1.Effects of the human 21 .5 kDa MBP gene silencing on the proliferation and apoptosis of glioma cells
Ruimin TIAN ; Hanyan WANG ; Fang YI ; Xiaoming WANG ; Jianye CHEN
Chongqing Medicine 2015;(1):10-13
Objective To investigate the effects of silencing of the human cerebral 21 .5 kDa myelin basic protein (MBP) gene on the proliferation and apoptosis of the glioma U251 cells .Methods The 21 .5 kDa MBP sequence‐specific short hair‐pin RNA (shR‐NA) recombinant plasmids pGenesil‐1‐MBP‐3 were transfected into the human glioma cell line(U251) ,the cells of U251 was used as MBP silencing group ,the cells transfected with negative control plasmids used as negative control group ,and the cells transfected with liposomes used as blank control group .Real‐time PCR and Westernblot were used to detect the expression levels of the 21 .5 kDa MBP mRNA and protein in each group ,and the cell proliferation curve was measured by CCK‐8 assay ,the apoptosis rate was a‐nalysised by Flow cytometry .Results Both the mRNA and the protein expression levels of the 21 .5 kDa MBP of MBP silencing group were significantly lower than those in the control groups (P<0 .05);the cellular proliferation activity of the MBP silencing group decreased significantly (P<0 .05)while the cellular apoptotic rate increased significantly (P<0 .05) .Conclusion Silencing of the human cerebral 21 .5 kDa MBP gene may playa dual role in the inhibition of proliferation and the promotion of apoptosis of the glioma U251 cells .
2.Correlation Between Breast Imaging Reporting and Data System of Ultrasound and 18F-lfuorodeoxyglucose PET/CT in Diagnosis of Breast Diseases
Peng YU ; Ruimin WANG ; Baixuan XU ; Jiahe TIAN
Chinese Journal of Medical Imaging 2014;(10):730-734
Purpose To evaluate the breast imaging reporting and data system (BI-RADS) of ultrasound and 18F-lfuorodeoxyglucose (FDG) PET/CT in diagnosis of breast tumor, and to analyze the correlation between them. Materials and Methods 18F-FDG PET/CT images and ultrasound images of 103 patients with suspected breast cancer were retrospectively analyzed to get correlation between the maximum standardized uptake values (SUVmax) and BI-RADS. Sensitivity, speciifcity, positive predictive value, negative predictive value were compared with histology or follow-up results as golden standard. Results Of the 103 lesions, 46 were benign and 57 were malignant. Pearson correlation coefifcient was 0.464 (P<0.01). The sensitivity, speciifcity, positive predictive value, and negative predictive value of PET/CT were 89.47%, 73.91%, 80.95%and 84.99%, respectively;those of BI-RADS were 94.70%, 69.60%, 79.42%and 91.38%, respectively. The sensitivity, speciifcity, positive predictive value, and negative predictive value in patients with BI-RADS 3-4 were 88.90%, 71.40%, 66.65%, and 90.91%, respectively. The sensitivity, speciifcity, positive predictive value, and negative predictive value for BI-RADS grading diagnosis were 88.90%, 46.40%, 51.60%and 86.67%, respectively. Conclusion There is no signiifcant correlation between SUVmax and BI-RADS. BI-RADS has low speciifcity for patients with BI-RADS grade 3-4, while PET/CT can make up this shortcoming. Combined diagnosis in the breast disease can be potentially recommended in clinics.
3.The study on the value of stagnation rate at 15 min of indocyanine green for assessment of hepatic reserve function
Ruimin TIAN ; Guowang LIU ; Wei WEI ; Kecheng TANG
Tianjin Medical Journal 2016;44(9):1136-1138
Objective To observe the clinical value of indocyanine green retention rate at 15 minutes (ICGR15) in the evaluation of hepatic functional reserve. Methods A total of 185 patients with liver disease, including 45 cases of liver failure, 90 cases of cirrhosis (child A, B and C, respectively), 20 cases of acute hepatitis, 30 cases of chronic hepatitis (mild, moderate). Expression levels of ICGR15 were compared between groups. Values of ICGR15, total bilirubin (TBIL), albumin (ALB), blood coagulation time (PT) were compared before treatment and one month after treatment in hepatic failure group. Levels of alanine aminotransferase (ALT), aspertate aminotransferase (AST), TBIL and ICGR15 were compared before treatment and 1 month after treatment in acute hepatitis group. Results Levels of ICGR15 (%) were 56.3±14.7, 28.9±9.6, 22.4±6.8 and 13.7±2.3 in liver failure group, liver cirrhosis group, acute hepatitis group and chronic hepatitis group, which showed a gradual downward trend (F=125.317, P<0.05). Among them, the levels of ICGR15 (%) were 17.3±5.4, 25.7±7.5 and 34.5±7.3 in Child A, B and C groups of liver cirrhosis group, which showed a gradual upward trend (P<0.05). After one month treatment, levels of TBIL, PT and ICGR15 were significantly lower than T helper 17 cells; intima-media thickness before the treatment in liver failure group. The levels of ALT, AST, TBIL and ICGR15 were significantly lower after treatment than those before treatment in acute hepatitis group (P<0.05). Conclusion ICGR15 can reflect hepatic reserved function, which is not affected by the application of albumin and fresh plasma, and makes up the deficiency of PT and ALB detection.
