Search

Vitamin D (VD) plays an important role in infectious and autoimmune diseases. We investigated the association between the serum VD levels and active human cytomegalovirus (HCMV) infections in pediatric systemic lupus erythematosus (SLE) patients. From January 2015 to June 2021, 120 children were diagnosed with SLE, and 100 healthy children were enrolled. The serum 25-hydroxyvitamin D [25(OH)D] levels were detected using ELISA. The serum anti-HCMV IgM antibodies were measured by a chemiluminescence immunoassay. Comparisons of the 25(OH)D levels between SLE patients and healthy children were performed, as well as subgroups of SLE patients with or without active HCMV infections. The serum 25(OH)D levels of SLE patients were significantly lower than those of healthy children (35.3 ± 12.9 vs. 49.3 ± 15.3, P0.001). The VD deficiency ratio was higher in SLE patients (89.2%) than that in healthy children (52.0%). The serum 25(OH)D levels in the positive anti-HCMV IgM group were significantly lower than those of the negative anti-HCMV IgM group (30.6 ± 12.3 vs. 38.2 ± 12.5, P0.001). A severe VD deficiency ratio was significantly higher in HCMV-IgM(+)-SLE patients (42.2%) than that in HCMV-IgM(-)-SLE patients (13.3%). This study suggested that the serum VD level is associated with active HCMV infections in pediatric SLE patients.

Human metapneumovirus (HMPV), which is distributed worldwide, is a significant viral respiratory pathogen responsible for causing acute respiratory tract infections (ARTIs) in children. The aim of the present study was to investigate the epidemiological and genetic characteristics of HMPV in pediatric patients in Hangzhou China following the peak of onset of coronavirus disease 2019 (COVID-19). A total of 1442 throat swabs were collected from the pediatric patients with a diagnosis of ARTI from November 2020 to March 2021. The following viruses were detected by real-time polymerase chain reaction analysis: HMPV, RSV, adenovirus, hPIV1-3, influenza A, and influenza B. A two-step method was used to amplify the F genes of the HMPV-positive samples. Following sequencing, phylogenetic analyses were conducted using the MEGA version 7 software package. Among the 1442 samples, 103 (7.14%) were positive for HMPV. No significant differences were observed in the gender distribution. The highest incidence of HMPV occurred in children older than 6 years and the lowest was noted in children younger than 6 months. Lower respiratory tract infections were diagnosed at a higher rate than upper respiratory tract infections in HMPV-infected children. Only 10 HMPV-infected children (5.41%) were inpatients compared with 93 outpatients (7.39%). Co-infection was observed in 31 HMPV-positive samples including 24 samples of double infection and seven samples of triple infection. A total of 61F gene fragments of HMPV, which were approximately 727 bp in length were successfully sequenced. All the HMPVs belonged to the genotype B and were clustered into subgenotypes B1 (1.6%, 1/61) and B2 (98.4%, 60/61). A total of four specific amino acid substitutions were noted as follows: aa280, aa296, aa392, and aa396. These substitutions were present between sequences derived from the subgenotypes B1 and B2 in the fusion open reading frame from position 244 to 429. In conclusion, the present study provided significant information regarding the epidemiological and genetic characteristics of HMPV in children living in Hangzhou. Following the first peak of the COVID-19 pandemic, HMPV was considered an important viral respiratory pathogen present in children with ARTI.

The novel coronavirus 2019 (COVID-19) caused by SARS-CoV-2 spread rapidly worldwide, posing a severe threat to public life and health. It is significant to realize rapid testing and timely control of epidemic situations under the condition of limited resources. However, laboratory-based standardized nucleic acid detection methods have a long turnaround time and high cost, so it is urgent to develop convenient methods for detecting COVID-19. This paper summarizes the point-of-care testing (POCT) developed for novel coronavirus from three aspects: nucleic acid extraction, nucleic acid amplification, and detection methods. This paper introduces a commercial real-time detection system that integrates the abovementioned three steps and the matters needing attention in use. The primary purpose of this review is to provide a reference for emergency response and rapid deployment of COVID-19 and some other emerging infectious diseases.

