1.Effect of LncRNA RMRP on ferroptosis induced by oxygen glucose deprivation/reperfusion in mouse HL-1 cardiomyocytes by regulating miR-766-5p
Lei He ; Xinglan Sun ; Yingxing Wu ; Yuan Xu ; Xiang Peng ; Chenkai Hu
Acta Universitatis Medicinalis Anhui 2025;60(12):2207-2214
Objective:
To investigate the effect and mechanism of long non-coding RNA RMRP(LncRNA RMRP) on oxygen-glucose deprivation/reperfusion(OGD/R)-induced ferroptosis in mouse HL-1 cardiomyocytes by regulating miR-766-5p.
Methods:
HL-1 cells were cultured in vitro, and OGD/R models were established. The expression levels of LncRNA RMRP in HL-1 cells at various reperfusion time points were subsequently quantified using qRT-PCR. The LncRNA RMRP small RNA interference fragment(si-RMRP) and its corresponding negative control(si-NC), as well as the miR-766-5p inhibitor and its respective negative control(inhibitor-NC), were transfected into HL-1 cells. Subsequently, the cells were subjected to OGD/R treatment. CCK-8 assay was employed to evaluate cell viability. Assay kits were employed to measure the levels of lactate dehydrogenase(LDH) in the cell supernatant, as well as the intracellular levels of malondialdehyde(MDA), superoxide dismutase(SOD), glutathione(GSH), and ferrous ion(Fe2+). qRT-PCR analysis was conducted to assess the expression levels of LncRNA RMRP and miR-766-5p. Western blot analysis was conducted to assess the expression levels of proteins associated with ferroptosis including GPX4, SLC7A11, and FTH1. Dual-luciferase reporter assays were performed to investigate the sponge adsorption relationship between LncRNA RMRP and miR-766-5p.
Results:
As reperfusion time extended, the expression level of LncRNA RMRP in cells progressively increased(P2+ levels within the cells, and decreased the activities of SOD and GSH in cells(P2+ levels within the cells, and promoting SOD and GSH activities in cells(PP < 0. 01) . Furthermore , silencing LncRNA RMRP upregulated the protein expression levels of GPX4 , SLC7A11 , and FTH1(P < 0. 01) . The dual-luciferase reporter assay confirmed that LncRNA RMRP could regulate the expression of miR-766- 5p through a sponge adsorption mechanism. Partial inhibition of miR-766-5p inhibitor expression could mitigate the improvement effect caused by LncRNA RMRP silencing on OGD/R-induced ferroptosis in HL-1 cells.
Conclusion
Silencing LncRNA RMRP inhibits OGD/R-induced ferroptosis in HL-1 cells , potentially through the sponge-mediated regulation of miR-766-5p expression.
2.Prognostic efficacy of pericoronary fat attenuation index and fibrous plaque index in patients with acute coronary syndrome
Cong HUANG ; Feng WEN ; Xinglan WANG ; Chen LIU ; Hongqin LIANG ; Xi YANG ; Chengwei MOU ; Jian WANG
Journal of Army Medical University 2025;47(17):2106-2114
Objective To explore the predictive value of fat attenuation index(FAI)and fibrous plaque index(FPI)for the prognosis of patients with acute coronary syndrome(ACS).Methods A retrospective cohort study was conducted on 334 ACS patients undergoing percutaneous coronary intervention(PCI)in the First Affiliated Hospital of Army Military Medical University and Yongchuan Hospital of Chongqing Medical University from March 2021 to July 2023.All patients received coronary computed tomography angiography(CCTA)to measure FAI and FPI.According to the occurrence of major adverse cardiovascular events(MACE)with 1 year of follow-up,they were divided into MACE group(n=108)and non-MACE group(n=226).The baseline data,CCTA data and results of laboratory tests were collected and compared between the 2 groups.Multivariate logistic regression analysis was used to analyze the relationship of FAI and FPI with the prognosis of ACS patients,and ROC curve was drawn to evaluate its predictive efficiency.Results Among the 334 ACS patients,108(32.34%)experienced MACE.When compared with the non-MACE group,the MACE group exhibited significantly larger proportions of diabetes(72.22%vs 31.86%)and left main coronary artery disease(18.52%vs 7.08%),but lower success rate of operation(79.63%vs 93.81%,P<0.05).Radiologic results showed that the proportion of severe stenosis(20.37%vs 