4.The diagnostic value of serum sialic acid for glioma
Ruimin MA ; Guojun ZHANG ; Yan CHEN ; Hui ZHAO ; Meng TIAN ; Xixiong KANG
Chinese Journal of Laboratory Medicine 2016;39(3):201-204
Objective To discuss the diagnostic value of serum sialic acid for glioma.Methods A retrospective study was conducted.The levels of sialic acid in serum samples of 95 glioma patients, 175 patients with brain benign tumor and 400 normal persons from October 2014 to March 2015 were detected by automatic biochemistry analyzer using enzymic method.The SNK-q test and analysis of variance were used to compare the difference of the groups.By making receiver operating characteristic ( ROC) curve, the cut-off value, sensitivity, specificity, positive predictive value and negative predictive value were calculated to assess the diagnostic value of serum salivary acid.Then the cut-off value was validated by using the serums of 30 glioma patients and 30 normal persons who were out-patients and healthy controls.Results The levels of serum sialic acid in patients with gliomas, patients with brain benign tumor and healthy individuals were (0.66 ±0.14 ) g/L, (0.61 ±0.09 )g/L, (0.54 ±0.07 )g/L.The serum salivary acid of glioma patients were higher than brain benign tumor patients (q=6.74,P<0.05) and normal persons (q=16.42,P<0.05), and there was no significant difference (F=1.67,P>0.05) among the glioma patients of different grades (8 of gradeⅠ,32 of gradeⅡ,24 of grade Ⅲ,31 of grade Ⅳ).There was no significant difference between the low grade patients (gradeⅠandⅡ) and the high grade patients (gradeⅢandⅣ) (t=0.55, P>0.05), but the level of serum sialic acid of high grade group had an increasing trend than the low grade group.The area under the ROC curves was 0.79.The cut-off value of serum salivary acid for diagnosing glioma was 0.61 g/L.The sensitivity was 67.74%, and the specificity was 80.60%.The positive predictive value was 44.68%, and the negative predictive value was 90.76%.Then the cut-off value was used as a diagnostic criteria, and the detected results of 30 glioma patients and 30 normal persons showed that the sensitivity was 63.30% and the specificity was 83.30%.Conclusions The serum sialic acid has good specificity and negative predictive value for diagnosing glioma.It may be a valuable diagnostic marker.
5.Proteomic fingerprinting of N-linked glycoproteins involved in hepatocellular carcinoma
Jin MA ; Yijun QI ; Ruimin LIU ; Ming WANG ; Tian ZHANG ; Han ZHU ; Yuanfang MA
Acta Anatomica Sinica 2014;(4):493-499
Objective To identify differentially expressed N-linked glycoproteins between hepatocellular carcinoma ( HCC) and adjacent non-tumorous liver tissues .Methods N-linked glycoproteome was extracted by multi-lectin affinity chromatography comprising concanavalin A (ConA), lentil lectin (LCH), and snowdrop lectin (GNA) and subsequently subjected to two-dimensional electrophoresis ( 2DE ) and mass spectrometry ( MS ) for identification of differential glycoproteins between 10 pairs of HCC and adjacent non-cancer tissue .Western blotting was used to verify different expression of human liver carboxylesterase 1 (hCE1), haptoglobin (HP)and cathepsin D (CD).Invasion potential in vitro was examined after si-RNA mediated CD gene scilencing .Results LC-ESI-MS/MS identified a total of 28 differentially expressed glycoproteins (14 up-regulation and 14 down-regulated).Western blotting detected consistent down-regulation of hCE1 and HP, and up-regulation of pro-cathepsin D (pCD) in HCC.Up-regulation of ConA-binding CD (ConA-CD), however , was verified in HCC only after ConA-CD enrichment by ConA chromatography .Down-regulation of CD expression mediated by CD-siRNA markedly inhibited the in vitro invasive potential of SNU449 and SNU473.Conclusion Dysregulation of HP , hCE1 expression and alteration of glycans linked to CD may play crucial roles in pathogenesis of HCC.
