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Membrane proteins tend to interact with each other in the cell membranes to form protein clusters and perform the corresponding physiological functions. However, because channel proteins are involved in many biological functions, their distribution and nano-organization in these protein clusters are unclear. To study the distribution patterns and relationships between the different channel proteins, we identified the locations of glucose transporter 1 (Glut1) and Band3 (anion transporter 1) precisely in the topography of the cytoplasmic side of the human red blood cell (hRBC) membranes using combined atomic force microscopy (AFM) and single-molecule localization microscopy (SMLM). The AFM results revealed that membrane proteins interacted with each other and aggregated into protein islands. The SMLM results showed that Glut1 and Band3 tended to form protein clusters in the hRBC membranes, and there was a strong colocalization between the two proteins. The results of the combined AFM and SMLM method indicated that the protein clusters of Glut1 and Band3 were mainly located in the protein islands of topography, and the protein islands in topography also interacted with each other to assemble into larger protein clusters or functional microdomains.

Background Lung cancer has the highest case fatality rate among cancers because of uncontrolled proliferation and early metastasis of cancer cells in the lung tissue. This study aimed to clarify the role of the non-SMC condensin I complex, subunit G (NCAPG) in lung adenocarcinoma (LUAD), explore the mechanisms of its progression, and lay the foundation for the search for new biological markers. Methods We analyzed overlapping differentially expressed genes (DEGs) from three datasets; a protein–protein interaction (PPI) network was subsequently constructed and analyzed using Cytoscape. We then selected NCAPG for validation because of its poor prognosis and because it has not been sufficiently studied in the context of LUAD. Immunohistochemical analysis was used to detect the expression of NCAPG in LUAD tissues, and the relationships between NCAPG and clinical parameters were analyzed. In vitro and in vivo experiments were conducted to verify the role of NCAPG in LUAD. Finally, we studied the specific mechanism of action of NCAPG in LUAD. Results Through comprehensive analysis of the GSE43458, GSE75037, and The Cancer Genome Atlas databases, we identified 517 overlapping DEGs. Among them, NCAPG was identified as a hub gene. Immunohistochemical analysis revealed that NCAPG was strongly associated with the clinical stage, M-classification, and N-classification. Univariate and multivariate Cox regression analyses indicated that NCAPG was a prognostic risk factor for LUAD, while the in vitro experiments showed that NCAPG overexpression promoted proliferation, migration, invasion, and epithelial-mesenchymal transition. Furthermore, knockdown of NCAPG inhibited LUAD progression, both in vitro and in vivo. Mechanistically, NCAPG overexpression increased p-Smad2 and p-Smad3 expressions in the transforming growth factor β (TGF-β) signaling pathway. Additionally, rescue experiments indicated that TGF-β signaling pathway inhibitors could restore the effect of NCAPG overexpression in LUAD cells. Conclusions NCAPG may promote proliferation and migration via the TGF-β signaling pathway in LUAD.

Hong Xue,1,2 Baosong Xie,1,2 Nengluan Xu,1,2 Hongru Li,1,2 Qianshun Chen,2,3 Weiping Xie,4 Hong Wang4 1Department of Respiratory and Critical Care Medicine, Fujian Provincial Hospital, Fuzhou, Fujian, People’s Republic of China; 2Provincial School of Clinical Medicine, Fujian Medical University, Fuzhou, Fujian, People’s Republic of China; 3Department of Thoracic Surgery, Fujian Provincial Hospital, Fuzhou, Fujian, People’s Republic of China; 4Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital with Nanjing Medical University, Nanjing, Jiangsu, People’s Republic of ChinaCorrespondence: Baosong Xie; Hong XueDepartment of Respiratory and Critical Care Medicine, Fujian Provincial Hospital, No. 134, East Street, Fuzhou, Fujian, 350000, People’s Republic of ChinaTel +86 591-88217531Email xiebaos_baosong@163.com; xuehong_xhx@163.comPurpose: Etanercept (ETN), a tumor necrosis factor-α (TNF-α) inhibitor, has been applied in the treatment of many diseases. However, whether it has effects on chronic obstructive pulmonary disease (COPD) and its interaction with tumor necrosis factor receptor 1 (TNFR1) remained unknown.Methods: Histopathological analysis of lung tissues from non-smokers and smokers with or without COPD was conducted using hematoxylin–eosin (H&E) staining, Van Gieson (VG) staining, and terminal transferase-mediated biotin dUTP nick end labeling (TUNEL). TNF-α content was measured using Immunohistochemistry. Correlation analysis among apoptosis rate, smoke index, the FEV1/FVC ratio, and TNF-α-positive cells was performed. After ETN treatment and transfection of overexpressed or silenced TNFR1, levels of inflammatory cytokines, apoptosis and related genes expressions in cigarette smoke extract (CSE)-treated human pulmonary artery endothelial cells (HPAECs) were detected using enzyme-linked immunosorbent assay (ELISA), Hoechst 33342 staining, flow cytometry, quantitative real-time PCR (qRT-PCR) and Western blot.Results: Pulmonary arterial remodeling and increased apoptotic and TNF-α+ HPAECs were found in lung tissue of smokers with or without COPD, with higher degrees in smokers with COPD. The numbers of apoptotic and TNF-α+ HPAECs were positively correlated with smoke index, while the FEV1/FVC ratio was negatively correlated with apoptotic HPAECs. In HPAECs, ETN downregulated the expressions of proteins related to CSE-induced apoptosis and the TNF receptor family, decreased CSE-induced cell apoptosis and inflammatory cytokine levels, and inhibited TNFR1 expression and p65 phosphorylation. Overexpressed TNFR1 reversed the effects of ETN on CSE-treated HPAECs, whereas silencing TNFR1 did the opposite.Conclusion: ETN protected HPAECs against CSE-induced inflammation and apoptosis via downregulating TNFR1, thus providing a potential therapy for smoking-induced COPD.Keywords: chronic obstructive pulmonary disease, etanercept, apoptosis, cigarette smoke extract, human pulmonary artery endothelial cells, tumor necrosis factor receptor 1

Blood glucose (BG) prediction is an effective approach to avoid hyper- and hypoglycemia, and achieve intelligent glucose management for patients with type 1 or serious type 2 diabetes. Recent studies have tended to adopt deep learning networks to obtain improved prediction models and more accurate prediction results, which have often required significant quantities of historical continuous glucose-monitoring (CGM) data. However, for new patients with limited historical dataset, it becomes difficult to establish an acceptable deep learning network for glucose prediction. Consequently, the goal of this study was to design a novel prediction framework with instance-based and network-based deep transfer learning for cross-subject glucose prediction based on segmented CGM time series. Taking the effects of biodiversity into consideration, dynamic time warping (DTW) was applied to determine the proper source domain dataset that shared the greatest degree of similarity for new subjects. After that, a network-based deep transfer learning method was designed with cross-domain dataset to obtain a personalized model combined with improved generalization capability. In a case study, the clinical dataset demonstrated that, with additional segmented dataset from other subjects, the proposed deep transfer learning framework achieved more accurate glucose predictions for new subjects with type 2 diabetes.