10.62%),FAI(-80.12±6.41 HU vs-72.34±7.09 HU)and FPI(0.58±0.41 vs 0.26±0.12)were obviously increased in the MACE group than the non-MACE group(P<0.05).Laboratory tests indicated that there were statistical differences between the 2 groups in high-density lipoprotein-cholesterol(HDL-C,1.20±0.15 vs 1.09±0.16 mmol/L),miR-126(0.91±0.12 vs 0.96±0.15)and SST2(38.45±5.67 vs 34.30±4.89 ng/mL,P<0.05).Multivariate Logistic regression analysis revealed that FAI(OR=1.200,95%CI:1.136~1.268),FPI(OR=63.157,95%CI:14.126~282.374),moderate stenosis(OR=1.332,95%CI:1.024~1.859),severe stenosis(OR=1.480,95%CI:1.074~2.039),miR-126(OR=0.007,95%CI:0.001~0.077),and sST2(OR=1.192,95%CI:1.113~1.277)were independent predictors of MACE(P<0.05).ROC curve analysis displayed that stenosis degree(AUC=0.622,95%CI:0.561~0.683,P=0.001),FAI(AUC=0.790,95%CI:0.741~0.839,P=0.001)and FPI(AUC=0.700,95%CI:0.638~0.761,P=0.001),miR-126(AUC=0.646,95%CI:0.584~0.707,P=0.001),sST2(AUC=0.700,95%CI:0.638~0.761,P=0.001)had certain predictive values for ACS prognosis.Conclusion Coronary FAI and FPI can be used as independent prognostic indicators of ACS patients,and their numerical changes are closely related to plaque stability and inflammatory state.
3.Conventional ultrasound and contrast-enhanced ultrasound for predicting cervical lymph node metastasis in head and neck squamous cell carcinoma
Chuan LIU ; Wei MA ; Xinwei CHEN ; Lin CHEN ; Min PAN ; Yanshi LI ; Xinglan LIU ; Guohua HU
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2025;60(5):496-504
Objective:To evaluate the diagnostic efficacy of conventional ultrasound combined with contrast-enhanced ultrasound (CEUS) for detecting cervical lymph node metastasis in head and neck squamous cell carcinoma (HNSCC).Methods:This retrospective case control study included 259 patients (249 males and 10 females,aged 40-88 years with a median age of 63 years) who were pathologically diagnosed with HNSCC at the First Affiliated Hospital of Chongqing Medical University from April 2022 to August 2024. A total of 259 lymph nodes were assessed(78 pathologically positive nodes and 181 pathologically negative ones). Receiver operating characteristic (ROC) curves were plotted to compare the diagnostic efficacies of conventional ultrasound combined with CEUS and CT, MRI, or conventional ultrasound for cervical lymph node metastasis. Patients were randomly assigned to a training group ( n=208) and a validation group ( n=51) (8∶2). Univariate and multivariate Logistic regression were used to analyze the independent risk factors for lymph node metastasis prediction. A nomogram model was constructed based on independent risk factors. The model predictive efficacy was assessed by ROC curve, and model′s fit was evaluated by Hosmer-Lemeshow test. Results:The area under the curve (AUC) for conventional ultrasound combined with CEUS in predicting cervical lymph node metastasis in HNSCC was 0.923 (95% CI: 0.880-0.965), which was significantly higher than those for enhanced CT, cervical enhanced MRI, and conventional ultrasound [AUC were 0.820 (95% CI: 0.753-0.886), 0.802 (95% CI: 0.737-0.866), and0.836 (95% CI: 0.774-0.899), respectively].There were no statistically significant differences in clinical data between the two groups (all P>0.05). Univariate Logistic regression analysis showed that, among contrast-enhanced ultrasound features, the centripetal perfusion pattern, irregular perfusion defect type, and uneven distribution of contrast agent were significantly associated with cervical lymph node metastasis (all P<0.05). Among conventional ultrasound features, L/S ratio≤1.5, heterogeneous internal echoes, uneven cortex thickening, blurred corticomedullary boundary, loss of lymphatic gate structure, and peripheral or mixed type on color Doppler flow imaging were significantly associated with cervical lymph node metastasis (all P<0.05). Multivariate Logistic regression analysis indicated that the centripetal perfusion pattern, heterogeneous internal echoes, and uneven cortical thickening were independent risk factors for cervical lymph node metastasis (all P<0.05). A nomogram model was constructed based