1.Intra- and interobserver reproducibility in the assessment of coronary artery disease: evaluation with invasive coronary angiography and CT coronary angiography
Mingli SUN ; Bin Lü ; Runze WU ; Shiguo LI ; Zhicheng JING ; Lei HAN ; Yanmin HUO ; Fangfang YU ; Shiliang JIANG ; Ruping DAI ; Jianhua LU ; Zhihui HOU ; Yang GAO ; Huili CAO ; Yongjian WU ; Yuejin YANG ; Shubin QIAO
Chinese Journal of Radiology 2012;46(2):104-109
Objective To investigate the intra- and interobserver repeatability of coronary artery disease (CAD) diagnosis based on invasive coronary angiography (ICA) and CT coronary angiography (CTCA).Methods Two readers with comparable experience ( over 10 years) independently evaluated ICA results of 42 consecutive patients with a blind method. After 30 days,one of them reviewed the same patients again.Another two comparable-experience (over 10 years) readers evaluated the results of CTCA (prospectively ECG-triggering) from the same 42 patients in the same way.The inter-reader and intra-reader repeatability of ICA and CTCA were analyzed by performing Kappa test and calculating the percentage of the segments with agreement on stenotic degree.Using ICA as reference,the accuracy of CTCA in diagnosing CAD was studied by comparing the area under ROC. Results The Kappa between readers for ICA and CTCA were 0.91 and 0.81.Intra-reader Kappa were 0.92 and 0.83 respectively (x2 =509.4 and 432.5,all P <0.01 ).The percentage of the segments with agreement between readers on the degree of stenosis were 80.8% (494/611) in ICA and 75.2% (469/624) in CTCA ( x2 =2.75,P =0.10),and within the same reader,86.9% (531/611)in ICA and 81.9% (511/624) in CTCA(x2 =3.76,P =0.053).With≥ 50%narrowing as a CAD diagnosis criterion,the agreement rates for two readers were 96.6% (590/611 ) in ICA and 94.4% (589/624) in CTCA( x2 =3.36,P =0.07),and for the same reader,97.4% (595/611) in ICA,95.4% (595/624) in CTCA ( x2 =3.62,P =0.06).Using ICA as reference,two readers of CTCA results achieved a sensitivity and specificity of 84.9% (530/624)and 98.1% (612/624).The area under ROC was 0.94 (95% CI 0.91-0.97).Conclusions Both ICA and CTCA demonstrate good repeatability in diagnosing CAD.The repeatability of ICA is superior to that of CTCA.A certain discrepancy exists in two readings from the same reader or two readers.
2.Expression of lncRNA MIR210HG in preeclampsia placental tissue and its functional analysis
Xueli LI ; Lu ZHANG ; Bo HOU ; Shunfu PIAO ; Qian TANG ; Mei DONG ; Shiguo LIU ; Caixia CAO
Chinese Journal of Obstetrics and Gynecology 2021;56(6):425-433
Objective:To investigate the differential expression of long non-coding RNA (lncRNA) in placental tissues of women with preeclampsia (PE) and the effect of MIR210HG on the biological function of HTR8/SVneo cells.Methods:A total of 39 cases of PE women (PE group) and 39 cases of normal pregnant women (CTL group) admitted to the Affiliated Hospital of Qingdao University from July 2018 to July 2019 were collected. (1) Transcriptome sequencing (RNA-seq) was used to analyze the differentially expressed lncRNAs in the placental tissues of the two groups. (2) The expression level of MIR210HG, one of the differentially expressed lncRNAs, in the placental tissues of the two groups was detected by real-time quantitative PCR. And the correlations between the expression level of MIR210HG and systolic blood pressure, diastolic blood pressure and neonatal birth weight were analyzed. (3) The constructed small interfering RNA and negative control (NC) RNA were transfected into the HTR8/SVneo cells. The cells were divided into MIR210HG knockdown (KD) group and NC group. The effects of living cell counting (CCK-8) and transwell assay on the proliferation and migration of HTR8/SVneo cells were detected. (4) RNA interacting with MIR210HG was predicted using the Encyclopedia of RNA Interactomes (ENCORI) database. Gene Ontology (GO) functional annotation, Kyoto Encyclopedia of Gene and Genomes (KEGG) and BioCarta pathway enrichment analysis were performed.Results:(1) A total of 26 significantly differentially expressed lncRNAs were found by RNA-seq, among which 21 lncRNAs were up-regulated and 5 lncRNAs were down-regulated. (2) The relative expression level of MIR210HG in the PE group was significantly higher than that in the CTL group (9.30±1.90 and 1.10±0.20, respectively; t=4.425, P<0.01). The relative expression level of MIR210HG had positive linear correlation with systolic blood pressure ( r2=0.234, P<0.05) and diastolic blood pressure ( r2=0.190, P<0.05), but had a negative linear correlation with newborn birth weight ( r2=0.157, P<0.05). (3) Compared with the NC group, the proliferation and migration ability of HTR8/SVneo cells in the KD group were increased (all P<0.05). (4) A total of 38 RNAs that might interact with MIR210HG were predicted by ENCORI database. GO functional annotation analysis showed that MIR210HG might be involved in the functions of 27 pathways, including the regulation of production of molecular mediator of immune response, etc; KEGG pathway analysis showed that MIR210HG might be involved in the function of 8 pathways including allograft rejection, etc; Biocarta pathway analysis showed that MIR210HG may be involved in the functions of 8 pathways, including the eukaryotic initiation factor (eIF) pathway, etc. Conclusion:The expression of MIR210HG is up-regulated in the placental tissue of PE women, and MIR210HG might be a regulator of the biological behavior of trophoblast cells.
