Family History of Type 2 Diabetes and Its Association with Beta Cell Function and Lipid Profile in Newly Diagnosed Pediatric Patients with Type 1 Diabetes.

OBJECTIVE: The objective of this study was to explore the associations between a family history of type 2 diabetes (T2D) and beta-cell function, as well as lipid profile, in pediatric patients newly diagnosed with type 1 diabetes (T1D). METHODS: A retrospective analysis was conducted on children under 14 years of age who were newly diagnosed with T1D at the Children's Hospital of Zhejiang University between August 2018 and August 2022. Clinical features, metabolic profiles, beta-cell function, and lipid profile were evaluated. RESULTS: A total of 316 children were diagnosed with new-onset T1D. Among them, 28.2% had a family history of T2D. Patients with T1D who had a family history of T2D experienced a later onset of the disease (p = 0.016), improved HOMA2-%B levels (p = 0.003), and increased concentrations of HDL-C (p = 0.005). In addition, no statistically significant differences in age at onset, HOMA2-%B levels, or HDL-C were found when assessing the interaction between family history of T2D and type of diabetes mellitus (autoimmune T1D/idiopathic T1D). CONCLUSION: A family history of T2D may contribute to the heterogeneity of T1D patients in terms of HOMA2-%B levels and lipid profile. This highlights the significance of taking into account T2D-related factors in the diagnosis and treatment of T1D.

The chronic consumption of dietary fructose promotes the gut Clostridium species imbalance and bile acid alterations in developing nonalcoholic fatty liver disease.

Excessive fructose intake is associated with the rising prevalence of nonalcoholic fatty liver disease (NAFLD). The gut microbiome (GM) and bile acids (BAs) are involved in the pathogenesis of NAFLD, but the impact of fructose on their cross-talk is unclear. In this study, adult male C57BL/6J mice were fed a normal diet with tap water (ND) or with 4% fructose in the drinking water (Fru), 60% high-fat diet with tap water (HF) or with 4% fructose solution (HFF) for 12 weeks. Targeted BA analysis was performed in five anatomical sites including the liver, ileum contents, portal serum, cecum contents, and feces. Metagenomic sequencing was performed to explore gut dysbiosis. Within 12 weeks, the 4% fructose diet could initially stimulate gut dysbiosis and BA upregulation in the ileum, portal serum, and cecum when the intestinal and hepatic transport system remained stable without hepatic lipid accumulation. However, the chronic consumption of fructose promoted HF-induced NAFLD, with significantly increased body weight, impaired glucose tolerance, and advanced liver steatosis. BA transporters were inhibited in HFF, causing the block of internal BA circulation and increased BA secretion via cecum contents and feces. Notably, lithocholic acid (LCA) and its taurine conjugates were elevated within the enterohepatic circulation. Meanwhile, the Clostridium species were significantly altered in both Fru and HFF groups and were closely associated with fructose and BA metabolism. In summary, excessive fructose caused gut dysbiosis and BA alterations, promoting HF-induced NAFLD. The crosstalk between Clostridium sp. and LCA species were potential targets in fructose-mediated NAFLD.

Molecular epidemiology and phylogenetic analyses of human adenovirus in pediatric patients with acute respiratory infections from Hangzhou during COVID-19 pandemic.

