[Study on transcriptome characteristics of respiratory syncytial virus bronchiolitis in children by RNA sequencing].

To explore the biological characteristics related to the pathogenesis and severity of respiratory syncytial virus (RSV) bronchiolitis by RNA sequencing of white blood cells in children with RSV bronchiolitis. This study is a case-control study. A total of 87 children diagnosed with bronchiolitis and RSV antigen positive and/or RSV nucleic acid positive in the pediatric respiratory department of the Second Affiliated Hospital of Wenzhou Medical University from October 2019 to April 2022 were selected as the case group. The case group was divided into three groups based on the condition: mild, moderate, and severe, and there were two groups according to the presence or absence of atopic symptoms: the atopic group and the non-atopic group, forty healthy children in the same period were selected as the control group. The whole blood leukocyte RNA of the children in the case group and the control group was extracted for RNA sequencing, and the data were analyzed to obtain differentially expressed genes (DEGs). Then, the immunobiological pathways and genes related to the pathogenesis, disease condition, and atopy were screened through Gene Ontology (GO) annotation, Kyoto Gene and Genome Encyclopedia (KEGG) annotation, and protein interaction network (PPI) construction methods. Construct the weighted gene co-expression network analysis (WGCNA) module to identify potential biological indicators related to disease severity.Compared with the control group, the case group had a total of 1 782 DEGs, including 1 586 upregulated genes and 196 downregulated genes. The GO pathway enrichment of DEGs is mainly enriched in molecular functions such as peroxidase activity and oxidoreductase activity. In the cytological components, it is mainly enriched in cytoplasmic vesicle lumen and secretory granule lumen. In biological processes, it is mainly enriched in processes such as neutrophil activation involved in immune responses, neutrophil degranulation, and neutrophil activation. KEGG analysis is mainly concentrated in the signal pathway of the viral protein interaction with cytokine and cytokine receptor. A PPI network was constructed to screen four genes at the core position, including CCL2, IL-10, MMP9 and JUN. The DEGs obtained by comparing different disease groups with the control group are mainly enriched in retrograde endocannabinoid signaling and cell apoptosis pathways. WGCNA analysis showed that the brown module related to oxygen saturation was most closely related to the disease, and its gene was mainly enriched in the RNA helicase retinoic acid inducible gene-I (RIG-I) like receptor signal pathway. There are 230 specific DEGs in the atopic group and 444 in the non-atopic group. KEGG enrichment analysis results show that both groups are enriched to NF-κB signaling pathway, the characteristic does not cause significant changes in immune response and transcriptome characteristics in children with RSV bronchiolitis. In conclusion, neutrophil activation, degranulation pathway and signal pathway of interaction between viral protein and cytokine and cytokine receptor are involved in the immune response of RSV bronchiolitis host. CCL2, IL-10, MMP9 and JUN genes may be associated with the pathogenesis. They might be potential biomarkers related to disease severity in RIG-I like receptors, cell apoptosis, and endogenous cannabinoid related signaling pathways.

Ultra-early electroacupuncture rehabilitation for intravenous thrombolysis-induced cerebral infarction.

OBJECTIVE: The purpose of this research was to investigate whether it is possible to perform ultra-early interventional electroacupuncture on individuals who had experienced intravenous thrombolysis prior to receiving therapy for acute cerebral infarction. PATIENTS AND METHODS: Patients who have undergone intravenous thrombolysis between July 2019 and March 2021 were eligible for participation in this study. The participants were divided into two groups; one group received electroacupuncture therapy 24 hours after their condition became stable, while the other group received treatment 48 hours after their condition became stable. Both groups received the same therapy for their respective forms of rehabilitation. The Fugl-Meyer Motion Assessment Scale (FMA) was used to assess the patients' motor function before and after therapy, as well as two weeks and one month after treatment. The scores of the FMA were recorded before and after treatment. RESULTS: After therapy, the FMI scores were higher in both groups (p<0.05), and the researchers found that the ultra-early electroacupuncture intervention was related to higher FMI ratings 2 weeks and 1 month after treatment (p<0.05). In neither of the two study groups was there any sign of a major adverse response or consequence (p>0.05). CONCLUSIONS: This research offers evidence that ultra-early interventional electroacupuncture rehabilitation therapy may be an effective and safe method of treatment for individuals who have had a cerebral infarction after receiving intravenous thrombolysis. The results lend credence to the notion that this kind of therapy should be taken into consideration as an adjunctive model for rehabilitation in patients of this type.