on the above independent risk factors, and offered an AUC of 0.933 (95% CI:0.890-0.976) in the training group, which was significantly higher than that for enhanced CT, enhanced MRI, conventional ultrasound, and conventional ultrasound combined with CEUS [AUC were 0.818 (95% CI:0.747-0.890), 0.807 (95% CI:0.739-0.875), 0.842 (95% CI: 0.777-0.908), and 0.921 (95% CI:0.876-0.967), respectively]. In the validation group, the AUC was 0.866 (95% CI:0.703-1.000), which was higher than that for cervical enhanced CT, enhanced MRI, and conventional ultrasound [AUC were 0.820 (95% CI:0.613-1.000), 0.711 (95% CI:0.478-0.943), and 0.748 (95% CI:0.515-0.982), respectively]. Conclusion:The combination of conventional ultrasound and CEUS has significant clinical application value in predicting cervical lymph node metastasis in HNSCC.
4.Diagnostic value of fetal echocardiography for ventricular septal defect and reasons for missed diagnosis and misdiagnosis
Wenying YUAN ; Qichang ZHOU ; Lin ZHANG ; Xinglan HUANG ; Shuizhen LIU ; Ling HUANG ; Fang LIU
Chinese Journal of Perinatal Medicine 2025;28(7):593-597
Objective:To explore the diagnostic value of fetal echocardiography (ECHO) for ventricular septal defect (VSD) and the causes of missed diagnosis and misdiagnosis by analyzing the standard of ECHO sections and image quality.Methods:A retrospective analysis was conducted on 94 VSD fetuses diagnosed by ECHO at Xiangtan Maternal and Child Health Care Hospital from January 2022 to June 2024. Clinical data including the anatomic subtypes, associated anomalies, chromosomal karyotype results, pregnancy outcomes, and neonatal ECHO results were collected and analyzed. Besides, by reviewing the ultrasound images from the institution's ultrasound workstation, a descriptive analysis was conducted on the ECHO section criteria and image quality in the missed and misdiagnosed cases.Results:Among 10 984 fetuses undergoing ECHO, a total of 94 cases of fetal VSD were detected (0.7%), including 45 (48%) isolated VSD and 49 (52%) non-isolated VSD cases. Anatomic distribution revealed perimembranous type predominance (67, 71%), followed by muscular (24, 26%), subarterial types (2, 2%), and perimembranous+muscular type (1, 1%). Among 29 missed diagnosis cases (fetal ECHO-negative but neonatal ECHO-confirmed VSDs), all were isolated VSD, muscular types accounted for 19 (66%), perimembranous types for 8 (27%), and subarterial types for 2 (7%). The causes of missed diagnosis include: technical limitation in 14 cases (48%), restricted cardiac acoustic window in 11 cases (38%), and operator-dependent factors in four cases (14%). All 18 misdiagnosed cases occurred in isolated VSD, with 15 cases of perimembranous and three cases of muscular types. The causes of misdiagnosis include: nine cases due to non-perpendicular beam alignment to perimembranous septum, six cases due to excessively high color flow gain,and three cases due to the misinterpretation of the right ventricular outflow tract flow above the aortic valve as VSD septal perforation blood flow. The sensitivity and specificity of fetal ECHO for diagnosing VSD were 76.42% and 99.83%. Among the 94 cases of fetal VSD, 38 cases (40%) underwent chromosomal karyotype analysis and chromosomal microarray analysis, which identified chromosomal karyotype abnormalities or pathogenic gene variants in 4/16 non-isolated VSDs (all perimembranous), while no anomalies were detected in 22 isolated VSDs. Pregnancy outcomes showed 31 terminations due to associated structural or chromosomal abnormalities versus 63 live births. In the live births, who underwent echocardiography in the neonatal period, 19 cases (30%) showed no VSD and were considered to have undergone spontaneous in utero closure, and persistent defects were found in 44 (70%).Conclusions:Fetal ECHO demonstrates high sensitivity and specificity in the diagnosis of fetal VSD. Critical quality control measures include individualized parameter adjustment and systematic multiplanar continuous scanning to minimize diagnostic errors. Isolated small and medium-sized muscular and perimembranous VSD usually have a good prognosis.