Background: Acute Respiratory Infections (ARIs) are a major cause of morbidity and mortality worldwide. Human Adenovirus (HAdV), responsible for 5%-10% of children's ARIs, is one of the most prevalent pathogens. Our study aimed to analyze the epidemiology and phylogenesis of HAdV in pediatric patients with ARIs in Hangzhou during the COVID-19 pandemic. Method: Between November 2020 and March 2021, we collected 1,442 nasopharyngeal swabs from children with ARIs at Children's Hospital, Zhejiang University School of Medicine. Epidemiological statistics, phylogenetic and amino acid (AA) mutation analysis were conducted. Results: Our findings revealed that 386 (26.77%) samples tested positive for HAdV, with the highest rate in children aged 6-18 years and the lowest in children aged 0-1 year, indicating a different age preference of HAdV compared with pre-pandemic period. Outpatients had a significantly higher positive rate than inpatients. Moreover, patients with HAdV-coinfection exhibited more severe clinical symptoms than those with HAdV-single infection. Our phylogenetic analysis demonstrated that species HAdV-C (type 1, 2, 6) were the predominant circulating strains in Hangzhou during the COVID-19 pandemic. Further AA mutation analysis identified seventeen mutations of particular concern for biological characterization. Conclusion: In conclusion, our study provides valuable epidemiological and molecular data that will aid in epidemiological surveillance, antiviral therapies and the development of specific vaccine types, leading to improve public health.

Bile Acid Alterations Characterize the Early Onset and Progression of Nonalcoholic Fatty Liver Disease in Young Mice Fed with High Fat and Fructose Diet.

SCOPE: Bile acids (BAs) are closely associated with obesity and non-alcoholic fatty liver disease (NAFLD). How BAs change within the enterohepatic circulation during the onset and progression of NAFLD as biomarkers deserves to be explored. METHODS AND RESULTS: Four-week-old young mice were fed with high-fat diet plus 4% v/w fructose drinking water (HFF) or normal diet with tap water (ND) for 4 and 12 weeks. In comparison, eight-week-old adult mice were fed with HFF or ND for 12 weeks. BAs were measured in six different anatomical sites to evaluate the systematic changes of BAs within the enterohepatic circulation. The dysregulated BA metabolism had occurred in HFF after 4-week intervention, represented by increased primary BAs and decreased hyocholic acid (HCA) species. After 12 weeks, the impact was more significant with increased secondary BA synthesis and excretion, particularly for lithocholic acid (LCA) species. More interestingly, the BA changes were more significant in younger mice in response to 12-week diet intervention. CONCLUSIONS: The enterohepatic circulation of BAs changed with the development of NAFLD, and the younger mice were more susceptible to the unhealthy diet. HCA and LCA species may be potential biomarkers for predicting the development of NAFLD.

Profiles and predictive value of cytokines in children with human metapneumovirus pneumonia.

BACKGROUND: Human metapneumovirus (HMPV) is an important cause of respiratory tract infections in young children. Early innate immune response to HMPV is focused on induction of antiviral interferons (IFNs) and other pro-inflammatory cytokines that are critical for the formation of adaptive immune responses. To evaluate the predictive value of Th1/Th2 cytokines which include IL-2, IL-4, IL-6, IL-10, INF-γ and TNF-α in pneumonia caused by HMPV. METHODS: A retrospective study was performed among 59 pneumonia pediatric patients with HMPV infection and 33 healthy children as the control cohort, which was detected by the immunofluorescence assay, and the Th1/Th2 cytokines were measured by flow cytometry. 131 children infected with Influenza virus A (IVA) and 41 children infected with influenza virus B (IVB) were detected by RT-PCR assay in throat swabs. RESULTS: When compared with the healthy children, children who were infected with HMPV pneumonia had a significantly lower level of IL-2 (p < 0.001) and higher levels of IL-4 (p < 0.001), IL-6 (p = 0.001), IL-10 (p < 0.001), and IFN-γ (p < 0.001). Compared with patients diagnosed with IVA or IVB infection, HMPV-positive patients had significantly higher levels of IL-4 (p < 0.001 and < 0.001), IFN-γ (p < 0.001 and < 0.001), and TNF-α (p < 0.001 and 0.016). Moreover, compared with IVA patients, HMPV-positive patients had a significantly lower level of IL-6 (p = 0.033). Finally, when comparing cytokine levels among the patients with HMPV pneumonia, IL-6 and TNF-α levels were found to be significantly higher in the severe group than the mild group (p = 0.027 and 0.049). The IL-6 and TNF-α were used to differentiate between mild symptoms and severe symptoms in children diagnosed with HMPV pneumonia with an AUC of 0.678 (95% CI 0.526-0.829) and 0.658 (95% CI 0.506-0.809), respectively. CONCLUSION: Our study indicated that difference in cytokine trends depending on the virus species. The levels of IL-4, TNF-α and IFN-γ were significantly distinguished in children infected with HMPV versus IVA and IVB. IL-6 and TNF-α may be helpful in assessing the severity and prognosis of HMPV infection.

