1, 25-Dihydroxyvitamin D3 suppresses cell cycle progression and thus growth of prostate cancer cells by inducing expression of limb bud and heart development (LBH)

Purpose: To investigate the function of limb bud and heart development (LBH) in 1α, 25-dihydroxyvitamin D3 (1,25D)-mediated inhibitory effect on proliferation of prostate cancer cells.Methods: The inhibitory effect of 1,25D on growth and cell cycle progression of lymph node carcinoma of the prostate (LNCaP) cells was determined using cell counting kit-8 (CCK-8) assay, 5-ethynyl-2'-deoxyuridine (EdU) assay and flow cytometry, while the expression levels of LBH in response to treatment with 1,25D were determined by quantitative reverse-transcription PCR (qRT-PCR) and western blottingting. The expression levels of LBH in cells were down/up regulated by transfection with siRNA or overexpression plasmids, and then cell growth and cell cycles were measured using the CCK-8 assay, EdU assay, and flow cytometry. Finally, the growth inhibitory effect of 1,25D on LBH knockdown cells were determined using CCK-8 and EdU assays.Results: Treatment with 1,25D arrested LNCaP cells in G0/G1 phase of cell cycle, suppressed the growth of the cells and induced the expression of LBH. Overexpression/knockdown of LBH in LNCaP cells suppressed/promoted cell growth and accumulated/decreased cells in the G0/G1 phase. Moreover, knockdown of LBH reversed the inhibitory effect of 1,25D on cell proliferation of LNCaP cells.Conclusion: Inhibitory effect of 1,25D on cell cycle progression and cell proliferation might be via LBH.Keywords: Cell proliferation, Limb bud and heart development, Prostate cancer, 1α, 25-Dihydroxyvitamin D

HSPA12A Attenuates Lipopolysaccharide-Induced Liver Injury Through Inhibiting Caspase-11-Mediated Hepatocyte Pyroptosis via PGC-1α-Dependent Acyloxyacyl Hydrolase Expression

Liver dysfunction is strongly associated with poor survival of sepsis patients. Cytosolic lipopolysaccharide (LPS) sensing by Caspase-4/5/11 for pyroptosis activation is a major driver of the development of sepsis. Studies in macrophages and endothelial cells have demonstrated that LPS is inactivated by acyloxyacyl hydrolase (AOAH) and leading to desensitizing Caspase-4/5/11 to LPS. However, little is known about the cytosolic LPS-induced pyroptosis in hepatocytes during sepsis. Heat shock protein 12A (HSPA12A) is a novel member of the HSP70 family. Here, we report that LPS increased HSPA12A nuclear translocation in hepatocytes, while knockout of HSPA12A (Hspa12a−/−) in mice promoted LPS-induced acute liver injury. We also noticed that the LPS-induced Caspase-11 activation and its cleavage of gasdermin D (GSDMD) to produce the membrane pore-forming GSDMDNterm (markers of pyroptosis) were greater in livers of Hspa12a−/− mice compared with its wild type controls. Loss- and gain-of-function studies showed that HSPA12A deficiency promoted, whereas HSPA12A overexpression inhibited, cytosolic LPS accumulation, Caspase-11 activation and GSDMDNterm generation in primary hepatocytes following LPS incubation. Notably, LPS-induced AOAH expression was suppressed by HSPA12A deficiency, whereas AOAH overexpression reversed the HSPA12A deficiency-induced promotion of LPS-evoked and Caspase-11-mediated pyroptosis of hepatocytes. In-depth molecular analysis showed that HSPA12A interacted directly with peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) and increased its nuclear translocation, thereby inducing AOAH expression for cytosolic LPS inactivation, which ultimately leading to inhibition of the Caspase-11 mediated pyroptosis of hepatocytes. Taken together, these findings revealed HSPA12A as a novel player against LPS-induced liver injury by inhibiting cytosolic LPS-induced hepatocyte pyroptosis via PGC-1α-mediated AOAH expression. Therefore, targeting hepatocyte HSPA12A represents a viable strategy for the management of liver injury in sepsis patients

Metabolic Labeling of Peptidoglycan with NIR-II Dye Enables in vivo Imaging of Gut Microbiota.

Deepening our understanding of mammalian gut microbiota has been greatly hampered by the lack of a facile, real-time and in vivo bacterial imaging method. To address this unmet need in microbial visualization, we herein report the development of a second near-infrared (NIR-II)-based method for in vivo imaging of gut bacteria. Using D-propargylglycine in gavage and then click reaction with an azide-containing NIR-II dye, gut microbiota of a donor mouse was strongly labeled with NIR-II fluorescence on their peptidoglycan. The bacteria could be readily visualized in recipient mouse gut with high spatial resolution and deep tissue penetration under NIR irradiation. We then adopted this chemical strategy to image different bacterial species, which expanded its applicability in microbiology. Moreover, by employing this method, we found that the biogeography of gut microbiota was dramatically affected by host’s gastrointestinal motilities. The NIR-II-based metabolic labeling strategy reported here, to our knowledge, provides the first protocol for facile in vivo visualization of gut microbiota within deep tissues, and offers an instrumental tool for deciphering the complex biology of these gut "dark matters"

