Case report: A severe clinical phenotype of pontocerebellar hypoplasia type 7 with compound heterozygous variants of TOE1.

Pontocerebellar Hypoplasia (PCH) is a rare autosomal recessive hereditary neurological degenerative disease. To elaborate upon the clinical phenotypes of PCH and explore the correlation between TOE1 gene mutations and clinical phenotype, we analyze the clinical and genetic features of a Chinese infant afflicted with pontocerebellar dysplasia accompanied by gender reversal with bioinformatics methods. The main clinical features of this infant with TOE1 gene mutation included progressive lateral ventricle widening, hydrocephalus, severe postnatal growth retardation, and hypotonia, and simultaneously being accompanied by 46, XY female sex reversal. Whole exome sequencing revealed a compound heterozygous mutation in the TOE1 gene (c.299T > G, c.1414T > G), with the protein homology modeling-generated structure predicting a pathogenic variation, which is closely related to the clinical manifestations in the patient. The new mutation sites, c.299T > G and c.1414T > G, in the TOE1 gene are pathogenic variants of pontocerebellar hypoplasia type 7.

Effect of aqueous extract of Millettia speciosa Champ on intestinal health maintenance and immune enhancement of Cyprinus carpio.

Millettia speciosa Champ (MSP) is a natural Chinese herb that improves gastrointestinal health and enhances animal immunity. An 8-week feeding trial with different MSP levels (0, 150, 300, and 600 mg/kg) was conducted to evaluate the promotive effects of MSP in Cyprinus carpio. Results indicate that MSP improved intestinal immunity to some extent evidenced by the immuno-antioxidant parameters and the 16S rRNA in the Illumina MiSeq platform. With the analysis of transcriptome sequencing, 4685 differentially expressed genes (DEGs) were identified, including 2149 up-regulated and 2536 down-regulated. According to the GO and KEGG enrichments, DEGs were mainly involved in the immune system. Transcriptional expression of the NOD-like signaling pathway and key genes retrieved from the transcriptome database confirmed that innate immunity was improved in response to dietary MSP administration. Therefore, MSP could be used as a feed supplement that enhances immunity. This may provide insight into Chinese herb additive application in aquaculture production.

Whole genome sequencing and evolution analyses of Human metapneumovirus.

Human metapneumovirus (HMPV) is a major pathogen of acute respiratory tract infections (ARTIs) in children. Whole genome sequence analyses could help understand the evolution and transmission events of this virus. In this study, we sequenced HMPV whole genomes to improve the identification of molecular epidemiology in Beijing, China. Nasopharyngeal aspirates of hospitalized children aged < 14 years old with ARTIs were screened for HMPV infection using qPCR. Fourteen pairs of overlapping primers were used to amplify whole genome sequences of HMPV from positive samples with high viral loads. The epidemiology of HMPV was analysed and 27 HMPV whole genome sequences were obtained. Sequence identity and the positional entropy analyses showed that most regions of HMPV genome are conserved, whereas the G gene contained many variations. Phylogenetic analysis identified 25 HMPV sequences that belonged to a newly defined subtype A2b1; G gene sequences from 24 of these contained a 111-nucleotide duplication. HMPV is an important respiratory pathogen in paediatric patients. The new subtype A2b1 with a 111-nucleotide duplication has become predominate in Beijing, China.

Effects of TLR2/4 signalling pathway in western mosquitofish (Gambusia affinis) after Edwardsiella tarda infection.

Gambusia affinis is regarded as an important animal model. Edwardsiella tarda is one of the most serious pathogens affecting aquaculture. The study explores the effects of TLR2/4 partial signalling pathway in G. affinis of E. tarda infection. The study collected the brain, liver, and intestine after E. tarda LD50 and 0.85% NaCl solution challenge at different times (0 h, 3 h, 9 h, 18 h, 24 h, and 48 h). In these three tissues, the mRNA levels of PI3K, AKT3, IRAK4, TAK1, IKKß, and IL-1ß were substantially enhanced (p < .05) then returned to normal levels. Additionally, Rac1 and MyD88 in liver had different trend with other genes in brain and intestine, which displayed significantly indifference. The overexpression of IKKß, and IL-1ß indicated that E. tarda also caused immune reaction in intestine and liver, which would be consistent with delayed edwardsiellosis, which causes intestinal lesions and liver and kidney necrosis. Additionally, MyD88 plays a smaller role than IRAK4 and TAK1 in this signalling pathways. This study could enrich the understanding of the immune mechanism of the TLR2/4 signalling pathway in fish and might help to prescribe preventive measures against E. tarda to prevent infectious diseases in fish.

