Development of an immunoinflammatory indicator-related dynamic nomogram based on machine learning for the prediction of intravenous immunoglobulin-resistant Kawasaki disease patients.

BACKGROUND: Approximately 10-20% of Kawasaki disease (KD) patients suffer from intravenous immunoglobulin (IVIG) resistance, placing them at higher risk of developing coronary artery aneurysms. Therefore, we aimed to construct an IVIG resistance prediction tool for children with KD in Shanghai, China. METHODS: Retrospective analysis was conducted on data from 1271 patients diagnosed with KD and the patients were randomly divided into a training set and a validation set in a 2:1 ratio. Machine learning algorithms were employed to identify important predictors associated with IVIG resistance and to build a predictive model. The best-performing model was used to construct a dynamic nomogram. Moreover, receiver operating characteristic curves, calibration plots, and decision-curve analysis were utilized to measure the discriminatory power, accuracy, and clinical utility of the nomogram. RESULTS: Six variables were identified as important predictors, including C-reactive protein, neutrophil ratio, procalcitonin, CD3 ratio, CD19 count, and IgM level. A dynamic nomogram constructed with these factors was available at https://hktk.shinyapps.io/dynnomapp/. The nomogram demonstrated good diagnostic performance in the training and validation sets (area under the receiver operating characteristic curve = 0.816 and 0.800, respectively). Moreover, the calibration curves and decision curves analysis indicated that the nomogram showed good consistency between predicted and actual outcomes and had good clinical benefits. CONCLUSION: A web-based dynamic nomogram for IVIG resistance was constructed with good predictive performance, which can be used as a practical approach for early screening to assist physicians in personalizing the treatment of KD patients in Shanghai.

Identification of key signaling pathways and hub genes related to immune infiltration in Kawasaki disease with resistance to intravenous immunoglobulin based on weighted gene co-expression network analysis.

Background: Kawasaki disease (KD) is an acute vasculitis, that is, the leading cause of acquired heart disease in children, with approximately 10%-20% of patients with KD suffering intravenous immunoglobulin (IVIG) resistance. Although the underlying mechanism of this phenomenon remains unclear, recent studies have revealed that immune cell infiltration may associate with its occurrence. Methods: In this study, we downloaded the expression profiles from the GSE48498 and GSE16797 datasets in the Gene Expression Omnibus database, analyzed differentially expressed genes (DEGs), and intersected the DEGs with the immune-related genes downloaded from the ImmPort database to obtain differentially expressed immune-related genes (DEIGs). Then CIBERSORT algorithm was used to calculate the immune cell compositions, followed by the WGCNA analysis to identify the module genes associated with immune cell infiltration. Next, we took the intersection of the selected module genes and DEIGs, then performed GO and KEGG enrichment analysis. Moreover, ROC curve validation, Spearman analysis with immune cells, TF, and miRNA regulation network, and potential drug prediction were implemented for the finally obtained hub genes. Results: The CIBERSORT algorithm showed that neutrophil expression was significantly higher in IVIG-resistant patients compared to IVIG-responsive patients. Next, we got differentially expressed neutrophil-related genes by intersecting DEIGs with neutrophil-related module genes obtained by WGCNA, for further analysis. Enrichment analysis revealed that these genes were associated with immune pathways, such as cytokine-cytokine receptor interaction and neutrophil extracellular trap formation. Then we combined the PPI network in the STRING database with the MCODE plugin in Cytoscape and identified 6 hub genes (TLR8, AQP9, CXCR1, FPR2, HCK, and IL1R2), which had good diagnostic performance in IVIG resistance according to ROC analysis. Furthermore, Spearman's correlation analysis confirmed that these genes were closely related to neutrophils. Finally, TFs, miRNAs, and potential drugs targeting the hub genes were predicted, and TF-, miRNA-, and drug-gene networks were constructed. Conclusion: This study found that the 6 hub genes (TLR8, AQP9, CXCR1, FPR2, HCK, and IL1R2) were significantly associated with neutrophil cell infiltration, which played an important role in IVIG resistance. In a word, this work rendered potential diagnostic biomarkers and prospective therapeutic targets for IVIG-resistant patients.

Automated deep neural network-based identification, localization, and tracking of cardiac structures for ultrasound-guided interventional surgery.