5.Conventional ultrasound and contrast-enhanced ultrasound for predicting cervical lymph node metastasis in head and neck squamous cell carcinoma
Chuan LIU ; Wei MA ; Xinwei CHEN ; Lin CHEN ; Min PAN ; Yanshi LI ; Xinglan LIU ; Guohua HU
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2025;60(5):496-504
Objective:To evaluate the diagnostic efficacy of conventional ultrasound combined with contrast-enhanced ultrasound (CEUS) for detecting cervical lymph node metastasis in head and neck squamous cell carcinoma (HNSCC).Methods:This retrospective case control study included 259 patients (249 males and 10 females,aged 40-88 years with a median age of 63 years) who were pathologically diagnosed with HNSCC at the First Affiliated Hospital of Chongqing Medical University from April 2022 to August 2024. A total of 259 lymph nodes were assessed(78 pathologically positive nodes and 181 pathologically negative ones). Receiver operating characteristic (ROC) curves were plotted to compare the diagnostic efficacies of conventional ultrasound combined with CEUS and CT, MRI, or conventional ultrasound for cervical lymph node metastasis. Patients were randomly assigned to a training group ( n=208) and a validation group ( n=51) (8∶2). Univariate and multivariate Logistic regression were used to analyze the independent risk factors for lymph node metastasis prediction. A nomogram model was constructed based on independent risk factors. The model predictive efficacy was assessed by ROC curve, and model′s fit was evaluated by Hosmer-Lemeshow test. Results:The area under the curve (AUC) for conventional ultrasound combined with CEUS in predicting cervical lymph node metastasis in HNSCC was 0.923 (95% CI: 0.880-0.965), which was significantly higher than those for enhanced CT, cervical enhanced MRI, and conventional ultrasound [AUC were 0.820 (95% CI: 0.753-0.886), 0.802 (95% CI: 0.737-0.866), and0.836 (95% CI: 0.774-0.899), respectively].There were no statistically significant differences in clinical data between the two groups (all P>0.05). Univariate Logistic regression analysis showed that, among contrast-enhanced ultrasound features, the centripetal perfusion pattern, irregular perfusion defect type, and uneven distribution of contrast agent were significantly associated with cervical lymph node metastasis (all P<0.05). Among conventional ultrasound features, L/S ratio≤1.5, heterogeneous internal echoes, uneven cortex thickening, blurred corticomedullary boundary, loss of lymphatic gate structure, and peripheral or mixed type on color Doppler flow imaging were significantly associated with cervical lymph node metastasis (all P<0.05). Multivariate Logistic regression analysis indicated that the centripetal perfusion pattern, heterogeneous internal echoes, and uneven cortical thickening were independent risk factors for cervical lymph node metastasis (all P<0.05). A nomogram model was constructed based on the above independent risk factors, and offered an AUC of 0.933 (95% CI:0.890-0.976) in the training group, which was significantly higher than that for enhanced CT, enhanced MRI, conventional ultrasound, and conventional ultrasound combined with CEUS [AUC were 0.818 (95% CI:0.747-0.890), 0.807 (95% CI:0.739-0.875), 0.842 (95% CI: 0.777-0.908), and 0.921 (95% CI:0.876-0.967), respectively]. In the validation group, the AUC was 0.866 (95% CI:0.703-1.000), which was higher than that for cervical enhanced CT, enhanced MRI, and conventional ultrasound [AUC were 0.820 (95% CI:0.613-1.000), 0.711 (95% CI:0.478-0.943), and 0.748 (95% CI:0.515-0.982), respectively]. Conclusion:The combination of conventional ultrasound and CEUS has significant clinical application value in predicting cervical lymph node metastasis in HNSCC.