Amino-Functionalized Single-Lanthanide Metal-Organic Framework as a Ratiometric Fluorescent Sensor for Quantitative Visual Detection of Fluoride Ions.

Excessive content of fluoride ions (F-) in water will lead to water pollution and endanger human health, so the research on the method of low-cost, rapid, and efficient detection of F- is of particular significance. In this work, an amino-functionalized ligand with an appropriate triplet energy excited state, 2'-amino-[1,1':4',1″-terphenyl]-3,3″,5,5″-tetracarboxylic acid (H4TPTC-NH2), was selected to construct a luminescent single-lanthanide metal-organic framework, EuTPTC-NH2, with uncoordinated amino groups for the detection of F-. Based on host-guest interactions, that is, hydrogen bonds formed between the free amino groups and F- ions, EuTPTC-NH2 was developed as a ratiometric fluorescence probe for F- detection with good anti-interference ability, low detection limit, high water stability, and selectivity. It was found that EuTPTC-NH2 has an excellent linear response to F- in the concentration range of 0-80 µM with high sensitivity and a low detection limit of 11.26 µM. A hydrogel membrane based on the combination of EuTPTC-NH2 and agarose was also prepared for the quantitative visual detection of F- in water.

Cytokine Gene Polymorphisms in Chinese Children with Idiopathic Nephrotic Syndrome.

BACKGROUND: Cytokines play a role in the progression of idiopathic-nephrotic syndrome (INS). OBJECTIVES: To investigate the association of different cytokine genes polymorphisms with INS incidence and response to steroid therapy in Chinese children. METHODS: 182 children with INS and 100 healthy controls were enrolled in this study. Blood genomic DNAs were used to analyze20 single nucleotide polymorphisms (SNPs) in 8 cytokine genes includingIL-21, IL-18, IL-6, IFN-γ, IL-4, IL-10, IL-17F, IL-17A d by multi-PCR with next-generation sequencing. RESULTS: Among 182 children with INS, 89 (48.6%) were steroid-sensitive (SS), 73 (39.9%) were steroid-dependent (SD) and 21 (11.5%) were steroid-resistant (SR). In 20 SNPs, IL-4-rs2243283 exhibited a significantly different genotype distribution between INS and the healthy controls (CC is a risk genotype: 66.5% of INS VS 51% of the control; OR=1.91, p=0.012). Patients carrying AG genotype (rs2275913, IL-17A) had a significantly higher risk of steroid-dependent response (69.1% of SD VS 46.4% of SS; OR=2.58, p=0.014). Similarly, patients carrying A allele of IL-10-rs1800872 (39.0% of SD VS 26.7% of SS; OR=1.76, p=0.018) and C allele of IL-10-rs1800896 (12.3% of SD VS 3.9% of SS; OR=3.44, p=0.004) had a higher risk of steroid-dependent response. However, none of these 20 SNPs showed a significant difference between SS group and SR group. CONCLUSION: Among the 20 cytokine gene SNPs, IL-4-rs2243283 might increase the susceptibility to INS in Chinese children; rs2275913 of IL-17A, rs1180972, and rs1800896 of IL-10 show association with the steroid -response in Chinese INS children.

Water-Stable Eu6-Cluster-Based fcu-MOF with Exposed Vinyl Groups for Ratiometric and Fluorescent Visual Sensing of Hydrogen Sulfide.