Human metapneumovirus in hospitalized children with acute respiratory tract infections in Beijing, China.

BACKGROUND: This study aims to described the epidemiology and genotypic diversity of Human metapneumovirus (HMPV) and the impact of SARS-CoV-2 on the prevalence of HMPV in hospitalized children with Acute respiratory tract infections (ARTIs) in Beijing, China. METHODS: From April 2018 to March 2019 and from September 2020 to August 2021, nasopharyngeal aspirates (NPAs) from hospitalized children with ARTIs in Beijing were collected and subjected to real-time polymerase chain reaction tests for HMPV. Then genotyping, detection of 15 common respiratory viruses and clinical characteristics were analyzed on HMPV positive samples. RESULTS: 7.9% (124/1572) enrolled pediatric patients were identified as having HMPV infection, and the majority of children under the age of 5 (78.2%, 92/124), From April 2018 to March 2019. The detection rate of HMPV in spring and winter is significantly higher than that in summer and autumn. The co-infection rate were 37.1% (46/124), the most common co-infected virus were parainfluenza virus type 3 (HPIV-3). The main diagnosis of HMPV infection was pneumonia (92.7%,115/124), most patient have cough and fever. Of 78 HMPV-positive specimens, A2b (82.1%,64/78) were the main epidemic subtypes. Hospitalized children with HMPV genotype A infection had a higher viral load compared to genotype B. During the COVID-19 outbreak, Among 232 samples, only 4 cases were HMPV-positive. After statistical test, the detection rate of HMPV during the COVID-19 pandemic has decreased significantly compared with that before the epidemic (p = 0.001). CONCLUSIONS: HMPV is an important cause of ARTIs in children under 5 years old. The epidemic peak is generally in winter and spring, and the A2b subtype is the most common. However, under the prevention and control of the COVID-19 pandemic, the HMPV infection of hospitalized children with ARTIs has decreased significantly.

The impact of deleting the mitfa gene in zebrafish on the intestinal microbiota community.

To explore the relationship between the mitfa gene and intestinal microbiota, the 16S rRNA gene amplicon sequencing was performed to compare the intestinal microbiota composition of the mitfa knockout zebrafish line (CKO group) and the wild-type zebrafish (WT group) in this study. The results showed that the Fusobacteria and Firmicutes were significantly decreased and the Dependentiae and Patescibacteria were significantly increased in the CKO group at the phylum level. Furthermore, the relative abundance of Citrobacter, Gordonia, Mesorhizobium, Legionella, and Bradyrhizobium were extremely higher in the CKO group, whereas the other four genera Nocardia, Pannonibacter, Shinella, and Cetobacterium were significantly declined in the CKO group at the genus level. Due to these changed intestinal microbiota appear to be related to lipid metabolism and immunity, eight lipid metabolism-related genes and nine inflammation-related genes were detected in the intestinal. The results showed that the expression levels of these genes were significant differences between the CKO and WT group. These results indicated that the deletion of mitfa can affect the expression levels of immune and metabolism-related genes, and causing changes in the composition of the intestinal microbiota.

S100A9 blockade improves the functional recovery after spinal cord injury via mediating neutrophil infiltration.