Purpose This study set out to determine the antimicrobial resistance trends of Haemophilus influenzae isolates from pediatric hospitals in Mainland China, which would provide basis for clinical treatment. Methods The Infectious Disease Surveillance of Pediatrics (ISPED) collaboration group conducted this study. H. influenzae strains isolated from nine pediatric hospitals in Mainland China were included. Disk diffusion method was used for antimicrobial susceptibility test. Cefinase disc was used for detection of β-lactamase. Results In total, 13810 H. influenzae isolates were included during 2017–2019: 93.17% of which were from respiratory tract specimens, 4.63% from vaginal swabs, 1.10% from secretion, and 1.10% from others. Of all strains, 63.32% isolates produced β-lactamase; 8.22% isolates were β-lactamase-negative and ampicillin-resistant (BLNAR). The resistance to sulfamethoxazole-trimethoprim was 70.98%, followed by resistance to ampicillin (69.37%), cefuroxime (51.35%), ampicillin-sulbactam (38.82%), azithromycin (38.21%), amoxicillin-clavulanate (35.28%). More than 90% of H. influenzae isolates were susceptible to ceftriaxone, cefotaxime, meropenem, levofloxacin and chloramphenicol. The resistance rate of ampicillin and azithromycin in H. influenzae showed an increasing trend through the years. Statistically significant differences in antibiotic-resistance rates of all the antibiotics except chloramphenicol were found in different regions. The major Multi-Drug Resistance pattern was resistant to β-lactams, macrolides, and sulfonamides. Conclusions There is a rising trend of resistance rate of ampicillin and azithromycin in H. influenzae. Antimicrobial resistance of H. influenzae deserves our ongoing attention. Third-generation cephalosporin could be the preferred treatment option of infections caused by ampicillin-resistant H. influenzae.

IL-10 is thought to play an important role in preventing inflammatory bowel disease (IBD), although its efficacy is limited in IBD inflammation treatment. The purpose of this study is to investigate the association between SNP polymorphism in the promoter region of the IL-10 gene and Crohn's disease (CD).In 86 children with CD disease and 142 healthy controls, polymorphisms of three SNPs (rs3790622, rs1800872, and rs1800896) in the IL-10 promoter region were successfully identified. The risk alleles, genotypes, and haplotypes were also analyzed in the CD patient group and the control group. 2 × 2 chi-square test was used to identify whether there is a statistically significant association between CD risk and SNP polymorphisms.According to the chi-square test results, only the polymorphism of rs1800872 was associated with pediatric CD. T allele in rs1800872 showed a high risk for pediatric CD (Pearson χOur data suggested that T allele, TT genotype, and haplotype GTT in rs1800872 were associated with the susceptibility to pediatric CD in China.

Objective This study aimed to investigate characteristics of human cytomegalovirus (HCMV) glycoprotein H (gH) genotypes in urine, throat swab, and serum from children and breast milk from children's mothers. Methods Fresh urine samples, throat swabs, or serum samples from children and breast milk samples from children's mothers were collected for HCMV DNA detection. The positive samples of HCMV DNA were further detected by fluorescent quantitative polymerase chain reaction (PCR) with gH typing. Results Of 1,703 HCMV DNA-positive samples, the highest proportion (83.3%, 85/102) of children aged between 21 days and 3 months was detected positive in breast milk samples (p = 0.002), and the highest proportion (70.5%, 110/156) of children aged above 3 months was detected positive in throat swab samples (p = 0.002). HCMV in throat swab specimens is mainly high copy (p Conclusion gH1 is the predominant genotype of HCMV in each kind of sample in China. However, the distribution of the HCMV gH genotype is different among different samples.

To investigate the interaction between long non-coding RNA4.9 (lncRNA4.9) and transforming growth factor-β1 (TGF-β1) in the model of HCMV DNA. The experiment was divided into 5 groups: NC group, PCDNA-lncRNA4.9 group, SH-lncRNA4.9 group, LV-TGF-β1) Group, and SH-lncRNA4.9 group. Compared with the NC group, overexpression of lncRNA4.9 could significantly increase (P P P P P

Toll-like receptor 4 (TLR4) is considered to be involved in the pathogenesis and progression of atopic dermatitis (AD). In the present study, we evaluated the relationship between TLR4 gene polymorphisms and the susceptibility or severity of AD among Chinese Han children.A total of 132 AD patients and 100 healthy controls were enrolled in this study. Four single-nucleotide polymorphisms (rs19277914, rs11536891, rs7869402, and rs11536889) of the TLR4 gene were genotyped by multiplex PCR combined with next-generation sequencing.Our results showed that a significantly reduced risk for AD was associated with C allele [p = 0.008; odds ratio (OR) = 0.41, C vs. T], TC genotype (p = 0.022; OR = 0.41, TC vs. TT), and TC + CC genotype (p = 0.010; OR = 0.39, TC + CC vs. TT) of TLR4 rs11536891. The frequency of the haplotype GCCG (rs1927914-rs11536891-rs7869402-rs11536889) in AD patients was lower than that in the controls (p = 0.010; OR = 0.38). Moreover, the results indicated that a higher risk of severe AD was related to the T allele (p = 0.019; OR = 2.97, T vs. C) and the TC genotype (p = 0.021; OR = 3.34, TC vs. CC) of TLR4 rs7869402. A risk haplotype of TLR4 (GTTG) was found in severe AD patients (p = 0.010; OR = 5.26).Our data suggested that TLR4 rs11536891 polymorphism was associated with the susceptibility to AD in Chinese Han children. And TLR4 rs7869402 might confer the severity of pediatric AD patients.

