1.Assessment and preliminary clinical application of a domestic nucleic acid detection reagent for hepatitis D virus
Yongcong LI ; Rongchen YUAN ; Kuanhui XIANG ; Guomin OU ; Tianxun HUANG ; Fangming CHENG ; Zhenchao TIAN ; Xiongwei LIU ; Xiaozhong WANG ; Feng GUO ; Yahong MA ; Jing ZHOU ; Erhei DAI ; Bangning CHENG ; Tong LI ; Tao SHEN ; Chungen QIAN
Chinese Journal of Laboratory Medicine 2024;47(3):239-244
Objective:This study aims to evaluate the quality and explore the preliminary clinical applications of a domestically developed hepatitis D virus nucleic acid quantification reagent (abbreviated as"domestic HDV RNA reagent").Methods:The sensitivity and accuracy of the reagent were evaluated in accordance with the WHO HDV RNA international standard, employing the Bio-Rad CFX Opus 96 real-time fluorescence quantitative PCR analysis system. Serial dilutions of pseudo-viruses or cell culture-derived virus were used to determine the linear range of the domestic HDV RNA reagent. Specificity was assessed using positive samples of HAV, HBV, HCV infection, and HEV national reference materials. Precision was evaluated with samples at both high and low concentrations. In a comparative analysis, 30 HDV IgG positive samples were tested using both the domestic HDV RNA reagent and the RoboGene HDV RNA kit based on the ABI 7500 FAST DX system. The Pearson correlation coefficient (r) was used to examine the correlation between the two reagents.Results:The domestic HDV RNA reagent demonstrated a high sensitivity of up to 6 IU/ml, consistent with that of the comparator reagent. The calibration curve for WHO HDV RNA standards had a slope of -3.286, with an amplification efficiency of 101.6%. The linear detection range spanned from 10 to 10 8 IU/ml for eight HDV genotypes. The domestic HDV RNA reagent exhibited exceptional specificity, without cross-reactivity observed with HAV, HBV, HCV, or HEV. Accuracy assessments at five concentration levels met the required standards, with intra-assay precision coefficient of variation ( CV) ranging from 1.20% to 4.20%, and inter-assay precision CV from 1.20% to 7.90%. The detection results for HDV IgG positive samples were highly correlated with the comparator reagent ( r=0.984, P<0.001), achieving a diagnostic accuracy of 100% compared to sequencing results. Conclusion:In this study, the domestic HDV RNA reagent possesses excellent specificity, accuracy, precision, and a broad linear range, attaining a sensitivity level on par with international reagents of the same type.
2.The diagnostic value of copeptin in the cardiorenal syndrome rats and the association with heart and kidney impairment
Xiaosheng SHENG ; Mingxing DING ; Fangming GUO ; Zhangping YU ; Xian HUA ; Li LIN
Chinese Journal of Internal Medicine 2018;57(7):518-521
To explore the diagnostic value of copeptin (CPP) in cardiorenal syndrome (CRS) in rats and the association between CPP and impairment of heart and kidney, 60 male SD rats were randomly divided into blank control group (CK group), kidney failure group (SNX group), heart failure group (MI group), and CRS group. Heart and kidney function and their histology changes in rats from each group were detected. The correlation between serum CPP and heart and kidney function indexes was performed with Pearson correlation analysis. The HE staining of heart and kidney showed that the tissue lesion was more severe in CRS group than in SNX group and MI group. There was a significant positive correlation between serum CPP and brain natriuretic peptide (BNP) (r=0.638, P<0.05). No correlation was observed between serum CPP and cardiac function index (left ventricular systolic pressure, left ventricular diastolic pressure, left ventricular end-diastolic pressure) or renal function index (serum creatinine, urine creatinine, blood urea nitrogen) (r=0.512, 0.189, -0.063, 0.207, 0.290, 0.595, respectively, all P>0.05). The CPP level is associated with the degree of heart and kidney damage in CRS rats.