As an efflux pump, P-glycoproteins (P-gps) are over-expressed in many cancer cell types to confer them with multi-drug resistance. Many studies have focused on elucidating their molecular structure or protein expression; however, the relationship between the molecular assembly and dysfunction remains unclear. Super-resolution microscope is an excellent imaging tool to reveal the molecular biological details, but its high-quality imaging often suffers from the labeling method currently available. In this work, by exploiting its specificity and small size, tariquidar (specific inhibitor of P-gp) was modified by TAMRA to form a small chemical probe of P-gp. By direct stochastic optical reconstruction microscopic (dSTORM) imaging, tariquidar-TAMRA was first revealed to possess a higher labeling superiority and high binding specificity. Then, with the application of tariquidar-TAMRA labeling, we found that P-gps accumulate into larger and denser clusters on cancer cells and drug-resistant cells than on normal cells and drug-sensitive cells, indicating that P-gps can facilitate the pumping efficiency by aggregating together to form functional platforms. Moreover, these specific distribution patterns might serve as potential biomarkers for tumor and drug therapy screening.

Protein tyrosine kinase-7 (PTK7), as an important membrane receptor, regulates various cellular activities, including cell polarity, movement, migration, and invasion. Although lots of research studies focused on revealing its functions from the aspect of the expression of the gene and protein are present, the relationship between the spatial distribution at the single-molecule level and the function remains unclear. Through combining aptamer probe labeling and super-resolution imaging technology, after verifying the specificity and superiority of the aptamer probe, a more significant clustering distribution of PTK7 is found on the MCF10A cell basal membrane than on the apical membrane, which is thought to be related to their specific functions on different membranes. By exploring the relationship between the assembly of PTK7 and lipid rafts, actin cytoskeleton, and carbohydrate chains on the membrane, the unique distribution of PTK7 on disparate membranes is revealed to be probably because of the varied dominant position of these three factors. These findings present the detailed spatial information of PTK7 and the related potential organization mechanism on the cell membrane, which will facilitate a better understanding of the relationship between the molecular assembly and its function, as well as the overall structure of the cell membrane.

Lymphangioleiomyomatosis (LAM) is a rare disease affecting young women, which occurs sporadically or in patients with tuberous sclerosis complex (TSC). The main manifestations of TSC in the kidney include cysts and angiomyolipoma (AML). Although renal cell carcinoma (RCC) is not a manifestation of TSC, it has a 2-4% incidence in TSC patients. Furthermore, LAM is rare in patients with RCC. Herein, we present a case of a 40-year-old woman with LAM and RCC in the right kidney. We checked for mutations in the TSC1 and TSC2 genes from both blood and kidney lesions and found a heterozygous mutation of c.1717-30G> A in intron 16 of TSC2 gene. In TSC patients, the diagnosis of RCC is challenging because the cancer is rare, and it is often difficult to distinguish it from AML with conventional imaging techniques. Therefore, it is recommended that patients with TSC undergo renal imaging follow-ups annually for kidney masses.

With the stagnancy of antibiotics development, polymyxins have become the last defense for treatment of multidrug-resistant (MDR) Gram-negative bacteria, whereas the effect of polymyxin monotherapy is limited by resistance. The objective of this study was to evaluate the effects of polymyxin B (PMNB)-vorinostat (SAHA) combination therapy against Gram-negative pathogens in vitro and in vivo. The antibacterial activities of PMNB and SAHA were evaluated by susceptibility testing. The synergistic effect was assessed by checkerboard tests and time-killing kinetics experiments. Cellular morphology studies and reactive oxygen species (ROS) assay were conducted to explore potential mechanisms. Also, Galleria mellonella models were made to evaluate the antibacterial effects in vivo. PMNB-SAHA had the synergistic effect against all tested isolates, reducing >2 log10 colony-forming units (CFU)/mL at 40 minutes, and showed more powerful antibacterial effects than PMNB alone in the 24-hour window. Cellular morphology study showed the change of membrane and disruption of integrity. ROS assay showed more oxidative stress in combination than PMNB or SAHA monotherapy. In animal models, PMNB-SAHA showed a higher survival rate than that of monotherapy. This study is the first to report the synergistic antibacterial effect of PMNB-SAHA therapy against MDR Gram-negative bacteria. Further clinical research is needed to confirm the results.

Aims The emergence of carbapenem‐resistant Klebsiella pneumoniae (CRKP) strains has led to increased mortality and morbidity rates. Tigecycline, a new class of broad‐spectrum glycyl‐tetracycline antibiotics, has been used to target multi‐ and pan‐drug‐resistant bacterial infections. This study aimed to assess the molecular characteristics of CRKP in a tertiary hospital, and its susceptibility to tigecycline, to create a reference for hospital infection control and clinical drug use. Methods and Results We retrieved patient clinical information and CRKP characterization from medical records and detected the MIC of tigecycline using the micro‐broth dilution method. Multi‐locus sequence typing was performed, and antibiotic resistance genes associated with CRKP were detected by qPCR. A total of 166 CRKP strains were detected in the sputum, urine and blood among intensive care unit patients (average age, 69·6 years). The most infrequently observed resistance genes were amikacin resistance genes, followed by tobramycin resistance genes. KPC ‐2, CTX‐M9 and CTX‐M1 were the most frequently detected resistance genes. Conclusions No strain was resistant to tigecycline (MIC ≥ 8 µg ml−1). Twenty‐four sequence types were identified, with ST11 being the most common type. Significance and Impact of the Study Clinicians and infection control experts should be aware of CRKP prevalence to facilitate clinical treatment and improve nosocomial infection control.

The Wingless-type (Wnt) signaling pathway plays an important role in the development and progression of cancer. This study aimed to evaluate the relationship between single nucleotide polymorphisms (SNPs) in the Wnt pathway and the risk of bone metastasis in patients with non-small cell lung cancer (NSCLC). We collected 500 blood samples from patients with NSCLC and genotyped eight SNPs from four core genes (WNT2, AXIN1, CTNNB1 and APC) present within the WNT pathway. Moreover, we assessed the potential relationship of these genes with bone metastasis development. Our results showed that the AC/AA genotype of CTNNB1: rs1880481 was associated with a decreased risk of bone metastasis. Polymorphisms with an HR of < 1 had a cumulative protective impact on the risk of bone metastasis. Furthermore, patients with the AC/AA genotype of CTNNB1: rs1880481 was associated with Karnofsky performance status score, squamous cell carcinoma antigen and Ki-67 proliferation index. Lastly, patients with the AC/AA genotype of CTNNB1: rs1880481 had significantly longer median progression free survival time than those with the CC genotype. In conclusion, SNPs within the Wnt signaling pathway are associated with a decreased risk of bone metastasis, and may be valuable biomarkers for bone metastasis in patients with NSCLC.

Community-acquired pneumonia (CAP) is a lower respiratory tract infection. It is one of the commonest infectious diseases and the third leading cause of death worldwide. However, the epidemiological profiles of CAP in southeastern China are unknown. Data of inpatients and outpatients diagnosed with CAP from January 1, 2015 to December 31, 2020 were obtained from the National Healthcare Big Data in Fuzhou (Fuzhou Database). This database covers medical data from 37 hospitals and 159 community health service stations. The incidence rate, treatment pattern, and direct medical costs of CAP were assessed using clinical data. A total of 8,156,237 patients were enrolled, with a mean age of 33.72 ± 20.88 years. The overall incidence rate of CAP was 3.13 (95% confidence interval [CI]: 3.11–3.15) per 1000 person-years (PY), with 15.97 (95% CI: 15.85–16.08) per 1000 PY in children below 5 years old and 2.62 (95% CI: 2.57–2.66) per 1000 PY in the elderly ≥60 years. The cost per outpatient was $242.83 ± 341.62, and the cost per inpatient was $4,530.4 ± 9,151.68. The three most used therapeutic drugs in patients with CAP are cefotaxime, moxifloxacin, and azithromycin. In addition, despite the ability of both imported and domestic pneumococcal conjugate vaccines to reduce the incidence rate of CAP, the current vaccination coverage rates were relatively low. We suggest that more attention should be paid to the disease burden of CAP, especially due to its great economic burden in China.