[Expression of heterogeneous nuclear ribonucleo-protein A2B1 in mouse cerebellar development and human medulloblastoma].

Objective: To investigate the expression and potential role of heterogeneous nuclear ribonucleo-protein A2B1 (HNRNPA2B1) in mouse cerebellar development and the significance of HNRNPA2B1 in human medulloblastoma. Methods: The data of HNRNPA2B1 RNA expression in mouse and human cerebella were obtained from databases. Western blot and immunohistochemical staining were performed to detect the protein level of HNRNPA2B1 in mouse cerebella at different ages. The expression level of HNRNPA2B1 in control human cerebellum and medulloblastoma was detected by immunohistochemical staining. m6A-IP-qPCR method was applied to confirm whether HNRNPA2B1 RNA in Daoy cells was modified with m6A.Western blot was used to detect the effect of MG132 treatment on the HNRNPA2B1 protein level in Daoy cells. Results: The level of HNRNPA2B1 protein in postnatal mouse cerebella was higher than that in adult mouse cerebella, with weak HNRNPA2B1 staining in external granular cells while strong staining in mature Purkinje cells and molecular layer. Compared with control normal human cerebella, the RNA expression level of HNRNPA2B1 increased in medulloblastoma, while immunohistochemical staining showed that the mean intensity of HNRNPA2B1 decreased in medulloblastoma. HNRNPA2B1 RNA in medulloblastoma and Daoy cells was modified by m6A. The HNRNPA2B1 protein level in Daoy cells increased upon MG132 treatment. Conclusions: HNRNPA2B1 is dynamically expressed during mouse cerebellar development. Compared with normal human cerebella, HNRNPA2B1 is significantly up-regulated at transcriptional level but obviously down-regulated at translational level in medulloblastoma. These results indicate that HNRNPA2B1 may be involved in cerebellar development process and medulloblastoma tumorigenesis. The m6A methylation in HNRNPA2B1 transcript and protein ubiquitin-proteasome pathway may account for the down-regulation of HNRNPA2B1 at protein level.

Molecular mapping and candidate gene analysis for resistance to powdery mildew in Cucumis sativus stem.

Powdery mildew (PM) of cucumber (Cucumis sativus), caused by Podosphaera xanthii, is a major foliar disease worldwide and resistance is one of the main objectives in cucumber breeding programs. The resistance to PM in cucumber stem is important to the resistance for the whole plant. In this study, genetic analysis and gene mapping were implemented with cucumber inbred lines NCG-122 (with resistance to PM in the stem) and NCG-121 (with susceptibility in the stem). Genetic analysis showed that resistance to PM in the stem of NCG-122 was qualitative and controlled by a single-recessive nuclear gene (pm-s). Susceptibility was dominant to resistance. In the initial genetic mapping of the pm-s gene, 10 SSR markers were discovered to be linked to pm-s, which was mapped to chromosome 5 (Chr.5) of cucumber. The pm-s gene's closest flanking markers were SSR20486 and SSR06184/SSR13237 with genetic distances of 0.9 and 1.8 cM, respectively. One hundred and fifty-seven pairs of new SSR primers were exploited by the sequence information in the initial mapping region of pm-s. The analysis on the F2 mapping population using the new molecular markers showed that 17 SSR markers were confirmed to be linked to the pm-s gene. The two closest flanking markers, pmSSR27and pmSSR17, were 0.1 and 0.7 cM from pm-s, respectively, confirming the location of this gene on Chr.5. The physical length of the genomic region containing pm-s was 135.7 kb harboring 21 predicted genes. Among these genes, the gene Csa5G623470 annotated as encoding Mlo-related protein was defined as the most probable candidate gene for the pm-s. The results of this study will provide a basis for marker-assisted selection, and make the benefit for the cloning of the resistance gene.

