A Piezoelectric-Piezoresistive Coupling Electric Field Sensor for Large Dynamic Range AC Electric Field Measurements.

We propose a piezoelectric-piezoresistive coupling electric field sensor capable of performing large dynamic range AC electric field measurements. The electric field sensor utilizes direct coupling between piezoelectric (PE) materials and piezoresistive (PR) strain gauges, in conjunction with an external signal conditioning circuit, to measure AC electric fields effectively. We verified the feasibility of the scheme using a finite element simulation, fabricated a prototype of the electric field sensor, and characterized the properties of the prototype. The testing results indicate that the sensor exhibits an ac resolution of 50 V/m and a linear measurable electric field range of 0 to over 200 kV/m, which keeps the linearity at less than 0.94% from 1 Hz to over 5 kHz. Furthermore, the sensor also has advantages, such as a small size and low power consumption. The sensor can enhance the comprehensive observability and measurability of digital power grids.

Study on pyroptosis-related genes Casp8, Gsdmd and Trem2 in mice with cerebral infarction.

Objective: Cerebral infarction is the main cause of death in patients with cerebrovascular diseases. Our research aimed to screen and validate pyroptosis-related genes in cerebral infarction for the targeted therapy of cerebral infarction. Methods and results: A total of 1,517 differentially expressed genes (DEGs) were obtained by DESeq2 software analysis. Gene set enrichment analysis results indicated that genes of middle cerebral artery occlusion (MCAO) mice aged 3 months and 18 months were enriched in pyroptosis, respectively. Differentially expressed pyroptosis-related genes (including Aim2, Casp8, Gsdmd, Naip2, Naip5, Naip6 and Trem2) were obtained through intersection of DEGs and genes from pyroptosis Gene Ontology Term (GO:0070269), and they were up-regulated in the brain tissues of MCAO mice in GSE137482. In addition, Casp8, Gsdmd, and Trem2 were verified to be significantly up-regulated in MCAO mice in GSE93376. The evaluation of neurologic function and triphenyltetrazolium chloride staining showed that the MCAO mouse models were successfully constructed. Meanwhile, the expressions of TNF-α, pyroptosis-related proteins, Casp8, Gsdmd and Trem2 in MCAO mice were significantly up-regulated. We selected Trem2 for subsequent functional analysis. OGD treatment of BV2 cell in vitro significantly upregulated the expressions of Trem2. Subsequent downregulation of Trem2 expression in OGD-BV2 cells further increased the level of pyroptosis. Therefore, Trem2 is a protective factor regulating pyroptosis, thus influencing the progression of cerebral infarction. Conclusions: Casp8, Gsdmd and Trem2 can regulate pyroptosis, thus affecting cerebral infarction.

Innovation driver and county air pollution: cost-benefit analysis perspective.

Innovation is the first power to drive the county's green and low-carbon. It is crucial to explore the impact of innovation on air pollution from the perspective of counties at the bottom of the administrative division hierarchy. The article is aimed at exploring the direct impact effects, spatial spillover effects, impact mechanism pathways, non-linear relationships, and cost-benefits of innovation drive on air pollution in counties. To this end, based on the collection of county-level data from 2007 to 2020 in mainland China, the article constructs a fixed-effects model, a dynamic panel model, and a spatial Durbin model for analysis. For every 1% increase in the quantity of innovation, the county SO2 emission concentration decreases by 0.2% on average; for every 1% increase in the quality of innovation, the county SO2 emission concentration decreases by 0.3% on average. When the county innovation quantity driver increases by one standard deviation, the county SO2 concentration decreases by an average of 0.29%; when the county innovation quality driver each standard deviation increases, the county SO2 concentration is reduced by 0.33% on average. The significant entry of high-end factors, the increased frequency of regulation by the environmental protection department, and the increasing efficiency of energy use are the important mechanism pathways for innovation-driven reduction of air pollution in counties. There is no significant "(inverted) U-shaped" relationship between innovation-driven air pollution in the county samples. There is a negative spatial spillover effect of the innovation quality drive on air pollution control in all Chinese county samples. Innovation to drive the declining size of the county's sulfur dioxide can bring about one billion yuan (about 139.81 million U.S. dollars) in comprehensive economic benefits. In the coming period, county governments should build a new pattern of "blue sky and white clouds" with neighboring regions in terms of spatial agglomeration of high-end elements, green transformation and utilization of energy, and intelligent monitoring and supervision of pollution.