The present study aimed to assess the diagnostic significance of serum bone metabolic parameters in children with growing pains (GPs).All patients diagnosed with GP and healthy controls matched with age and gender were recruited at the outpatient clinic of Children's Hospital at Zhejiang University School of Medicine from August 2016 to August 2021. In all subjects, serum levels of calcium (Ca), phosphorus (P), procollagen type-I N-terminal (PINP), parathormone (PTH), 25-hydroxyvitamin D (25-(OH)D), osteocalcin (OC), N-terminal cross-linked telopeptides of type-I collagen (CTX), and tartrate-resistant acid phosphatase type 5b (TRACP5b) were investigated. The univariate analysis, multivariate logistic regression analysis, and receiver operating characteristic (ROC) curve were used to identify the bone metabolic parameters factors for diagnosing GP.We enrolled 386 children with GP and 399 healthy controls in present study. The mean age of GP group was 5.319 years, and, primarily, the subjects were preschool-age children. The gender ratio (male-to-female) was 1.27 in GP group. After adjusting for age and gender, we identified that the serum levels of Ca (p 0.001, OR: 25.039), P (p = 0.018, OR: 2.681), PINP (p 0.001, OR: 1.002), and PTH (p = 0.036, OR: 0.988) were independent diagnostic factors associated with GP. Area under curve (AUC) of the ROC curves was in the order: PINP (0.612) Ca (0.599) P (0.583) PTH (0.541). A combination of independent diagnostic factors and multivariable logistic regression analysis provided a refined logistic regression model to improve the diagnostic potential, of which the AUC had reached 0.655.Serum levels of Ca, P, PINP, and PTH could be independent diagnostic factors associated with GP. The logistic model was significantly superior to bone metabolic parameters for diagnosing GP.

Chronic active Epstein-Barr virus (CAEBV) infection is a very rare and potentially life-threatening illness caused by long-term EBV infection. Globally, the prevalence is the highest among young children and adolescents with increased frequency in Asians and Native Americans, whereas it is sporadically encountered in European countries. Typically, patients present with nonspecific symptoms such as fever, lymphadenopathy, hepatosplenomegaly, and liver dysfunction. However, the complications of CAEBV and its treatment are quite complex and require great care. We report a case of a 3-year-old boy with CAEBV infection who was later diagnosed with pancreatic mass and recurrent pancreatitis. A multidisciplinary board was consulted for correct diagnosis and treatment plan making. The treatment included pharmaceutical and surgical (duodenum-preserving pancreatic head resection) approaches. The patient showed tremendous improvement following the third discharge from the hospital and is still free of any symptoms. In this case report, we discuss differential diagnosis and comprehensively examined the possibility of the development of pancreatitis caused by EBV infection, the possibility of autoimmune pancreatitis, and the possibility of hereditary pancreatitis. To confirm or rule out the first two etiologies, laboratory and pathology results were studied. We also performed exon sequencing using Agilent exome capture kit to rule out hereditary pancreatitis. The clinical course of this disease and the way it was handled deserves attention so that similar cases receive prompt treatment.

BACKGROUND: In recent years, B-cell dysfunction has been found to play an important role in the pathogenesis of primary nephrotic syndrome (PNS). B cells play a pathogenic role by secreting antibodies against their target antigens after transforming into plasma cells. Therefore, this study aimed to screen the autoantibodies that cause PNS and explore their pathogenic mechanisms. METHODS: Western blotting and mass spectrometry were employed to screen and identify autoantibodies against podocytes in children with PNS. Both in vivo and in vitro experiments were used to study the pathogenic mechanism of PNS. The results were confirmed in a large multicenter clinical study in children. RESULTS: Annexin A(2) autoantibody was highly expressed in children with PNS with a pathological type of minimal change disease (MCD) or focal segmental glomerulosclerosis without genetic factors. The mouse model showed that anti-Annexin A(2) antibody could induce proteinuria in vivo. Mechanistically, the effect of Annexin A(2) antibody on the Rho signaling pathway was realized through promoting the phosphorylation of Annexin A(2) at Tyr24 on podocytes by reducing its binding to PTP1B, which led to the cytoskeletal rearrangement and damage of podocytes, eventually causing proteinuria and PNS. CONCLUSIONS: Annexin A(2) autoantibody may be responsible for some cases of PNS with MCD/FSGS in children.