Background: The increase in the use of ultrasound-guided interventional therapy for cardiovascular diseases has increased the importance of intraoperative real-time cardiac ultrasound image interpretation. We thus aimed to develop a deep learning-based model to accurately identify, localize, and track the critical cardiac structures and lesions (9 kinds in total) and to validate the algorithm's performance using independent data sets. Methods: This diagnostic study developed a deep learning-based model using data collected from Fuwai Hospital between January 2018 and June 2019. The model was validated with independent French and American data sets. In total, 17,114 cardiac structures and lesions were used to develop the algorithm. The model findings were compared with those of 15 specialized physicians in multiple centers. For external validation, 516,805 tags and 27,938 tags were used from 2 different data sets. Results: Regarding structure identification, the area under the receiver operating characteristic curve (AUC) of each structure in the training data set, optimal performance in the test data set, and median AUC of each structure identification were 1 (95% CI: 1-1), 1 (95% CI: 1-1), and 1 (95% CI: 1-1), respectively. Regarding structure localization, the optimal average accuracy was 0.83. As for structure identification, the accuracy of the model significantly outperformed the median performance of the experts (P<0.01). The optimal identification accuracies of the model in 2 independent external data sets were 89.5% and 90%, respectively (P=0.626). Conclusions: The model outperformed most human experts and was comparable to the optimal performance of all human experts in cardiac structure identification and localization, and could be used in the external data sets.

Effects of immunoglobulin plus prednisolone in reducing coronary artery lesions in patients with Kawasaki disease: study protocol for a phase III multicenter, open-label, blinded-endpoints randomized controlled trial.

BACKGROUND: Kawasaki disease (KD) is an acute systemic vasculitis of unclear etiology that mainly affects infants and young children. Strategies to reduce the incidence and severity of coronary artery lesions (CALs), the determinant factor in the long-term prognosis of KD, are currently a focus of studies on KD. Corticosteroids, preferred in the treatment of the majority of vasculitides, are controversial in the treatment of acute KD. In this trial, we will evaluate whether the addition of prednisolone to standard intravenous immunoglobulin (IVIG) plus aspirin therapy can reduce the occurrence of CAL in Chinese patients with KD. METHODS: This is a multicenter, prospective, open-label, randomized controlled trial, which is expected to be conducted in more than 20 hospitals in China and aims to assess the efficacy and safety of IVIG + prednisolone treatment versus standard treatment. Patients with KD who fulfill the inclusion and exclusion criteria will be recruited and randomized (1:1) to receive either a large dose of IVIG (2 g/kg over 12-24 h with a maximum dose of 60 g) + aspirin 30 mg/kg/d or IVIG (2 g/kg over 12-24 h) + aspirin 30 mg/kg/d + prednisolone (2 mg/kg/d with a maximum dose of 60 mg tapered over 15 days after normalization of C-reactive protein concentration). The primary outcome will be the occurrence of CAL at 1 month of illness. The follow-up duration for each participant will be set as 1 year. Patients and treating physicians will be unmasked to group allocation. DISCUSSION: This will be the first multicenter randomized controlled trial to evaluate the efficacy of IVIG + aspirin + prednisolone in Chinese pediatric patients with KD, which may provide high-level evidence for improving the initial treatment for acute KD. TRIAL REGISTRATION: ClinicalTrials.gov NCT04078568 . Registered on 16 August 2018.

Construction of Smart City Street Landscape Big Data-Driven Intelligent System Based on Industry 4.0.

With the application of engineering management in smart city construction under Industry 4.0, the intelligent design of urban street landscape has attracted extensive attention. Affected by the low intelligent level of traditional landscape design, the existing urban landscape composite system has difficulty in meeting the needs of smart city construction. Therefore, this paper proposes the construction of street landscape big data-driven intelligent decision support system based on Industry 4.0. Based on the complex network theory, this paper analyzes the structure, links, nodes, driving forces, and functional requirements of urban street landscape and then puts forward the construction content and implementation method of urban street landscape intelligent decision support system. The system consists of four aspects: intelligent infrastructure, service, protection and maintenance, and management and evaluation system. Its implementation not only reflects the cooperation and effective application of intelligent technology in each stage of street landscape construction, but also provides reference for the application of engineering management in other fields under Industry 4.0.

Metabolomics analysis of the serum from children with urolithiasis using UPLC-MS.