Therapeutic effects of Buyang Huanwu Tang combined with RT-PA intravenous thrombolysis on stroke of Qi deficiency and blood stasis type and its impact on Keap1-Nrf2/ARE pathway antioxidant stress.

This study investigated the impact of combining traditional Chinese medicine, Buyang Huanwu Tang, with intravenous thrombolysis using alteplase (rt-PA) in treating ischemic stroke patients with qi deficiency and blood stasis. A single-center clinical randomized trial involved 117 ischemic stroke patients treated with rt-PA in the neurology department from January 2019 to December 2021. Patients were randomly divided into two groups: the control group (58 patients) received rt-PA alone, while the combined group (59 patients) received rt-PA along with Buyang Huanwu Tang. Neurological deficit scores (NIHSS) were assessed before and after treatment, along with hemorheological indicators, vascular endothelial growth factor (VEGF), matrix metalloproteinase-9 (MMP-9), and Keap1-Nrf2/ARE pathway oxidative stress indicators (Keap1, Nrf2, ARE, and NQO1 proteins). Before treatment, there were no significant differences between the groups. After treatment, the combination group exhibited lower NIHSS scores at 4, 8, and 12 weeks, indicating significant improvement compared to the control group. Additionally, the combination group demonstrated reduced plasma viscosity, low and high shear viscosity, and improved red blood cell aggregation compared to the control group after 8 weeks of treatment. Furthermore, the combination group showed elevated MMP-9 levels and reduced VEGF levels, suggesting favorable outcomes. Regarding the Keap1-Nrf2/ARE pathway, Nrf2 and NQO1 protein expression levels were higher in the combination group after 8 weeks of treatment. Clinical efficacy assessment revealed that the combined treatment group had a significantly better overall treatment response. In conclusion, combining Buyang Huanwu Tang with rt-PA intravenous thrombolysis effectively mitigated oxidative stress damage in the Keap1-Nrf2/ARE pathway among ischemic stroke patients with qi deficiency and blood stasis. This approach promoted neurological function recovery and improved overall treatment outcomes.

Dimeric Pillar[5]arene as a Novel Fluorescent Host for Controllable Fabrication of Supramolecular Assemblies and Their Photocatalytic Applications.

A dimeric fluorescent macrocycle m-TPE Di-EtP5 (meso-tetraphenylethylene dimeric ethoxypillar[5]arene) is synthesized based on the meso-functionalized ethoxy pillar[5]arene. Through the connectivity of two pillar[5]arenes by CC double bond, the central tetraphenylethylene (TPE) moiety is simultaneously formed. The resultant bicyclic molecule not only retains the host-guest properties of pillararenes but also introduces the interesting aggregation-induced emission properties inherent in the embedded TPE structure. Three dinitrile derivatives with various linkers are designed as guests (G1, G2, and G3) to form host-guest assemblies with m-TPE Di-EtP5. The morphological control and fluorescence properties of the assemblies are successfully realized. G1 with a shorter alkyl chain as the linker completely threads into the cavities of the host. G2, due to its longer chain length, forms a linear supramolecular polymer upon binding to m-TPE Di-EtP5. G3 differs from G2 by possessing a bulky phenyl group in the middle of the chain, which can be further assembled with m-TPE Di-EtP5 to form supramolecular layered polymer and precipitated out in solution, and can be efficiently applied to photocatalytic reactions.

Comparative analysis of the clinical and epidemiological characteristics of human influenza virus versus human respiratory syncytial virus versus human metapneumovirus infection in nine provinces of China during 2009-2021.