Enteroviruses (EVs) are the major cause of hand, foot, and mouth disease (HFMD) and herpangina in children. In this study, we conducted a molecular investigation of EVs in throat swab samples from children in Hangzhou, China with a diagnosis of HFMD or herpangina. EVs were detected using one-step real-time RT-PCR, and their serotypes were determined based on partial VP1 gene sequences. The molecular typing results revealed the presence of six different EV serotypes in HFMD cases, including coxsackievirus (CV) A16 (20/30, 66.7%), CVA4 (3/30, 10.0%), CVA6 (3/30, 10.0%), EVA71 (2/30, 6.7%), CVB4 (1/30, 3.3%), and CVB5 (1/30, 3.3%). Eleven different EV serotypes were detected in herpangina cases, among which CVA4 was the most frequently detected serotype (105/170, 61.8%), followed by CVA16 (30/170, 17.6%), CVB4 (9/170, 5.3%), CVA6 (6/170, 3.5%), CVB3 (5/170, 2.9%), CVA10 (3/170, 1.8%), EVA71 (4/170, 2.4%), Echo9 (3/170, 1.8%), CVA9 (2/170, 1.2%), CVB1 (3/170, 1.8%) and CVA5 (1/170, 0.6%). The nucleotide sequence identity of EV strains from the same subtype ranged from 80.7% to 100%, and most of the EVs were closely related to virus strains found in Australia and mainland China. In conclusion, CVA 16 and CVA 4 were the main serotypes causing HFMD and herpangina, respectively, in children in Hangzhou in 2016. Most of these EVs were closely related to virus strains from Australia and mainland China.

Objective: To evaluate the clinical diagnostic value of colloidal gold assay for detection of influenza A and B virus. Methods: Results were compared for colloidal gold assay for influenza antigen detection and real-time RT-PCR for RNA detection. Results: By performing the colloidal gold assay, the positive rate was 25.67% (105/409) for influenza A virus and 8.56% (35/409) for influenza B virus. While tested by real-time RT-PCR, 107(26.2%) were positive for influenza A virus and 35(8.6%) for influenza B virus. Using real-time RT-PCR as the gold standard, the sensitivity and specificity of the colloidal gold assay were 84.1% and 95.0%, respectively, for influenza A virus and 85.7% and 98.7% for influenza B virus. Conclusion: This colloidal gold assay may be one of the viable tools for the rapid screening for influenza viruses in an outpatient clinical practice.

Objectives: We evaluated the epidemiological features and various inflammatory markers in hospitalized children with influenza virus infection in China. Methods: The real-time RT-PCR assay was performed for detection and genotyping of influenza A and B virus. Th1/Th2 cytokines, WBC, and CRP were determined in influenza virus positive children. Results: H1N1 and Yamagata were the prevalent genotypes of influenza A and B virus in Hangzhou, respectively. IL-2, IL-10, and CRP were significantly increased and IFN-γ was decreased in children with severe Influenza A virus infection, and TNF-α and IFN-γ levels were found to be significantly lower in children with severe Influenza B virus infection. Conclusion: Increased IL-2, IL-10, and CRP with decreased IFN-γ may indicate a severe influenza A virus infection, and decreased TNF-α and IFN-γ may indicate a severe influenza B virus infection in children.

ObjectiveThis study aims to analyze the molecular epidemiology, resistance, and pathogenicity of Salmonella enterica subsp. diarizonae isolated from children.MethodsWhole genome sequencing was carried out, and molecular serotypes, sequence types, resistance genes, and virulence genes of S. enterica subsp. diarizonae isolates were analyzed. Antimicrobial susceptibility test was determined by commercialized microdilution method.ResultsA total of three isolates of S. enterica subsp. diarizonae were isolated during 2015 to 2020. The molecular serotypes of the three strains were 61:c:z35, 61:l,v:1,5,7:[z57], and 65:k:z, respectively, and the sequence types were ST1845, ST233, and ST1263. All the three isolates were susceptible to ceftriaxone, ceftazidime, cefepime, amoxycillin/clavulanic acid, piperacillin/tazobactam, ertapenem, imipenem, levofloxacin, and trimethoprim/sulfamethoxazole. No other resistant gene was detected except aac(6’)-Iaa. There were no resistant plasmids detected in all the three isolates. A total of 76 genes were present in all isolates, containing 49 genes of Type III Secretion System (T3SS) mediated by SPI-1and SPI-2, 13 genes of adherence (type 1 fimbriae, Agf, and MisL-related genes), 11 genes of iron uptake (Yersiniabactin), two genes of magnesium uptake, and one gene of typhoid toxin(cdtB).ConclusionThe serotypes and sequence types of S. enterica subsp. diarizonae isolates were rarely reported in children; all the S. enterica subsp. diarizonae isolates were susceptible to detected antibiotics; T3SS, adherence, iron uptake, magnesium uptake, and typhoid toxin were responsible for pathogenicity of the S. enterica subsp. diarizonae isolates in children.