3.The predictive value of AGEs in the risk and prognosis of coronary heart disease in diabetic patients
Xiaosheng SHENG ; Mingxing DING ; Xian HUA ; Zhangping YU ; Xiaoxia YE ; Fangming GUO ; Beiwei YU ; Wenwei XU
Journal of Chinese Physician 2017;19(11):1665-1667,1671
Objective To predict the value of advanced glycation end products (AGEs) in the risk and severity of coronary heart disease in diabetic patients.Methods Totally 120 cases were divided into 3 groups.Group A had no diabetes mellitus (DM) and no coronary atherosclerotic heart disease (CAD).Group B had DM without CAD.Group C had DM with CAD.The levels of AGEs,low density lipoprotein cholesterol (LDL-C),glycosylated hemoglobin,and the severity of coronary stenosis were detected.Receiver operating characteristic (ROC) curve was used to evaluate the sensitivities and specificities of AGEs for the diagnosis of DM with CAD.Results The highest level of AGEs,LDL-C,glycosylated hemoglobin and carotid plaque was in the C group,with a statistically significant difference (P < 0.05).The severity of AGEs was significantly correlated with the severity of CAD and the Gensini score of AGEs (r =0.445).ROC curve showed that the sensitivity,specificity and area under ROC curve of AGEs determination of DM with CAD were 80.0%,75.0%,and 0.86,respectivity.Conclusions The level of AGEs is significantly correlated with atherosclerosis and prognosis in DM.The severity of CAD was higher in the patients with higher AGEs,and the incidence and severity of restenosis increased.
4.Correlation between the serum ferritin levels and the post-stroke depression at 3 months after ischemic stroke
An MAO ; Haiyun LEI ; Chunyan KANG ; Fangming GUO ; Mengmeng YAN ; Zi YU ; Zhongyuan LU ; Zhongming QIU
International Journal of Cerebrovascular Diseases 2016;24(5):412-417
Objective To investigate the correlation between the serum ferritin levels and the post-stroke depression (PSD). Methods From July 2014 to October 2015, the inpatients with the first-ever acute ischemic stroke were colected consecutively. Chemiluminescence microparticle immune assay was used to measure the serum ferritin levels within 24 h after admission. Depressive symptoms were screened by using the 17-item Hamilton depression scale (HAMD-17) at 3 months after onset. In patients with a HAMD-17 score ≥7, the depression was further diagnosed according to The Diagnostic and Statistical Manual of Mental Disorders, 4th edition. Results A total of 200 patients with the first-ever acute ischemic stroke were enroled, 55 (27. 5% ) of them were diagnosed as PSD. There were significant differences in the body mass index (BMI), years of education, waist circumference, high sensitive-C-reactive protein, homocysteine, National Institutes of Health Stroke Scale score (at baseline, discharge, and day 90), mRs score (at discharge and day 90), BI (at discharge and day 90), and the proportions of widowed or solitary patients between the PSD group and the non-PSD group (al P < 0. 05 ). The serum ferritin level in the PSD group was significantly higher than that in the non-PSD group ( median [ interquartile range], 261. 90[142. 10-364. 90] μg/L vs. 164. 40[132. 50- 195. 10] μg/L; Z = - 4. 814, P < 0. 001 ). Multivariate logistic regression analysis adjusted for confounding factors showed that the baseline serum ferritin level >136. 375 μg/L was an independent risk factor for PSD (odds ratio 1. 041 per 1-quartile increase, 95%confidence interval 1. 009-1. 239; P = 0. 045). Conclusions The elevated baseline serum ferritin level is associated with PSD.
5.Construction, expression, and bio-activity assay of an anti-IL-1βscfv and TNFR1 fusion protein
Fangming KAN ; Guiping REN ; Mo GUO ; Yang HAN ; Jianying QI ; Yu ZHANG ; Yakun ZHANG ; Deshan LI
Chinese Journal of Microbiology and Immunology 2012;(10):855-860
Objective To express the anti-IL-1βscfv and soluble TNF receptor 1 (sTNFR1),and analyze their bio-activities.Methods sTNFR1 was obtained by RT-PCR from the total RNA of HeLa cells,and fused with IL-1βscfv by the hinge fragment of IgG molecule.The fusion gene IL-1scfv:TNFR1 was cloned into the expression vector pET27b(+).The fusion protein was expressed and purified from inclusion bodies.Results The ELISA analysis showed that the fusion protein could bind hIL-1β and hTNF-α respectively in a dose-dependent manner,indicating that scfv and sTNFR in the fusion protein can form the correct spatial configuration.The dolt-blot analysis showed that the fusion protein could concurrently bind with hIL-1β and hTNF-α,indicating that the combination of the two parts of the fusion protein does not influence each other for binding to their target molecules.The bioactivity assay showed that the fusion protein could inhibit both the cytotoxicity of hTNF-α on L929 cells and hIL-1β-induced proliferation of L929 cells,indicating that the fusion protein has the ability to neutralize hTNF-α and hIL-1β.Conclusion A bispecific fusion protein IL-1scfv:TNFR1 was successfully constructed.The fusion protein has the ability to inhibit the biological activity of hTNF-α and hIL-1β,and provides a drug candidate for the treatment of rheumatoid arthritis.