Spinal cord injury (SCI) refers to damage to the spinal cord resulting from trauma, disease or degeneration. Controlling the inflammatory process and restoring neural homeostasis is hypothesized to prevent injury aggravation. S100 calcium-binding protein A9 (S100A9) is a pro-inflammatory alarm protein, which is expressed in and released by activated neutrophils. However, whether S100A9 could serve as an effective target for the treatment of SCI has not been reported to date. In the present study, a T10 spinal cord contusion injury model was established in Sprague-Dawley rats. S100A9 expression level was determined in the serum and injured spinal cord tissue via ELISA, reverse transcription-quantitative PCR (RT-qPCR) and western blotting. The S100A9-specific blocker, ABR-238901 (ABR), was administered during the inflammatory phase of SCI, as a form of treatment. Subsequently, the morphological structure, neuronal viability and inflammatory levels of injured spinal cord were observed by histopathology, immunohistochemistry and RT-qPCR. In the obtained results, S100A9 was found to be highly expressed in the injured spinal cord and serum in the first 3 days after SCI. However, at 28 days after surgery, ABR treatment significantly improved motor function, reduced the cavity formation and neutrophil infiltration in the lesion, which was verified via H&E staining and immunohistochemistry for myeloperoxidase. Furthermore, ABR treatment was found to effectively improve the survival and viability of neurons, as shown via Nissl staining and immunofluorescence of the synaptic plasticity markers, microtubule associated protein 2 and neurofilament 200. Moreover, S100A9 blockade effectively upregulated the mRNA expression level of the anti-inflammatory genes, IL-4 and IL-10 and downregulated the mRNA expression level of the pro-inflammatory factors, IL-1ß, IL-6 and TNF-α. In addition, S100A9 blockade notably alleviated the apoptosis level of the injured nerve cells. Therefore, the findings of the present study revealed that S100A9 may be a useful target for the treatment of SCI.

Tunable microwave-optical entanglement and conversion in multimode electro-opto-mechanics.

We study tunable double-channel microwave-optical (M-O) entanglement and coherent conversion by controlling the quantum interference effect. This is realized in a two-mechanical-mode electro-opto-mechanical (EOM) system, in which two mechanical resonators (MRs) are coupled with each other by phase-dependent phonon-phonon interaction, and link the interaction between the microwave and optical cavity. It's demonstrated that the mechanical coupling between two MRs leads to the interference of two pathways of electro-opto-mechanical interaction, which can generate the tunable double-channel phenomena in comparison with a typical three-mode EOM system. In particular, by tuning of phonon-phonon interaction and couplings between cavities with MRs, we can not only steer the switch from the M-O interaction with a single channel to that of the double-channel, but also modulate the entanglement and conversion characteristics in each channel. Moreover, our scheme can be extended to an N-mechanical-mode EOM system, in which N discrete channels will be observed and controlled. This study opens up prospects for quantum information transduction and storage with a wide bandwidth and multichannel quantum interface.

Nanoplastics Induce More Serious Microbiota Dysbiosis and Inflammation in the Gut of Adult Zebrafish than Microplastics.

Microplastics (MPs) (< 5 mm) and nanoplastics (NPs) (< 100 nm) are emerging environmental pollutants and have been proved could cause a series of toxicity in aquatic organisms. In this study, the effects on gut microbiota of adult zebrafish exposed for 21 days to 10 µg/L and 1 mg/L of MPs (8 µm) and NPs (80 nm) were evaluated. We analyzed the intestinal microbial community of zebrafish using high throughput sequencing of the 16S rRNA gene V3-V4 region and also performed transcriptional profiling of the inflammation pathway related genes in the intestinal tissues. Our results showed that both spherical polystyrene MPs and NPs could induce microbiota dysbiosis in the gut of zebrafish. The flora diversity of gut microbiota significantly increased under a high concentration of NPs. At the phylum level, the abundance of Proteobacteria increased significantly and the abundance of Fusobacteria, Firmicutes and Verrucomicrobiota decreased significantly in the gut after 21-day exposure to 1 mg/L of both MPs and NPs. Furthermore, interestingly, the abundance of Actinobacteria decreased in the MPs treatment groups but increased in the NPs treatment groups. At the genus level, revealed that the relative abundance of Aeromonas significantly increased both in the MPs and NPs treatment groups. Moreover, it was observed that NPs increased mRNA levels of il8, il10, il1ß and tnfα in the gut, but not in MPs exposure group, indicating that the NPs may have a more serious effect on the gut of zebrafish than MPs to induce microbiota dysbiosis and inflammation in the gut.

Pathomechanism of Lower-level Discogenic Groin Pain and Clinical Outcomes of Percutaneous Endoscopic Discectomy for the Treatment of Discogenic Groin Pain.

