Three-dimensional architecture of granulosa cell derived from oocyte cumulus complex, revealed by FIB-SEM.

The oocyte cumulus complex is mainly composed of an oocyte, the perivitelline space, zona pellucida and numerous granulosa cells. The cumulus granulosa cells (cGCs) provide a particularly important microenvironment for oocyte development, regulating its growth, maturation and meiosis. In this study, we studied the internal structures and cell-to-cell connections of mouse cGCs using focused ion beam scanning electron microscopy (FIB-SEM). We reconstructed three-dimensional models to display characteristic connections between the oocyte and cGCs, and to illustrate various main organelles in cGCs together with their interaction relationship. A special form of cilium identified in granulosa cell was never reported in previous literature.

SENP1 modulates chronic intermittent hypoxia-induced inflammation of microglia and neuronal injury by inhibiting TOM1 pathway.

Chronic intermittent hypoxia (CIH) is a characteristic pathophysiological change of obstructive sleep apnea syndrome (OSAS). Inflammation of microglia induced by CIH, plays a vital role in OSAS-associated cognitive dysfunction. SUMO-specific proteases 1 (SENP1) has been implicated in tumor inflammatory microenvironment and cells migration. However, the role of SENP1 in CIH-induced neuroinflammation remains unknown. We aimed to investigate the effect of SENP1 on neuroinflammation and neuronal injury. After the preparation of SENP1 overexpression microglia and SENP1 knockout mouse, CIH microglia and mice were established using an intermittent hypoxia device. Results showed that CIH reduced the level of SENP1 and TOM1, induced the SUMOylation of TOM1, and promoted microglial migration, neuroinflammation, neuronal amyloid-beta 42 (Aß42) deposition and apoptosis in vitro and in vivo. After SENP1 overexpression in vitro, the enhanced SUMOylation of TOM1 was inhibited; the level of TOM1 and microglial migration were enhanced; neuroinflammation, neuronal Aß42 deposition and apoptosis were significantly reduced. However, the administration of siRNA-TOM1 suppressed microglial migration, neuroinflammation, neuronal Aß42 deposition and apoptosis. After SENP1 knockout in vivo, the SUMOylation enhancement of TOM1 was accelerated, microglial migration was inhibited. Neuroinflammation, neuronal Aß42 deposition and apoptosis, cognitive impairment was significantly exacerbated. Overall, the results demonstrated that SENP1 promoted microglial migration by alleviating the de-SUMOylation of TOM1, thus contributing to attenuate neuroinflammation, neuronal Aß42 deposition and neuronal apoptosis induced by CIH.

Effect of Sperm 26S Proteasome on Embryo Formation and Development in In Vitro Fertilization.

The aim of this study was to evaluate the effects of sperm proteasome activity on fertilization outcome and embryo development after IVF. Following density gradient centrifugation for IVF purpose, the spermatozoa of 84 infertile patients with tubal factor were evaluated by luciferase enzymatic activity to assess the proteasome quantity. The mean age of patients was 33.8 years, and the mean concentration of human spermatozoa 26S proteasome was 674.53 ng/ml. After IVF, the embryos were scored for morphology. The spermatozoa proteasome activity was both positively correlated with fertilization rate in vitro (P = 0.0003) and 2PN rate (P = 0.0007). Compared to low fertilization rate group, the high fertilization rate group showed a significantly higher level of spermatozoa proteasome activity (P = 0.002). In conclusion, sperm proteasome activity provides additional data on sperm functional capacity in terms of fertilization during IVF.

Molecular epidemiology and clinical characterization of human rhinoviruses circulating in Shanghai, 2012-2020.