A comparative analysis of confirmed cases of human influenza virus (HIFV), human respiratory syncytial virus (HRSV), and human metapneumovirus (HMPV) was conducted to describe their clinical and epidemiological characteristics. During 2009-2021, active surveillance of acute respiratory infections (ARIs) was performed in nine provinces of China. Clinical and epidemiological information and laboratory testing results of HIFV, HRSV, and HMPV were analyzed. Among 11591 ARI patients, the single-infection rates of HIFV, HRSV, and HMPV were 15.00%, 9.59%, and 2.24%, respectively; the coinfection rate of these three viruses was 0.64%. HIFV infection was mainly in adults aged 15-59 years, accounting for 39.10%. HRSV and HMPV infections were mainly in children under 5 years old, accounting for 87.13% and 83.46%, respectively. Patients with HRSV infection were younger than HMPV. HRSV and HMPV had high similarities in clinical manifestations, presenting with lower respiratory symptoms. HIFV mainly presented with an upper respiratory infection. The epidemic peak of HRSV was earlier than that of HIFV, and that of HMPV was later than those of HRSV and HFIV. A total of 85.14% of coinfection cases were children under 5 years old. Coinfection might increase the risk of pneumonia in HIFV cases. During 2020-2021, the positive rates and seasonal patterns of these three viruses changed due to the impact of the COVID-19 pandemic. Certain clinical and epidemiological features were observed in HIFV, HRSV, and HMPV infections, which could be beneficial for guiding clinical diagnosis, treatment, and prevention of these three viruses in China.

Can the Adjustment and Renovation Policies of Old Industrial Cities Reduce Urban Carbon Emissions?-Empirical Analysis Based on Quasi-Natural Experiments.

Based on a literature review and theoretical mechanism, this paper takes the implementation point of the adjustment and transformation policy for old industrial cities as the breakthrough point, and uses a regression model to explore the impact of the adjustment and transformation policy of these old industrial cities on urban carbon emissions. This paper also robustly tests the effective mechanisms and environmental hypotheses. Overall, the implementation of the adjustment and renovation policy has significantly reduced the carbon emissions of old industrial cities by about 0.068 units. Compared with the control group cities, the pilot cities reduced carbon emissions by an average of about 310,000 tons after the implementation of the policy. Based on a summary of the excellent Chinese case experience and an empirical analysis, it can be concluded that improvements in the green innovation capacity of old industrial cities, the agglomeration of high-end service industries, and the strengthening of ecological restoration are important mechanisms that lead to reduced carbon emissions. There is no subsequent exacerbation of the carbon intensity of neighboring cities, and there is insufficient evidence to prove pollution via neighboring transfers and use of the beggar-thy-neighbor policy. The extended analysis shows that the "inverted U-shaped" CO2 Kuznets environmental curve hypothesis is significantly present in the sample of old industrial cities, but most cities do not cross the threshold. In 2013, about 60% of the urban sample economic growth and carbon emissions showed signs of tapping into potentials and increasing efficiency (absolute decoupling) and intensive expansion (relative decoupling). In old industrial cities, the proportion of relative decoupling shows a fluctuating upward trend. In the future, the government should accurately select its own development orientation and actively seek the "best balance" between economic growth and a green and low-carbon path.

Cornuside ameliorated experimental autoimmune encephalomyelitis by limiting the recruitment of CD4+ T lymphocytes in the spinal cord

We conducted this study to determine whether cornuside could improve the neurological deficit symptoms of experimental autoimmune encephalomyelitis (EAE) rats, as well as determine the potential involvement of CD4+ T lymphocytes, vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and tumor necrosis factor-α (TNF-α). Altogether, 32 Lewis rats were randomly divided into control, EAE, EAE/prednisolone, and EAE/cornuside, wherein their neurological function was assessed every day. CD4+ T lymphocyte recruitment into the spinal cord (SC) was evaluated using immunohistochemistry. The VCAM-1, ICAM-1 and TNF-α mRNA expressions in the SC were determined by real-time quantitative PCR, and the VCAM-1 and ICAM-1 proteins were determined by western blotting. Compared to the control group, the EAE group rats with neurological deficits had enhanced CD4+ T lymphocyte infiltration and higher expression levels of VCAM-1, ICAM-1, and TNF-α in the SC. Meanwhile, compared with the EAE group, the EAE/cornuside and EAE/prednisolone groups had lower neurological scores, less CD4+ T lymphocyte infiltrations, and lower expression levels of VCAM-1, ICAM-1, and TNF-α in the SC. Thus, cornuside ameliorated EAE, which could be owed to the inhibition of CD4+ T lymphocyte recruitment and VCAM-1, ICAM-1, and TNF-α expressions in the SC