The temporal bone microdissection of miniature pigs as a useful large animal model for otologic research.

CONCLUSION: Compared with traditional animal models, the miniature pig may be a better model for biomedical research because its morphology has many similarities with that of humans. OBJECTIVE: To investigate the suitability of the miniature pig as an animal model for otological research as regards morphology. METHODS: Microdissection of the temporal bone of 10 miniature pigs was performed and recorded on photographs. RESULTS: The morphology and measurements of the external, middle, and inner ear, and the lateral recess of the miniature pigs were completed by microdissection. The temporal bone structures, including the external, middle, inner ear, and the lateral recess, were similar in the miniature pig and humans.

Use of the translabyrinthine approach to repair congenital spontaneous cerebrospinal fluid leakage in five Chinese patients with Mondini dysplasia.

OBJECTIVES: To introduce a more effective surgical technique for the repair of spontaneous incurable cerebrospinal fluid (CSF) leakage (CSF otorrhea). METHODS: The cases of five Chinese patients with Mondini dysplasia and CSF leakage were reviewed. All patients presented with rhinorrhea or otorrhea from childhood and recurrent meningitis attacks. CSF leakage was repaired surgically using the translabyrinthine approach. RESULTS: Facial paralysis occurred in the first two patients, but was cured 2 months later. The other three patients recovered well and had no facial paralysis or complication. CSF leakage did not recur in any patient after follow-up periods of 0.5-3 years. CONCLUSION: Mondini dysplasia with CSF leakage is rare. Adequate diagnosis and treatment are required to avoid life-threatening complications, such as meningitis. We recommend the translabyrinthine approach as an alternative surgical technique that reduces recurrence.

The diagnosis and surgical treatment of occult otogenic CSF leakage.

CONCLUSIONS: The diagnosis of occult otogenic cerebrospinal fluid (CSF) leakage is challenging and it can easily be misdiagnosed. Some characteristics of clinical presentation can supply important clues and confirmed diagnosis should be obtained according to these clues and suitable imaging studies before meningitis develops. Different surgical techniques should be adopted to treat the CSF leakage according to different leakage etiologies, and good results can be obtained. OBJECTIVE: The aim of the study was to evaluate the diagnosis and surgical treatment of occult otogenic CSF leakage, including the characteristics of clinical presentation, imaging studies, and operation methods in order to decrease the rate of misdiagnosis and obtain a good curative effect. METHODS: We performed a retrospective review of 11 cases of CSF leakage that were all misdiagnosed and accompanied by meningitis, operated in our department from 2007 to 2012 after a mean follow-up of 3 years. In this context, the characteristics of clinical presentation, imaging studies, and management of CSF leakage were studied. RESULTS: The CSF leakage had arisen traumatically (n = 9) or congenitally (n = 2). The medical history and special clinical presentation such as repeated otorrhea or rhinorrhea, fever, headache, and unilateral deafness can supply important diagnostic clues. Imaging studies including high-resolution noncontrast CT (HRCT), CT cisternography, and magnetic resonance imaging (MRI) are very important diagnostic methods. The surgical repairs were performed via a transmastoid approach (n = 8), packing the vestibule (n = 1) or a translabyrithine approach (n = 2). Recurrent leakage did not occur.

Acute scrotum in Henoch-Schönlein purpura.

Henoch-Schönlein purpura (HSP) is a systemic vasculitis with manifestations usually involving the skin, gastrointestinal tract, kidney and joints. Epididymitis is rarely seen as a complication of HSP. It is easily misdiagnosed as testicular torsion, causing the patient to undergo unnecessary surgery, because the patient may have complained of severe scrotal pain and swelling. We report a 5-year-old boy who was suffering from HSP associated with acute scrotal pain and swelling of the left testicle. No gastrointestinal signs were noted but severe joint pain, swelling and palpable skin lesions in the lower limbs and the buttocks were found. Prednisolone was prescribed and the boy recovered without surgical intervention.