Human rhinoviruses (HRVs) cause acute upper and lower respiratory tract infections and aggravation of asthma and chronic obstructive pulmonary disease. The 5' untranslated region (5' UTR) and the VP4/VP2 region are widely used for genotyping of HRVs. Members of the species Rhinovirus A and Rhinovirus C have been reported to be more frequently associated with severe disease than members of the species Rhinovirus B. We report the clinical and molecular epidemiological characteristics of HRVs circulating from 2012 to 2020 in Shanghai. A total of 5832 nasopharyngeal swabs from patients with acute respiratory infections were collected. A real-time reverse transcription polymerase chain reaction assay was used for virus detection. The 5' untranslated region and VP4/VP2 region were amplified and sequenced for genotyping and phylogenetic analysis. The overall rate of rhinovirus detection was 2.74% (160/5832), with members of species A, B, and C accounting for 68.13% (109/160), 20.00% (32/160), and 11.88% (19/160) of the total, respectively. A peak of HRV infection was observed in autumn (5.34%, 58/1087). Patients in the 3- to 14-year-old age group were the most susceptible to HRV infection (χ2 = 23.88, P = 0.017). Influenza virus and Streptococcus pneumoniae were detected more frequently than other pathogens in cases of coinfection. Recombination events were identified in 10 strains, which were successfully genotyped by phylogenetic analysis based on the 5' UTR-VP4/VP2 region but not the 5' UTR region alone. We observed a high degree of variability in the relative distribution of HRV genotypes and the prevalence of HRV infection in Shanghai and found evidence of recombination events in the portion of the genome containing the 5' UTR and the VP4/VP2 region between HRV-C strains and HRV-A-like strains. This study is important for surveillance of the spread of HRVs and the emergence of new variants.

SENP1 modulates microglia-mediated neuroinflammation toward intermittent hypoxia-induced cognitive decline through the de-SUMOylation of NEMO.

Intermittent hypoxia (IH)-induced cognition decline is related to the neuroinflammation in microglia. SUMOylation is associated with multiple human diseases, which can be reversed by sentrin/SUMO-specific proteases 1 (SENP1). Herein, we investigated the role of SENP1 in IH-induced inflammation and cognition decline. BV-2 microglial cells and mice were used for inflammatory response and cognition function evaluation following IH treatment. Biochemical analysis and Morris water maze methods were used to elaborate the mechanism of SENP1 in IH impairment. Molecular results revealed that IH induced the inflammatory response, as evidenced by the up-regulation of NF-κB activation, IL-1ß and TNF-α in vitro and in vivo. Moreover, IH decreased the expression of SENP1, and increased the SUMOylation of NEMO, not NF-κB P65. Moreover, SENP1 overexpression inhibited IH-induced inflammatory response and SUMOylation of NEMO. However, the inhibitions were abolished by siRNA-NEMO. In contrast, SENP1 depletion enhanced IH-induced inflammatory response and SUMOylation of NEMO, accompanying with increased latency and reduced dwell time in mice. Overall, the results demonstrated that SENP1 regulated IH-induced neuroinflammation by modulating the SUMOylation of NEMO, thus activating the NF-κB pathway, revealing that targeting SENP1 in microglia may represent a novel therapeutic strategy for IH-induced cognitive decline.

SENP1-mediated NEMO de-SUMOylation inhibits intermittent hypoxia induced inflammatory response of microglia in vitro.

Among the seven small ubiquitin-like modifier (SUMO)-specific proteases (SENPs), our previous work showed that SENP1 suppressed nuclear factor kappa B (NF-κB) activation and alleviates the inflammatory response in microglia. However, the mechanism is still largely unknown. In this study, western blot analysis and enzyme-linked immunosorbent assay were utilized for evaluating the extent of NF-κB activation and expression of proinflammatory cytokines. qPCR and western blot analysis were performed to detect SENP1 expression. Coimmunoprecipitation followed by western blot analysis was applied to measure the changes in SUMOylation of NF-κB essential modulator (NEMO) and P65 in microglia with or without overexpression of SENP1. As the results, we found that intermittent hypoxia (IH) triggered the activation of NF-κB and upregulated the expression levels of tumor necrosis factor-α and interleukin-6. Interestingly, our data indicated that the SUMOylation of NEMO was enhanced by IH while SUMOylation of P65 was not affected. Further, our data showed that overexpression of SENP1 could decrease the extent of NF-κB activation and inhibit the inflammatory response of microglia through regulating the SUMOylation of NEMO. Collectively, this study presents the first report of the SENP1-controlled de-SUMOylation process of NEMO and its critical role in regulating NF-κB activation and proinflammatory cytokines secretion in microglia cells. This study would benefit for clarifying the role of SENP1 in IH-induced activation of microglia, thus providing potential therapeutic targets for obstructive sleep apnea treatment.