6.Diagnostic value of fetal echocardiography for ventricular septal defect and reasons for missed diagnosis and misdiagnosis
Wenying YUAN ; Qichang ZHOU ; Lin ZHANG ; Xinglan HUANG ; Shuizhen LIU ; Ling HUANG ; Fang LIU
Chinese Journal of Perinatal Medicine 2025;28(7):593-597
Objective:To explore the diagnostic value of fetal echocardiography (ECHO) for ventricular septal defect (VSD) and the causes of missed diagnosis and misdiagnosis by analyzing the standard of ECHO sections and image quality.Methods:A retrospective analysis was conducted on 94 VSD fetuses diagnosed by ECHO at Xiangtan Maternal and Child Health Care Hospital from January 2022 to June 2024. Clinical data including the anatomic subtypes, associated anomalies, chromosomal karyotype results, pregnancy outcomes, and neonatal ECHO results were collected and analyzed. Besides, by reviewing the ultrasound images from the institution's ultrasound workstation, a descriptive analysis was conducted on the ECHO section criteria and image quality in the missed and misdiagnosed cases.Results:Among 10 984 fetuses undergoing ECHO, a total of 94 cases of fetal VSD were detected (0.7%), including 45 (48%) isolated VSD and 49 (52%) non-isolated VSD cases. Anatomic distribution revealed perimembranous type predominance (67, 71%), followed by muscular (24, 26%), subarterial types (2, 2%), and perimembranous+muscular type (1, 1%). Among 29 missed diagnosis cases (fetal ECHO-negative but neonatal ECHO-confirmed VSDs), all were isolated VSD, muscular types accounted for 19 (66%), perimembranous types for 8 (27%), and subarterial types for 2 (7%). The causes of missed diagnosis include: technical limitation in 14 cases (48%), restricted cardiac acoustic window in 11 cases (38%), and operator-dependent factors in four cases (14%). All 18 misdiagnosed cases occurred in isolated VSD, with 15 cases of perimembranous and three cases of muscular types. The causes of misdiagnosis include: nine cases due to non-perpendicular beam alignment to perimembranous septum, six cases due to excessively high color flow gain,and three cases due to the misinterpretation of the right ventricular outflow tract flow above the aortic valve as VSD septal perforation blood flow. The sensitivity and specificity of fetal ECHO for diagnosing VSD were 76.42% and 99.83%. Among the 94 cases of fetal VSD, 38 cases (40%) underwent chromosomal karyotype analysis and chromosomal microarray analysis, which identified chromosomal karyotype abnormalities or pathogenic gene variants in 4/16 non-isolated VSDs (all perimembranous), while no anomalies were detected in 22 isolated VSDs. Pregnancy outcomes showed 31 terminations due to associated structural or chromosomal abnormalities versus 63 live births. In the live births, who underwent echocardiography in the neonatal period, 19 cases (30%) showed no VSD and were considered to have undergone spontaneous in utero closure, and persistent defects were found in 44 (70%).Conclusions:Fetal ECHO demonstrates high sensitivity and specificity in the diagnosis of fetal VSD. Critical quality control measures include individualized parameter adjustment and systematic multiplanar continuous scanning to minimize diagnostic errors. Isolated small and medium-sized muscular and perimembranous VSD usually have a good prognosis.