Pediatric urolithiasis is a common urologic disease with high morbidity and recurrence rates. Recent studies have shown that metabolic dysfunction plays a vital role in the pathogenesis of urolithiasis, especially in children, but the specific mechanism is still unclear. Metabolomics is an ideal technology for exploring the mechanism of metabolic disorders in urolithiasis. In the present study, a serum metabolomics based on ultra-performance liquid chromatography mass spectrometry was performed. A total of 50 children subjects were recruited for the study, including 30 patients with kidney stones and 20 normal controls (NCs). Principal component analysis and orthogonal partial least-squares determinant analysis were carried, and 40 metabolites were found to be significantly altered in patients with kidney stones, mainly involving retinol metabolism, steroid hormone biosynthesis, and porphyrin and chlorophyll metabolism. The kidney stone group appeared to have a lower serum level of bilirubin, but a relative higher level of retinal, all-transretinoic acid, progesterone, and prostaglandin E2 compared with those of the NC group. All the findings suggest that patients with urolithiasis have several metabolic characteristics, which are related to stone formation or compensation. These metabolites and pathways are very likely associated with development of kidney stones and should be considered as potential novel targets for treatment and prevention.

Uric acid and sphingomyelin enhance autophagy in iPS cell-originated cardiomyocytes through lncRNA MEG3/miR-7-5p/EGFR axis.

This study aimed to determine the metabolites associated with ventricular septal defect (VSD) and the underlying mechanisms. Blood samples and thymus tissues were collected from VSD patients to perform LC-MS-based metabolomics assay and generate iPS cell-derived cardiomyocytes, respectively. VSD rat model was used in vivo study. RT-PCR, western blotting, immunohistochemistry, luciferase activity assay, GFP-LC3 adenovirus and GFP and RFP tfLC3 assay, and transmission electron microscopy were performed to investigate the underlying mechanisms. The metabolites uric acid (UA) and sphingomyelin (SM) increased in the serum of VSD patients, along with enhanced autophagy. The combination of UA and SM treatment could promote autophagy and inhibit EGFR and AKT3 expressions. Overexpression of EGFR and AKT3 suppressed autophagy in UA and SM-treated cardiomyocytes, respectively. Also, lncRNA MEG3 knockdown and overexpression could enhance and inhibit autophagy in UA and SM-treated cardiomyocytes, respectively, through targeting miR-7-5p. Moreover, miR-7-5p mimics and inhibitors promoted and inhibited autophagy in UA and SM-treated cardiomyocytes, respectively, via target EGFR. In VSD rat model, upregulation of MEG3 could reverse high level of autophagy and decrease serum UA and SM. In conclusion, UA and SM are essential VSD-associated metabolic biomarkers and MEG3/miR-7-5p/EGFR axis is critical to the regulation of autophagy in cardiomyocytes.

Generation of a Urine-Derived Ips Cell Line from a Patient with a Ventricular Septal Defect and Heart Failure and the Robust Differentiation of These Cells to Cardiomyocytes via Small Molecules.

BACKGROUND/AIMS: Ventricular septal defects (VSDs) are one of the most common types of congenital heart malformations. Volume overload resulting from large VSDs can lead to heart failure (HF) and constitutes a major cause of pediatric HF with a series of often-fatal consequences. The etiology of VSD with HF is complex, and increasing evidence points toward a genetic basis. Indeed, we identified an L2483R mutation in the ryanodine receptor type 2 (RyR2) in a 2-month-old male patient with VSD with HF. METHODS: We generated integration-free induced pluripotent stem cells from urine samples (UiPSCs) of this patient using Sendai virus containing the Yamanaka factors and characterized these cells based on alkaline phosphatase activity, pluripotency marker expression, and teratoma formation. Then, we induced the derived UiPSCs to rapidly and efficiently differentiate into functional cardiomyocytes through temporal modulation of canonical Wnt signaling with small molecules. Real-time PCR and immunofluorescence were used to verify the expression of myocardium-specific markers in the differentiated cardiomyocytes. The ultrastructure of the derived myocardial cells was further analyzed by using transmission electron microscopy. RESULTS: The established UiPSC lines were positive for alkaline phosphatase activity, retained the RyR2 mutation, expressed pluripotency markers, and displayed differentiation potential to three germ layers in vivo. The UiPSC-derived cells showed hallmarks of cardiomyocytes, including spontaneous contraction and strong expression of cardiac-specific proteins and genes. However, compared with cardiomyocytes derived from H9 cells, they had a higher level of autophagy, implying that autophagy may play an important role in the development of VSD with HF. CONCLUSION: The protocol described here yields abundant myocardial cells and provides a solid platform for further investigation of the pathogenesis, pharmacotherapy, and gene therapy of VSD with HF.