BACKGROUND: Groin pain can be induced by high-level (L1-L2 or L2-L3) lumbar disc herniation. However, 4.1% of patients with lower-level (L4-L5 or L5-S1) lumbar disc herniation also complained of groin pain. The pathomechanism of groin pain occurring due to lumbar disc herniation at and below the L4-5 levels is still unclear. OBJECTIVE: To investigate the afferent pathways of lower-level lumbar disc herniation induced groin pain. And evaluate the clinical results of transforaminal endoscopic discectomy treatment for discogenic groin pain. STUDY DESIGN: This retrospective observational study used an experimental design (institutional review board: HROH 201-C2-100). SETTING: The research took place in the Laboratory Research Center and spine center at The First Affiliated Hospital of Harbin Medical University. METHODS: Firstly, 14 adult Wistar rats were randomly divided into 2 groups: control group (the paravertebral sympathetic trunks were preserved) and experimental group (the paravertebral sympathetic trunks were resected). All Wistar rats were intraperitoneally anesthetized, and then 1 (mu)L of fast blue was injected into the dorsal rami of L2 spinal nerves on the right side. Forty hours later, 2 (mu)L of nuclear yellow was injected into the right posterior portion of the L5-L6 intervertebral disc. The L1 and L2 spinal ganglia were sectioned 8 hours later to observe fluorescently double-labeled cells and the effect of paravertebral sympathetic trunk resection. Secondly, 14 adult Wistar rats were anesthetized, and the right posterior portion of the L5-L6 intervertebral disc was electrostimulated to observe potential changes in the genitocrural nerve in the ipsilateral inguinal region. To evaluate the clinical outcomes of transforaminal endoscopic discectomy for the treatment of discogenic groin pain, between September 2015 and May 2017, transforaminal endoscopic discectomy was performed on 30 patients with lower-level discogenic groin pain. Outcomes were analyzed utilizing the visual analog scale, Oswestry disability index, and MacNab Criteria. RESULTS: The total proportion of cells in the right L1 and L2 spinal ganglia with fast blue/nuclear yellow double labeling was 3.33% and 3.41% (48 and 56), respectively. The number of fluorescently double-labeled cells in the resected paravertebral sympathetic trunk group was significantly less (P < 0.01). Electrical stimulation of the right posterior portion of the L5-L6 intervertebral disc could elicit action potentials in the ipsilateral genitofemoral nerve. All patients were followed for 12 months, and the visual analog scale score at 1 week, 1 month, 3 months, 6 months, and 12 months after the operation was 0.79 ± 0.55, 0.54 ± 0.55, 0.47 ± 0.65, 0.51 ± 0.65, and 0.69 ± 0.55, respectively, showing a significant decrease compared with the preoperative visual analog scale score (P < 0.01). Based on the MacNab scoring system, the effective rate was 100%, and the rate of good and excellent results was 93.3%. LIMITATIONS: A relatively small number of patients and a short follow-up period. CONCLUSIONS: Discogenic groin pain is transmitted by sympathetic nerves and appears in the area segmentally innervated by the anterior rami of the L1 and L2 spinal nerves. Posterolateral percutaneous transforaminal endoscopic discectomy and radiofrequency thermal annuloplasty are effective minimally invasive alternative treatments for discogenic groin pain.

miRNA-221 Regulates Spinal Cord Injury-Induced Inflammatory Response through Targeting TNF-α Expression.

OBJECTIVES: To investigate the roles of miR-221 in spinal cord injury (SCI) as well as the underlying mechanism. METHODS: A mouse model of SCI was generated and used to examine dynamic changes in grip strength of the mouse upper and lower limbs. The expression of miR-221 and tumor necrosis factor-α (TNF-α) was detected by RT-qPCR and Western blot. Levels of inflammation and oxidative stress in microglia cells of the injured mice overexpressing miR-221 were then measured by ELISA. Bioinformatics analysis and dual-luciferase reporter assay were conducted to identify the miR-221 target. RESULTS: We successfully constructed SCI mouse model. The results of qRT-PCR showed that miR-221 was gradually upregulated in the spinal cord tissue of mice in the SCI group with the prolonged injury time. At the same time, the mRNA and protein of TNF-α gradually decreased. We further confirmed through cell experiments that the inflammatory factors TNF-α and IL-6, as well as iNOS and eROS, were upregulated in spinal cord microglia cells of SCI mice, and upregulation of miR-122 can inhibit their expression. Finally, the luciferase reporter experiment confirmed that miR-122 targeted TNF-α. CONCLUSIONS: We present evidence that miR-221 promotes functional recovery of the injured spinal cord through targeting TNF-α, while alleviating inflammatory response and oxidative stress.