ObjectiveTo evaluate the necessity of Covid-19 vaccination in children aged < 12 y by comparing the clinical characteristics in unvaccinated children aged < 12 y with vaccinated patients aged ≥ 12y during the Delta surge (B.1.617.2) in Putian, Fujian, China.MethodsA total of 226 patients with SARS-Cov-2 Delta variant (B.1.167.2; confirmed by Realtime PCR positive and sequencing) were enrolled from Sep 10th to Oct 20th, 2021, including 77 unvaccinated children (aged < 12y) and 149 people aged ≥ 12y, mostly vaccinated. The transmission route was explored and the clinical data of two groups were compared; the effect factors for the time of the nucleic acid negativization (NAN) were examined by R statistical analysis.ResultsThe Delta surge in Putian spread from children in schools to factories, mostly through family contact. Compared with those aged ≥ 12y, patients aged < 12y accounted for 34.07% of the total and showed milder fever, less cough and fatigue; they reported higher peripheral blood lymphocyte counts [1.84(1.32,2.71)×10^9/L vs. 1.31(0.94,1.85)×10^9/L; pp< 0.05]. Upon admission, their COVID19 antibodies (IgM and IgG) and IgG in convalescence were lower [0.13(0.00,0.09) vs. 0.12(0.03,0.41), pp pp=0.13).ConclusionChildren aged < 12y may be critical hidden spreaders, which indicates an urgent need of vaccination for this particular population.

Human epidermal growth factor receptor-2 (HER2) is a member of the epidermal growth factor receptor (HER) family that is involved in various biological processes such as cell proliferation, survival, differentiation, migration and invasion. It generally functions in the form of homo-/hetero-dimers or oligomers with other HER family members. Although its essential roles in cellular activities have been widely recognized, questions concerning the spatial distribution of HER2 on the membranes and the interactions between it and other ErbB family members remain obscure. Here, we obtained a high-quality dSTORM image of HER2 nanoscale clusters recognized by peptide probes, and found that HER2 forms clusters containing different numbers of molecules on cell membranes. Moreover, we found that HER2 and EGFR formed hetero-oligomers on non-stimulated cell membranes, whereas EGF stimulation reduced the degree of heteromerization, suggesting that HER2 and EGFR hetero-oligomers may inhibit the activation of EGFR. Collectively, our work revealed the clustered distribution of HER2 and quantified the changes of the interaction between HER2 and EGFR in the resting and active states at the single molecular level, which promotes a deeper understanding of the protein-protein interaction on cell membranes.

IntroductionMining knowledge from continuous glucose monitoring (CGM) data to classify highly heterogeneous patients with type 2 diabetes according to their characteristics remains unaddressed. A refined clustering method that retrieves hidden information from CGM data could provide a viable method to identify patients with different degrees of dysglycemia and clinical phenotypes.Research design and methodsFrom Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, we selected 908 patients with type 2 diabetes (18–83 years) who wore blinded CGM sensors (iPro2, Medtronic, California, USA). Participants were clustered based on CGM data during a 24-hour period by our method. The first level extracted the knowledge-based and statistics-based features to describe CGM signals from multiple perspectives. The Fisher score and variables cluster analysis were applied to fuse features into low dimensions at the second level. The third level divided subjects into subgroups with different clinical phenotypes. The four subgroups of patients were determined by clinical phenotypes.ResultsFour subgroups of patients with type 2 diabetes with significantly different statistical features and clinical phenotypes were identified by our method. In particular, individuals in cluster 1 were characterized by the lowest glucose level factor and glucose fluctuation factor, and the highest negative glucose factor and C peptide index. By contrast, cluster 2 had the highest glucose level factor and the lowest C peptide index. Cluster 4 was characterized by the greatest degree of glucose fluctuation factor, was the most insulin-sensitive, and had the lowest insulin resistance. Cluster 3 ranked in the middle concerning the CGM-derived metrics and clinical phenotypes compared with those of the other three groups.ConclusionA novel multilevel clustering approach for knowledge mining from CGM data in type 2 diabetes is presented. The results demonstrate that subgroups are adequately distinguished with notable statistical and clinical differences.

Purpose The mammalian target of rapamycin complex 1 (mTORC1) signaling pathway plays a vital role in cancer development and progression. This study aimed to investigate the relationship between genotype variants in mTORC1 pathway and the risk of brain metastasis (BM) in patients with non-small cell lung cancer (NSCLC). Methods We extracted genomic DNA from blood samples of 501 NSCLC patients and genotyped eight single-nucleotide polymorphisms (SNPs) in three core genes [mammalian target of rapamycin (mTOR), mammalian lethal with sec-13 protein 8 (mLST8) and regulatory-associated protein of mTOR (RPTOR)] of the mTORC1 pathway. The associations between these SNPs and the risk of BM development were assessed. Results The AG/GG genotype of mLST8:rs26865 and TC/CC genotype of mLST8:rs3160 were associated with an increased risk of BM [hazard ratios (HR) 2.938, 95% confidence interval (CI) 1.664–5.189, p p p 2), high level of squamous cell carcinoma (SCC) antigen and Ki-67 proliferation index. Moreover, patients with AG/GG genotype of mLST8:rs26865 had significantly lower median overall survival than those with AA genotype (12.1 months versus 21.6 months, p = 0.04). Conclusions Our results indicate that polymorphisms in mTORC1 pathway were significantly associated with increased risk of BM and may be valuable biomarkers to identify NSCLC patients with a high risk of BM.

Rationale Our pilot study suggested that noninvasive ventilation (NIV) reduced the need for intubation compared with conventional administration of oxygen on patients with “early” stage of mild acute respiratory distress syndrome (ARDS, PaO2/FIO2 between 200 and 300). Objectives To evaluate whether early NIV can reduce the need for invasive ventilation in patients with pneumonia-induced early mild ARDS. Methods Prospective, multicenter, randomized controlled trial (RCT) of NIV compared with conventional administration of oxygen through a Venturi mask. Primary outcome included the numbers of patients who met the intubation criteria. Results Two hundred subjects were randomized to NIV (n = 102) or control (n = 98) groups from 21 centers. Baseline characteristics were similar in the two groups. In the NIV group, PaO2/FIO2 became significantly higher than in the control group at 2 h after randomization and remained stable for the first 72 h. NIV did not decrease the proportion of patients requiring intubation than in the control group (11/102 vs. 9/98, 10.8% vs. 9.2%, p = 0.706). The ICU mortality was similar in the two groups (7/102 vs. 7/98, 4.9% vs. 3.1%, p = 0.721). Multivariate analysis showed minute ventilation greater than 11 L/min at 48 h was the independent risk factor for NIV failure (OR, 1.176 [95% CI, 1.005–1.379], p = 0.043). Conclusions Treatment with NIV did not reduce the need for intubation among patients with pneumonia-induced early mild ARDS, despite the improved PaO2/FIO2 observed with NIV compared with standard oxygen therapy. High minute ventilation may predict NIV failure. Trial registration NCT01581229. Registered 19 April 2012 Electronic supplementary material The online version of this article (10.1186/s13054-019-2575-6) contains supplementary material, which is available to authorized users.