Systematic review of inhaled corticosteroid withdrawal effects in chronic obstructive pulmonary disease, and comparison with two "real-life" studies.

Chronic obstructive pulmonary disease (COPD) is common and preventable. The long-term safety of inhaled corticosteroid (ICS) use in COPD is still unclear and requires further investigation. This systematic review aimed to determine the effect of withdrawal of ICS use in COPD by examining randomized controlled trials (RCTs) and comparing their findings with those of "real-life" studies. Two independent reviewers searched for RCTs in the PubMed, EMBASE, and Cochrane databases and in CINAHL. Searches were conducted by using controlled vocabulary and free-text aliases for corticosteroids, COPD, and RCTs in each database. Data extraction was also conducted by the two reviewers. The main outcomes were exacerbations, lung function, health-related quality of life, symptoms, and exercise capacity. To assess the effect of ICS withdrawal more comprehensively, we also searched for "real-life" studies, and explored the reasons for different results among different trials. We located five RCTs, which met the inclusion criteria, and two "real-life" studies. Due to definitional and other discrepancies among trials, we could not perform a meta-analysis. In the RCTs, exacerbation was reported as an outcome in four out of five RCTs. Only one study showed that the risk of exacerbation did not increase after ICS withdrawal. Decrease in lung function from baseline was found in the withdrawal group in four trials, but only three trials found a statistically significant difference. All five trials compared differences in health-related quality of life. Two trials did not find significant changes, while a small but statistically significant difference in favor of the ICS group was observed in the other trials. In contrast, in the two real-life studies, no differences in forced expiratory volume in the first second (FEV1) values and exacerbation rate were observed between patients who were and were not withdrawn from ICS treatment. We concluded that the effect of ICS withdrawal on patients with COPD may be dependent on disease severity, use of background long-acting bronchodilator medication, and whether COPD is combined with airway hyper-responsiveness, among other factors.

Differences in airway remodeling and airway inflammation among moderate-severe asthma clinical phenotypes.

BACKGROUND: To identify asthma clinical phenotypes using cluster analysis and improve our understanding of heterogeneity in asthma. METHODS: Clustering approaches were applied to 203 patients who were diagnosed with asthma in XinHua Hospital (January 2012 to December 2015). One hundred and twenty patients underwent multi-slice spiral computed tomography (MSCT) examination and 30 underwent bronchial mucosal biopsy for evaluation of airway remodeling and airway inflammation among the phenotypes. RESULTS: Four groups were identified. Patients in cluster 1 (n=52) had early onset atopic asthma and patients in cluster 2 (n=65) had small airway obstruction and atopic asthma. Cluster 3 (n=52) was a unique group of patients with late-onset and non-atopic asthma. Patients in cluster 4 (n=34) had severe airflow obstruction and obvious airway remodeling as observed on MSCT (P<0.05). According to the immunohistochemistry of IL-5 and IL-17 (P<0.05), the results of clusters 1 and 2 may be attributable to the Th2 immune response, whereas those of clusters 3 and 4 to the Th17 immune response. CONCLUSIONS: Four distinct clinical phenotypes of asthma were identified by cluster analysis. The results of the MSCT and pathological examinations may suggest specific pathogeneses among the phenotypes.

Primary malignant melanoma of the lung: A case report and literature review.

RATIONALE: Primary malignant melanoma of the lung (PMML) is an extremely rare neoplasm with a dismal prognosis. The diagnosis of PMML is very difficult and is based on several clinical, radiological, and histopathological criteria. PATIENT CONCERNS: A 61-year-old women was admitted with a 2-month history of a productive cough and chest pain provoked by breathing and coughing. Computed tomography (CT) scans of the chest showed a large, solid tumor in the right middle lobe of the lung. Puncture biopsy of the right lung lesion was performed using B-ultrasound guidance, and immunohistochemical tests were performed. DIAGNOSES: The diagnosis of PMML was histopathologically confirmed by puncture biopsy with B-ultrasound guidance of the right lung lesion. INTERVENTIONS: The patient refused to receive surgery, adjuvant chemotherapy, or radiation therapy. OUTCOMES: The patient died 6 months after the diagnosis. LESSONS: The clinical manifestation and imaging features of PMML are not specific, and it does not differ from the more common primary bronchogenic carcinoma. In addition, it cannot be discriminated from other forms of primary melanoma according to its histology and immunohistochemistry. The treatment of choice is an aggressive surgical approach, combined with radiation therapy, chemotherapy, and immunotherapy.