Epidemiology and genotypic diversity of human metapneumovirus in paediatric patients with acute respiratory infection in Beijing, China.

BACKGROUND: Acute respiratory tract infections (ARTIs) causes high amounts of morbidity and mortality worldwide every year. Human metapneumovirus (HMPV) is a major pathogen of ARTIs in children. In this study, we aimed to investigate the epidemiology and genotypic diversity of HMPV in children hospitalized with ARTIs in Beijing, China. METHODS: Hospitalized children aged < 14 years with ARTIs were enrolled from April 2017 to March 2018; nasopharyngeal aspirates were collected and subjected to real-time polymerase chain reaction tests for HMPV. HMPV-positive samples were genotyped based on a partial N gene. Whole genome sequences were determined for samples with high viral loads. RESULTS: 4.08% (52/1276) enrolled paediatric patients were identified as having HMPV infection. The epidemic season is winter and early spring, children aged ≤ 4 years were more susceptible to HMPV infection (47/52, 90.38%). The co-infection rate were 36.54% (19/52), the most common co-infected virus were influenza and respiratory syncytial virus. The main diagnoses of HMPV infection were pneumonia (29/52, 55.77%) and bronchitis (23/52, 44.23%), while the main clinical manifestations were cough, fever, rhinorrhoea, and sneeze. Among 48 HMPV-positive specimens, A2b (19/48, 39.58%) and B1 (26/48, 54.17%) were the main epidemic subtypes. Patients with HMPV genotype A infection had a higher viral load compared to genotype B patients (6.07 vs. 5.37 log10 RNA copies/ml). Five complete sequences of HMPV were obtained. This is the first report of a whole genome sequence of HMPV-B1 isolated in China. CONCLUSIONS: HMPV is an important respiratory pathogen in paediatric patients. Cases of HMPV infection could burden hospitals in the epidemic season. HMPV viral loads and genotypes have no correlation with co-infection or clinical characteristics.

Isolation and characterization of WUPyV in polarized human airway epithelial cells.

BACKGROUND: Washington University polyomavirus (WUPyV) is a novel human polyomavirus detected in childwith acute respiratory infection in 2007. However, the relationship between WUPyV and respiratory diseases has yet to be established for lacking of a suitable in vitro culture system. METHODS: To isolate WUPyV with human airway epithelial (HAE) cells, the positive samples were incubated in HAE, and then the nucleic acid, VP1 protein and virions were detected using real-time PCR, immunofluorescence and electron microscopy respectively. RESULTS: The result showed that WUPyV could replicate effectively in HAE cells and virions with typical polyomavirus characteristics could be observed. Additionally, the entire genome sequence of the isolated strain (BJ0771) was obtained and phylogenetic analysis indicated that BJ0771 belongs to gene cluster I. CONCLUSIONS: Our findings demonstrated clinical WUPyV strain was successfully isolated for the first time in the world and this will help unravel the etiology and pathogenic mechanisms of WUPyV in respiratory infection diseases.

Percutaneous Endoscopic Interlaminar Lumbar Discectomy with Local Anesthesia for L5-S1 Disc Herniation: A Feasibility Study.