Cornuside alleviates experimental autoimmune encephalomyelitis by inhibiting Th17 cell infiltration into the central nervous system.

The present study was conducted to clarify the therapeutic effect of cornuside on experimental autoimmune encephalomyelitis (EAE) and its influence on T helper 17 (Th17) cell and regulatory T (Treg) cell infiltration into the central nervous system. Rats were randomly placed into four treatment groups: control, EAE, EAE+cornuside, and EAE+prednisolone. The neurological function scores of rats were assessed daily. On the second day after EAE rats began to show neurological deficit symptoms, the four groups were treated with normal saline, normal saline, cornuside (150 mg/kg), and prednisolone (5 mg/kg), respectively. The treatment was discontinued after two weeks, and the spinal cord was obtained for hematoxylin and eosin (H&E) and luxol fast blue staining, as well as retinoic acid receptor-related orphan receptor γ (RORγ) and forkhead box protein P3 (Foxp3) immunohistochemical staining. Blood was collected for Th17 and Treg cell flow cytometry testing, and the serum levels of interleukin (IL)-17A, IL-10, transforming growth factor-ß (TGF-ß), IL-6, IL-23, and IL-2 were measured via enzyme-linked immunosorbent assay (ELISA). Compared with rats in the EAE group, rats in the EAE+cornuside and EAE+prednisolone groups began to recover from neurological deficits earlier, and had a greater degree of improvement of symptoms. Focal inflammation, demyelination, and RORγ-positive cell infiltration were reduced by cornuside or prednisolone treatment, whereas the Foxp3-positive cell numbers were not significantly different. Meanwhile, the number of Th17 cells and the IL-17A, IL-6, and IL-23 levels were lower in the blood after cornuside or prednisolone treatment, whereas the number of Treg cells or the levels of IL-10, TGF-ß, and IL-2 were not markedly different. Cornuside can alleviate symptoms of EAE neurological deficits through its anti-inflammatory and immunosuppressive effects, and Th17 cells may be one of its therapeutic targets.

Association of Multiple Dopamine D3 Receptor Gene 3'UTR Polymorphisms with Susceptibility to Parkinson's Disease and Clinical Efficacy of Piribedil Therapy.

Objective: To investigate the correlation between the Dopamine D3 receptor (DRD3) 3'untranslated region (3'UTR) gene polymorphism and susceptibility to Parkinson's disease (PD) and the clinical effect of the DRD2 and DRD3 agonist piribedil treatment. Methods: Sanger sequencing was used to analyze the single nucleotide polymorphisms (SNPs) within the 3'UTR rs76126170, rs9868039, rs9817063, and rs3732790 loci of the DRD3 gene in 284 PD patients and 284 controls. PD patients were treated with piribedil sustained-release tablets (50 mg) combined with levodopa and benserazide hydrochloride tablets, three times daily (patients with first-diagnosed PD were only administrated with piribedil sustained-release tablets) for 3 months. The Unified Parkinson's Disease Rating Scale (UPDRS) and the Hoehn and Yahr disease stage were evaluated at baseline and after 3 months of treatment. Results: The T allele carriers of the DRD3 gene rs76126170 locus were more susceptible to PD than the C allele carriers (odds ratio [OR] = 3.44, 95% confidence interval [CI]: 2.46-4.80, p < 0.01). Carriers of the rs9868039 A allele had a decreased risk of PD compared to those with G allele (OR = 0.67, 95% CI: 0.53-0.86, p < 0.01). C allele carriers at rs9817063 were less likely to develop PD than those with T allele (OR = 0.74, 95% CI: 0.58-0.94, p = 0.02). No significant correlation was observed between the alleles or genotypes of the rs3732790 locus and PD susceptibility (p > 0.05). The various genotypes of the DRD3 gene loci rs76126170, rs9868039, and rs9817063 in PD patients were associated with significant differences with regard to reduction of UPDRS scores and Hoehn and Yahr stage after 3 months of treatment (p < 0.05). Conclusion: The alleles and genotypes of the DRD3 gene 3' UTR SNP loci rs76126170, rs9868039, and rs9817063 are associated with PD susceptibility and the clinical efficacy of piribedil treatment.

