iliac veins simultaneouly.14-28 days after the operation.the 6 swine were killed to remove the TIPS and vascular stent and the pathological examinations were performed on the tissues within the shunt and stent.The similarities and difierences of the tissues within the shunt and stent were analvzed with Kruskal Wallis test. Results Restenosis of TIPS occurred in 4 models and complete occlusion were seen in 2,while all vascular stents were patent and coated with a thin layer of intimal tissueElectron micmscopic results showed that the tissues in restenotic TIPS were loose and with more extra matrix and fibers.and less smooth muscle,fibroblastic and myofibroblastic cells with different and irregular shape and rich secretory granules.The tissues in patent,TIPS contained more extra fibers,smooth muscle and fibmblastic cells with normal organdie.The intimal tissues in vascular stent contained more fibers and fibroblasts cells.less smooth muscle cells.On immunohistoehemical staining,the tissues in restenotlc and Datent TIPS as well as the intimal tissues in vascular stent had strong positive expression for anti-SMCactin-α.the expression were gradually weakened for PCNA,the intimal tissues in vascular stent had a strong positive expression for vimentin,while the expression of the tissues in restenotic and patent TIPS were weakened gradually.For myoglobulin,the tissues in restenotic TIPS had weakly positive expression,the expression in patent TIPS and vascular stent were almost negative. Western blot results for TGF-β showed that the absorbance ratios of the intima tissues in vascular stent,normal vascular tissues,normal liver tissues.the tissues in restenotic and patent TIPS were 0.23,0,0,0.57 and 0.30 respectively,and they were significantiy different (H=27.8,P＜0.01).Conclusions The tissues in restenotic TIPS mainly contain proliferated SMCs which have positive expression for anti-SMC-actin-α,strong proliferation and movement but unstability.The tissues in patent TIPS and intimal tissues in Vascular stent mainly contain fibroblastie cells which have positive expression for vimentin and stability.
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Objective To investigate the trans-arterial delivery of p53 gene transfection efficiency and therapy effect on hepatic carcinoma in combination with transferrin mediated by liposome. Methods Twenty-five VX2 experimental rabbits were randomly divided into five groups, and the different doses of transferrin combined with p53-LipofectAMINE complex were delivered into the hepatic arteries of the VX2 hepatic carcinoma models. The tissue protein of the carcinoma was extracted after 48 h and the transfection efficiency and expression rate of p53 gene were analyzed by western blot and immune histochemical techniques, to inspect the expression proportion of p53 with different doses transferring. Another ten VX2 were divided into two groups, recombinant plasmid p53-LipofectAMINE complex and transferrin-p53-LipofectAMINE complex were delivered into the hepatic arteries in two groups respectively. The liver function, size of the tumor and survival time of the animals was compared between the two groups, and results were analyzed statistically. Results Semiquantitative analysis by Western Blot showed that the transfection and expression efficiency of p53 gene combined with transferrin were higher than those without it. By immune histochemieal techniques, the p53 gene's positive rate of highly expression with various doses of transferrin were found to be different, and there was remarkable difference between the groups with and without transferring. They were 58. 33%, 69. 44%, 80. 00%, 83.33%, 81.67% respectively, there was remarkable difference between the groups with and without transferring ( Totality: x2 = 42. 37, P < 0. 01 ). The p53 gene's positive rate of expression increased gradually as the doses of transferrin increasing from 0 up to 200 μg, but the differences of positive rate had no statistical significance as the doses of transferrin increasing from 200 up to 400 μg ( x2 section : 3 groups as former x2 = 4. 82, P < 0. 05,3 groups as latter x2 =0. 67 ,P <0. 05). There was no statistical difference in the liver function at points of time between VX2 rabbits with and without transferring. But the tumors' sizes had significant difference at various points of time. Conclusion Liposome-mediated p53 gene on treating hepatic carcinoma by trans-arterial gene delivery combined with transferrin is safe, and it can markedly enhance transfection efficiency and improve the therapy effect of p53 gene.
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Objective To evaluate the diagnostic significance of full-field digital mammography (FFDM)for atypieal breast cancer findings.Methods Seven hundred-eighteen cases with breast cancer were examined using FFDM and atypical mammographic findings were found in 134 cases.Craniocaudal (CC)view and mediolateral oblique(MLO)view were conducted for each patient.Mediolateral view or spot view Was achieved if necessary.Preoperative localization Was conducted for the patients with nonpalpable breast cancer8.Results (1)The masses with well.circumscribed margin on mammography were more common in infiltrating duetal carcinoma(22/106),mucinous carcinoma(8/10),medullary carcinoma (5/6),and increased with age and reduction of the mass density.(2)Long spieulation,architectural distortion,patchy high density were mainly found in infiltrating ductal carcinoma and 30-40(24 cases),41-50(34 cases)years old patients.Long speculation wag mainly found in 30-40 years old patients (10/30).(3)Hish homogenous density and subcutaneous edema in the entire breast and mass-like area were most ffequendy found in infiltrating ductal carcinoma at 30-40(2 cases)and 41-50(5 cases)years old.High density and subcutaneous edema were only found in dense breast(8 cases).Conclusion The atypical findings of breast cancer in foil-field digital mammography are associated with the pathological type of cancer.patient age and the gland density of the breast.
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Objective To study retrospectively the curative effect of emergent transcatheter arterial embolization(TAE)on ruptured liver cancer with hemorrhage. Methods Twenty-three patients with rupture of liver cancer were emergent treated with TAE for massive hemorrhage. Results Twenty-two patients were treated by TAE effectively with good hemostasis and without recurrence of bleeding as well as liver failure, except one referred for surgery due to anomalous vascular route leading to failure of catheterization. Nine of 22 patients accepted more than one time of TAE according to the curative effect and progress of the disease, but reduction of tumor size or necrosis could be achieved in all these patients. Conclusions Emergent TAE is an effective treatment for ruptured liver cancer with hemorrhage, having good effect of hemostasis and providing the possibility for subsequent treatment.