Detection of H2S in the biological system has attracted enormous attention in recent years. In this work, a new vinyl-functionalized metal-organic framework (MOF), [(Me2NH2)2] [Eu6(µ3-OH)8(BDC-CH═CH2)6(H2O)6] (Eu-BDC-CH═CH2, BDC-CH═CH2 = 2-vinylterephthalic acid), was synthesized under solvothermal conditions. The vinyl groups in the ligands can not only modulate the "antenna effect" of the ligand on Eu3+ ions but also serve as an exposed reactive site to allow for the quantitative detection of H2S by Eu-BDC-CH═CH2. The ratiometric fluorescent probe has the advantages of water stability, acid-base stability (pH = 2-11), fast response (<2 min), high selectivity, and sensitivity (LOD = 38.4 µM). We also used Eu-BDC-CH═CH2 to detect and analyze H2S in tap and lake waters, demonstrating the potential of the probe for biological and environmental applications. In addition, the MOF-based agarose hydrogel film allows for the visual detection of H2S via a smartphone by identifying the RGB values. The vinyl-functionalized MOF can thus be a powerful sensing platform for H2S.

Urinary Sediment mRNA Level of CREBBP and CYBA in Children With Steroid-Resistant Nephrotic Syndrome.

Background: This study aimed to evaluate gene expression patterns in urinary sediment samples of children with steroid-resistant nephrotic syndrome (SRNS). Methods: The messenger RNA (mRNA) levels of 770 immune-related genes were detected using a NanoString nCounter platform. To verify the NanoString results, quantitative analysis of nine gene mRNAs was performed using real-time RT-PCR in more samples. Results: Firstly, compared with the steroid-sensitive nephrotic syndrome (SSNS) group (n=3), significant changes were observed in the mRNA level of 70 genes, including MAP3K14, CYBA, SLC3A2, CREB-binding protein (CREBBP), CD68, forkhead box P1 (FOXP1), CD74, ITGB2, IFI30, and so forth, in the SRNS group (n=3). A total of 129 children with idiopathic nephrotic syndrome (INS), 15 with acute glomerulonephritis, and 6 with immunoglobulin A nephropathy (IgAN) were enrolled to verify the NanoString results. Compared with patients with IgAN, those with INS had significantly lower levels of FOXP1 (P=0.047) and higher levels of CREBBP (P=0.023). Among SSNS, the mRNA level of ITGB2 was significantly lower in the non-relapse group than in the non-frequent relapse and frequent-relapse groups (P=0.006). Compared with the SSNS group, CREBBP was significantly elevated in the SRNS group (P=0.02). Further, CYBA significantly decreased in the SRNS group (P=0.01). The area under the curve (AUC) for CREBBP and CYBA was 0.655 and 0.669, respectively. CREBBP had a sensitivity of 83.3% and a specificity of 49.4% and CYBA had a sensitivity of 58.3% and a specificity of 83.1% to rule out SSNS and SRNS. The diagnosis value was better for CREBBP+CYBA than for CREBBP or CYBA alone, indicating that the combination of CREBBP and CYBA was a more effective biomarker in predicting steroid resistance (AUC=0.666; sensitivity=63.9%; specificity=76.4%). Conclusions: This study was novel in investigating the urinary sediment mRNA level in children with INS using high-throughput NanoString nCounter technology, and 70 genes that may relate to SRNS were found. The results revealed that the urinary sediment mRNA level of ITGB2 was significantly lower in the non-relapse group than in the non-frequent relapse and frequent-relapse groups. Meanwhile, CREBBP was significantly elevated and CYBA was significantly lowered in the SRNS group compared with the SSNS group.

Molecular epidemiology of rotavirus A and adenovirus among children with acute diarrhea in Hangzhou, China.

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.

Hemolytic specimens in complete blood cell count: Red cell parameters could be revised by plasma free hemoglobin.