Genetic diversity of Haemonchus contortus isolated from sympatric wild blue sheep (Pseudois nayaur) and sheep in Helan Mountains, China.

BACKGROUND: Haemonchus contortus is known among parasitic nematodes as one of the major veterinary pathogens of small ruminants and results in great economic losses worldwide. Human activities, such as the sympatric grazing of wild with domestic animals, may place susceptible wildlife hosts at risk of increased prevalence and infection intensity with this common small ruminant parasite. Studies on phylogenetic factors of H. contortus should assist in defining the amount of the impact of anthropogenic factors on the extent of sharing of agents such as this nematode between domestic animals and wildlife. METHODS: H. contortus specimens (n = 57) were isolated from wild blue sheep (Pseudois nayaur) inhabiting Helan Mountains (HM), China and additional H. contortus specimens (n = 20) were isolated from domestic sheep that were grazed near the natural habitat of the blue sheep. Complete ITS2 (second internal transcribed spacer) sequences and partial sequences of the nad4 (nicotinamide dehydrogenase subunit 4 gene) gene were amplified to determine the sequence variations and population genetic diversities between these two populations. Also, 142 nad4 haplotype sequences of H. contortus from seven other geographical regions of China were retrieved from database to further examine the H. contortus population structure. RESULTS: Sequence analysis revealed 10 genotypes (ITS2) and 73 haplotypes (nad4) among the 77 specimens, with nucleotide diversities of 0.007 and 0.021, respectively, similar to previous studies in other countries, such as Pakistan, Malaysia and Yemen. Phylogenetic analyses (BI, MP, NJ) of nad4 sequences showed that there were no noticeable boundaries among H. contortus populations from different geographical origin and population genetic analyses revealed that most of the variation (94.21%) occurred within H. contortus populations. All phylogenetic analyses indicated that there was little genetic differentiation but a high degree of gene flow among the H. contortus populations among wild blue sheep and domestic ruminants in China. CONCLUSIONS: The current work is the first genetic characterization of H. contortus isolated from wild blue sheep in the Helan Mountains region. The results revealed a low genetic differentiation and high degree of gene flow between the H. contortus populations from sympatric wild blue sheep and domestic sheep, indicating regular cross-infection between the sympatrically reared ruminants.

Is obstructive sleep apnea associated with cardiovascular and all-cause mortality?

BACKGROUND: Studies have reported inconsistent findings regarding the association between obstructive sleep apnea (OSA) and future risks of cardiovascular and all-cause mortality. We conducted a meta-analysis to investigate whether OSA is an independent predictor for future cardiovascular and all-cause mortality using prospective observational studies. METHODS: Electronic literature databases (Medline and Embase) were searched for prospective observational studies published prior to December 2012. Only observational studies that assessed baseline OSA and future risk of cardiovascular and all-cause mortality were selected. Pooled hazard risk (HR) and corresponding 95% confidence intervals (CI) were calculated for categorical risk estimates. Subgroup analyses were based on the severity of OSA. RESULTS: Six studies with 11932 patients were identified and analyzed, with 239 reporting cardiovascular mortality, and 1397 all-cause mortality. Pooled HR of all-cause mortality was 1.19 (95% CI, 1.00 to 1.41) for moderate OSA and 1.90 (95% CI, 1.29 to 2.81) for severe OSA. Pooled HR of cardiovascular mortality was 1.40 (95% CI, 0.77 to 2.53) for moderate OSA and 2.65 (95% CI, 1.82 to 3.85) for severe OSA. There were no differences in cardiovascular mortality in continuous positive airway pressure (CPAP) treatment compared with healthy subjects (HR 0.82; 95% CI, 0.50 to 1.33). CONCLUSIONS: Severe OSA is a strong independent predictor for future cardiovascular and all-cause mortality. CPAP treatment was associated with decrease cardiovascular mortality.