BACKGROUND: General anesthesia (GA), which is routinely applied in patients who undergo percutaneous endoscopic interlaminar lumbar discectomy (PEILD) of L5-S1 disc herniation, is closely associated with postoperative cognitive dysfunction (POCD) in the elderly. Local anesthesia (LA) is an alternative pain control protocol that has not yet been fully evaluated. OBJECTIVES: To evaluate the feasibility of LA in PEILD compared with GA. STUDY DESIGN: A retrospective study. SETTING: This study took place at the First Affiliated Hospital of Harbin Medical University. METHODS: A total of 120 patients (aged 60-85 years) diagnosed with L5-S1 disc herniation and with American Society of Anesthesiologists fitness grade I or II between March 2016 and August 2017 were enrolled in the current study. Patients were randomly divided into LA group and GA group. For LA, 0.25% lidocaine was injected layer-by-layer into skin, subcutaneous tissue, fasciae, lumbar facet joint, muscle, and ligamentum flavum followed by injection of 1.33% lidocaine into epidural space; for GA, propofol, sufentanil, and cisatracurium were infused intravenously at 1 to 2 mg/kg, 0.3 µg/kg, and 0.15 mg/kg, respectively. Visual Analog Scale (VAS), Oswestry Disability Index (ODI), and MacNab Criteria (MNC) evaluated the feasibility of LA as pain control protocol in comparison to GA before and after operation. The development of POCD was assessed by the Mini-Mental State Examination 1 and 7 days postsurgery. Feasibility of LA as a pain control protocol was also evaluated by patient's willingness to receive the same surgical procedure immediately and 24 hours after the surgery, and intraoperative fluoroscopy use, blood loss, surgery duration, postoperative bed confinement, and duration and cost of hospital stay were also evaluated. RESULTS: Patients in both LA and GA groups had comparable VAS grade, ODI, and MNC pre- and post-PEILD, with significant pain reduction after operation. However, POCD developed only in GA group but not in LA group. In addition, compared with GA, LA group did not require postoperative bed confinement, had significantly shorter hospital stay, and lower hospital cost. Low intraoperative VAS grade and willingness to receive the same procedure reflected the acceptance of LA by patients. LIMITATIONS: The development of POCD was examined only 7 days after operation. The follow-up should be extended to 3 months and 2 years postoperation. CONCLUSIONS: LA has satisfactory pain control and low-risk of POCD in PEILD and is well accepted by patients. The benefits of LA are no postoperative bed confinement, faster recovery, shorter hospital stay, and lower hospital cost. KEY WORDS: L5-S1 disc herniation, older patients, percutaneous endoscopic interlaminar lumbar discectomy, local anesthesia, general anesthesia, postoperative cognitive dysfunction, American Society of Anesthesiologists grade, Oswestry Disability Index, MacNab Criteria, Mini-Mental State Examination.

Human adenovirus among hospitalized children with respiratory tract infections in Beijing, China, 2017-2018.

BACKGROUND: Human adenoviruses (HAdVs) cause a wide range of diseases. However, the genotype diversity and epidemiological information relating to HAdVs among hospitalized children with respiratory tract infections (RTIs) is limited. Here, we describe the epidemiology and genotype distribution of HAdVs associated with RTIs in Beijing, China. METHODS: Nasopharyngeal aspirates (NPA) were collected from hospitalized children with RTIs from April 2017 to March 2018. HAdVs were detected by a TaqMan-based quantitative real-time polymerase chain reaction (qPCR) assay, and the hexon gene was used for phylogenetic analysis. Epidemiological data were analyzed using statistical product and service solutions (SPSS) 21.0 software. RESULTS: HAdV was detected in 72 (5.64%) of the 1276 NPA specimens, with most (86.11%, 62/72) HAdV-positives cases detected among children < 6 years of age. HAdV-B3 (56.06%, 37/66) and HAdV-C2 (19.70%, 13/66) were the most frequent. Of the 72 HAdV-infected cases, 27 (37.50%) were co-infected with other respiratory viruses, most commonly parainfluenza virus (12.50%, 9/72) and rhinovirus (9.72%, 7/72). The log number of viral load ranged from 3.30 to 9.14 copies per mL of NPA, with no significant difference between the HAdV mono- and co-infection groups. The main clinical symptoms in the HAdV-infected patients were fever and cough, and 62 (86.11%, 62/72) were diagnosed with pneumonia. Additionally, HAdVs were detected throughout the year with a higher prevalence in summer. CONCLUSIONS: HAdV prevalence is related to age and season. HAdV-B and HAdV-C circulated simultaneously among the hospitalized children with RTIs in Beijing, and HAdV-B type 3 and HAdV-C type 2 were the most frequent.