Congenital cytomegalovirus infection (cCMVi) in the newborn is one of the most common congenital infections. The children who are infected will present different clinical manifestations. The mild symptoms may be asymptomatic. However, severe cases may have permanent sequelae and even endanger their life. This paper would review the research progresses in the following fields including the epidemiology, the route of transmission, clinical manifestation, screening methods , narrative perspective of prevention and control means. Then the prevention strategies would be summarized.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is raging worldwide. The COVID-19 outbreak caused severe threats to the life and health of all humans caused by SARS-CoV-2. Clinically, there is an urgent need for an in vitro diagnostic product to detect SARS-CoV-2 nucleic acid quickly. Under this background, commercial SARS-CoV-2 nucleic acid POCT products came into being. However, how to choose these products and how to use these products in a standardized way have brought new puzzles to clinical laboratories. This paper focuses on evaluating the performance of these commercial SARS-CoV-2 nucleic acid POCT products and helps the laboratory make the correct choice. At the same time, to standardize the use of this kind of product, this paper also puts forward corresponding suggestions from six elements of total quality management, namely, human, machine, material, method, environment, and measurement. In addition, this paper also puts forward some ideas on the future development direction of POCT products.

Coronavirus disease 2019 (COVID-19) is a grade B infectious disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). In pace with the spreading of the disease, biosafety risk of the biological specimen preservation in biobanks has been significantly increased and biosafety protection during biological specimen preservation become increasingly important. According to the related national rules and the corresponding guidelines of Chinese Medical Association, this paper introduced the etiology about SARS-CoV-2, epidemiology about COVID-19, and the biosafety protection principles of individuals and biological specimen storage places in the process of personal protection, protection of collection, transport, handling, preservation, detection, post-detection disposal and emergencies of biological specimen. Emphasized to carry out a strict biosafety-risk assessment on biological specimen basing on virus load information, infectivity, and sample type (possible contact transmission, aerosol transmission, and fecal oral transmission).

The rapid spread of the epidemic has aroused widespread concern in the international community. Severe acute respiratory syndrome coronavirus 2 (SARS‐COV‐2) originated from Wuhan's Huanan wholesale seafood market, with bats as the likely original hosts and pangolins as potential intermediate hosts. The current source of the disease is mainly patients infected with SARS‐COV‐2. Patients in the incubation period may also become sources of infection. The virus is mainly transmitted via respiratory droplets and contact, and the population is generally susceptible. The epidemic has progressed through the local outbreak stage and community transmission stage due to exposure at Wuhan's Huanan wholesale seafood market and is now in the stage of large‐scale transmission due to the spread of the epidemic. The basic productive number (R0) at the beginning of the epidemic was 2.2, with an average incubation period of 5.2 days. The proportion of critically ill patients was 23.4%, the mortality rate was lower than those of SARS and Middle East respiratory syndrome, and 96.5% of deaths occurred in Hubei Province, where the outbreak occurred first. Among them, elderly men with underlying diseases had a higher mortality rate. Chinese medical staff have summarized a set of effective strategies and methods in the diagnosis and treatment of this disease that are worthy of reference for their international counterparts. With powerful government intervention and the efforts of Chinese medical staff, China's outbreak has gradually improved., Research Highlights The current source of the disease is mainly patients infected with SARS‐COV‐2. Patients in the incubation period may also become sources of infection.The virus is mainly transmitted via respiratory droplets and contact, and the population is generally susceptible.The proportion of critically ill patients was 23.4%, the mortality rate was lower than those of SARS and Middle East respiratory syndrome. Among them, elderly men with underlying diseases had a higher mortality rate.

To establish a clinical prediction rule for acute bilirubin encephalopathy (ABE) in term/near-term neonates with extreme hyperbilirubinemia. A retrospective cohort study was conducted between January 2015 and December 2018. Six hundred seventy-three out of 26,369 consecutive neonates with extreme hyperbilirubinemia were enrolled in this study. Data included demographic characteristics, total serum bilirubin (TSB), albumin, bilirubin/albumin ratio (B/A), direct antiglobulin test, glucose-6-phosphate deficiency, asphyxia, sepsis, acidosis. ABE was defined as a bilirubin induced neurological dysfunction score of 4 to 9. We used stepwise logistic regression to select predictors of ABE and devised a prediction score. Of the 673 eligible infants, 10.8% suffered from ABE. Our prediction score consisted of 3 variables: TSB (as a continuous variable; odds ratio [OR] 1.16; 95% confidence interval [CI], 1.02–1.31; logistic coefficient 0.15), B/A (as a continuous variable; OR 1.88; 95% CI, 1.19–2.97; logistic coefficient 0.67), and sepsis (OR 3.78; 95% CI, 1.40–10.21; logistic coefficient 1.19). Multiplying the logistic coefficients by 10 and subtracting 75, resulted in the following equation for the score: Score = 12 × (if sepsis) + 1.5 × (TSB) + 7 × (B/A) − 75. The model performed well with an area under the curve of 0.871. The risk of ABE can be quantified according to TSB, B/A, and sepsis in term/near-term neonates with extreme hyperbilirubinemia.