7.Huaiqihuang Granules Affect Differentiation of Th17 Cells in IgA Vasculitis Nephritis Mice by Regulating AMPK/ACC Pathway
Xinglan YE ; Keying LI ; Jiaxuan LI ; Juan BAI ; Wencheng XU ; Hong LIU ; Xue XUE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(11):78-86
ObjectiveTo observe the intervention effect of Huaiqihuang granules (HQH) on immunoglobulin A vasculitis nephritis (IgAVN) mice and explore the underlying therapeutic mechanism. MethodFifty SPF-grade male Kunming mice were randomly divided into a normal group, an IgAVN model group, a dexamethasone group (2.5 mg·kg-1·d-1), a low-dose HQH group (4 g·kg-1·d-1), and a high-dose HQH group (8 g·kg-1·d-1). The mouse model was established using oral administration of gliadin combined with intravenous injection of India ink. After successful modeling, the mice were euthanized after 4 weeks of gastric gavage according to groups. The 24 h urinary total protein (24 h UTP), urine β2-microglobulin (β2-MG), serum total protein, albumin, IgA, etc. were detected in each group. Flow cytometry was used to determine the proportion of T helper 17 (Th17) cells in spleen cell suspension. Western blot was employed to detect the expression of adenosine 5'-monophosphate-activated protein kinase α (AMPKα), phosphorylated AMPKα (p-AMPKα), acetyl-CoA carboxylase 1 (ACC1), and phosphorylated ACC1 (p-ACC1) in Th17 cells. Pathological changes in the spleen and kidneys were observed. ResultCompared with the normal group, the IgAVN model group showed significant increases in 24 h UTP, urine β2-MG, total cholesterol (P<0.05), serum interleukin-17 (IL-17), IgA, Th17 proportion in the spleen cell suspension, and IL-17 expression in the spleen tissue (P<0.01), and significantly decreased serum total protein, albumin, p-AMPKα/AMPKα, and p-ACC1/ACC1 expression of Th17 cells (P<0.01). Compared with the IgAVN model group, in the 4th week, the 24 h UTP, urine β2-MG, serum IL-17, IgA levels, and renal IgA deposition were significantly reduced in each treatment group (P<0.01), and the Th17 proportion and IL-17 expression in spleen tissue were significantly decreased (P<0.05, P<0.01). Serum albumin levels significantly increased (P<0.05). Compared with the IgAVN model group, the dexamethasone group and the high-dose HQH group showed increases in serum total protein (P<0.01), p-AMPKα/AMPKα, and p-ACC1/ACC1 expression of Th17 cells (P<0.05, P<0.01). The high-dose HQH group showed a significant decrease in total cholesterol level (P<0.05). Various treatment groups showed different degrees of improvement in spleen and kidney pathological changes. ConclusionHQH may affect Th17 cell differentiation by regulating the AMPK/ACC pathway, correcting immune inflammatory disorders, and exerting therapeutic effects on IgAVN.
8.Efficacy and Safety of Rivaroxaban and Dabigatran in Anticoagulant Therapy for Patients with Non-valvular Atrial Fibrillation combined with Diabetes Mellitus
Bo CAO ; Min CHEN ; Lan XU ; Xinglan YAO ; Yueyuan ZHUANG
Herald of Medicine 2023;42(12):1835-1840
Objective To investigate the efficacy and safety of new oral anticoagulants(NOACs)rivaroxaban and dabigatran versus warfarin in the treatment of the patients with non-valvular atrial fibrillation combined with diabetes mellitus.Methods A retrospective study was performed on 119 cases of patients with non-valvular atrial fibrillation combined with diabetes mellitus from outpatient and inpatient of the Ninth People's Hospital of Suzhou City from Jan 2019 to June 2021.According to the use of anticoagulants,patients were divided into rivaroxaban 10 mg group(Group A,n=25),rivaroxaban 15 mg group(Group B,n=30),dabigatran group(Group C,n=29),and warfarin group(Group D,n=35).All patients were treated continuously for at least 6 months.The incidence of embolism and bleeding events,the changes of coagulation