Metabolomics analysis of the protective effect of rubusoside on palmitic acid-induced lipotoxicity in INS-1 cells using UPLC-Q/TOF MS.

Diabetes is one of the most severe chronic diseases worldwide. It is widely accepted that apoptosis of the pancreatic beta cell is an important cause for the induction of hyperglycemia and high levels of free fatty acids (FFAs), also called lipotoxicity associated with pancreatic beta cell dysfunction. Lipotoxicity has been proven to be an important pathogenic factor of diabetes. However, until now, the mechanism of FFA-induced lipotoxicity in INS-1 cells has not been fully understood. Current anti-diabetic drugs that protect islet cells are often toxic to healthy cells, resulting in negative side effects. Thus, there is an urgent need to identify more effective and safer anti-diabetic agents to protect pancreatic islet cells. Rubusoside (RUB) is a major ingredient in the leaves of Rubus suavissimus S. Lee, which decreases blood glucose levels by protecting pancreatic islet cells. However, the exact mechanism of this effect is unknown. In this study, metabolomics experiments based on UPLC-Q/TOF MS characterized a total of 15 metabolites as potential biomarkers associated with lipotoxicity induced by palmitic acid in INS-1 cells. According to the metabolic pathway analysis, pentose and glucuronate interconversions, and glycerophospholipid metabolism were recognized as the most influenced metabolic pathways associated with lipotoxicity. Unexpectedly, deviations of 14 metabolites in lipotoxic INS-1 cells were regulated by RUB, suggesting synergistic mediation of the abnormal metabolic pathways. The metabolomics strategy based on UPLC/Q-TOF MS analysis provides a new insight into the mechanisms of lipotoxicity induced by palmitic acid and the anti-lipotoxic activity of RUB in INS-1 cells.

miR-328-3p mediates the anti-tumor effect in osteosarcoma via directly targeting MMP-16.

BACKGROUND: Increasing reports demonstrated that dysregulated expression of microRNAs (miRNAs) leads to the progression of various tumors. Previous studies revealed that miR-328-3p exhibited dysregulated expression in various types of tumors. However, its function and underlying mechanism in osteosarcoma (OS) are still unexplored. METHODS: The expression of miR-328-3p in the tissues and OS cell lines was detected by qRT-PCR analysis. The effects of miR-328-3p in the proliferation were analyzed by MTT assay. The proliferation and apoptosis of OS cells were examined by colony formation assay and TUNEL staining respectively. The migration and tumor formation ability of OS cells were measured by wound healing assay and xenograft in vivo mice assay. Furthermore, the regulatory roles of miR-328-3p/MMP16 were determined by western blot and luciferase reporter assay. RESULTS: The expression of miR-328-3p was significantly decreased in OS tissues and cell lines. Furthermore, overexpression of miR-328-3p inhibited the cell proliferation and migration, but promoted the apoptosis of OS cells in vitro. Moreover, the analysis in vivo showed that miR-328-3p effectively suppressed the formation of tumors. According to the results of western blot analysis and luciferase reporter assay, we identified matrix metalloproteinase-16 (MMP-16) acted as a direct target of miR-328-3p. Moreover, the expression level of MMP-16, which participates in the occurrence and development of many cancers, was negatively correlated with the miR-328-3p expression in OS cells. CONCLUSION: miR-328-3p inhibited the proliferation, migration but accelerated the apoptosis of OS by directly inhibiting MMP-16. And miR-328-3p/MMP-16 axis may be one of the mechanisms of OS development and a novel potential method for the treatment of OS in clinic.

Quantitative detection of human Malawi polyomavirus in nasopharyngeal aspirates, sera, and feces in Beijing, China, using real-time TaqMan-based PCR.