6.Relationships between tau and brain atrophy in Alzheimer′s disease based on 18F-THK5317 PET/MR
Liping FU ; Xiaojun ZHANG ; Teng XIE ; Ruimin WANG ; Fang YI ; Jinming ZHANG ; Luning WANG ; Hengge XIE ; Baixuan XU ; Jiahe TIAN
Chinese Journal of Nuclear Medicine and Molecular Imaging 2021;41(6):321-326
Objective:To investigate the neuroimaging relationship between tau protein deposition and brain atrophy, and assess their relationships with cognitive decline in Alzheimer′s disease (AD) patients.Methods:From April 2017 to October 2019, 26 AD patients (12 males, 14 females, age (70.7±12.2) years) and 19 cognitively normal controls (CN; 9 males, 10 females, age (65.6±8.1) years) in Chinese PLA General Hospital were retrospectively enrolled. All subjects received (S)-6-[(3- 18F-fluoro-2-hydroxy)propoxy]-2-(4-methylaminophenyl)quinoline ( 18F-THK5317) PET/MR and the standardized uptake value ratio (SUVR) and gray matter volume (GMV) were measured. General linear model (GLM) was used to evaluate the differences of SUVR and GMV between two groups. Pearson correlation analysis was used to assess the relationships between SUVR and GMV, and relationships of SUVR and GMV with Mini-Mental State Examination (MMSE) scores in AD patients. Results:Compared with CN, the AD patients showed significantly increased 18F-THK5317 retention in lateral temporal, frontal, posterior cingulated/precuneus and occipital cortex with significant differences of SUVR between two groups (2.18±0.54 vs 1.78±0.09, 2.13±0.50 vs 1.82±0.06, 2.03±0.45 vs 1.69±0.08, 2.18±0.57 vs 1.76±0.10, t values: 2.58-6.57, all P<0.001). The AD patients also showed decreased GMV in medial temporal, posterior cingulated/precuneus and orbitofrontal cortex ( t values: 3.67-8.85, all P<0.001). In AD patients, SUVR was negatively associated with GMV in bilateral lateral temporal cortex, pre-frontal cortex and orbital frontal cortex ( r values: from -0.52 to -0.43, all P<0.05). Both SUVR ( r=-0.599, P=0.001) and GMV ( r=0.443, P=0.023) were significantly correlated with MMSE scores in AD patients. Conclusion:AD patients have neocortical 18F-THK5317 abnormal uptake and GMV reduction, which are significantly correlated with cognitive decline.
7.Diagnosis of brain tumor with multi-modality imaging of 11C-MET PET combined with functional MRI
Haodan DANG ; Changbin LIU ; Ruimin WANG ; Jiajin LIU ; Zhiwei GUAN ; Liping FU ; Can LI ; Jinming ZHANG ; Jiahe TIAN ; Baixuan XU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2017;37(9):527-531
Objective To evaluate the diagnostic efficiency of the parameters obtained from PET/MR in brain tumors.Methods In this prospective study,28 patients (21 males,7 females,age range: 6-82 years) with clinical suspicion of brain tumor from November 2012 to September 2015 underwent PET/MR multi-modality imaging.The examination of PET/MR included 11C-MET PET and multiple MR sequences.The qualities of images were estimated firstly.The ROC curve and the accuracy of SUVmax,ADCmean,rCBF and NAA/(Cho+Cre) ratio were calculated.The pathology or final clinical diagnosis was taken as the standard.The diagnostic efficiency of the multi-modality imaging was determined based on the cutoff values of the four parameters.Two-sample t test was used to analyze the differences of parameters between glioma group and inflammatory group.Results SUVmax,ADCmean,rCBF and NAA/(Cho+Cre) ratio were validated to be effective parameters in diagnosing brain tumors with the diagnostic accuracy of 89.3%(25/28),82.1%(23/28),78.6%(22/28) and 75.0%(21/28),respectively.The SUVmax exhibited the highest diagnostic accuracy,while the diagnostic efficiency of the combination of four parameters was superior to the separate parameter.The values of SUVmax,rCBF and NAA/(Cho+Cre) ratio were significantly different between glioma group(n=10) and inflammatory group(n=11;t values:-2.31,-3.11,-2.77,all P<0.05).Conclusions PET/MR can provide a one-stop examination with multi-modality imaging of brain.The obtained parameters SUVmax,ADCmean,rCBF and NAA/(Cho+Cre) ratio,especially their combination,have effective diagnostic values on brain tumor.
