MiR-331-5p suppresses gastric cancer cell proliferation, migration, invasion, and glycolysis via targeting PFKFB3

Purpose: To examine the role of microRNAs (miRNAs), miR-331-5p, in gastric cancer (GC). Methods: The mRNA level of miR-331-5p and protein level of 6-phosphofructo-2-kinase/fructose-2,6- bisphosphatase-3 (PFKFB3) were determined using quantitative real-time reverse transcription– polymerase chain reaction (qRT–PCR) and western blotting, respectively. The cell viability and proliferation of the two GC cell lines (AGS and MKN45) were evaluated using Cell Counting Kit-8 (CCK8) and bromodeoxyuridine (BrdU)  ssays. Cell migration and invasion of AGS and MKN45 were evaluated using wound healing and invasion assays, respectively. Potential interactions between miR331-5p and PFKFB3 were assessed by luciferase activity assay, while the effects of the interactions on cell physiology and metabolism were investigated in cells  verexpressing both miR-331-5p and PFKFB3. Results: MiR-331-5p overexpression inhibited cell proliferation, suppressed migration and invasion, and inhibited glycolysis in AGS and MKN45 cells. The mRNA for the glycolytic regulatory enzyme PFKFB3 was shown to be a direct target of miR-331-5p and modulated by miR-331-5p. In rescue experiments, PFKFB3 reversed the miR-331-5p-induced inhibition of proliferation, migration, invasion, and glycolysis in AGS cells. Conclusion: This work supports a role for miR-331-5p through the modulation of PFKFB3 activity in GC in vivo, thus providing insight into novel potential therapies for the treatment of GC

Antitussive efficacy of the current treatment protocol for refractory chronic cough: our real-world experience in a retrospective cohort study

BACKGROUND: The management of refractory chronic cough (RCC) is a great challenge. Neuromodulators have long been used for RCC with imperfect efficacy. OBJECTIVES: We summarized the outcomes of the current treatments used at our specialist cough clinic, which provides a guideline-led service and real-world experience for the future management of RCC. DESIGN: This is a single-centre retrospective observational cohort study. METHODS: Consecutive RCC patients (the first clinic visit between January 2016 and May 2021) were included into this observational cohort study. Medical records in the Chronic Cough Clinical Research Database were fully reviewed using uniform criteria. The included subjects were followed-up for at least 6 months after the final clinic visit via instant messages with the link to self-scaled cough-associated questionnaires. RESULTS: Overall, 369 RCC patients were analysed with a median age of 46.6 years and a cough duration of 24.0 months. A total of 10 different treatments were offered. However, 96.2% of patients had been prescribed at least one neuromodulator. One-third of patients had alternative treatments prescribed given the poor response to the initial therapy and 71.3% favourably responded to at least one of the treatments. Gabapentin, deanxit, and baclofen had comparable therapeutic efficacy (56.0%, 56.0%, and 62.5% respectively; p = 0.88) and overall incidences of adverse effects (28.3%, 22.0%, and 32.3% respectively; p = 0.76). However, 19.1 (7.7-41.8) months after the last clinic visit, 65.0% reported improvement (24.9%) or control of their cough (40.1%); 3.8% reported a spontaneous remission and 31.2% still had a severe cough. Both HARQ (n = 97; p < 0.001) and LCQ (n = 58; p < 0.001) demonstrated marked improvement. CONCLUSION: Trying different neuromodulators is a pragmatic strategy for RCC, which helped around two-thirds of patients. Relapse is common on withdrawal or reduction of dosage. Novel medication for RCC is an urgent clinical need. PLAIN LANGUAGE SUMMARY: This is the first report that fully represented a guideline-led treatment protocol for refractory chronic cough (RCC) based on a large series of patients, which evaluated the short- and long-term effects of the currently available treatments for RCC. We found that the therapeutic trial of different neuromodulators is a pragmatic strategy, which helped around two-thirds of patients. Gabapentin, deanxit (flupentixol/melitracen), and baclofen had similar therapeutic outcomes. This study may offer real-world experience for the future management of RCC

Improving the expression of recombinant pullulanase by increasing mRNA stability in Escherichia coli