The pandemic influenza A (H1N1)pdm09 virus continues to be a threat to human health. Low doses of mannan-binding lectin (MBL) (

Background: Respiratory syncytial virus (RVS) is the main cause of morbidity and mortality in the pediatric population. Objectives: The objective of the study was to compare the diagnostic accuracies of the colloidal-gold-Genesis and newly developed real-time RT-PCR-IngeniGen with an admitted real-time RT-PCR-HuaRuiAn as the gold standard for respiratory syncytial virus detection. Methods: These three assays were performed on collected specimens. Results: Altogether 258 specimens were studied. Among them, real-time RT-PCR-HuaRuiAn detected the respiratory syncytial virus in 166 samples, while colloidal-gold-Genesis detected 171 samples and real-time RT-PCR-IngeniGen detected 168 samples as positive. The overall sensitivity for the detection of the respiratory syncytial virus in comparison to real-time RT-PCR-HuaRuiAn was 95.2% for colloidal-gold-Genesis and 100% for real-time RT-PCR-IngeniGen. The specificity was 85.9% for colloidal-gold-Genesis and 97.8% for real-time RT-PCR-IngeniGen. The positive likelihood ratio was 6.75 for colloidal-gold-Genesis and 45.45 for real-time RT-PCR-IngeniGen. The negative likelihood ratio was 0.06 for colloidal-gold-Genesis and 0.00 for real-time RT-PCR-IngeniGen. The cycle threshold value of real-time RT-PCR-IngeniGen was higher than that of real-time RT-PCR-HuaRuiAn. Conclusions: Colloidal-gold-Genesis is a good alternative for fast screening of respiratory syncytial virus detection.

Background Rotavirus A (RVA) and adenovirus (Adv) are important causes of acute diarrhea in children. RVAs are classified into G and P genotypes based on viral proteins (VP)7 and VP4 gene and Adv contains over 70 genotypes based on hexon and fiber gene. This study aimed to characterize the molecular epidemiology of RVA and Adv in children with acute diarrhea during 2017–2018 in Hangzhou. Methods The stool samples were collected and tested for RVA and Adv by reverse transcription-quantitative PCR (RT-qPCR) assay. The RVA positive samples were detected by RT-PCR for VP7(G) and VP4([P]) genotypes, and the Adv positive samples were detected by PCR for genotyping by the target to hexon gene. Results Among 228 RVA-positive samples, G9 was detected as the most frequent genotype (195/228, 85.5%), followed by G3 (20/228, 8.8%), G2 (7/228, 3.1%) and G1 (6/228, 2.6%). G9 strains were closely related to strains from China and neighboring countries, as well as the USA. On the other hand, P[8] strains were detected in 219 (96.1%) samples with most closely related to one strain from Malawi, and P[4] in 9 (3.9%) samples. G9P[8] (84.6%, 193/228) was the most prevalent rotavirus A strains, followed by G3P[8] (8.8%, 20/228), G2P[4] (3.1%, 7/228), G1P[8] (2.6%, 6/228) and G9P[4] (0.9%, 2/228). Of 167 Adv-positive cases, 2 different genotypes were identified with 152 (91.0%) of Adv-41and 15 (9%) of Adv-40. All Adv strains were closely related to prototype strains of Adv types 40 and 41 in India. Conclusions G9P[8] of RVA and Adv-41 were the most common genotypes that caused children’s acute diarrhea in Hangzhou, 2017–2018.

Wilson’s disease (WD), also named hepatolenticular degeneration, is an autosomal-recessive disorder in which abnormal copper metabolism leads to copper excretion disorder and deposition in target organs. WD has a high mortality rate and disability rate, however, it is one of the treatable hereditary diseases. Irreversible tissue injury can be prevented if WD was diagnosed and treated before the development of clinical symptoms. Thus it is necessary to screen WD in the family members of the proband. First-degree relatives of a proband with WD should be screened. First-degree relatives should include the previous generation, siblings and the next generation. If available, genetic testing can be used as the primary screening method. Although the relatives of a proband are more likely to be patients with WD, the diagnosis should be based on sufficient evidence to avoid unnecessary lifelong treatment.

Objectives: Interleukin-33 (IL-33) was one of the members of IL-1 family, and it was reported that the single nucleotide polymorphisms (SNPs) in the IL-33 gene was contribute to the susceptibility to immune related diseases, including rheumatoid arthritis, asthma, Behcet’s disease, systemic lupus erythematosus and ankylosing spondylitis. However, the potential association between SNPs in IL-33 gene and Henoch-Schonlein purpura (HSP) was not explored. Methods: We designed a case-control study, a total of 181 HSP patients and 202 healthy pediatric controls were included, which aimed to study the relationship between the SNPs in the IL-33 gene (rs1929992, rs7044343) and HSP with a Chinese pediatric cohort. Results: There were no evidence for the association of rs1929992 and rs7044343 polymorphism in IL-33 gene with pediatric HSP patients (C versus T, P = 0.622, OR = 0.931, 95% CI: 0.7 - 1.238; TT versus CC, P = 0.742, OR = 0.908, 95% CI: 0.513 - 1.609; CT versus CC, P = 0.158, OR = 0.714, 95% CI: 0.446 - 1.141 and C versus T, P = 0.920, OR = 0.986, 95% CI: 0.742 - 1.309; TT versus CC, P = 0.939, OR = 0.978, 95% CI: 0.549 - 1.741; CT versus CC, P = 0.416, OR = 0.817, 95% CI: 0.501 - 1.331, respectively). Moreover, association studies were performed on the correlation between IL-33 gene polymorphisms and HSP nephritis patients as well as patients with gastrointestinal manifestation and joint involvement. However, there were no significant association regarding the distribution of allele and genotype frequencies between HSP patients with different system involvement and healthy controls either. Conclusions: The present findings indicated that both of rs1929992 and rs7044343 in gene IL-33 polymorphism were not related to the susceptibility to HSP and the different system involvement of HSP patients.