A novel ZIC3 gene mutation identified in patients with heterotaxy and congenital heart disease.

Heterotaxy syndrome (HTX) is characterized by left-right (LR) asymmetry disturbances associated with severe heart malformations. However, the exact genetic cause of HTX pathogenesis remains unclear. The aim of this study was to investigate the pathogenic mechanism underlying heterotaxy syndrome. Targeted next-generation sequencing (NGS) was performed for twenty-two candidate genes correlated with LR axis development in sixty-six HTX patients from unrelated families. Variants were filtered from databases and predicted in silico using prediction programs. A total of twenty-one potential disease-causing variants were identified in seven genes. Next, we used Sanger sequencing to confirm the identified variants in the family pedigree and found a novel hemizygous mutation (c.890G > T, p.C297F) in the ZIC3 gene in a male patient that was inherited from his mother, who was a carrier. The results of functional indicated that this ZIC3 mutation decreases transcriptional activity, affects the affinity of the GLI-binding site and results in aberrant cellular localization in transfected cells. Moreover, morpholino-knockdown experiments in zebrafish demonstrated that zic3 mutant mRNA failed to rescue the abnormal phenotype, suggesting a role for the novel ZIC3 mutation in heterotaxy syndrome.

The Efficacy and Safety of Mainstream Medications for Patients With cDMARD-Naïve Rheumatoid Arthritis: A Network Meta-Analysis.

Background: The mainstream medications for rheumatoid arthritis (RA) include conventional disease-modifying antirheumatic drugs (cDMARDs), which mostly are methotrexate (MTX), and biologic agents such as adalimumab (ADA), certolizumab (CZP), etanercept (ETN), golimumab (GOL), infliximab (IFX), and tocilizumab (TCZ). This network meta-analysis was aimed at evaluating the efficacy and safety of the medications above and interventions combining cDMARDs and biologic agents for patients with RA. Methods: PubMed, EMBASE, Cochrane Library, and ClinicalTrials.gov were searched systematically for eligible randomized controlled trials (RCTs). Outcomes concerning efficacy and safety were evaluated utilizing odds ratios (ORs) and 95% credible intervals (CrI). The outcomes of efficacy would be evaluated through remission and American College of Rheumatology (ACR) scores. The surface under the cumulative ranking curve (SUCRA) was calculated to rank each treatment on each index. Results: A total of 20 RCTs with 9,047 patients were included, and the efficacy and safety of the concerning interventions for RA were evaluated. Compared with cDMARDs alone, TCZ+MTX, ETN+MTX, IFX+MTX, TCZ, and ADA+MTX showed significant statistical advantage on ACR20, ACR50, and ACR70. Apart from that, as for remission, TCZ+MTX, IFX+MTX, TCZ, and CZP+MTX performed better compared to cDMARDs alone. The SUCRA ranking also indicated that TCZ+MTX was the intervention with best ranking in the entire four efficacy indexes followed by ETX+MTX and IFX+MTX. However, there was no obvious difference among these medications compared with cDMARDs when it comes to safety, which need more specific studies on that. Conclusion: TCZ+MTX was potentially the most recommended combination of medications for RA due to its good performance in all outcomes of efficacy. ETX+MTX and IFX+MTX, which also performed well, could be introduced as alternative treatments. However, considering the adverse events, the treatments concerning should be introduced with caution.

Placenta­derived mesenchymal stem cells improve airway hyperresponsiveness and inflammation in asthmatic rats by modulating the Th17/Treg balance.