Gefitinib sensitization of cisplatin-resistant wild-type EGFR non-small cell lung cancer cells.

PURPOSE: The usual first-line strategy of wild-type EGFR (wtEGFR) non-small cell lung cancer (NSCLC) remains cisplatin-based chemotherapy. However, cisplatin often loses effectiveness because most tumors acquire drug resistance over time. As EGFR is the most important pro-survival/proliferation signal receptor in NSCLC cells, we aimed at investigating whether cisplatin resistance is related to EGFR activation and further evaluating the combined effects of cisplatin/gefitinib (EGFR-tyrosine kinase inhibitor, EGFR-TKI) on cisplatin-resistant wtEGFR NSCLC cells. MATERIALS AND METHODS: EGFR activation was analysed in parental and cisplatin-resistant wtEGFR NSCLC cell lines (H358 and H358R, A549 and A549R). Cellular proliferation and apoptosis of H358R/A549R cells treated with cisplatin or gefitinib, alone or in combination were investigated, and the related effector protein was detected by western blot analysis. Anti-tumor effect of two drugs combined was evaluated in animal models of H358R xenografts in vivo. RESULTS: EGFR was significantly phosphorylated in cisplatin-resistant wtEGFR NSCLC cells H358R and A549R than their parental cells. In H358R and A549R cells, anti-proliferative ability of gefitinib was further improved, and gefitinib combined with cisplatin enhanced inhibition of cellular survive/proliferation, and promotion of apoptosis in vitro. The combined effects were also associated with the inhibition of EGFR downstream effector proteins. Similarly, in vivo, gefitinib and cisplatin in combination significantly inhibited tumor growth of H358R xenografts. CONCLUSION: Abnormal activation of EGFR may induce wtEGFR NSCLC cell resistance to cisplatin. The combined effects of cisplatin/gefitinib suggest that gefitinib, as a combination therapy for patients with cisplatin-resistant wtEGFR NSCLC should be considered.

Behavior and Hippocampal Epac Signaling to Nicotine CPP in Mice.

Tobacco use is a major challenge to public health in the United States and across the world. Many studies have demonstrated that adult men and women differ in their responses to tobacco smoking, however neurobiological studies about the effect of smoking on males and females were limited. Exchange protein directly activated by cAMP (Epac) signaling participates in drug addictive behaviors. In this study, we examined the hippocampal Epac signaling in nicotine-induced place conditioning mice. Nicotine at 0.2 mg/kg and 0.4 mg/kg induced a conditioned place preference (CPP) in male and female mice, respectively. After CPP, male mice presented less anxiety-like behavior as demonstrated by an open-field test. The hippocampal Epac2 protein was elevated in both male and female nicotine place conditioning mice. However, Rap1 protein was elevated and CREB phosphorylation was reduced in female nicotine place conditioning mice. Our data provide direct evidence that hippocampal Epac signaling is altered in nicotine-induced CPP mice. Pharmacology manipulation Epac signaling may open a new avenue for the treatment of nicotine abuse and dependence.

BEZ235 increases sorafenib inhibition of hepatocellular carcinoma cells by suppressing the PI3K/AKT/mTOR pathway.