Externally controlling the excitation of a neuronal subset through ion channels activation can modulate the firing pattern of an entire neural circuit in vivo. As nanovalves in the cell membrane, ion channels can be opened by light (optogenetics) or ultrasonic (sonogenetics) means. A thoroughly analyzed force sensor is the Escherichia coli mechano sensitive channel of large conductance (MscL). Here we expressed MscL in rat hippocampal neurons in a primary culture and showed that it could be activated by low-pressure ultrasound pulses. The gain-of-function mutation, I92L, sensitized MscL's sonic response, triggering action potentials at a peak negative pressure as low as 0.25 MPa. Further, the I92L MscL reliably elicited individual spikes by timed brief pulses, making excitation programmable. Because MscL opens to tension in the lipid bilayer, requiring no other proteins or ligands, it could be developed into a general noninvasive sonogenetic tool to manipulate the activities of neurons or other cells and potential nanodevices.

Background: Human cytomegalovirus (HCMV) infection is very common and latency can be reactivated in the future. And it can alter the intracellular environment, similar to other herpesviruses, for viral replication and survival. The aim of this study was to investigate the influence of HCMV infection on autophagy in human acute monocytic leukemia cell line (THP-1 cells). Methods: Reverse transcription polymerase chain reaction (RT-PCR), western blot, and transmission electron microscope (TEM) were used to examine autophagy level. The concentrations of autophagy-related proteins Beclin 1, Atg5, and the light chain three (LC3) were counted when compared with actin level. Results: The expression levels of Beclin1, Atg5, and LC3II mRNAs increased gradually between 1 and 5 days(d) postinfection (p.i.) and subsequently decreased little by little when compared with the control THP-1 cells. However, results of western blot analysis displayed that the level of LC3II increased gradually after 1 day p.i. and decreased at 7 days after infection. But the levels of Atg5 and Beclin1 increased gradually after 2 days p.i. and began to decrease at 5 days after infection, respectively. Conclusion: These results suggested that HCMV infection can facilitate the autophagy and autophagy level may decrease in latent phase. More studies on the relationship between HCMV latency and autophagy are needed to determine the role of autophagy in HCMV latent infection that may help find out a therapeutic approach for clinical treatment.

Background: Human herpes viruses, as common viruses, not only affect mainly the skin, mucosa, and nervous tissue, but also can cause a variety of serious diseases in children. Objectives: The aim of this study was to determine the sensitivity and specificity of multiplex PCR-based DNA microarray technology in comparison with PCR method and IgM ELISA. Methods: A total of 108 blood samples from children with viral infections were collected and analyzed by multiplex PCR-based DNA microarray technology, PCR method, and IgM ELISA. Results: Of 108 specimens, 16 were positive which gave a positive rate of 14.8%. Most of the patients were infected with EBV and HCMV. The sensitivity and specificity of this technology for detecting human herpes viruses were 100% when compared to PCR method. The crude agreement between multiplex PCR-based DNA microarray technology and IgM ELISA for detecting human herpes viruses was 95.4%. Conclusions: The results indicated that multiplex PCR-based DNA microarray technology is a rapid auxiliary diagnostic method for simultaneous detection of the seven common herpes viruses with high sensitivity and specificity.

Mycoplasma pneumoniae (M. pneumoniae) infection can cause community acquired pneumonia in children. A real-time method of simultaneous amplification and testing of M. pneumoniae (SAT-MP) was developed to diagnose M. pneumoniae targeting a region of the ribosomal RNA. The SAT-MP assay can accurately identify M. pneumoniae with a detection range from 101 to 107 CFU/ml. In this study, the specimens from 315 children with pneumonia were collected and analyzed by SAT-MP in parallel with real-time PCR method and IgM ELISA assay. The positive rates of these specimens examined by SAT-MP assay, real-time PCR method and IgM ELISA assay were 16.51%, 15.56% and 12.70% respectively. While there was statistical significance (p = 0.04) between SAT-MP assay and IgM ELISA assay, no statistical significance (p = 0.25) was found between SAT-MP assay and real-time PCR method and these two methods had high consistency (kappa value = 0.97). These findings indicate that the newly developed SAT-MP assay is a rapid, sensitive and specific method for identifying M. pneumoniae with potential clinical application in the early diagnosis of M. pneumoniae infection.