Pyomyositis in childhood: a case report.

Pyomyositis is a primary infection of skeletal muscle. We report the case of a previously healthy six-year-old who suffered from pyomyositis in the right lower back. He presented with lower back pain and low-grade fever for one week. After a series of laboratory and imaging studies, the diagnosis of right multifidus muscle pyomyositis with abscess formation was made. The patient recovered rapidly after incision and drainage therapy, accompanied by antibiotic treatment. Methicillin-resistant Staphylococcus aureus was cultured from the abscess discharge. It was strongly suspected that herbal medicines and common cold medication the patient had been prescribed before admission to our hospital produced a masking effect that delayed the diagnosis.

Anti-FD4 idiotypic antibody mimicking human gastric cancer-associated antigen.

Rabbits were immunized with the Fab fragment of a murine monoclonal antibody (McAb) PD4 against human gastric cancer to produce anti-PD4-idiotypic antibody (alpha PD4-Ab2). The alpha PD4-Ab2 could not only competitively inhibit binding of McAb PD4 to gastric cancer cell MGC803, but also induce delayed-type hypersensitivity (DTH) to MGC803 in mice. Spleen cells of mice immunized with alpha PD4-Ab2 were fused with myeloma cell SP2/0 to form hybridoma secreting Ab3 which could be bound to target cell MGC803. McAb C7-Ab3, one of the Ab3, could selectively react with a 40 kD tumor-associated antigen located on MGC803 cell membrane, as well as McAb PD4. The results indicate that alpha PD4-Ab2 possesses determinants (internal image antigen) similar to those on MGC803, and could mimic human gastric cancer-associated antigen.

Molecular cloning and nucleotide sequence analysis of a transforming gene from human gastrocarcinoma cell line.

Mouse and rat fibroblasts were transfected with total DNA from human gastrocarcinoma cell line BGC-823. It was shown by hybridization assay that the genome of one of the rat secondary foci contains transforming genes from the human gastrocarcinoma cell line, which are homologous to the protooncogene c-Ha-ras in the normal cells. The genomic library of the rat secondary foci was constructed, using lambda phage EMBL3 as the vector. The transforming gene Ha-ras of the human gastrocarcinoma cell was thus cloned by screening the library with the probes of human Alu repeat sequence and c-Ha-ras. The nucleotide sequences of the first and second exons were analysed by M13-dideoxy method. The result shows that the nucleotide sequence of the transforming gene is the same as that of the normal protooncogene except one nucleotide difference in the first exon.

Activated c-Ha-ras oncogene with a guanine to thymine transversion at the twelfth codon in a human stomach cancer cell line.

The rat fibroblast cell line Rat 1 was transfected with total DNA of a gastrocarcinoma cell line, BGC-823. The transforming gene was cloned from the genomic library of the secondary transformants using in situ hybridization with a probe of the human Alu repeat sequence. This cloned gene is homologous to the protooncogene c-Ha-ras. The activation lesion of the transforming gene was identified by sequence analysis as a single nucleotide substitution of thymine for guanine in the 12th codon. This results in the substitution of valine for glycine at the 12th amino acid of the Mr 21,000 protein.

[DNA transformation activity of a human gastrocarcinoma cell line].

DNAs of three cell lines of human gastrocarcinoma (MGC-803, BGC-823 and PACM-82) and two fresh solid tumors of human stomach cancer were used to transfect NIH3T3 and Rat-1 cells. The transformed cells were selected with high concentration of glucose and low concentration of serum, or with medium containing Geneticin (G418) after co-transfection of pSVneo and DNAs of stomach cancer cell line or primary transformants. From the second round transfection, we had obtained transformants which could grow with high colony forming efficiency in soft agarose and were tumorigenic in nude mice. The southern blot analysis showed that the cellular DNA of the transformants contained human Alu repeat sequence and the transformed gene from stomach cancer cell line (BGC-823) and was homologous to proto-oncogene c-Ha-ras. The transforming gene is able to induce neoplastic transformation of NIH3T3 and Rat-1 cells.