BACKGROUND: Sorafenib is an oral multi-kinase inhibitor that inhibits hepatocellular carcinoma (HCC) via the Ras/Raf/MAPK pathway. However, sorafenib loses effectiveness because most tumors acquire drug resistance over time. As the PI3K/AKT/mTOR signaling pathway is also activated abnormally in HCC, we evaluated the effect of sorafenib, in combination with a dual PI3K/mTOR inhibitor, BEZ235, on HCC cell proliferation and survival in vitro. MATERIALS AND METHODS: Biological phenotypes were analysed in HCC cell lines, parental and sorafenib-resistant HepG2 cells (HepG2 and HepG2R), treated with sorafenib or BEZ235, alone or in combination. HCC cellular proliferation and apoptosis were investigated, and perturbations of the Ras/Raf/MAPK and PI3K/AKT/mTOR signaling/survival pathways were evaluated by western blot analysis. RESULTS: BEZ235 enhanced sorafenib inhibition of cellular proliferation, migration, and promotion of apoptosis in HepG2 and HepG2R cells. The combined effects were associated with inhibition of phosphorylation of AKT, mTOR and S6K in the PI3K/AKT/mTOR pathway, whereas the combination of sorafenib and BEZ235 did not significantly alter the Ras/Raf/MAPK pathway compared with the effect of sorafenib alone. CONCLUSION: Sorafenib/BEZ235 combination has potent anti-HCC cell activity. This anti-tumor activity is most likely multi-factorial, mainly involving PI3K down-regulation and AKT, mTOR and S6K dephosphorylation. Combined inhibition of PI3K/AKT/mTOR and Ras/Raf/MAPK pathways enhances sorafenib inhibition of HCC. The results of these in vitro studies suggest that trials of combined sorafenib and BEZ235 in the treatment of HCC should be considered.

Vitamin D enhances resistance to aspergillus fumigatus in mice via inhibition of excessive autophagy.

The role of vitamin D in the regulation of lung immune defense and inflammatory response has attracted more and more attention. Vitamin D deficiency is closely related to respiratory tract infections. However, few studies have elucidated the mechanism of vitamin D deficiency on host pulmonary resistance to Aspergillus fumigatus (A. fumigatus). In this paper, the role of autophagy and Treg regulation in the treatment of rat models of A. fumigatus infection with vitamin D was investigated. We intratracheally injected the A. fumigatus spores into Mice fed with sufficient vitamin D (VitD+) or deficient diets (VitD-). Mortality, fungal load and weight changes were evaluated. The conidia of lung tissue were isolated for analysis of viability. Alveolar macrophages (AMs) were stimulated with a viable A. fumigatus conidia for determining the formation of lysosomes in vitro. The autophagy-related proteins dectin-1, ROS and LC3BII expression in AMs were measured. Fluorescence and Western blot were performed to evaluate the autophagic flux and Treg cells were detected by flow cytometry. After inoculation with A. fumigatus, the vitamin D deficient mice exhibited a higher rate of death, more fungal growth, and more weight loss than its sufficient peers. The viability of A. fumigatus conidia in VitD+ mice was significantly lower than that in VitD- mice. In the case of A. fumigatus infection, vitamin D delays the formation of lysosomes against A. fumigatus through autophagy. The autophagy flow measurement experiment also found that the vitamin D group lowered autophagy levels in cells and a small number of Treg cells. In conclusion, Vitamin D deficiency can lead to impaired lung defense in mice, which may be associated with the formation of excessive autophagy-induced lysosomes and increased counts of Treg cells.

[Protective effect of curcumin on dopamine neurons in Parkinson's disease and its mechanism].