INTRODUCTION: Hemolysis is the main cause of unqualified clinical samples. In this study, we established a method for detecting and evaluating hemolysis in whole blood test. We used a mathematical formula for correcting the influence of hemolysis on complete blood cell count (CBC) so as to avoid re-venipuncture and obtain more accurate parameters of red blood cell detection, reduce the burden of patients, and improve the efficiency of diagnosis and treatment. METHODS: Hemolytic samples were selected and then corrected using the new formula. Plasma free hemoglobin (fHB) was used as the criterion to determine the degree of hemolysis; the uncertainty of measurement is acceptable as the limit value of deviation between the measured value and the revised value. Hemolysis simulation analysis in vitro and continuous monitoring of clinical patients were used to verify the correction effect. RESULTS: A total of 83 clinical samples with hemolysis were collected and analyzed; fHB 1.4 g/L was selected as the unacceptable value for clinical hemolysis detection. In hemolytic samples, the red blood cell parameters corrected by formula are significantly different from those uncorrected and had a good consistency with those before hemolysis. CONCLUSION: The results show that the hemolysis phenomenon of CBC has a significant impact on routine blood testing. By using the new formula, the influence of hemolysis on erythrocyte and related parameters can be quickly and easily corrected, thus avoiding venipuncture again for re-examination, reducing diagnostic errors, and saving medical resources.

Evaluation of a commercial latex agglutination test for detecting rotavirus A and human adenovirus in children's stool specimens.

OBJECTIVES: Rotavirus A and human adenovirus are the two most common causes of infantile diarrhea; thus, it is of great importance to find out a rapid and accurate diagnostic method. This study aimed to evaluate the diagnostic significance of latex agglutination test for detection of rotavirus A and human adenovirus. METHODS: A prospective study was conducted on 214 diarrhea children from September 2018 to March 2019 in our hospital. Fresh stool samples were collected for detection of rotavirus A and human adenovirus by latex agglutination test and quantitative reverse transcription polymerase chain reaction (RT-qPCR). Then, the consistency of results detected by these two methods was analyzed． RESULTS: With performing the latex agglutination test, it was revealed that positive rates for detecting rotavirus A virus and human adenovirus were 23.83% (51/214) and 25.24% (54/214), respectively. Meanwhile, results of RT-qPCR showed that positive rates for detecting rotavirus A virus and human adenovirus were 58 (27.10%) and 59 (27.57%), respectively. Using RT-qPCR as the gold standard, the sensitivity and specificity of the latex agglutination test for detecting rotavirus A were 81.03% and 97.44%, and the corresponding values for detecting human adenovirus were 76.27% and 94.19%, respectively. CONCLUSION: This latex agglutination test showed a satisfactory consistency with RT-qPCR for detecting rotavirus A and human adenovirus. The mentioned commercial assay may be highly appropriate for rapid screening of rotavirus A and human adenovirus.

Distribution Characteristics of Hepatitis B Serological Markers in Hospitalized Children and Adolescents in Zhejiang, China between 2006 and 2010.

BACKGROUND/AIMS: To investigate serological patterns of hepatitis B based on electrochemiluminescent immunoassays and the distribution characteristics of these patterns in hospitalized children and adolescents in Zhejiang, China between 2006 and 2010. METHODS: Five serological markers, including hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg), antibody to hepatitis B surface antigen (anti-HBs), antibody to hepatitis B e antigen (anti-HBe), and antibody to hepatitis B c antigen (anti-HBc), were chosen as a routine panel to monitor hepatitis B virus (HBV) infection and vaccination efficacy. A total of 33,187 children (21,187 boys and 12,000 girls) were selected using the following exclusion criteria: a previous diagnosis of hepatitis, age >16 years or an address outside of Zhejiang. RESULTS: The average HBV vaccination coverage rates among 20,766 boys and 11,782 girls were 98.62% and 98.68%, respectively. Seventeen serological patterns of hepatitis B were found, and the dominant pattern was 'anti-HBs (+) alone' (62.03%) followed by 'negative pattern' (23.46%). The rates of the other 15 patterns ranged from 8.14% to 0.003%. Of 236 HBsAg-positive patients, the overall rate of seropositivity was 0.71%. The anti-HBs levels were grouped into 3 ranges (10-100 mIU/mL, 100-1,000 mIU/mL, and >1,000 mIU/mL) for all anti-HBs-positive children (36.08%, 43.43%, and 20.49%, respectively). CONCLUSIONS: A low HBsAg carrier rate and a relatively high anti-HBs positive rate are present in hospitalized children and adolescents in Zhejiang. The distribution of serological patterns is associated with age but is mostly independent of gender.

