Objective:This study aimed to introduce droplet digital PCR (ddPCR) for monitoring Epstein-Barr virus (EBV) infection after allogeneic hematopoietic stem cell transplantation (alloHSCT) in children and assess its viability as a complementary detection method in clinical settings.Methods:A total of 290 blood samples from 47 children undergoing alloHSCT were collected. Both ddPCR and real-time quantitative PCR (qPCR) were employed to detect EBV DNA load in plasma, with a comparison of detection efficiencies between the two methods. Continuous monitoring of 39 children was conducted to observe dynamic changes in EBV DNA load in plasma and analyze the merits and drawbacks of both methods.Results:The EBV positive detection rate of ddPCR was significantly higher than that of qPCR ( χ2=20.25, P<0.001), particularly in samples with low viral loads. Among the children monitored continuously for EBV DNA, 14 out of 39 exhibited positive ddPCR result. Notably, in two cases where patients displayed rash and fever symptoms with positive ddPCR result but negative qPCR result, ddPCR demonstrated heightened sensitivity in early EBV infection detection. Conclusions:ddPCR holds certain advantages in monitoring EBV infection post-alloHSCT in children, especially for samples with low viral loads. However, as this method is still in the exploratory stage of clinical application, further research and practice are needed to validate its utility.

Background::This study aims to observe the dynamic changes of renal artery (RA) disease and cortical blood perfusion (CBP) evaluated by contrast-enhanced ultrasound (CEUS) after percutaneous transluminal renal artery stenting (PTRAS) in patients with severe atherosclerotic renal artery stenosis (ARAS) and to analyze the relationship between CBP and prognosis.Methods::This was a single-center retrospective cohort study. A total of 98 patients with unilateral severe ARAS after successful PTRAS in Beijing Hospital from September 2017 to September 2020 were included. According to renal glomerular filtration rate (GFR) detected by radionuclide imaging at 12 months after PTRAS, all patients were divided into the poor prognosis group ( n = 21, GFR decreased by ≥20% compared with baseline) and the control group ( n = 77, GFR decreased by < 20% or improved compared with baseline). Renal artery stenosis was diagnosed by digital subtraction angiography, and renal CBP was evaluated by CEUS using TomTec Imaging Systems (Germany) before PTRAS, at 6 months and 12 months after discharge. The receiver operating characteristic (ROC) curve with area under the curve (AUC) was used to analyze the predictive value of CBP parameters, including area under ascending curve (AUC1), area under the descending curve (AUC2), rising time (RT), time to peak intensity (TTP), maximum intensity (IMAX), and mean transit time (MTT) for poor prognosis. Results::Among the 98 patients, there were 52 males (53.1%), aged 55-74 years old, with an average age of 62.1 ± 8.7 years, and an average artery stenosis of 82.3 ± 12.9%. The poor prognosis group was associated with significantly increased incidence of diabetes (76.2% vs. 41.6%), and lower levels of GFR of the stenotic kidney (21.8 mL/min vs. 25.0 mL/min) and total GFR (57.6 mL/min vs. 63.7 mL/min) (all P < 0.05), compared with the control group ( P < 0.05). In addition, the rate of RA restenosis was significantly higher in the poor prognosis group than in the control group (9.5% vs. 0, χ2 = 9.462, P = 0.002). Compared with the control group, the poor prognosis group was associated with significantly decreased baseline AUC1 and AUC2, and extended duration of TTP and MTT ( P < 0.05). At 6 months and 12 months of follow-up, patients in the control group were associated with markedly increased AUC1, AUC2, and IMAX, and shorter duration of RT and MTT ( P < 0.05). The ROC curve showed that the predictive values of AUC1, AUC2, RT, TTP, IMAX, and MTT for poor prognosis were 0.812 (95% CI: 0.698-0.945), 0.752 (95% CI: 0.591-0.957), 0.724 (95% CI: 0.569-0.961), 0.720 (95% CI: 0.522-0.993), 0.693 (95% CI: 0.507-0.947), and 0.786 (95% CI: 0.631-0.979), respectively. Conclusions::Preoperative renal CBP in severe ARAS patients with poor prognosis is significantly reduced, and does not show significant improvement after stent treatment over the first year of follow-up. The parameter AUC1 may be a good predictor for renal dysfunction after PTRAS in severe ARAS patients.Trial Registration::ChiCTR.org.cn, ChiCTR1800016252.