Herpangina, usually caused by coxsackie virus A, is prevalent in children spreading through the fecal-oral transmission and the respiratory droplets dissemination. Also, it is mostly asymptomatic and self-limiting. In our study, we found that large outbreak of herpangina in children occurred in the summer of 2015 in Hangzhou, China. From May 1th to August 31th, a total of 10 210 children were diagnosed with herpangina in Children’s Hospital of Zhejiang University School of Medicine. 2 310 throat swabs were collected and tested for enterovirus detection by real-time RT-PCR, while 1 651 cases were positive with the rate of 71.5%. Based on VP1 gene or 5′UTR region sequences, Coxsackievirus A2, A4, A6, A10, B2, B4 and echovirus 30 were detected in these cases. More importantly, Coxsackievirus A2 may be the major subtype of enterovirus resulting in children with herpangina in hangzhou, China.

Human cytomegalovirus (HCMV) has evolved several immune evasion strategies. One strategy is controlling the movement of peripheral blood mononuclear cells (PBMCs) by encoding homologues of chemokines. Our aim was to determine whether HCMV open reading frame (ORF) UL128 could encode a protein that attracts PBMCs like a β-chemokine. The recombinant UL128 protein was synthesized by construction of a stably transfected CHO-UL128 cell line, and a chemotaxis assay showed that UL128 was able to attract PBMCs with a potency equal to that of MIP-1α in vitro. We hypothesize that UL128 protein may act as a β-chemokine homologue in viral pathogenesis.

Objectives: To investigate the possible target genes of methyl-CpG-binding protein 2 (MeCP2) that contribute to Rett syndrome (RTT).Methods: Brain tissues were taken from Mecp2308/Y mice or control mice and then subjected to real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR), immunohistochemical staining, and Western blot analysis for connexin (Cx)43, Cx45, Cx40, Cx32, 2,3-cyclic nucleotide 3-phosphohydrolase (CNPase), and glial fibrillary acidic protein (GFAP).Results: The expression of CNPase in subcortical white matter and hippocampi was lower in RTT mice compared to control mice at both mRNA and protein levels. In contrast, the expression of Cx43, Cx40, Cx45, Cx32, or GFAP was not altered in RTT mice compared to control mice.Conclusion: The downregulation of CNPase expression in the brain may be a possible consequence of MECP2 gene mutation, and the indicated dysfunction of the oligodendrocytes in the subcortical white matter and hippocampi may be involved in RTT p...

Human cytomegalovirus (HCMV) is a common pathogen that causes persistent infections in immune deficient patients and results in significant morbidity and mortality, particularly among transplant recipients and children. Different HCMV glycoprotein H (gH) genotypes may cause different diseases and affect the severity of these diseases. To develop a sensitive quantitative real-time PCR assay that could rapidly distinguish between two HCMV gH genotypes, primers were designed to target the conserved region of the gH gene. gH1 and gH2 probes were designed to target the two variable regions. Standard HCMV strains (AD169 and TOWNE) and 203 clinical urine samples from HCMV infected children were used for the present study. Based on the primer-probe set used to detect the target gH gene segment of HCMV, our quantitative real-time PCR assay specifically discriminated between HCMV gH1 and gH2 with a detection limit of approximately 10(2) viral copies/ml. Among the 203 clinical urine samples tested, 145 were gH1 positive, 56 were gH2 positive, and 2 were positive for both. Thus, we developed a gH gene-based real time-PCR method that could rapidly, stably, and specifically distinguish between two HCMV gH genotypes. We found HCMV gH1 to be common among children examined in Zhejiang, China.

RATIONALE The presence of purpura is a compulsory criteria for the diagnosis of Henoch-Schonlein purpura (HSP). Typical purpura of HSP is distributed symmetrically over the extensor surfaces of the lower limbs, buttocks, and forearms with the occasional involvement of trunk and face in children. It occurs only involving the bottom of the feet has never been reported. PATIENT CONCERNS A 7-year-old girl was admitted to the hospital with abdominal pain, vomiting, and fever. DIAGNOSES Combining clinical manifestations with results of radiologic examinations, acute appendicitis was suspected and a laparotomy was considered. Purpura was found on the bottom of her feet when she was in the operating room and HSP was diagnosed. INTERVENTIONS The patient was treated with glucocorticosteroids, antibiotics, cimetidine, and restriction of feeding. OUTCOMES The abdominal pain and purpura resolved at discharge and there were no recurrences in the subsequent 3-, 6-, and 12-month follow-ups. LESSONS Careful examination of skin including the bottom of the feet can help to direct the diagnostic workup for children with abdominal pain.