OBJECTIVE: To investigate the effect of curcumin on dopamine neurons in Parkinson's disease (PD) and its mechanism. METHODS: SH-SY5Y human neuroblastoma cells were treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to establish the PD cell model. The model cells were treated with curcumin and/or autophagy inhibitor 3-MA. After 48 h of drug treatment, the number of surviving dopamine neurons was detected by tyrosine hydroxylase immunofluorescence method. Western blotting was used to detect protein expression of α-Synuclein (α-Syn), transcription factor EB (TFEB) and autophagy-related proteins lysosome-associated membrane protein 2A (LAMP2A) and microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ); RT-PCR was used to detect mRNA expression of α-Syn. RESULTS: Compared with MPTP model group, curcumin increased the number of surviving dopamine neurons(P<0.01), decreased both protein expression and mRNA expression of α-Syn (all P<0.01), and increased protein expression of TFEB, LAMP2A and LC3-Ⅱ (all P<0.01). When curcumin and 3-MA were given concurrently, the number of surviving dopamine neurons, protein expression of TFEB, LAMP2A and LC3-Ⅱ increased (P<0.05 or P<0.01), and both protein expression and mRNA expression of α-Syn decreased (P<0.05 or P<0.01) compared with MPTP model group; but the number of surviving dopamine neurons and protein expression of LAMP2A and LC3-Ⅱ decreased compared with curcumin group (all P<0.05). CONCLUSIONS: Curcumin exerts protective effect on dopamine neurons in PD, which may be associated with enhancing autophagy and promoting the clearance of α-Syn.

Emergence of BA9 genotype of human respiratory syncytial virus subgroup B in China from 2006 to 2014.

A study was conducted to investigate the circulation of HRSV subgroup B (HRSVB) in China in recent years. HRSVB sequences from 365 samples collected in 1991, 2004 and 2008-2014 in China, together with 332 Chinese HRSVB sequences obtained from GenBank were analyzed to determine the geographic and yearly distribution of HRSVB. Phylogenetic analysis revealed these HRSVB sequences clustered into 4 genotypes with different frequencies: BA (83%), CB1 (11%), SAB (3.0%) and GB3 (0.7%). Between 2005 and 2013, there was a co-circulation of BA and non-BA genotypes in China. Genotypes BA9 and BA10 were two of the main BA genotypes detected in this study. Genotype BA9 was first detected in China in 2006 and became the predominant HRSVB genotype circulating in China from 2008 to 2014. Three different lineages were detected for both genotypes BA9 and BA10. Time to the most recent common ancestor for genotypes BA9 and BA10 was estimated for years 1997 and 1996, respectively. Results of this study not only contribute to the understanding of the circulation pattern, but also the phylogenetic pattern and evolution of HRSVB in China from 1991 to 2014.

Long noncoding RNA eosinophil granule ontogeny transcript inhibits cell proliferation and migration and promotes cell apoptosis in human glioma.

Glioma is the most common primary brain tumor and represents one of the most aggressive and lethal types of human cancer. Recent advances have implicated long noncoding RNAs (lncRNAs) as crucial mediators of cancer development and progression. The present study aimed to investigate the role of a newly-discovered lncRNA, termed eosinophil granule ontogeny transcript (EGOT), in the aggressive abilities of cells in human glioma. It was initially found that the relative transcription level of EGOT in glioma cancerous tissues was significantly lower than that in adjacent non-cancerous tissues. EGOT was differentially expressed in a series of glioma cell lines, with its lowest level in high aggressive U251 and U87 cells. When EGOT was overexpressed by an expression plasmid, cell viability was significantly inhibited in U251 and U87 cells. Furthermore, with EGOT overexpression, the cell cycle was arrested at G0/G1 phase and consequently, cell apoptosis was significantly promoted along with the activities of caspase-3 and caspase-9. The migration abilities of EGOT-overexpressed cells were inhibited by 71.4% in U251 cells and by 69.5% in U87 cells. These data suggest that overexpression of EGOT inhibits cell proliferation and migration, and promotes cell apoptosis in glioma. Therefore, EGOT has potent anticancer activity and may function as a tumor suppressor in human glioma.

[ESAT-6 inhibits autophagy in macrophages and promotes the growth of BCG].