Objective: Ciprofloxacin resistance (CIPR) for Shigella isolates is becoming more prevalent. This study systematically investigated the antibacterial activity of ciprofloxacin (CIP)/fosfomycin (FOS) combination in vitro and in vivo against CIPR S. flexneri isolates. Method: Eighty CIPR S. flexneri isolates were selected for synergy studies by the microtiter plate checkerboard assay. Two S. flexneri isolates ({"type":"entrez-nucleotide","attrs":{"text":"GN120471","term_id":"227482297","term_text":"GN120471"}}GN120471, CIPRFOSR; GN120454, CIPRFOSS) were used to investigate the efficacy of the CIP/FOS combination by the time-kill methodology. Clinically relevant concentrations (CIP, 0.5, 1, or 2.5 μg/mL; FOS, 30, 150, or 300 μg/mL) were combined, and the colony counts were conducted at 3, 5, 8, and 24 hours. The in vivo activity of the CIP/FOS combination was assessed using a Galleria mellonella larvae model. Results: In checkerboard assays, 31 strains (38.75%) showed synergy for the CIP/FOS combination. For the isolate {"type":"entrez-nucleotide","attrs":{"text":"GN120471","term_id":"227482297","term_text":"GN120471"}}GN120471, monotherapy with CIP or FOS at all concentrations produced little or no bacterial killing, while the CIP/FOS combination produced enhanced bacterial killing with FOS concentrations of 150 and 300 μg/mL, especially when combined with CIP at 2.5 μg/mL. For the isolate GN120454, the CIP/FOS combination at all concentrations produced more rapid and extensive killing (up to 5log10 colony forming units (CFU)/mL with many combinations) than with either antibiotic alone. Mortality at 96 hours was around 80% at approximately 104 CFU/larva for {"type":"entrez-nucleotide","attrs":{"text":"GN120471","term_id":"227482297","term_text":"GN120471"}}GN120471 and GN120454. When CIP at 2.5 μg/mL was combined with FOS at 150 μg/mL for the bactericidal activity in vivo, the survival rates for CIP/FOS combination against {"type":"entrez-nucleotide","attrs":{"text":"GN120471","term_id":"227482297","term_text":"GN120471"}}GN120471-infected and GN120454-infected larvae were significantly higher than that of CIP (68.75% vs 25%, P=0.013; 81.25% vs 37.5%, P=0.012, respectively). Conclusion: Against CIPR S. flexneri isolates, the CIP/FOS combination induced synergy, and increased bacterial killing in vitro and in a simple invertebrate model of infection.

Objective:The relationship between OCT4 and clinicopathological features in lung cancer is shown to be controversial in recent publications. Therefore, we conducted this meta-analysis to quantitatively investigate the prognostic and clinicopathological characteristics of OCT4 in lung cancer.Methods:A comprehensive literature search of the PubMed, EMBASE, Cochrane Library, WOS, CNKI and Wanfang databases was performed to identify studies. Correlations between OCT4 expression and survival outcomes or clinicopathological features were analyzed using meta-analysis methods.Results:Twenty-one studies with 2523 patients were included. High OCT4 expression showed a poorer overall survival (OS) (univariate: HR= 2.00, 95% CI = (1.68, 2.39), pConclusion:OCT4 over-expression in lung cancer was strongly related to poorer clinicopathological features and worse survival outcomes, which suggests that OCT4 could be a valuable prognostic marker in lung cancer.

BackgroundEarly alarm of hypoglycemia, detection of asymptomatic hypoglycemia, and effective control of blood glucose fluctuation make a great contribution to diabetic treatment. In this study, we designed a multi-level hypoglycemia early alarm system to mine potential information in Continuous Glucose Monitoring (CGM) time series and improve the overall alarm performance for different clinical situations.MethodsThrough symbolizing the historical CGM records, the Prefix Span was adopted to obtain the early alarm/non-alarm frequent sequence libraries of hypoglycemia events. The longest common subsequence was used to remove the common frequent sequence for achieving the hypoglycemia early alarm in different clinical situations. Then, the frequent sequence pattern libraries with different risk thresholds were designed as the core module of the proposed multi-level hypoglycemia early alarm system.ResultsThe model was able to predict hypoglycemia events in the clinical dataset of level-I (sensitivity 85.90%, false-positive 23.86%, miss alarm rate 14.10%, average early alarm time 20.61 min), level-II (sensitivity 80.36%, false-positive 17.37%, miss alarm rate 19.63%, average early alarm time 27.66 min), and level-III (sensitivity 78.07%, false-positive 13.59%, miss alarm rate 21.93%, average early alarm time 33.80 min), respectively.ConclusionsThe proposed approach could effectively predict hypoglycemia events based on different risk thresholds to meet different prevention and treatment requirements. Moreover, the experimental results confirm the practicality and prospects of the proposed early alarm system, which reflects further significance in personalized medicine for hypoglycemia prevention.

Background: The membrane lipid ceramide plays important roles in regulating tumor growth, chemotherapy drug resistance, and apoptosis. However, the mechanisms through which ceramide induces apoptosis are still unclear. In this study, we explored the biological functions and underlying mechanism of C6-ceramide in brain metastasis (BM) arising from non-small cell lung cancer (NSCLC).Methods: The effects of C6-ceramide on cell apoptosis were studied by flow cytometry. Cell Counting Kit-8 assays, wound-healing assays, and flow cytometry were performed to investigate the biological functions of C6-ceramide. An in vitro blood-brain barrier (BBB) model was constructed, and its effectiveness and availability were tested by evaluating horse radish peroxidase activity and junction-related protein expression. The underlying signaling pathways of C6-ceramide were detected by western blotting, and further verification was performed using pathway inhibitors. RNA sequencing was used to confirm the involvement of C6-ceramide and associated pathways in BM arising from NSCLC.Results: C6-ceramide induced apoptosis in NSCLC cells, and the optimal working concentration of C6-ceramide was 50 μM. C6-ceramide not only suppressed cell proliferation and migration but also arrested cells at the G1/S transition. An in vitro BBB model constructed using human umbilical vein endothelial cells and human astrocytes cells showed the lowest permeability and the strongest tight junction connections when cells were cocultured for 72 h. Moreover, C6-ceramide suppressed the passage of cells through the BBB model. C6-ceramide at least partially influenced NSCLC biological functions by downregulating the phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) signaling pathway, which was further confirmed by rescue assays and RNA sequencing. Conclusions: C6-ceramide functioned as a tumor suppressor by inducing NSCLC cell apoptosis and BM through the PI3K/AKT/mTOR signal pathway. Thus, this pathway may serve as a novel potential therapeutic target for patients with BM arising from NSCLC.