Inhibition of hepatitis B virus (HBV) by LNA-mediated nuclear interference with HBV DNA transcription.

Silencing target genes with small regulatory RNAs is widely used to investigate gene function and therapeutic drug development. Recently, triplex-based approaches have provided another attractive means to achieve targeted gene regulation and gene manipulation at the molecular and cellular levels. Nuclear entry of oligonucleotides and enhancement of their affinity to the DNA targets are key points of such approaches. In this study, we developed lipid-based transport of a locked-nucleic-acid (LNA)-modified oligonucleotide for hepatitis B virus (HBV) DNA interference in human hepatocytes expressing HBV genomic DNA. In these cells, the LNA-modified oligonucleotides passed efficiently across the cell membrane, and lipid-coating facilitated translocation from the cytoplasm to the nucleus. The oligonucleotide specifically targeting HBV DNA clearly interfered with HBV DNA transcription as shown by a block in pregenomic RNA (pgRNA) production. The HBV DNA-targeted oligonucleotide suppressed HBV DNA replication and HBV protein production more efficiently than small interfering RNAs directed to the pgRNA. These results demonstrate that fusion with lipid can carry LNA-modified oligonucleotides to the nucleus where they regulate gene expression. Interfering with HBV DNA transcription by LNA-modified oligonucleotides has strong potential as a new strategy for HBV inhibition.

Hepatitis B virus X protein stimulates IL-6 expression in hepatocytes via a MyD88-dependent pathway.

BACKGROUND & AIMS: Hepatitis B virus (HBV) X protein (HBx) has been implicated in HBV-associated carcinogenesis by activating signal transduction pathways and influencing gene transcription in liver cells. We aimed to investigate the underlying mechanisms for HBx-induced production of interleukin-6 (IL-6), one of the major inflammatory mediators that stimulate hepatocellular carcinoma development. METHODS: HBx was overexpressed in hepatic and hepatoma cell lines and IL-6 expression levels were measured by quantitative RT-PCR and ELISA. The activation of IRAK-1, ERKs/p38, and NF-κB was determined by Western blotting using specific anti-phosphoprotein antibodies. The role of MyD88 in these processes was analyzed by MyD88 RNAi and expression of an inactive MyD88 mutant. RESULTS: Expression of HBx in hepatic and hepatoma cells led to a dramatic enhancement of IL-6 synthesis and secretion. Dysfunction of MyD88 in these cells prevented the HBx-triggered IL-6 production. HBx expression also activated downstream signaling proteins of MyD88 including IRAK-1, ERKs/p38, and NF-κB. Inactivation of these signaling molecules blocked IL-6 synthesis as well. HBx-stimulated the expression of MyD88. CONCLUSIONS: In hepatocytes and hepatoma cells, HBx stimulates the production of IL-6 in a MyD88-dependent manner, indicating that parenchymal liver cells are an additional source of high levels of IL-6 in the HBV-infected liver microenvironment. HBx could be involved in HBV-mediated liver carcinogenesis, through this mechanism of action.

[Role of HBx in hepatocellular carcinoma development].

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in China, which is mainly caused by hepatitis B virus (HBV) infection. The X gene product (HBx) of HBV has extensive trans-activating functions. HBx affects the signal transduction, apoptotic cell death and cell cycle through interaction with variety intracellular proteins in infected hepatocytes. In view of the importance of HBx in HBV replication and in hepatic cell functions, the role of HBx in HCC development has been attracting great attention.