indicator,blood glucose,liver and kidney function indexes were compared before and after treatment among the four groups.Results Thromboembolic events:1 cases(4.00%)of stoke or thromboembolic events occurred in Group A;2 cases(6.67%)occurred in Group B;2 cases(6.90%)occurred in Group C,and 5 cases(14.28%)occurred in Group D.Bleeding events:1 cases(4.00%)of bleeding events occurred in Group A;1 cases(3.33%)occurred in Group B;2 cases(6.90%)occurred in Group C,and 8 cases(22.85%)occurred in Group D.There was no statistically significant difference among the four groups of stoke or thromboembolic events(P>0.05).The incidence of bleeding events in Group A and B were both statistically lower than Group D(P<0.05),and the risk of bleeding events of Group C was similar to Group D(P>0.05)and there was also no statistically significant difference among Group A,B and C(P>0.05).The activated partial thromboplastin time(APTT)of all patients was significantly prolonged after treatment compared with before treatment(P<0.05),but none of them exceeded the upper limit of the normal value by 2 times.The APTT and international normalized ratio(INR)values of Group A and B,INR values of Group C were all statistically better than those of Group D(P<0.05)after the treatment.There were no statistically significant difference in the comparison of blood glucose,liver and kidney function index as well(P>0.05).Conclusion The new oral anticoagulants rivaroxaban and dabigatran showed similar effects in prevention of stroke or thromboembolic events,but better safety profiles with lower risks of bleeding events compared to warfarin in patients with non-valvular atrial fibrillation combined with diabetes mellitus.Dabigatran is comparable in efficacy and safety when compared with rivaroxaban and deserves to be promoted for clinical use.
9.Tidal breathing nasal nitric oxide in preschool children aged 3 to 5 years
Xiang ZHANG ; Xinglan WANG ; Shunying ZHAO
Chinese Journal of Applied Clinical Pediatrics 2023;38(4):271-274
Objective:To explore the threshold of tidal breathing nasal nitric oxide (TB-nNO) in diagnosing primary ciliary dyskinesia (PCD) in children aged 3 to 5 years.Methods:Retrospective study.The TB-nNO values were examined of 165 healthy children aged 3-5 in a kindergarten in Xicheng District, Beijing, from March 27 to March 29, 2018, which were also measured in children aged 3-5 years who were diagnosed as PCD, cystic fibrosis, bronchiolitis obliterans, bronchiectasis caused by other diseases and asthma in the Second Department of Pediatric Pneumology, Beijing Children′s Hospital, Capital Medical University from January 2018 to December 2021.Relevant factors associated with TB-nNO in normal children were screened by a multiple linear regression model.The cut-off value of TB-nNO in diagnosing PCD in preschool children aged 3-5 years was determined by calculating the maximum area under the receiver operating characteristic (ROC) curve.Results:TB-nNO value in healthy children aged 3, 4 and 5 years were (94.8±36.4) nL/min, (103.3±50.7) nL/min and (106.9±61.5) nL/min, respectively.The mean TB-nNO value in 9 children with PCD was (18.9±10.8) nL/min.TB-nNO values in 49 children with asthma, 19 children with bronchiolitis obliterans, 17 children with bronchiectasis and 6 children with cystic fibrosis were (97.7±51.1) nL/min, (93.2±49.2) nL/min, (93.7±75.3) nL/min and (45.4±18.2) nL/min, respectively.Using 30 nL/min of TB-nNO as the cut-off point, the sensitivity and specificity of TB-nNO in diagnosing PCD were 88.9% (8/9) and 96.9%, respectively.The area under the ROC curve was 98.3% (95% CI: 95.3%-100.0%). Conclusions:TB-nNO value of 30 nL/min can be used as the cut-off point in the diagnosis screening of PCD in children aged 3-5 years.Its diagnostic value in this age group should be further evaluated.