6.Therapeutic effects of human umbilical cord mesenchymal stem cell transplantation on acute liver failure in rats
Hongying GUO ; Hui ZHU ; Zhengguo ZHANG ; Lihong QU ; Hong XIAO ; Yanling FENG ; Ming XIANG ; Fangming LI ; Xiang HU ; Jiefei WANG
Chinese Journal of Infectious Diseases 2011;29(5):261-266
Objective To study the therapeutic effects of human umbilical cord mesenchymal stem cells (hUCMSCs) on acute liver failure ( ALF) induced by D-galactosamine (D-gal) and lipopolysaccharide (LPS) in rats. Methods The ALF model was obtained through intraperitoneal injection of D-gal(300 mg/kg)and LPS (20μg/kg)in Wister rats. The hUCMSCs were transplanted after intoxication. All rats were divided into four groups, and each group received either hUCMSCs or 0.9% NaCl solution through intraperitoneal or tail-intravenous injection. To evaluate the liver function of each group, the levels of alanine aminotransferase (ALT), total bilirubin (TBil) and serum albumin (Alb) were measured on the day of hUCMSCs transplantation and the following 1, 2, 3, 5 and 7 days. All rats were then sacrificed to examine the liver histology at day 7. Analyses were done by using Fisher's exact test, unpaired t test and Mann-Whitney U test. Results There were no significant differences of survival rates among four groups (Fisher's exact test, both P = 1. 00). The levels of ALT, TBil and Alb in group receiving hUCMSCs intraperitoneally were (804. 9 ± 88. 0) U/L,(17. 4±2. 7) μmol/L and (20. 9±0. 8) g/L, respectively after 2 days of injection, whereas in the corresponding control group, those were (1294. 3± 171. 4) U/L, (32. 3±5. 5) μmol/L and (16. 1±0. 9) g/L, respectively, which indicated that hUCMSCs transplantation significantly improved the liver function (t = 2. 640, P =0.020;t=2.529, P = 0. 025;t= - 3. 833, P = 0. 002). Both of hUCMSCs-transplanted groups showed no significant differences. Liver histological data showed that transplantation of hUSMSCs through either intraperitoneal or tail-intravenous injection alleviated liver damage (U=4. 500, P = 0. 005;U=4. 500, P = 0. 008) and the mitotic index also increased in hUCMSCs-transplanted groups (U=4. 000, P = 0. 005; U=5. 500, P = 0. 013). Conclusions The levels of ALT, TBil and Alb can rapidly normalize in ALF rats after injected with hUCMSCs either intraperitoneally or tail-intravenously. hUCMSCs application raises the mitotic index, enhances hepatocellular regeneration and improves histological status.
7.Risk factors of acute myocardial infarction following primary percutaneous coronary intervention among elderly patients
Fangming GUO ; Xiaohuan WANG ; Guangping LI ; Xin CHEN ; Yuguang JIN
Journal of Geriatric Cardiology 2009;6(2):67-70
Background and Objective Large randomized controlled trials have demonstrated that percutaneous coronary intervention (PCI) with the routine use ofdrug-eluting stents is safe and effective, however, the patients older than 75 years undergoing PCI are at increased risk for major adverse cardiac events, so that the patients are usually excluded from this trial. The aim of the present study was to assess the early clinical outcome and risk factors in old patients with acute ST elevation myocardial infarction (STEMI) following primary PCI. Methods We analyzed the outcome after stenting in 136 patients older than 60 years in our coronary care unit with acute STEMI, and the patients were further classified in 2 age groups: patients≥75 years and <75 years. Results Though the older group had a higher prevalence of adverse baseline characteristics and lower final TIMI flow than those of the younger, the procedural success had no difference between two groups. The main adverse clinical events (MACE) for the old group was a little higher comparing with the younger in 12-month following up. Conclusions Our study suggest that drug-eluting stent implantation in elderly patients with acute ST elevation myocardial infarction has high initial procedural success rates despite having more severe baseline risk characteristics, and to shorten the time form symptom onset to PC1 and improve final TIMI flow strategy may decrease MACE among old patients following PCI.