Clinical and pathological features and imaging manifestations of bronchial anthracofibrosis: the findings in 15 patients.

BACKGROUND: Bronchial anthracofibrosis (BAF) has been defined as a luminal narrowing associated with anthracotic pigmentation on bronchoscopy without a relevant history of pneumoconiosis or smoking. The aim of the study is to study the clinical features and imaging manifestations of BAF, and to promote the awareness of this disease. METHOD: Between October 2006 and January 2012, 15 patients were diagnosed at our department as BAF that showed a narrowing or obliteration of lobar or segmental bronchi with anthracotic pigmentation in the overlying mucosa on bronchoscopy. The medical records including clinical features, imaging manifestations, electronic bronchoscopic findings, and pathological features were analysed, and the literature was reviewed. RESULTS: A total of 15 patients were analyzed; 13 were female (86.7%) and two were male (13.3%) and the age range was from 62 to 86 years with a mean age of 74.5 years. Three cases (20.0%) had a history of tuberculosis. The most common clinical symptoms of BAF were cough (100%), expectoration (73.3%), dyspnea (60.0%), and fever (46.7%). Twelve cases displayed mild to moderate obstructive ventilatory dysfunction. In the electronic bronchoscopic evaluation, the most common findings were black bronchial mucosal pigmentation, bronchial stenosis, bronchial occlusion, and bronchial mucosal folds. Pathological evaluation revealed chronic inflammation of the bronchial mucosa, submucosal carbon particle deposition, and mucosal or submucosal fibrosis. Chest CT scans showed that 15 patients had bronchial stenosis or obstruction (direct signs) with the right middle lobe being the most common site (11 cases, 73.3%). The indirect sign was mainly the presence of bronchial obstructive diseases (including secondary infection), represented by 11 cases of pulmonary consolidation (73.3%), seven cases of atelectasis (46.7%), and five cases of nodules (33.3%). The CT mediastinal window showed bronchial lymph node lesions, mediastinal lymph node calcification (12 cases, 80.0%), and enlargement of multiple mediastinal lymph nodes. CONCLUSIONS: The diagnosis of BAF was mainly based on bronchoscopic evaluation. Its pathogenesis is currently unclear, although it may be related to tuberculosis or bio-fuel inhalation. The diagnosis of BAF has important clinical significance, and improved awareness of this disease will contribute to prevention of unnecessary thoracotomies.

Cigarette smoke-induced skeletal muscle atrophy is associated with up-regulation of USP-19 via p38 and ERK MAPKs.

Ubiquitin-specific proteases (USPs) deubiquitinate ubiquitin-protein conjugates in the ubiquitin-proteasome system. Previous research shows that ubiquitin-specific protease-19 (USP-19) is up-regulated in mammalian skeletal muscle in some degradative conditions, such as including fasting, diabetes, dexamethasone treatment, and cancer, and its function is associated with muscle atrophy. However, it is still unclear whether USP-19 is involved in muscle atrophy induced by chronic obstructive pulmonary disease. Rats exposed to chronic cigarette smoke and L6 myotubes incubated with cigarette smoke extract (CSE) were studied here. Using western blot analysis and quantitative real-time polymerase chain reaction (qPCR), we observed over-expression of USP-19 and down-regulation of myosin heavy chain (MHC) in both models. Moreover, CSE exposure inhibited myogenic differentiation and myotube formation in L6 myotubes. To explore the mechanism underlying these effects, we investigated the levels of phosphorylated mitogen-activated protein kinases (MAPKs) and total MAPKs. Exposing myotubes to CSE resulted in the general activation of MAPKs such as p38, JNK, and ERK1/2. The ERK inhibitor PD98059 and the p38 inhibitor SB203580 significantly blocked the increase in USP-19 gene expression induced by CSE. Our findings suggest that USP-19 is associated with muscle atrophy in response to cigarette smoke and is a potential therapeutic target. CSE promotes myotube wasting in culture partly by inhibiting myogenic differentiation and acts via p38 and ERK MAPK to stimulate expression of USP-19 in vitro.