Objective:To evaluate the performance of the Cepheid Xpert Xpress Flu/RSV assay (Xpert) in the detection of children infected with influenza virus (Flu) or respiratory syncytial virus (RSV).Methods:Nasopharyngeal specimens were collected from children who showed symptoms of respiratory infection and tested FluA, FluB and RSV by Xpert and sequencing assay respectively side by side. Discordant result were tested with a laboratory-developed real-time PCR for resolution.Results:A total of 388 nasopharyngeal swabs (NAPs) from children with acute respiratory tract infection were analyzed and the result showed 91.75-94.85% agreement between two tests. The sensitivity of FluA, FluB and RSV detected by Xpert and sequencing assay were 99.21% (125/126) vs. 92.86% (117/126), 100.00% (109/109) vs. 84.40% (92/109) and 100.00% (52/52) vs. 40.38% (21/52), respectively. The specificity of FluA, FluB and RSV detected by Xpert were all lower than that of the sequencing assay: 95.42% (250/262) vs 99.24% (260/262), 99.28% (277/279) vs 99.64% (278/279) and 99.70% (335/336) vs 100.00% (336/336). The positive predictive values (PPV) of FluA, FluB and RSV detected by Xpert were lower than those of the sequencing assay: 91.30% (126/138) vs. 98.33% (118/120), 98.18% (108/110) vs. 98.92% (92/93) and 98.11% (52/53) vs. 100.00% (21/21), respectively. The negative predictive values (NPV) of FluA, FluB and RSV detected by Xpert were higher than those of the sequencing assay: 99.60% (251/252) vs. 96.67% (261/270), 99.64% (279/280) vs. 94.27% (280/297) and 100.00% (337/337) vs. 91.60% (338/369).Conclusions:The Cepheid Xpert Xpress Flu/RSV assay is a sensitive, reliable and rapid assay for the detection of FluA, FluB and RSV in pediatrics.

Objective:To analyze the dominant genotypes and epidemic characteristics of human rhinovirus (HRV) in pediatric community-acquired pneumonia (CAP) in China.Methods:Between June 2017 to December 2019, throat swabs or nasopharyngeal aspirates were collected from pediatric CAP patients hospitalized in 6 medical institutions in Southern and Northern China (bounded by Qinling and Huaihe River), respectively.A total of 16 species of common respiratory viruses were screened using respiratory pathogen detection kits.Samples with positive HRV were genotyped for further epidemiological analysis.Results:The total detection rate of HRV in pediatric CAP (2 913 cases) was 12.2%(356 cases) in this study, which was 10.3%(145/1 410 cases) and 14.0%(211/1 503 cases) in Northern and Southern China, respectively.The detection rate of HRV in the Southern region was significantly higher than that in the Northern region, the difference was statistically significant( χ2=9.562, P=0.002). Epidemiological analysis showed that the distribution of HRV-positive cases in the Northern region was similar in all age groups, while 90.5% of positive cases in the Southern region were infants under 3 years.Classified by seasonal distribution, HRV-positive cases in the Northern region were mainly distributed in summer and autumn, while those in the Southern region were mainly distributed in spring and autumn.The coinfection rate of HRV and other pathogens was 40.7%, which was significantly higher in the Southern region than that of Northern region (49.0% vs.28.0%), the difference was statistically significant( χ2=15.801, P<0.001). The most common pathogens mixed with HRV were respiratory syncytial virus (RSV), human boca virus (HBOV) and parainfluenza virus type Ⅲ (PIV3) in the Northern region, with the mixed infection rate of 5.0%, 5.0% and 4.0%, respectively, which were RSV, PIV3 and human metapneumovirus (HMPV) in the Southern region, with the mixed infection rate of 9.0%, 7.0% and 6.0%, respectively.The genotyping results showed that all three HRV genotypes were detected.HRV-A (118/220 cases) was the dominant genotype, followed by HRV-C(82/220 cases). Conclusions:The detection rate of HRV in pediatric CAP cases is 12.2% in this study.There are significant differences in age and seasonal distribution of HRV-positive cases in the Southern and Northern regions.Coinfection of HRV and other pathogens is detected.The coinfection rate is significantly higher in the Southern region than that in the Northern region.HRV-A is the dominant genotype, followed by HRV-C.