HIV-1 infection is enhanced by cell-cell adhesions between infected and uninfected T cells called virological synapses (VS). VS are initiated by the interactions of cell-surface HIV-1 envelope glycoprotein (Env) and CD4 on target cells and act as sites of viral assembly and viral transfer between cells. To study the process that recruits and retains HIV-1 Env at the VS, a replication-competent HIV-1 clone carrying an Env-sfGFP fusion protein was designed to enable live tracking of Env within infected cells. Using surface pulse-labeling of Env and fluorescence recovery after photobleaching (FRAP) studies, we observed targeted accumulation and sustained recycling of Env between the endocytic recycling compartment (ERC) and the VS. We observed dynamic exchange of Env at the VS while the viral structural protein, Gag, was largely immobile at the VS. The disparate exchange rates of Gag and Env at the synapse indicate that retention of Env is not likely to be maintained by entrapment into an immobile Gag lattice or through immobilizing interactions with CD4 on the target cell. A FRAP study of an Env endocytosis mutant showed that recycling is required for the rapid exchange of Env at the VS. We conclude that the mechanism of Env accumulation at the VS and incorporation into nascent particles involves continuous internalization and targeted secretion rather than irreversible interactions with the budding virus.

As continuous glucose monitoring (CGM) systems provide critical feedback information of blood glucose concentration to the artificial pancreas for patients with type 1 diabetes (T1D), faults in CGM may seriously affect the computation of insulin infusion rates which can lead to fatal consequences accompany with hypoglycemia or hyperglycemia. In the present work, the k-medoids clustering algorithm is modified by calculating cluster number with a Bayesian Information Criterion (BIC)-based cost function and the SAC (SSE-ASW Criterion) evaluation coefficient which considers both SSE (Sum of Square due to Error) and ASW (Average Silhouette Width) criteria. Then, the modified k-medoids clustering algorithm is proposed to detect sensor failures online with CGM measurements. Different from the qualitative model-based methods and quantitative model-based methods, sufficient clean data are the only requirement of the proposed method. During online monitoring, the new glycemic variability is then tracked against predefined confidence limits during training period to indicate abnormality. The feasibility of the proposed method is successfully assessed using CGM data collected from the UVa/Padova metabolic simulator.

Objectives: Carbapenem-resistant Klebsiella pneumoniae (CRKP) have undergone extensive dissemination in worldwide resulting in increased mortality. We performed a retrospective analysis of epidemiology and risk factors for CRKP infection in a general teaching hospital in China.Methods: A molecular and clinical study were conducted for 98 CRKP in a tertiary hospital from January 2013 to December 2016. Carbapenemase gene detection, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were performed. Logistic regression was also used to identify the risk factors associating with the 30-day mortality.Results: The producition of KPC cabapenemase was the main resistant mechanism and increased annually with a significant difference. However, the molecular outcome revealed the dominance and diversity in CRKP with 24 sequence types (STs) and 59 PFGE types (PTs ). The ST11 CRKP were documented as the predominant strains in our study, which showed a significant increasing trend year by year. Additionally, the predominant ST11 CRKP corresponding to PT10 and PT15 remained a typical fashion. Of note, the new advantage PT09 or PT16 were just discovered in 2016 which also corresponding to ST11. Meanwhile, the factors on 30-day mortality and ST11 proportionality with CRKP infection were assessed, which showed that ST11, appropriate empirical treatment, and hospital stays were independently associated with 30-day mortality. Conclusions: ST11 CRKP payed a dominant role in the process, but the homology of these strains was polymorphic and the advantage clusters would be changed by evolution. Additionally, besides appropriate empirical treatment and hospital stays, ST11 CRKP was independently associated with the 30-day mortality, first reported as we know.

Klebsiella (K.) pneumoniae is a common cause of pneumonia-derived sepsis in human and is associated with high morbidity and mortality. The microbiota promotes and maintains host immune homeostasis during bacterial infections. However, the mechanisms by which the gut microbiota affects immune responses in the lung still remain poorly understood. Here, we performed cecal metabolomics sequencing and fecal 16s rRNA sequencing in K. pneumoniae-infected mice and uninfected controls and showed that K. pneumoniae infection led to profound alterations in the gut microbiome and thus the cecal metabolome. We observed that the levels of Lactobacillus reuteri and Bifidobacterium pseudolongum were significantly decreased in K. pneumoniae-infected mice. Spearman correlation analysis showed that alterations in the richness and composition of the gut microbiota were associated with profound changes in host metabolite concentrations. Further, short-chain fatty acids (SCFAs), including acetate, propionate, and butyrate, were detected in cecal contents and serum by gas chromatography-mass spectrometry (GC-MS). We observed that the concentrations of these three SCFAs were all lower in the infected groups than in the untreated controls. Lastly, oral supplementation with these three SCFAs reduced susceptibility to K. pneumoniae infections, as indicated by lower bacterial burdens in the lung and higher survival rates. Our data highlight the protective roles of gut microbiota and certain metabolites in K. pneumoniae-pneumonia and suggests that it is possible to intervene in this bacterial pneumonia by targeting the gut microbiota.

BackgroundSince the end of 2019 to the present day, the outbreak of the coronavirus disease 2019 (COVID-19) has had an immense impact on China and on other countries worldwide. This outbreak represents a serious threat to the lives and health of people all around the world. The epidemic first broke out in Wuhan, where the Chinese government was unable to prevent the spread of the disease by implementing home quarantine measures. Mobile cabin hospitals were used to relieve pressure on hospitals due to the need for beds while also isolating the sources of the infection through a centralized quarantine and treatment of mild cases.MethodThis paper reviewed and summarized the treatment of patients with mild illness and symptoms during the period from the construction to the closing of the Hanyang Mobile Cabin Hospital in Wuhan, China, and presented the operational elements and possible improvements of running this hospital.ResultsMobile cabin hospitals helped China to curb the epidemic in only 2 incubation periods in 28 days.The basic conditions required for a normal operation of mobile cabin hospitals included the selection of the environment, medical staff to patient ratio, organizational structure, management model, admission criteria, treatment approaches, discharge process, livelihood guarantee, security, and other safeguarding measures. All of these components were performed carefully in Wuhan Hanyang Mobile Cabin Hospital, without medical staff being infected.ConclusionThe mobile cabin hospital compulsory quarantine for mild patients was an alternative method to combat COVID-19. It is hoped that the presented work in this manuscript can serve as a reference for the emergency prevention and control measures for global epidemic outbreaks.