3.Effect of knockdown of PRDX6 on adaptive expression of bile acid transporter in HepG2 cells induced by rifampicin
Liufang Qu ; Mengxue Huang ; Shiguo Cao ; Gang Chen ; Jianming Xu ; Weiping Zhang
Acta Universitatis Medicinalis Anhui 2024;59(4):660-665
Objective :
To investigate the role of knockdown of peroxiredoxin-6 ( PRDX6) in injury and adaptive expression of bile acid transporter in human hepatoellular carcinomas ( HepG2 ) cells induced by rifampicin (RFP) .
Methods :
Cells in logarithmic growth phase were uniformly inoculated in six-well plates , and HepG2 cells were transiently transfected with specific PRDX6-siRNA and control-siRNA to construct the knockdown group and control group . After 24 h of induction with 100 μmol/L RFP , Western blot and qRT-PCR were performed to detect the protein and gene expression levels of PRDX6 , multidrug resistance protein 1 (MDR1) , multidrug resist- ance-associated proteins 2 , 3 and 4 (MRP2 , MRP3 and MRP4) , and Na + /taurine taurocholate cotransporter pro- tein (NTCP) . Annexin V-FITC/PI double staining assay was used to detect the apoptosis rate of cells in each group ; CCK-8 assay was used to detect the changes of cell proliferation in each group; The relative contents of ala- nine aminotransferase ( ALT) , aspartate aminotransferase ( AST) , total bilirubin ( TBIL) , indirect bilirubin (IBIL) and total bile acid (TBA) in the supernatant of cell culture medium of each group were detected by kits .
Results :
RFP increased the protein and gene expression levels of MRP2 , MRP3 , MRP4 , MDR1 , NTCP and PRDX6 in HepG2 cells (P < 0. 05) , while the protein and gene expression levels of MRP2 , MRP3 , MRP4 , MDR1 and NTCP decreased to different degrees after PRDX6 knockdown (P < 0. 05) . In addition , PRDX6 knockdown re- sulted in increased apoptosis rate of HepG2 cells (P < 0. 05) , decreased cell proliferation ability (P < 0. 05) , and increased levels of cell injury markers (ALT , AST , TBIL , DBIL , TBA) in cell culture supernatants (P < 0. 05) .
Conclusion
RFP increased the protein and gene expression of bile acid transporter and PRDX6 to increase in HepG2 cells . However , following knockdown of PRDX6 and treatment with RFP , the protein and gene expression levels of the bile acid transporter decreased and cell injury was aggravated , suggesting that PRDX6 played a protec- tive role in RFP-induced adaptive response in HepG2 cells .
4.Effects of knockdown of MANF on adaptive expression of bile acid transporter in HepG2 cells induced by rifampicin
Shiguo Cao ; Qiong Dai ; Mengxue Huang ; Liufang Qu ; Jianming Xu ; Weiping Zhang
Acta Universitatis Medicinalis Anhui 2023;58(4):665-671
Objective :
To investigate the effect of mesencephalic astrocyte-derived neurotrophic factor (MANF) on the adaptive expression of bile acid transporter in human hepatoellular carcinomas (HepG2) induced by rifampicin (RFP) .
Methods:
The control group cell line (Y07) and the knockdown group cell line (Y25) were constructed by lentiviral stable transfection technology.The Y07 and Y25 cells were treated with RFP of 200 μmol / L for 48 h, and qRT-PCR and Western blot were used to detect the protein and gene expression levels of MANF,bile salt export pump ( BSEP) ,multidrug resistance-related proteins 2 /3 /4 ( MRP2 ,MRP3 ,MRP4) ,multidrug resistance protein 1 (MDR1) ,organic solute transporter a / β ( OSTα/ β) ,organic anion transporter ( OATP2B1) .The protein and gene expression levels of proliferating cell nuclear antigen ( PCNA) ,proliferating cell marker Ki67 were used to evaluate the proliferation of cells in each group changes in levels.Changes in the protein and gene expression levels of C / EBP homologous protein( CHOP) and cysteinyl aspartate specific proteinase-3 ( Caspase-3) were used to evaluate the apoptosis of cells in each group.The relative contents of alanine aminotransferase(ALT) ,aspartate aminotransferase(AST) ,alkaline phosphatase ( ALP) ,total bilirubin ( TBIL) ,indirect bilirubin ( IBIL) and total bile acid(TBA) in the supernatant of cell culture medium of each group were detected by kits.
Results:
RFP could induce the protein and gene expression of MANF,BSEP ,MRP2 ,MRP3 ,MRP4 ,MDR1 ,OSTα , OSTβ , OATP2B1 in HepG2 cells (P <0. 05 ) ,while the protein and gene expression levels of BSEP ,MRP2 ,MRP3, MRP4,MDR1,OSTα、OSTβ、OATP2B1 decreased after MANF knockdown(P<0. 05) .Moreover,under the action of RFP,the protein expression of PCNA and Ki67 in the knockdown group was still higher.The protein and gene levels of CHOP and Caspase-3 significantly increased after MANF knockdown(P<0. 05) .The levels of the hepatic cell injury markers in the cell supernatant increased significantly(P<0. 05) .
Conclusion
RFP can induce the expression of bile acid transporter such as BSEP,MRP2,MRP3,MRP4,MDR1,OSTα , OSTβ and OATP2B1 to increase in HepG2 cells(P<0. 05) ,but the expression of bile acid transporter of HepG2 after MANF knockdown will significantly decrease under the induction of rifampicin(P<0. 05) ,and cell indury is aggravated,indicating that MANF plays a protective role in RFP-induced adaptive responses by regulating the bile acid transporter.