8.Left atrial function and structure changes after catheter ablation in patients with atrial fibrillation
Xiaohuan WANG ; Guangping LI ; Fangming GUO ; Gang XU ; Yunshan CAO
Clinical Medicine of China 2009;25(5):485-487
Objective To investigate the left atrium (LA) function and structure changes in the paroxysmal atrial fibrillation (AF) patients after catheter ablation using tissue Doppler imaging. Methods After complete pul-monary vein, radiofrequency ablation guided by Ensite NavX System, LA systolic function and LA diameter, volume, mean mitral gradient and mitral annulus early and advanced diastolic peak velocity were assessed in 32 cases of par-oxysmal AF patients,which were compared with age-matched controls before and after 24 hours, 1 week, 1 month) AF ablation. Results AF did not occur again in 32 AF paroxysmal patients after isolation. LA diameter and volume in AF groups before ablation were larger than controls(P<0.01), which were also larger 24 hours after ablation than before (P>0.05 ), but LA volume was larger than before (P<0.05), and decreased in I week after ablation (P< 0.05), but had no significant difference compared with controls in 1 month(P>0.05 ) ;mitral annulus advanced di-astolic peak velocity decreased in AF groups before ablation (P<0.01 )and was lower than that 24 hours after cathe-ter ablation (P<0.05 ), but increased after 1 week (P<0.05 or P <0.01 ), and had no significance after 1 month. Conclusion Catheter ablation is the effective way to manage AF because LA is distended and atrial systolic func-tion is reduced within 24 hours after procedure, then gradually increased in a week and will nearly recover to that be-fore procedure in a month, which may be correlated with LA repair, implicating that postprocedural thromboembollc risk and procedure injury should be taken into consideration.
9.Clinical study on right ventricular function of pulmonary thromboembolism
Fangming GUO ; Guangping LI ; Yan TANG ; Xiuqin SONG ; Jun YANG
Clinical Medicine of China 2008;24(9):869-871
Objective To study the right ventricular function of patients with pulmonary thromboembolism by using color Doppler ultrasound.Methods 31 patients with acute pulmonary thromboembolism,compared with 31 vohnteers,were enrolled in this study.The right ventricular anterior wall movement,right ventricular end diastolic volume,right ventricular ejection fraction,and myocardial performance index were observed by echocardiography.Resuits The right atrium diameter,right ventricles diameter,right ventrieular end diastolic volume and pulmonary artery inner diameter in study group were much larger than that in control group (P<0.01),and the right ventricular anterior wall movement and right ventrieular ejection fraction decreased in study group (P<0.01).Tricuspidal annular E peak velocity tended to be decreased,isovolumie relaxation time and isovolumic contraction time were prolonged and myocardial performance index was increased (P<0.01).Right ventricular myocardial performance index showed significant correlation with right ventrieular ejection fraction (r=0.78,P<0.01),isovolumic relaxation time and isovolumic contraction time(rl=0.88,r2=0.57,P<0.01).Conclusion The right ventricular function in patients with pulmonary thromboembelism is decreased and myocardial performance index is a sensitive index which can be used to reflect right ventricular function in pulmonary thromboembolism.
10.Diagnosis of Congenital Dextrocardia with Complex Anomalies by Echocardiography
Fangming GUO ; Minzhong FAN ; Jun YANG
International Journal of Pediatrics 2007;0(04):-
0.05),the specificity of transthoracic echocardiography in diagnosing the atrial situs,ventricular structure,the connection of the ventricle and arteries or auricle,arterial situs was more than 90%.In mirror-image dextrocardia,majority had concordant segmental connection and could present anomalies with simple anomalies(42.9%) including atrial septal defect and ventricular septal defect,in addition,the major complex anomalies such as pulmonary stenosis were tetralogy of Fallot,double outlet of right ventricle(50%);In dextroversion,majority had discordant segmental connection and mostly presented anomalies with pulmonary stenosis or pulmonary atresia,including single ventricle,double outlet of right ventricle,transpo sition of great arteries,pulmonary atresia and so on(88.6%).Conclusion Echoca rdiography can exactly diagnose the most congenital dextrocardia with complex cardiovascular anomalies,and can be the first or alternative screening choice for diagnosing dextrocardia.

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