Objective:To understand the profile type of serum Epstein-Barr virus (EBV) antibodies in children with infectious mononucleosis (IM), and to analyze the significance of viral capsid antigen (VCA) IgG antibody affinity in the diagnosis of IM.Methods:Retrospective analysis was performed on the results of the serum anti-EBV antibody profile and plasma EBV nucleic acid test of 150 hospitalized children with IM diagnosed in Beijing Children′s Hospital, Capital Medical University, from May 2016 to May 2019.Anti-EBV antibody profiles, including anti-VCA-IgG, anti-VCA-IgM, anti-early antigen (EA) IgA, anti-EBV nuclear antigen (EBNA) IgG, and anti-VCA-IgG affinity, were detected by enzyme-linked immunosorbent assay (ELISA). Plasma EBV nucleic acids were detected by real-time quantitative PCR.Results:There were mainly two types of anti-EBV antibody profiles in 150 children with IM: (1)130 cases who were positive for anti-VCA-IgM/IgG, negative for anti-EBNA-IgG and positive for anti-VCA-IgG antibodies with low affinity, accounting for 86.7% (130/150 cases), of which 50 cases were positive for anti-early antigen IgA; (2)18 cases who were negative for anti-VCA-IgM, positive for anti-VCA-IgG, negative for anti-EBNA-IgG and positive for anti-VCA-IgG antibody with low affinity, accounting for 12.0% (18/150 cases), of which 5 cases were positive for anti-EA IgA.EBV DNA was measured in 132 children, with a posi-tive rate of 37.9% (50/132 cases).Conclusions:There were several types of serum EBV antibody profiles in children with IM, 12.0% of patients with IM in this study were negative for anti-VCA-IgM, and the diagnosis of IM was confirmed by the affinity of anti-VCA IgG.

Objective:To observe the effect of the cortical blood perfusion parameter of wash-in area under curve (iAUC) with contrast-enhanced ultrasound(CEUS) on the effect of short-term outcomes of stent implantation in patients with severe renal artery stenosis (RAS).Methods:Retrospective analysis was performed on 82 patients with unilateral severe RAS who received stent implantation in Beijing Hospital from October 2017 to December 2019. According to the baseline iAUC before CEUS, all patients were divided into the poorly-perfused group (iAUC<850.0 dB×s) (37 cases) and the well-perfused group (iAUC≥850.0 dB×s) (45 cases). Baseline and perioperative clinical-imaging data were analyzed between the two groups. Followed up for 10-12 (11.5±1.7) months, Kaplan-Meier survival curves and Log-rank test were used to analyze the rate of adverse cardiac and renal vascular events and hypertension control rates.Results:Compared with the well-perfused group, the poorly-perfused group showed a longer course of hypertension, more diabetic patients, higher systolic blood pressure, diastolic blood pressure, 24 h average systolic blood pressure, and 24 h average diastolic blood pressure, lower glomerular filtration rate, and severe renal artery stenosis. Besides, the iAUC, wash-out AUC and the peak intensity were lower, the average transit time was longer, and the hypoglycemic treatment rate was higher (all P<0.05). Kaplan-Meier survival curve and Log-rank test analysis showed that the occurrence of cardio-renal vascular events ( HR=0.361, 95% CI=0.144-0.907, P=0.012) and renal function deterioration rate ( HR=0.286, 95% CI=0.090-0.914, P=0.035) in the well-perfused group were significantly lower than those in the poorly-perfused group. The blood pressure results demonstrated that the effective rate of hypertension treatment in the well-perfused group was significantly higher than that in the poorly-perfused group (93.3% vs 59.5%, P<0.001), but the improvement rate of hypertension (60.0% vs 43.2%) and cure rate (28.9% vs 16.2%) were not statistically significant between the two groups(all P>0.05). Conclusions:Severe RAS patients with decreased baseline iAUC often have diabetes, longer duration of hypertension, significantly reduced glomerular filtration rate and more severe RAS, short-term outcomes are worse with stent implantation.