Objective The aims of this study were to describe azithromycin (AZM) susceptibility patterns among Shigella isolates in Anhui, China and identify predictors of resistance with mobile element-mediated genes. Methods A total of 517 non-duplicate Shigella isolates (449 S. flexneri and 68 S. sonnei) were collected in the Anhui Province of China from September 2011-September 2015, and screened for the plasmid-mediated genes of decreased susceptibility to AZM (DSA), using polymerase chain reaction amplification and sequencing. Conjugation experiments and pulsed-field gel electrophoresis were conducted for all mphA-positive DSA isolates. Results The DSA rate for 449 S. flexneri isolates was 33.6%, compared with 39.7% for 68 S. sonnei isolates. Among 161 DSA S. flexneri isolates, 93 (57.8%) carried the mphA gene. Among 27 DSA S. sonnei isolates, 11 (40.7%) carried the mphA gene. However, other plasmid-mediated DSA genes were not found in these isolates. A total of 89 transconjugants (95.7%) were obtained from 93 mphA-positive S. flexneri isolates through conjugation, and 10 transconjugants (90.9%) were obtained from 11 mphA-positive S. sonnei isolates. Furthermore, the minimum inhibitory concentrations (MICs) of AZM among 89 S. flexneri transconjugants ranged from 4 to 128 μg/mL, with an MIC50 of 8 μg/mL and MIC90 of 32 μg/mL. The MICs of AZM among 10 S. sonnei transconjugants ranged from 4 to 256 μg/mL, with an MIC50 of 8 μg/mL and MIC90 of 64 μg/mL. Thirteen clusters were found for mphA-positive S. flexneri, and five clusters were found for mphA-positive S. sonnei. Furthermore, 10 homologous isolates among 13 mphA-positive S. flexneri isolates with high-level DSA were from Sixian county and were multidrug-resistant strains. Of the 10 homologous S. flexneri isolates, eight were from children (≤5 years old), and two from the elderly (>60 years old). Conclusion Our study demonstrates that the DSA for Shigella isolates was severe, and the plasmid-mediated mphA gene was the most common macrolide resistance gene detected in Shigella isolates collected in Anhui, China. The mphA-positive S. flexneri isolates with high-level DSA facilitated clonal spread in children and the elderly. This finding is noteworthy and warrants further study.

Traffic accidents, falls, and many other events may cause traumatic spinal cord injuries (SCIs), resulting in nerve cells and extracellular matrix loss in the spinal cord, along with blood loss, inflammation, oxidative stress (OS), and others. The continuous development of neural tissue engineering has attracted increasing attention on the application of fibrin hydrogels in repairing SCIs. Except for excellent biocompatibility, flexibility, and plasticity, fibrin, a component of extracellular matrix (ECM), can be equipped with cells, ECM protein, and various growth factors to promote damage repair. This review will focus on the advantages and disadvantages of fibrin hydrogels from different sources, as well as the various modifications for internal topographical guidance during the polymerization. From the perspective of further improvement of cell function before and after the delivery of stem cell, cytokine, and drug, this review will also evaluate the application of fibrin hydrogels as a carrier to the therapy of nerve repair and regeneration, to mirror the recent development tendency and challenge.

Spinal cord injury (SCI) is a common issue in the clinic that causes severe motor and sensory dysfunction below the lesion level. FTY720, also known as fingolimod, has recently been reported to exert a positive effect on the recovery from a spinal cord injury. Through local delivery to the lesion site, FTY720 effectively integrates with biomaterials, and the systemic adverse effects are alleviated. However, the effects of the proper mass ratio of FTY720 in biomaterials on neural stem cell (NSC) proliferation and differentiation, as well as functional recovery after SCI, have not been thoroughly investigated. In our study, we fabricated electrospun poly (lactide-co-glycolide) (PLGA)/FTY720 scaffolds at different mass ratios (0.1%, 1%, and 10%) and characterized these scaffolds. The effects of electrospun PLGA/FTY720 scaffolds on NSC proliferation and differentiation were measured. Then, a rat model of spinal transection was established to investigate the effects of PLGA/FTY720 scaffolds loaded with NSCs. Notably, 1% PLGA/FTY720 scaffolds exerted the best effects on the proliferation and differentiation of NSCs and 10% PLGA/FTY720 was cytotoxic to NSCs. Based on the Basso, Beattie, and Bresnahan (BBB) score, HE staining and immunofluorescence staining, the PLGA/FTY720 scaffold loaded with NSCs effectively promoted the recovery of spinal cord function. Thus, FTY720 properly integrated with electrospun PLGA scaffolds, and electrospun PLGA/FTY720 scaffolds loaded with NSCs may have potential applications for SCI as a nerve implant.

HIV-1 transmission is usually initiated by a single viral strain called transmitted/ founder (T/F) virus. In in vitro models, HIV-1 can efficiently spread via cell-free and virological synapse (VS)-mediated cell-to-cell infection. Both modes of infection require the viral glycoprotein Envelope (Env). The efficiency with which T/F Envs initiate VS and mediate cell-to-cell infection has not been well characterized. Here we tested a panel of isogenic HIV-1 molecular clones that carry different Clade B T/F Envs. We found that despite variable infectivity among different Env clones in the two modes of infection, T/F Envs generally mediated efficient VS formation and subsequent cell-to-cell transfer. In contrast, in vitro infectivity of the T/F Env clones was more variable and strongly correlated with intrinsic fusogenicity of various Envs. We speculate that the conservation of cell-to-cell transfer by T/F Env is indicative of a biologically important function of Env.

The orderly organelle interaction network is essential for normal biological activity of cells. However, the mechanism of orderly organelle interaction remains elusive. In this report, we analyzed the structure characteristics of the cell membrane, endocytic vesicles, and the Golgi membrane through a high-resolution imaging technique and further comprehensively investigated the vesicle-transport process via epidermal growth factor receptor endocytosis and a recycling pathway using a real-time fluorescence tracing method. Our data suggest that orderly vesicle transport is due to protein protrusion from the outer surface of endocytic vesicles and that full membrane fusion between homotypic endocytic vesicles is a result of the rough outer surface. Finally, the kiss-and-run method, which is utilized by endocytic vesicles to communicate with the trans-Golgi network (TGN) is attributed to a dense protein layer at the outer surface of the TGN. In summary, by combining static structural analysis with dynamic tracing, we elucidate the mechanism of orderly vesicle transport from the overall structural features of the membrane. This work provides insight into the structural mechanisms underlying vital biological processes involving organelle interactions at the molecular level.

Background A novel coronavirus (CoV), severe acute respiratory syndrome (SARS)–CoV-2, has infected >75 000 individuals and spread to >20 countries. It is still unclear how fast the virus evolved and how it interacts with other microorganisms in the lung. Methods We have conducted metatranscriptome sequencing for bronchoalveolar lavage fluid samples from 8 patients with SARS–CoV-2, and also analyzed data from 25 patients with community-acquired pneumonia (CAP), and 20 healthy controls for comparison. Results The median number of intrahost variants was 1–4 in SARS–CoV-2–infected patients, ranged from 0 to 51 in different samples. The distribution of variants on genes was similar to those observed in the population data. However, very few intrahost variants were observed in the population as polymorphisms, implying either a bottleneck or purifying selection involved in the transmission of the virus, or a consequence of the limited diversity represented in the current polymorphism data. Although current evidence did not support the transmission of intrahost variants in a possible person-to-person spread, the risk should not be overlooked. Microbiotas in SARS–CoV-2–infected patients were similar to those in CAP, either dominated by the pathogens or with elevated levels of oral and upper respiratory commensal bacteria. Conclusion SARS–CoV-2 evolves in vivo after infection, which may affect its virulence, infectivity, and transmissibility. Although how the intrahost variant spreads in the population is still elusive, it is necessary to strengthen the surveillance of the viral evolution in the population and associated clinical changes.