BACKGROUND: Human Malawi polyomavirus (MWPyV) was discovered in 2012, but its prevalence and clinical characteristics are largely unknown. METHODS: We used real-time TaqMan-based PCR to detect MWPyV in the feces (n = 174) of children with diarrhea, nasopharyngeal aspirates (n = 887) from children with respiratory infections, and sera (n = 200) from healthy adults, and analyzed its clinical characteristics statistically. All the MWPyV-positive specimens were also screened for other common respiratory viruses. RESULTS: Sixteen specimens were positive for MWPyV, including 13 (1.47%) respiratory samples and three (1.7%) fecal samples. The samples were all co-infected with other respiratory viruses, most commonly with influenza viruses (69.2%) and human coronaviruses (30.7%). The MWPyV-positive children were diagnosed with bronchopneumonia or viral diarrhea. They ranged in age from 12 days to 9 years, and the most frequent symptoms were cough and fever. CONCLUSIONS: Real-time PCR is an effective tool for the detection of MWPyV in different types of samples. MWPyV infection mainly occurs in young children, and fecal-oral transmission is a possible route of its transmission.

Human polyomavirus type six in respiratory samples from hospitalized children with respiratory tract infections in Beijing, China.

BACKGROUND: HPyV6 is a novel human polyomavirus (HPyV), and neither its natural history nor its prevalence in human disease is well known. Therefore, the epidemiology and phylogenetic status of HPyV6 must be systematically characterized. METHODS: The VP1 gene of HPyV6 was detected with an established TaqMan real-time PCR from nasopharyngeal aspirate specimens collected from hospitalized children with respiratory tract infections. The HPyV6-positive specimens were screened for other common respiratory viruses with real-time PCR assays. RESULTS: The prevalence of HPyV6 was 1.7 % (15/887), and children ≤ 5 years of age accounted for 80 % (12/15) of cases. All 15 HPyV6-positive patients were coinfected with other respiratory viruses, of which influenza virus A (IFVA) (8/15, 53.3 %) and respiratory syncytial virus (7/15, 46.7 %) were most common. All 15 HPyV6-positive patients were diagnosed with lower respiratory tract infections, and their viral loads ranged from 1.38 to 182.42 copies/µl nasopharyngeal aspirate specimen. The most common symptoms were cough (100 %) and fever (86.7 %). The complete 4926-bp genome (BJ376 strain, GenBank accession number KM387421) was amplified and showed 100 % identity to HPyV6 strain 607a. CONCLUSIONS: The prevalence of HPyV6 was 1.7 % in nasopharyngeal aspirate specimens from hospitalized children with respiratory tract infections, as analyzed by real-time PCR. Because the coinfection rate was high and the viral load low, it was not possible to establish a correlation between HPyV6 and respiratory diseases.

Wnt4 signaling is associated with the decrease of proliferation and increase of apoptosis during age-related thymic involution.

The thymus is a central lymphoid organ that is responsible for T-lymphocyte development and maturation. Through negative and positive selection, lymphoid progenitor cells, which initiate from the bone marrow, develop into mature T cells in the thymus, and are subsequently involved in peripheral cell immunity. It has been reported that the Wnt signaling pathway exists widely in thymic epithelial cells and T lymphocytes. Wnt signaling affects the shape and function of thymic epithelial cells and has an important role in maintaining pro­T­cells, and in the subsequent T­cell differentiation. Previous studies have demonstrated that the Wnt signaling pathway participates in age­associated thymic involution. In the present study alterations in proliferation and apoptosis were investigated in murine thymic cells during aging. The results of the present study demonstrated that the aged thymus was characterized by markedly decreased cell numbers, as well as decreased proliferation and increased apoptosis. Concurrently, age­associated changes in thymic cell number and function were accompanied by a decrease in the transcription levels of Wnt4, and downregulation of forkhead box N1 and B­cell lymphoma­extra large, which are two target genes of the Wnt4 signaling pathway. In vitro studies demonstrated that activation of the Wnt4 signaling pathway promotes mouse thymus epithelial cell 1 (MTEC1) cell proliferation, and that Wnt4 signaling modulation alleviates dexamethasone­mediated MTEC1 cell apoptosis. These results suggest that normal expression levels of Wnt4 have a critical role in maintaining the balance between cell proliferation and apoptosis. Alterations in the Wnt signaling pathway may disrupt the epithelial network structure of the thymus, eventually leading to microenvironmental damage. Therefore, further studies regarding the effects of the Wnt signaling pathway on thymus development and age-related thymic involution, may be beneficial for improving the health conditions of the elderly.