Mesenchymal stem cells (MSCs) possess reparative and immunoregulatory properties, representing a hope for stem cell­based treatments. However, the mechanisms by which transplanted MSCs affect T helper (Th)17/regulatory T cell (Treg) balance in asthma patients remain unclear. The aim of the present study was to assess the therapeutic effects of human placenta MSCs (hPMSCs) in asthma, and explore the underlying mechanisms; in addition, the impact of hPMSCs transplantation on Th17/Treg balance in lymph and serum samples from asthmatic animals was evaluated. Sprague­Dawley rats were sensitized and challenged with ovalbumin (OVA). Administration of hPMSCs from human placenta resulted in increased Th17 and Treg in lymph samples compared with peripheral blood specimens. Enhanced pause values in OVA­treated animals were significantly higher than those in the control and hPMSCs treatment groups. The numbers of total cells, macrophages, neutrophils, and eosinophils were markedly increased in the OVA group compared with those of control + hPMSCs and control groups. In addition, interleukin 10, forkhead box P3 (Foxp3) and Treg levels in lymph, peripheral blood and lung tissue samples from asthma rats were increased significantly following hPMSC transplantation. Furthermore, Foxp3 protein levels increased, while those of RAR­related orphan receptor γ (RORγt) decreased after hPMSCs transplantation compared with the asthma group. Reduced IL­17, RORγt and Th17 levels were accompanied by reduced inflammatory cell infiltration, sub­epithelial smooth layer attenuation and mucus production in lung tissues. These results suggest that hPMSCs may improve airway hyperresponsiveness and inflammation by regulating the Th17/Treg balance in rats with asthma.

Complete Genome Sequence of a Porcine Endogenous Retrovirus Isolated from a Bama Minipig in Guangxi, Southern China.

A porcine endogenous retrovirus (PERV) strain, PERV-A-BM, was isolated from a Bama minipig in Guangxi, China. This is the first entire genome sequence of PERV isolated from Guangxi Bama minipigs. The isolate is closely related to isolates from Wuzhishan miniature pigs and distantly related to isolates from large white pigs.

Thymic derived iPs cells can be differentiated into cardiomyocytes.

Ventricular septal defect (VSD) is one of the common congenital heart malformations. Several factors lead to the development of VSD, including familial causes, exposure to certain drugs, infectious agents, and maternal metabolic disturbances. We considered that induced pluripotent stem (iPS) cells derived from VSD patients can be used to study the origin and pathogenesis of the VSD. Here, we show generation and cardiomyocyte differentiation potential of iPS cells from thymic epithelial cells of a patient with VSD (TECs-VSD) by overexpressing the four factors: OCT4, SOX2, NANOG, and LIN28 with lentiviral vectors. The self-renewal and pluripotency of the VSD-iPS cells was verified in iPS cells by in vitro expression of pluripotency markers and formation of teratoma in vivo. iPS cell lines from VSD patients differentiated into functional cardiomyocytes can serve as a model system for studying the pathophysiology and identifying etiology of VSD.

The expression profile analysis of NKX2-5 knock-out embryonic mice to explore the pathogenesis of congenital heart disease.

BACKGROUND: Mutation of NKX2-5 could lead to the development of congenital heart disease (CHD) which is a common inherited disease. This study aimed to investigate the pathogenesis of CHD in NKX2-5 knock-out embryonic mice. METHODS: The expression profile in the NKX2-5 knock-out embryonic mice (GSE528) was downloaded from Gene Expression Omnibus. The heart tissues from the null/heterozygous embryonic day 12.5 mice were compared with wild-type mice to identify differentially expressed genes (DEGs), and then DEGs corresponding to the transcriptional factors were filtered out based on the information in the TRANSFAC database. In addition, a transcriptional regulatory network was constructed according to transcription factor binding site information from the University of California Santa Cruz database. A pathway interaction network was constructed by latent pathways identification analysis. RESULTS: The 42 DEGs corresponding to transcriptional factors from the null and heterozygous embryos were identified. The transcriptional regulatory networks included five down-regulated DEGs (SP1, SRY, JUND, STAT6, and GATA6), and six up-regulated DEGs [POU2F1, NFY (NFYA/NFYB/NFYC), USF2 and MAX]. Latent pathways analysis demonstrated that ribosome, glycolysis/gluconeogenesis, and dilated cardiomyopathy pathways significantly interacted. CONCLUSION: The identified DEGs and latent pathways could provide new comprehensive view for understanding the pathogenesis of CHD.

[Enhanced expressions of Wnt5a/JNK signaling pathway-related molecules in the lung tissues of asthmatic rats].