Background: Pullulanase production in both wild-type strains and recombinantly engineered strains remains low. The Shine-Dalgarno (SD) sequence and stem-loop structure in the 5\u2032 or 3\u2032 untranslated region (UTR) are well-known determinants of mRNA stability. This study investigated the effect of mRNA stability on pullulanase heterologous expression. Results: We constructed four DNA fragments, pulA, SD-pulA, pulA-3t, and SD-pulA-3t,whichwere cloned into the expression vector pHT43 to generate four pullulanase expression plasmids. The DNA fragment pulA was the coding sequence (CDS) of pulA in Klebsiella variicola Z-13. SD-pulA was constructed by the addition of the 5\u2032 SD sequence at the 5\u2032 UTR of pulA. pulA-3t was constructed by the addition of a 3\u2032 stem-loop structure at the 3\u2032 UTR of pulA. SD-pulA-3t was constructed by the addition of the 5\u2032 SD sequence at the 5\u2032 UTR and a 3\u2032 stem-loop structure at the 3\u2032 UTR of pulA. The four vectors were transformed into Escherichia coli BL21(DE3). The pulA mRNA transcription of the transformant harboring pHT43-SD-pulA-3t was 338.6%, 34.9%, and 79.9% higher than that of the other three transformants, whereas the fermentation enzyme activities in culture broth and intracellularly were 107.0 and 584.1 times, 1.2 and 2.0 times, and 62.0 and 531.5 times the amount of the other three transformants (pulA, SD-pulA, and pulA-3 t), respectively. Conclusion: The addition of the 5\u2032 SD sequence at the 5\u2032 UTR and a 3\u2032 stem-loop structure at the 3\u2032 UTR of the pulA gene is an effective approach to increase pulA gene expression and fermentation enzyme activity

Single cell atlas for 11 non-model mammals, reptiles and birds.

The availability of viral entry factors is a prerequisite for the cross-species transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Large-scale single-cell screening of animal cells could reveal the expression patterns of viral entry genes in different hosts. However, such exploration for SARS-CoV-2 remains limited. Here, we perform single-nucleus RNA sequencing for 11 non-model species, including pets (cat, dog, hamster, and lizard), livestock (goat and rabbit), poultry (duck and pigeon), and wildlife (pangolin, tiger, and deer), and investigated the co-expression of ACE2 and TMPRSS2. Furthermore, cross-species analysis of the lung cell atlas of the studied mammals, reptiles, and birds reveals core developmental programs, critical connectomes, and conserved regulatory circuits among these evolutionarily distant species. Overall, our work provides a compendium of gene expression profiles for non-model animals, which could be employed to identify potential SARS-CoV-2 target cells and putative zoonotic reservoirs

Tracking the vortex motion by using Brownian fluid particles

In this paper, we propose a simple yet powerful vortex method to numerically approximate the dynamics of an incompressible flow. The idea is to sample the distribution of the initial vortices of the fluid flow in question and then follow vortex dynamics along Taylor's Brownian fluid particles. The weak convergences of this approximation scheme are obtained for both two-dimensional (2D) and three-dimensional (3D) fluid flows, though only for small time in 3D case. Based on our method, the simulation results are quite attracting

Aboveground Biomass Allometric Models for Evergreen Broad-Leaved Forest Damaged by a Serious Ice Storm in Southern China