Present coal seams in China are characterized by great gas pressure and content, low permeability features, and thus, the extraction of gas is difficult and gas disaster remains serious. To improve the permeability of coal seam, the hydraulic flushing is adopted, so the effect evaluation of stree relief and permeability enhancement will be necessary before gas extraction. In this paper, an electromagnetic radiation (EMR) experiment system of coal seam hydraulic flushing was established, and the EMR characteristics of coal during hydraulic flushing were studied. The EMR test of hydraulic flushing was carried out in the Liangbei coal mine, China, and the gas extraction flow and coal stress were measured simultaneously. And then, the variation laws of EMR and the effect evaluation of hydraulic flushing were analyzed. The results showed that coal deformation and fracture produced abundant EMR signals during hydraulic flushing. The variation of the EMR signals was synchronous with the variation of load, and the large the load variation was, the greater the EMR intensity and count were. Compared with before hydraulic flushing, the EMR signals changed significantly after hydraulic flushing, and the variation amplitude of the EMR intensity was consistent with the variation amplitude of the coal stress and gas extraction flow. So the EMR method achieved the objective of effect evaluation. It's feasible and dependable to evaluate the effect of coal seam hydraulic flushing by EMR.

Neuronal loss and the formation of glial scars are major pathological events that hinder axonal regrowth and neural regeneration after severe spinal cord injury (SCI). Therapeutic therapies that include both cell transplantation and reducing glial scarring is likely required to achieve substantial functional recovery after severe SCI. In this study, a dual-drug enhanced injectable hydrogel was designed and combined with neural stem cells (NSCs-cfGel) to enhance tissue regeneration in a rat complete transected SCI model. The injectable hydrogel was crosslinked by reversible hydrazone bonds, and the mechanical properties and degradation abilities were analyzed both in vitro and in vivo. When incorporated with two drugs (cetuximab and FTY720), the injectable hydrogel improved both the proliferation and neuronal differentiation and exhibited inhibitory astrocytic differentiation of NSCs in vitro. After transplantation into a rat complete transected SCI model, NSCs-cfGels not only significantly enhanced neuronal differentiation and integration of the grafted NSCs, but also inhibited glial scar formation, and promoted axonal regeneration and nerve circuit reconstruction. Consequently, SCI rats that received NSCs-cfGel implantation exhibited optimal functional improvement. Taken together, this study demonstrates a promising dual-drug enhanced injectable hydrogel for exogenous NSC delivery and thus provides a novel strategy for combination therapy of SCI.

A novel coronavirus (CoV), severe acute respiratory syndrome (SARS)-CoV-2, has infected75 000 individuals and spread to20 countries. It is still unclear how fast the virus evolved and how it interacts with other microorganisms in the lung.We have conducted metatranscriptome sequencing for bronchoalveolar lavage fluid samples from 8 patients with SARS-CoV-2, and also analyzed data from 25 patients with community-acquired pneumonia (CAP), and 20 healthy controls for comparison.The median number of intrahost variants was 1-4 in SARS-CoV-2-infected patients, ranged from 0 to 51 in different samples. The distribution of variants on genes was similar to those observed in the population data. However, very few intrahost variants were observed in the population as polymorphisms, implying either a bottleneck or purifying selection involved in the transmission of the virus, or a consequence of the limited diversity represented in the current polymorphism data. Although current evidence did not support the transmission of intrahost variants in a possible person-to-person spread, the risk should not be overlooked. Microbiotas in SARS-CoV-2-infected patients were similar to those in CAP, either dominated by the pathogens or with elevated levels of oral and upper respiratory commensal bacteria.SARS-CoV-2 evolves in vivo after infection, which may affect its virulence, infectivity, and transmissibility. Although how the intrahost variant spreads in the population is still elusive, it is necessary to strengthen the surveillance of the viral evolution in the population and associated clinical changes.

The outbreaks of 2019 novel coronavirus disease (COVID-19) caused by SARS-CoV-2 infection has posed a severe threat to global public health It is unclear how t

Background The increase in carbapenem-resistant Klebsiella pneumoniae (CRKP), especially the emergence of tigecycline-resistant K. pneumoniae (KP), is a serious public health concern. However, the underlying mechanism of tigecycline resistance is unclear. In this study, we evaluated the role of the CusS-CusR two-component system (TCS), which is associated with copper/silver resistance, in tigecycline resistance in CRKP. Methods Following the in vitro evolution of tigecycline-resistant KP, the minimum inhibitory concentrations of tigecycline were determined using the micro-broth dilution method. RNA sequencing and data analysis were performed to identify differentially expressed genes. Quantitative PCR (qPCR) was performed to verify the genes of interest. Genes associated with tigecycline resistance, such as ramR, tex (T), and tet (A), were detected by PCR, and then mutants were confirmed by sequencing. Additionally, the efflux pump-associated genes soxS, oqxA, oqxB, acrE, and acrF were also analyzed by qPCR. CusR was deleted and complemented by the suicide vector pKO3-Km plasmid and pGEM-T-easy plasmid, respectively. Results Nine strains of KP were evaluated in our study. Strains A2 and A3 were evolved from A1, B2, and B3 were evolved from B1, and C2 and C3 were evolved from C1. The tigecycline minimum inhibitory concentration for A1, B1, and C1 was 0.5 μg/mL; that for A2, B2, and C3 was 16.0 μg/mL; and that for A3, B3, and C3 was 32.0 μg/mL. RNA-sequencing and qPCR confirmed that the differentially expressed genes cusE, cusS, cusR, cusC, cusF, cusB, and cusA showed higher expression in C2 and C3 than in C1. Genes related to the efflux pump AcrAB-TolC showed higher expression in B2 and B3 than in B1. No mutants of ramR, tex (T), or tet (A) were detected. SoxS, oqxA, oqxB, acrE, and acrF did not show increased expression in any group. After deletion and complementation of cusR among C3, the MIC of tigecycline decreased to 4 μg/mL, and then recovered to 32 μg/mL. The expression of cusFBCA, correspondingly decreased and increased significantly. Conclusion In addition to its primary function in resistance to copper/silver, the CusS-CusR two-component system is associated with CRKP resistance to tigecycline.

Hydraulic flushing has the advantages of reducing coal stress and improving coal seam permeability, so it is widely used in high gas outburst coal seams in China. However, the effect evaluation of stress relief and permeability enhancement will be necessary before coal mining. In this paper, an acoustic emission (AE) experiment system of coal seam hydraulic flushing was established, and the AE characteristics of coal during hydraulic flushing were studied. The results show that the changes in AE signals are basically consistent with those in load and water pressure. In addition, the quantitative relationship between the signals and the damage of coal around the borehole is revealed by the theory of damage mechanics. The in-situ AE test of hydraulic flushing was performed in the Liangbei Coal Mine, by measuring gas flow and coal stress simultaneously. The variation of AE activities and the effect evaluation of hydraulic flushing were analyzed. The results reveal that the changes in coal seam stress are basically consistent with those of AE signals. After the flushing has been completed for 1&ndash, 3 h, the coal seam stress decreases rapidly, and the AE counts and intensity reach the peak. Therefore, this study proposes a new method for evaluating the effect of coal seam hydraulic flushing by AE, which has a very important practical significance.