OBJECTIVE: To observe the expression of Wnt5a/c-Jun N-terminal kinase (JNK) signaling pathway in the lung of asthmatic rats. METHODS: The experimental rats were randomly divided into two groups, the model group and the control group. After the rat models of asthma were established, airway pathological changes were observed by HE staining; the expression of Wnt5a mRNA in the lung tissues and peripheral blood cells was examined by real-time quantitative PCR; and the levels of phospho-c-Jun (p-c-Jun) and phospho-c-Jun N-terminal kinase (p-JNK) proteins in the lung tissues were measured by Western blotting. RESULTS: Compared with the control group, asthmatic rats presented with thickened bronchial mucosa with more obvious chrysanthemum fold, narrower airway, and more inflammatory cells infiltrated around trachea and pulmonary vessels. The inner airway wall and smooth muscle layer were much thicker in rats of asthma group than those of control group. The levels of Wnt5a mRNA, p-c-Jun and p-JNK proteins in the lung and blood of asthmatic rats increased as compared with the control group. CONCLUSION: The expressions of Wnt5a mRNA, p-JNK and p-c-Jun proteins were elevated in asthmatic rats.

Polycomb chromobox (Cbx) 7 modulates activation-induced CD4+ T cell apoptosis.

CD4(+) T cell polarization plays a critical role in a number of immune disorders; the pathogenesis is unclear. Chromobox homolog 7 (Cbx7) is involved in the gene transcription of several cell types. This study aims to investigate the mechanism by which Cbx7 modulates the CD4(+) T cell polarization. Expression of Cbx7 was assessed by quantitative RT-PCR and Western blotting. Apoptosis of CD4(+) T cell was analyzed by flow cytometry. The FasL promoter methylation was evaluated by the methylation specific PCR. The results showed that CD4(+) CD25(-) T cells express Cbx7 that was increased significantly after activation by exposing to anti-CD3/CD28 Ab, but suppressed by exposing to specific antigens. More apoptotic cells were detected in CD4(+) T cells with the Cbx7 gene knockdown. Exposure to insulin-like growth factor-1 up regulated the expression of Cbx7 in CD4(+) T cells. After antigen-specific TCR activation, Cbx7-deficient CD4(+) T cells expressed more FasL and showed the FasL gene promoter hyper demethylation than wild CD4(+) T cells. In addition, CD4(+) T cells with overexpression of Cbx7 showed lower levels of FasL gene promoter demethylation. We conclude that CD4(+) T cells express Cbx7; the latter prevents FasL expression and the activation-induced CD4(+) T cell apoptosis.

[Sequence analyses of HIRA gene 3'UTR region and related microRNA].

OBJECTIVE: To explore the HIRA gene sequences of 3'UTR region and elucidate the role of 3'UTR region of HIRA gene in the pathogenesis of tetralogy of Fallot (TOF). METHODS: Patients of TOF were confirmed by cardiac catheterization or surgery between April 2007 and December 2012 at our hospital. Mutations and single nucleotide polymorphisms (SNPs) were screened in 278 unrelated probands with isolated TOF and 515 controls. Target Scan was used to predict micro RNAs with possible combinations with 3'UTR region of HIRA gene. Dual-luciferase assay and real-time PCR were performed to detect the inhibition activity of micro RNAs on target genes. And χ(2) and t tests were used to analyze the results. RESULTS: Statistically significant change occurred in the alleleic frequencies of existing SNPs (rs:117447448) between TOF patients and control group (11.5% (32/278) vs 4.9% (25/515), P = 0.001) . The combining site of miR328 was predicted to be 10 bp upstream of SNP site. MiR328 was expressed in heart and it was related with myocardial infarction and atrial fibrillation. Dual-luciferase assay showed a decreased level of luciferase after co-transfection with miR328 (0.012 5 ± 0.000 6 vs 0.019 6 ± 0.003 8, P = 0.034). So was the expression of HIRA (1.039 6 ± 0.077 2 vs 1.608 7 ± 0.274 9, P = 0.037). However, the luciferase level was not affected by SNP (rs:117447448) (P = 0.380). CONCLUSIONS: The SNP (rs:117447448) of 3'UTR region of HIRA gene is related with TOF. HIRA is the target gene of miR328. Although SNP (rs:117447448) is not a major site of target gene HIRA for micro RNA328, it provides an important clue to in-depth studies of 3'UTR region of HIRA gene in the pathogenesis of TOF.

[An increased of Th17 cells and IL-23 and IL-17 in the lymph and blood of rats with bronchial asthma].