8.Cytotoxicity of human doppel protein and Dpl-like protein PrP?32-121 to SH-SY5Y cells
Kun XU ; Xin WANG ; Chan TIAN ; Song SHI ; Guirong WANG ; Qi SHI ; Ruimin ZHOU ; Huiying JIANG ; Yonglie CHU ; Xiaoping DONG
Journal of Xi'an Jiaotong University(Medical Sciences) 1982;0(01):-
Objective To observe the biological activities of human doppel(Dpl) protein transiently expressed and Dpl-like protein PrP?32-121 on a human neuroblastoma cell line SH-SY5Y.Methods Recombinant mammalian expression plasmids containing human PRND gene and truncated PrP?32-121 fragment were generated by PCR.The expression and location of Dpl and PrP?32-121 post-transfection were observed by IFA.The cytotoxicity was measured by MTT analysis.Cellular apoptosis was investigated by flow cytometry and Western blot.Results Both Dpl and PrP?32-121 protein were expressed and mainly located on the cell membrane.Remarkable cytotoxicity was detected on SH-SY5Y cells after 24 h transfection.Meanwhile,more Annexin V/PI positively-stained cells as well as lower levels of cellular pro-caspase-3 and Bel-2 were detected in the cells receiving Dpl and PrP?32-121 expressing plasmids.Conclusion Dpl protein transiently expressed and PrP?32-121 can lead to the similar neural cytotoxicity,probably triggering the cell apoptosis program.
9.Cytotoxicity of human doppel protein and Dpl-like protein PrPΔ32-121 to SH-SY5Y cells
Kun XU ; Xin WANG ; Chan TIAN ; Song SHI ; Guirong WANG ; Qi SHI ; Ruimin ZHOU ; Huiying JIANG ; Yonglie CHU ; Xiaoping DONG
Journal of Xi'an Jiaotong University(Medical Sciences) 2010;31(1):32-35,46
Objective To observe the biological activities of human doppel (Dpl) protein transiently expressed and Dpl-like protein PrPΔ32-121 on a human neuroblastoma cell line SH-SY5Y. Methods Recombinant mammalian expression plasmids containing human PRND gene and truncated PrPΔ32-121 fragment were generated by PCR. The expression and location of Dpl and PrPΔ32-121 post-transfection were observed by IFA. The cytotoxicity was measured by MTT analysis. Cellular apoptosis was investigated by flow cytometry and Western blot. Results Both Dpl and PrPΔ32-121 protein were expressed and mainly located on the cell membrane. Remarkable cytotoxicity was detected on SH-SY5Y cells after 24 h transfection. Meanwhile, more Annexin V/PI positively-stained cells as well as lower levels of cellular pro-caspase-3 and Bel-2 were detected in the cells receiving Dpl and PrPΔ32-121 expressing plasmids. Conclusion Dpl protein transiently expressed and PrPΔ32-121 can lead to the similar neural cytotoxicity, probably triggering the cell apoptosis program.
10.Construction of Luciferase Reporter Vector Containing High-mobility Group Box 1 Promoter in Mice and Characterization of Its Transcriptional Activity
Quan WEN ; Shaohui DU ; Ruimin TIAN ; Hui LI ; Ye BAI ; Fei LI ; Hang LEI ; Dongfeng CHEN ; Yiwei LI ; Jianhong ZHOU ; Saixia ZHANG ; Rudong DENG ; Yao YU
Journal of Guangzhou University of Traditional Chinese Medicine 2014;(5):810-813,820
Objective To construct luciferase reporter vector containing full-length high-mobility group box 1 ( HMGB1, GenBank NM-010439) promoter for the screening of medicine. Methods The full-length HMGB1 promoter was amplified by polymerase chain reaction ( PCR) , and then was inserted into GV238 vector to construct plasmid GV238-HMGB1-P-Luc. GV238-HMGB1-P-Luc combined with internal reference plasmid pRL was co-transfected into Hela cells ( GV238-HMGB1-P-Luc group, which served as positive control group) . Plasmid pGL3-basic combined with pRL was co-transfected into Hela cells (pGL3-basic group, which served as negative control group) . Additionally, lipopolysaccharides ( LPS, 0.2 μg/mL) was used as the activator for the positive control group (LPS group), and then sodium butyrate (SB, 10 mmol/L) was used as the inhibitor for LPS group ( SB group) . At the end of experiment hour 24, luciferase activity was detected. Results The results of digestion, amplification, sequencing and identification showed that the full length of HMGB1 promoter was 2 140 bp, and the DNA sequence was correct, without mutation. Luciferase activity in GV238-HMGB1-P-Luc group was increased as compared with that of the pGL3-basic group ( P<0.05) . Luciferase activity in the LPS group was increased ( P<0.01, compared with that of GV238-HMGB1-P-Luc group) , and then was decreased after the administration of SB ( P<0.01, compared with that of the LPS group) . Conclusion A model of luciferase reporter vector containing HMGB1 promoter has been successfully constructed. Its activity can be increased by LPS, and then is in hibited by SB. The model can be used for further screening of medicine with the activities of regulating HMGB1 promoter.