10.Expression features of cytochrome C oxidase subunit Ⅰ, BCL2 interacting protein 3 and interleukin-1β in MRL/lpr lupus mice liver tissue
Caifeng HUANG ; Yuqiong DENG ; Xinglan HUANG ; Xiaoqing ZHAO ; Sheng LIU ; Ping FAN ; Xinghua SONG ; Peng YAN ; Xiping CHENG
Chinese Journal of Rheumatology 2022;26(11):758-766,C11-3
Objective:To explore the expression features of cytochrome C oxidase subunit Ⅰ (MT-CO1), BCL2 interacting protein 3 (BNIP3) and interleukin (IL)-1β in the liver of MRL/lpr lupus mice.Methods:The mRNA and protein levels of MT-CO1, BNIP3, IL-1β, p16 and p21 in lupus mice and control mice were detected by polymerase chain reaction (PCR) and Western blot, the IL-1β expression site were detected by hematoxylin and eosin (HE) staining and immunohistochemical method, and themalondialdehyde (MDA) was detected by colorimetry. Hepatocytes and macrophages were stimulated with lipopolysaccharide (LPS), while hepatocytes were also cultured with supernatants obtained after macrophages stimulated with LPS, and the mRNA and protein levels of MT-CO1, BNIP3 and LC3B, as well as p16 and p21 expression, were determined by qPCR and Western blot. The expression of mitochondrial reactive oxygen species (mtROS) was detected by immunofluorescence. One way Analysis of Variance (ANOVA) was used to compare the mean of each group, and LSD method was used to compare the means of multiple samples, and Tamhane's T2 method was used to compare the means of multiple samples when the variance was uniform. Results:The results of PCR showed that the mRNA levels of MT-CO1 and BNIP3 in the liver tissue of the lupus group (0.14±0.04; 0.16±0.05) were significantly lower than those of the control group (0.11±0.04; 0.16±0.06), and the differences were statistically significant ( t=7.16, P<0.001; t=4.54, P<0.001). The expression levels of IL-1β, p16 and p21 in the lupus group (2.06±0.69; 0.37±0.14; 0.16±0.06) were significantly higher than those of the control group (0.23±0.06; 0.25±0.08; 0.11±0.04) ( t=9.58, P<0.001; t=24.35, P<0.001; t=22.36, P<0.001). The results of Western blot were consistent with those of PCR. HE staining showed lymphocyte infiltration in the liver tissue of lupus mice, and immunohistochemistry showed IL-1β in the liver tissue of lupus mice. The positive cells were mainly concentrated in the sinusoids, and the expression of hepatic parenchymal cells was not rearkable. The content of MDA in liver tissue of the lupus group (0.19±0.10) was higher than that of the control group (0.17±0.09), and the difference was statistically significant ( t=4.33, P=0.005). LPS directly stimulated AML12 hepatocytes (0.069±0.028; 0.17±0.07). The PCR results showed that compared with the control group (0.176±0.072; 0.08±0.03), the expression of MT-CO1, and BNIP3 were not significantly different ( t=1.01, P=0.337; t=0.88, P=0.399). The expression of IL-1β was significantly higher when incubated with the supernatants of LPS stimulated macrophages (0.28±0.09) compared than that of the control group (0.15±0.05) ( t=28.26, P<0.001). The results of PCR showed that the mRNA levels of MT-CO1 and BNIP3 in the LPS stimulated group (0.046±0.026; 0.17±0.05) were significantly lower than those in the control group (0.143±0.083; 0.18±0.06), and the differences were statistically significant ( t=7.52, P<0.001; t=4.24, P<0.001), The expression of p16 and p21 in LPS stimulated group (0.29±0.09; 0.27±0.09) were significantly higher than those in the control group (0.18±0.06; 0.22±0.07) ( t=13.54, P<0.001; t=8.69, P<0.001). The results of Western blot were consistent with those of PCR. Immunofluorescence showed that the fluorescence intensity of mtROS in LPS stimulated group (0.25±0.10) was higher than that in the control group (0.08±0.03), and the difference was statistically significant ( t= 4.86, P<0.001). Conclusion:Immune-mediated inflammation in the liver tissue of lupus mice can stimulate liver parenchymal cells to cause intracellular mitochondrial dysfunction. However, the mechanism of liver organ damage in lupus mice is not limited to the immune-mediated inflammation of immune active cells, but also include parenchymal cell mitochondrial dysfunction.


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