OBJECTIVE: To investigate whether immune response could change the number of CD4(+);IL-17(+);Th17 cells and the concentrations of IL-17 and IL-23 in the serum and lymph of asthmatic rats. METHODS: Both serum and lymph CD4(+);IL-17(+);Th17 cell numbers were measured by flow cytometry. The concentrations of IL-17 and IL-23 in the serum and lymph were assayed by ELISA. The expression of IL-23 p19 mRNA was examined by quantitative real-time PCR. RESULTS: Mononuclear cells (MNC) were collected from the blood and lymph of rats 0, 24 and 48 hours after final ovalbumin stimulation and were subsequently cultured for 18 hours. The serum and lymph of the asthma group produced higher amounts of CD4(+);IL-17(+);Th17 cells, and higher levels of IL-23p19 mRNA, than those of the healthy controls (P < 0.05). In addition, compared to lymph, serum had significantly lower amounts of CD4(+);IL-17(+);Th17 cells, decreased expression of IL-23p19 mRNA, and reduced IL-23 and IL-17 concentratons (P < 0.05). Cytokines and cell numbers of Th17 cells from the asthma group significantly increased compared with that of control group. CONCLUSION: These data suggested that asthma-driven CD4(+);IL-17(+);Th17-cell proliferation and release of IL-17 and IL-23 may be associated with different immune activities between the serum and lymph.

Roles of miR-1-1 and miR-181c in ventricular septal defects.

BACKGROUND: MicroRNAs (miRNAs) are endogenous noncoding RNAs of approximately 22 nucleotides in length that mediate post-transcriptional gene silencing by annealing to sequences in the 3'-untranslated region of target mRNAs. METHODS: In this study, we analyzed 25 candidate miRNAs selected based on microarray data for cardiac tissue from individuals with congenital heart defects (CHDs) and from healthy control tissue. RESULTS: This study identified specific changes in the miR-1-1 and miR-181c levels in human cardiac samples from individuals with ventricular septal defects (VSDs) relative to the levels in healthy control tissue. Increased levels of GJA1 and SOX9 were associated with the decreased expression of miR-1-1 in VSD patients, and increased miR-181c expression was correlated with downregulated BMPR2 levels. In addition, the results revealed that miR-1-1 and miR-181c directly regulate the expression of these predicted targets. CONCLUSIONS: miR-1-1 and miR-181c are associated with the pathogenesis of VSDs.

[Influence of anti-ICOS antibody on quantity and function of CD4(+);CD25(+);Foxp3(+);Treg from lymph and blood of rats with bronchial asthma].

AIM: To investigate the influences of anti-ICOS antibody (anti-ICOSAb) on quantity and function of CD4(+);CD25(+);Foxp3(+);Treg cells from lymph and peripheral blood of rats with bronchial asthma. METHODS: The mononuclear cells (MNC) from lymph and blood were co-cultured with anti-ICOSAb, and then the percentage of CD4(+);CD25(+);Foxp3(+);Treg cells were analyzed by flow cytometer (FCM) and the levels of IL-10 and TGF-ß1 in supernatants were determined by ELISA. RESULTS: The MNC were collected from lymph and blood at 0, 24 and 48 h after the last challenge, respectively, and the cells were cultured for 96 h in vitro. The percentage of CD4(+);CD25(+);Foxp3(+);Treg cells in the MNC from lymph was significantly higher than that from blood in each group (P<0.05); The percentage of CD4(+);CD25(+);Foxp3(+);Treg cells in the MNC from lymph and blood in asthma group was significantly lower compare with the normal control group (P<0.05); The percentage of CD4(+);CD25(+);Foxp3(+);Treg cells in the MNC from lymph and blood in the anti-ICOSAb group obviously decreased compare with the asthma group (P<0.05). At 0 h after the last challenge, the level of IL-10 in the supernatant of MNC from lymph and blood in the anti-ICOSAb group were significantly lower than that of the control and asthma groups (P<0.05), while there were no significant differences of TGF-ß1 expression in the supernatant of MNC from lymph and blood in each group at different time points. CONCLUSION: Blocking the ICOS/ICOSL signaling pathway by anti-ICOSAb could exacerbate the deficiency of CD4(+);CD25(+);Foxp3(+);Treg cells from lymph and blood in bronchial asthmatic rat, meanwhile inhibit the CD4(+);CD25(+);Foxp3(+);Treg cells secreting IL-10 at 0 h after the last challenge, but have no significant effect on the secretion of TGF-ß1.