Objective:To establish and validate a prognostic model of a contrast-enhanced ultrasound scoring(CEUS)system for evaluating renal artery stenosis(RAS)in the elderly.Methods:This was a single-center retrospective study.A total of 324 elderly RAS patients admitted to Beijing Hospital from October 2017 to July 2020 were randomly assigned into the model group(n=174)and the validation group(150)in a 1∶1 ratio.Clinical and imaging data of patients on admission including general conditions, previous medical history, blood pressure, blood creatinine, renal artery stenosis and cortical blood perfusion in the affected kidney and renal function(GFR)at 1-year follow-up were collected.Univariate and multivariate logistic regression was used to establish a model of the CEUS scoring system.The receiver operating characteristic(ROC)curve and area under the ROC curve(AUC)were used to evaluate prediction accuracy.Clinical application value of the CEUS scoring system model was evaluated via decision curve analysis using a nomogram.Results:Baseline clinical and radiomic data had no significant difference between the model group and the validation group( P>0.05). Multivariate logistic regression analysis results showed that age( OR=1.242, 95% CI: 1.081-1.427, P<0.01), diabetes( OR=1.545, 95% CI: 1.107-2.156, P<0.05), blood pressure( OR=1.328, 95% CI: 1.056-1.670, P<0.05), renal function( OR=2.374, 95% CI: 1.216-3.887, P<0.01)and cortical blood perfusion parameter( OR=2.646, 95% CI: 1.553-6.369, P<0.01)were risk factors for the deterioration of renal function during 1 year follow-up.Based on these results, a nomogram for the CEUS scoring system model was drawn, and its consistency index, the C-Index, was 0.725(95% CI: 0.653-0.776). The AUC of the CEUS scoring system was 0.824 and the Youden index was 0.711 in the model group, with a specificity of 0.774 and a sensitivity of 0.837.The AUC of the CEUS scoring system was 0.853 and the Youden index was 0.715 in the validation group, with a specificity of 0.684 and a sensitivity of 0.889.There was no significant difference in ROC curve between the two groups( D=1.387, P>0.05). In addition, calibration charts of the two models showed that the calibration curve of the CEUS scoring system was close to the standard curve, with no statistically significant difference( P>0.05). Conclusions:The CEUS scoring system model can be used to predict the risk of worsening renal function in elderly RAS patients during 1-year follow-up.

Epstein-Bar virus (EBV) infection is widespread within the human population with over 95% of adults being infected. In response to primary EBV infection, the host mounts an antiviral immune response comprising both innate and adaptive immune system. In healthy populations, the immune system can control EBV infection to a large extent. However, the virus cannot be cleared. Instead, EBV establishes a persistent latent infection in B lymphocytes characterized by limited viral gene expression. To establish a persistent infection efficiently, EBV have evolved a number of strategies to avoid immune elimination. In this review, we focus on the immune evasion mechanisms of EBV encoded immune-evasion proteins, microRNA and host exosome pathway.

The Epstein-Barr virus (EBV) is a gamma herpes virus associated with several types of malignancies. The EBV encodes viral microRNAs (miRNAs) that can target genes within cells. The EBV participates in signal transduction as well as the proliferation and differentiation of cells. How the target genes and functions of EBV-encoded miRNAs contribute to the pathogenesis of EBV is an important research topic. Some target genes have been validated since EBV-encoded miRNAs were discovered and, in this article, we summarize them and their functions.
Animals ; Epstein-Barr Virus Infections ; genetics ; metabolism ; virology ; Herpesvirus 4, Human ; genetics ; physiology ; Humans ; MicroRNAs ; genetics ; metabolism ; RNA, Viral ; genetics ; metabolism

OBJECTIVE To investigate the distribution of killer cell immunoglobulin-like receptors (KIR) and their specific ligands human leukocyte antigen-I (HLA-I) gene in northern China. METHODS One hundred and eighty-four unrelated northern Chinese Han individuals were recruited. Genotypes of the KIR and HLA-ABC genes were studied by sequence-specific primer polymerase chain reaction (SSP-PCR). RESULTS Sixteen KIR genes were detected among the 184 unrelated individuals. In all individuals, the four framework genes were present. The frequencies for those carrying the remaining 12 KIR genes have ranged from 16.3% to 99.5%. Twenty-four KIR genotypes were identified, for which half were detected in a single individual. A new genotype comprised of KIR2DL3, 3DL1, 2DP1 and the framework genes was detected in one subject. Respectively, 12, 27 and 11 specificities of HLA alleles were identified on the HLA-A, B, C loci. CONCLUSION The distribution of polymorphisms of KIR and its ligand HLA-ABC genes among northern Chinese Han population have been ascertained. The frequencies of 9 KIR/HLA combinations in the above population have been determined for the first time.
Child ; China ; ethnology ; Ethnic Groups ; Female ; Gene Frequency ; Histocompatibility Antigens Class I ; genetics ; Humans ; Male ; Receptors, KIR ; genetics