A catastrophic ice storm occurred in the spring of 2008, which severely destroyed nearly 13% of China&rsquo;s forests; among them, the broad-leaved forest suffered the most extensive damage. In this study, allometric models of the evergreen broad-leaved forests damaged at different recovery stages after the disaster were established to estimate the aboveground biomass of damaged trees. Plant plots were established and surveyed in damaged forests to determine species composition and diameter distribution, and finally a sample scheme was formulated that contained 47 trees from 13 species. The destructive measurements of aboveground biomass of trees selected according to the scheme were conducted in 2008, 2010, 2012 and 2016, respectively. Undamaged trees in the same region were also selected to measure the biomass in 2010. Linear regression of logarithmic transformation of the power function form was performed using Diameter at Breast Height (DBH) as predictor to develop biomass allometric models. The results showed that the ice storm caused tree aboveground biomass loss, which caused different parameters of the tree biomass models at different recovery stages. The models have a high accuracy in predicting trunk and total aboveground biomass, with high determination coefficients (R2, 0.913~0.984, mean 0.957), and have a relatively low accuracy in predicting the biomass of branches and leaves (R2, 0.703~0.892, mean 0.784). The aboveground biomass reduced by 35.0% on average due to the ice storm, and recovered to the same level of undamaged trees in the same diameter 8 years after the disturbance. The branches and leaves recovered very fast, and the biomass of these parts exceeded that of the undamaged trees, reaching the same diameter 2 years after the disaster, indicating an over compensatory growth. The trees with a smaller diameter were mostly composed of middle and late succession species, and recovered faster than other species, indicating that the ice storm may alter the forest structure and accelerate community succession. The biomass allometric models built in this study, combined with forest inventory data, can estimate forest biomass loss and recovery after disturbance, and offer an important sense of the assessment of forest damage and the formulation of forest post-disaster management strategies

Role of alveolar nitric oxide in gastroesophageal reflux-associated cough: prospective observational study

Background: Fractional exhaled nitric oxide (FeNO) measured at multiple exhalation flow rates can be used as a biomarker to differentiate central and peripheral airway inflammation. However, the role of alveolar nitric oxide (CaNO) indicating peripheral airway inflammation remains unclear in gastroesophageal reflux-associated cough (GERC). Objectives: We aimed to characterize the changes in alveolar nitric oxide (CaNO) and determine its clinical implication in GERC. Design: This is a single-center prospective observational study. Methods: FeNOs at exhalation flow rates of 50 and 200 ml/s were measured in 102 patients with GERC and 134 patients with other causes of chronic cough (non-GERC). CaNO was calculated based on a two-compartment model and the factors associated with CaNO were analyzed. The effect of anti-reflux therapy on CaNO was examined in 26 GERC patients with elevated CaNO. Results: CaNO was significantly elevated in GERC compared with that in non-GERC (4.6 ± 4.4 ppb versus 2.8 ± 2.3 ppb, p  5 ppb) had more proximal reflux events (24 ± 15 versus 9 ± 9 episodes, p  = 0.001) and a higher level of pepsin (984.8 ± 492.5 versus 634.5 ± 626.4 pg/ml, p  = 0.002) in sputum supernatant than those with normal CaNO. More GERC patients with high CaNO required intensified anti-reflux therapy (χ 2  = 3.963, p  = 0.046), as predicted by a sensitivity of 41.7% and specificity of 83.3%. Cough relief paralleled a significant improvement in CaNO (8.3 ± 3.0 versus 4.8 ± 2.6 ppb, p  < 0.001). Conclusion: Peripheral airway inflammation can be assessed by CaNO measurement in GERC. High CaNO indicates potential micro-aspiration and may predict a necessity for intensified anti-reflux therapy

Root Growth Was Enhanced in China Fir (<i>Cunninghamia lanceolata</i>) after Mechanical Disturbance by Ice Storm

Accurate estimation of forest biomass and its growth potential could be important in assessing the mitigation potential of forest for climate change. However, severe mechanical disturbance such as stem breakage imposed significant changes to tree individuals in biomass structure, which could bring new inaccuracy to biomass estimation. In order to investigate the influence of severe mechanical disturbance on tree biomass accumulation and to construct accurate models for biomass and carbon storage estimation, this paper analyzed the relationship between tree size and biomass for China fir (Cunninghamia lanceolata (Lamb.) Hook) which suffered stem breakage from, and survived, an ice storm. The performance of independent variables diameter (D) and height (H) of China fir, were also compared in biomass estimation. The results showed that D as an independent variable was adequate in biomass estimation for China fir, and tree height was not necessary in this case. Root growth was faster in China fir which had suffered breakage in the main stem by the ice storm, than China fir which were undamaged for at least 7 years after the mechanical disturbance, which, in addition to biomass loss in stem, caused changes in the allocation pattern of the damaged trees. This suggests biomass models constructed before severe mechanical disturbance would be less suitable in application for a subsequent period, and accurate estimations of biomass and forest carbon storage would take more effort