Objective To explore the interaction between 6 kD early secretory antigenic target (ESAT6) and autophagy in order to provide an experimental basis for the immune evasion mediated by ESAT6. Methods RAW264.7 cell lines (mouse macrophages) were treated with Earle's balanced salt solution (EBSS), and another cells were transfected by control plasmid pCMV-HA or recombinant plasmid pCMV-HA-ESAT6. Then we observed the growth of Bacillus Calmette-Guerin (BCG) in macrophages. Western blotting was used to detect the LC3 levels in RAW264.7 cells at 0, 8, 12, 32 hours after transducted by pCMV-HA-ESAT6. In RAW264.7 cells transfected with PCMV-HA, PCMV-HA-ESAT6, and treated with chloroquine (CQ) and CQ combined with pCMV-HA-ESAT6, which were lysed and cultured in Lowenstein-Jensen culture medium for BCG counting, LC3 was detected by Western blot analysis, and the number and size of lysosomes were observed by LysoTracker Red staining. Results Compared with control plasmid pCMV-HA transfected RAW264.7 cells, the number of BCG significantly increased in PCMV-HA-ESAT6-transfected cells, while decreased in EBSS-treated cells. PCMV-HA-ESAT6 transfection resulted in the increased transition of LC3 I to LC3 II in a time-depended manner. Compared with the controls, LysoTracker Red staining showed PCMV-HA-ESAT6, CQ and CQ plus PCMV-HA-ESAT6 transfections resulted in the increased number and size of lysosomes, and there were no differences among the three groups. Moreover, the growth potential of BCG was strong in the three transfection groups. Conclusion ESAT6 can inhibit the autophagy and promote the growth of BCG in RAW264.7 cells.

Loopback rolling circle amplification for ultrasensitive detection of Kras gene.

Mutations in Kras gene may be used as a diagnostic marker and a target for treatment of the broad spectrum of human cancers. In this study, we developed a new class of amplification assay, double-hairpin molecular beacon (DHMB)-based cascade rolling circle amplification (RCA), for ultrasensitive and selective detection of Kras gene in a homogenous solution. Specifically, target DNA can hybridize with DHMB and activate cyclical target strand-displacement polymerization (CTDP) and nicking-mediated strand-displacement polymerization (NMDP). The resulting nicked/displaced fragments substantially outnumber target DNA and cause the cascade rolling circle amplification (C-RCA) and nicked fragment-induced strand-displacement polymerization (NFDP). Even if four amplification processes are designed, only DHMB, padlock probe and polymerization primer are involved. Under optimized conditions, this screening system exhibits a linear range of 5 orders of magnitude (from 100fM to 20nM), and the detection limit is down to 16fM. Moreover, the developed biosensing system offers a high assay specificity for perfectly matched target DNA, and the measured data from practical samples demonstrated the potential application in the cancer diagnoses. As a proof-of-concept genetic assay, the novel signaling strategy, as well as desirable analytical capability, would significantly benefit the development of versatile amplification gene profiling platforms, revealing great promise in biological studies and medical diagnostics.

[SapM-induced fusion blocking of autophagosome-lysosome is depended on interaction with Rab7].

Objective To study the role of Rab7 in the blockage of autophagosome-lysosome fusion induced by secretory acid phosphatase (SapM), a virulence factor of mycobacterium tuberculosis. Methods The Raw264.7 cells were transfected with siRab7, and the P62 was detected using Western blotting. After transfected with mCherry-SapM, the co-localization of SapM and Rab7 in Raw264.7 cells was detected by immunofluorescence cytochemistry and the interaction of SapM with Rab7 was determined by co-immunoprecipitation. SapM mutants including SapM(δ ARCA), SapM(δ FRED) and SapM(δ CT) were used to transfect Raw264.7 cells, and their associations with Rab7 were analyzed. Results The treatment of siRab7 induced a significant increase of P62 in these cells. Immunofluorescence cytochemistry showed the intracellular co-localization of SapM and Rab7. Co-immunoprecipitation showed that SapM and Rab7 were precipitated by each other. Only SapM(δ CT) failed to interact with Rab7 among the three SapM mutants. Conclusion The inhibition of autophagosome-lysosome fusion induced by SapM is dependent on the interaction between SapM and Rab7.