Limb spasms are phenomena of hyperreflexia that occur after spinal cord injury. Currently, the clinical treatment is less than ideal. Our goal is to develop a combination therapy based on individualized medicine to reduce spasticity after spinal cord injury. In this study, rats received a severe contusive injury at the T9 segment of the spinal cord, followed by gene therapy with adenoassociated virus encoding human neurotrophic factor 3 (AAV-NT3) and a 2-week exercise program starting at 4 weeks after injury. We quantified the frequency of spasms during a swimming test at 4 and 6 weeks after injury and confirmed the results of the swimming test by measuring the H-reflex of the plantar muscle. We obtained weekly hind limb exercise scores to assess the effect of the interventions in hind limb motor function improvement. Then, we used immunofluorescence to observe the immunoreactivity of spinal motor neurons, synaptophysin, cholinergic interneurons, and GABAergic interneurons. We also measured the expression of KCC2 in the spinal cord by western blot. We found that AAV-NT3 gene therapy, exercise, and combination therapy all attenuated the frequency of spasms in the swimming test conducted at 6 weeks after spinal cord injury and increased rate-dependent depression of H-reflex. Combination therapy was significantly superior to AAV-NT3 alone in protecting motor neurons. Recovery of KCC2 expression was significantly greater in rats treated with combination therapy than in the exercise group. Combination therapy was also significantly superior to individual therapies in remodeling spinal cord neurons. Our study shows that the combination of AAV-NT3 gene therapy and exercise can alleviate muscle spasm after spinal cord injury by altering the excitability of spinal interneurons and motor neurons. However, combination therapy did not show a significant additive effect, which needs to be improved by adjusting the combined strategy.

The H9N2 avian influenza viruses infect poultry worldwide, and can potentially cause a human pandemic without adaptation. Vitamin D3 (D3) is increasingly being recognized for its extra-skeletal roles, such as the inflammatory and immune responses to infection. The aim of this study was to analyze the changes in vitamin D metabolizing enzymes and vitamin D receptor (VDR) in the lung tissues of mice infected with H9N2. The mice were intranasally inoculated with the appropriate dose of the virus, and various clinical indices were measured on days 3, 7, 14 and 21 post-infection. H9N2 infection significantly increased the expression levels of 1α-hydroxylase mRNA and protein, which is the activating enzyme of 25-hydroxyvitamin D (25(OH)D3), but had no significant effect on the 25(OH)D3 inactivating enzyme 24-hydroxylase, indicating that inactive D3 might be converted to its active form in the H9N2-infected lungs. Furthermore, a significant increase was also observed in the VDR mRNA and protein levels, suggesting enhanced responsiveness of the lung tissues to 1, 25(OH)2D3 post H9N2 infection. In addition, daily 25(OH)D3 injection from day 2–14 post-infection did not affect the clinical signs, virus replication and cytokine (IL-1β and TNF-α) production in the lungs of the infected mice. Given that the biological effects of D3 rely on its activation, and the binding of 1, 25(OH)2D3 to VDR in specific tissues, our findings provide novel insights into the possible role of vitamin D in the development and progression of influenza.

Objective.An electrocardiogram (ECG) is one of the most common means to diagnose arrhythmia according to different waveforms clinically. Although there are advanced classification methods such as deep learning, the single view feature cannot meet the demand of classification accuracy for new individuals. To this end, a classification model based on multiview fusion was proposed.Approach.First, handcrafted view features were extracted from heartbeats and then deep view features were obtained from the deep learning model. The features of two different perspectives were fused in the fully connected layer, and the random forest classifier was used instead of the Softmax classifier for classification. Notably, Bayesian optimization was utilized in the hyper-parameter tuning of the classifier. The proposed method employed the MIT-BIH database to classify five classes: normal heartbeat (N), left bundle branch block heartbeat (LB), right bundle branch block heartbeat (RB), atrial premature contraction (APC) and premature ventricular contraction (PVC).Main results.The experimental results achieved a higher average accuracy of 98.93%, average precision of 96.92%, average sensitivity of 96.46%, and average specificity of 99.33% in five types of heartbeat classification for inter-patient.Significance.The proposed framework improves the performance of ECG detection for new individuals. And it provides an feasible algorithmic model for single-lead wearable devices with multiview fusion.

This study investigated whether captopril can reverse drug resistance in metallo-β-lactamase (MBL)-producing carbapenem-resistant Klebsiella pneumoniae (CRKP) and increase their sensitivity to antimicrobial agents. And also aimed to further characterize the affinity of captopril for imipenemase 4 (IMP-4) to explore the drug resistance treatment of MBL-producing bacteria. Five clinically isolated MBL-producing strains of CRKP were screened and the combined effects of captopril and meropenem were examined in vitro and in vivo to analyze whether captopril can reverse antimicrobial resistance in drug-resistant bacteria. Additionally, enzyme inhibition kinetics was analyzed to characterize the affinity of captopril for IMP-4. In MBL-producing Klebsiella pneumoniae, combined treatment with captopril significantly reduced the minimum inhibitory concentration (MIC) of carbapenems to 1 μg/mL at least, and captopril inhibited New-Delhi metallo-β-lactamase 1 (NDM-1) and IMP-4 in a concentration-dependent manner in vitro. Following the infection of Galleria mellonella by IMP-expressing bacteria, the survival rates were significantly higher in the combination treatment group than in the monotherapy groups. And the bacterial load in the combination treatment group was significantly lower than those in the monotherapy groups and IMP-4–producing bacteria were more sensitive to the combination treatment than NDM-1–producing bacteria. Additionally, enzyme inhibition kinetics firstly illustrated that the half-maximal inhibitory concentration of captopril for IMP-4 was 26.34 μM, and the dissociation constant was 37.14 μM. In brief, captopril potentiated meropenem activity and restored its efficacy against MBL-producing CRKP. Additionally, analysis of enzyme inhibition kinetics confirmed that captopril has good inhibitory effects on IMP-4 activity. Therefore, captopril or its derivatives may have clinical utility for overcoming antibiotic resistance.

The purpose of this study was to validate the mutant selection window (MSW) hypothesis in vitro and in vivo with Escherichia coli and Pseudomonas aeruginosa exposed to fosfomycin. Two standard strains of Gram-negative bacteria, those are E. coli ATCC 25922 and P. aeruginosa ATCC 27853, were exposed to fosfomycin at concentrations below MIC, between the MIC and the mutant prevention concentration (MPC), and above the MPC in Luria-Bertani broth and in a tissue-cage infection model, respectively. With the in vitro time-kill studies, there were bacterial re-growth and emergence of resistance thereafter for both strains at antibiotic concentrations of × 4, × 8 and × 16 MIC. In our animal model, the loss in susceptibility of P. aeruginosa at fosfomycin concentrations fluctuated between the lower and upper boundaries of the MSW. In contrast, the emergence of resistant mutants of E. coli was not observed in vivo, regardless of fosfomycin dosage. Interestingly, the in vitro-isolated resistant mutants of E. coli showed a decreased growth rate compared with the susceptible parental strains, whereas no fitness cost in P. aeruginosa was observed. The emergence of antibiotic resistance is shaped by several factors. MSW theory may not apply to all antimicrobial-pathogen combinations. Before it can be used as a framework for the design of antimicrobial therapy, the existence of the window must be demonstrated not only in vitro but also in vivo.

We developed a high-throughput bead-based suspension array for simultaneous detection of 20 respiratory tract pathogens in clinical specimens. Pathogen-specific genes were amplified and hybridized to probes coupled to carboxyl-encoded microspheres. Fluorescence intensities generated via the binding of phycoerythrin-conjugated streptavidin with biotin-labeled targets were measured by the Luminex 100 bead-based suspension array system. The bead-based suspension array detected bacteria in a significantly higher number of samples compared to the conventional culture. There was no significant difference in the detection rate of atypical pathogensatypical pathogens or viruses between the bead-based suspension array and real-time PCR. This technology can play a significant role in screening patients with pneumonia.