Etiologic Diagnosis of Lower Respiratory Tract Bacterial Infections Using Sputum Samples and Quantitative Loop-Mediated Isothermal Amplification

Etiologic diagnoses of lower respiratory tract infections (LRTI) have been relying primarily on bacterial cultures that often fail to return useful results in time. Although DNA-based assays are more sensitive than bacterial cultures in detecting pathogens, the molecular results are often inconsistent and challenged by doubts on false positives, such as those due to system- and environment-derived contaminations. Here we report a nationwide cohort study on 2986 suspected LRTI patients across P. R. China. We compared the performance of a DNA-based assay qLAMP (quantitative Loop-mediated isothermal AMPlification) with that of standard bacterial cultures in detecting a panel of eight common respiratory bacterial pathogens from sputum samples. Our qLAMP assay detects the panel of pathogens in 1047(69.28%) patients from 1533 qualified patients at the end. We found that the bacterial titer quantified based on qLAMP is a predictor of probability that the bacterium in the sample can be detected in culture assay. The relatedness of the two assays fits a logistic regression curve. We used a piecewise linear function to define breakpoints where latent pathogen abruptly change its competitive relationship with others in the panel. These breakpoints, where pathogens start to propagate abnormally, are used as cutoffs to eliminate the influence of contaminations from normal flora. With help of the cutoffs derived from statistical analysis, we are able to identify causative pathogens in 750 (48.92%) patients from qualified patients. In conclusion, qLAMP is a reliable method in quantifying bacterial titer. Despite the fact that there are always latent bacteria contaminated in sputum samples, we can identify causative pathogens based on cutoffs derived from statistical analysis of competitive relationship

NDM-1-Positive K. pneumoniae at a Teaching Hospital in Southwestern China: Clinical Characteristics, Antimicrobial Resistance, Molecular Characterization, Biofilm Assay, and Virulence

Background. The emergence of the NDM-1-positive Klebsiella pneumoniae (K. pneumoniae) strains has led to limited therapeutic options for clinical treatment. Understanding the clinical characteristics, antimicrobial resistance, biofilm assay, and the virulence genes of these isolated strains is of great significance. Methods. The polymerase chain reaction (PCR) was used to screen isolated NDM-1-positive K. pneumoniae. The clinical information of the patients was collected from medical records. The NDM-1-positive K. pneumoniae isolates were subjected to antimicrobial susceptibility testing and multilocus sequence typing. Sixty strains of NDM-1-negative K. pneumoniae isolated during the same period were collected as the control group for the virulence analysis. The virulence phenotype of the strains was preliminarily evaluated by the string test and crystal violet semiquantitative biofilm formation experiment. PCR combined with gene sequencing was used to detect common high toxicity capsule genes (K1, K2, K5, K20, K54, and K57) and common virulence-related genes (entB, ybtS, ureA, ycf, WabG, FimH, uge, iutA, KfuB, aerobactin, rmpA, magA, Alls, IrnN, and VatD). Results. In the 30 nonduplicated NDM-1-positive K. pneumoniae isolates, 43.33% (13/30) of the patients had a history of a stay in the neonatal intensive care unit (NICU). All of the isolates exhibited multidrug resistance. Nine STs were identified, 77% (10/13) strains from the NICU were ST11. The NDM-1-positive K. pneumoniae string tests were all negative, and 35% (21/60) NDM-1-negative K. pneumoniae were positive. The ratios of NDM-1-positive K. pneumoniae isolates biofilm formation ability according to strong, medium, and weak classification were 67%, 23%, and 10%, respectively. NDM-1-negative K. pneumoniae isolates were 60%, 25%, and 15%, respectively. There was no statistical difference between the two groups (t = 0.61, P=0.2723). The virulence-associated genes with more than 80% of detection rates among the 30 NDM-1-positive K. pneumoniae isolates included entB (100%, 30/30), ybtS (93.33%, 28/30), ureA (90%, 27/30), ycf (83.33%, 25/30), and wabG (90%, 27/30). KfuB and iutA were detected at prevalence of 3.33% and 13.33%. vatD, allS, iroN, aerobactin, and rmpA were not detected. In the NDM-1-negative K. pneumoniae, all other 14 virulence genes except VatD were detected. After statistical analysis, FimH, WabG, ycf, iutA, kfuB, aerobactin, rmpA, and Alls virulence genes, P<0.005, there was a statistical difference. Conclusion. NDM-1-positive K. pneumoniae exhibited multidrug resistance, MLST typing is mainly ST11, there is small clonal dissemination in the NICU in the hospital, and the NDM-1-positive K. pneumoniae virulence genes carrier rate is lower than the NDM-1-negative K. pneumoniae virulence genes carrier rate

Design of Novel PLA/OMMT Films with Improved Gas Barrier and Mechanical Properties by Intercalating OMMT Interlayer with High Gas Barrier Polymers

Polymer/clay composites are an innovative class of materials. In this study, we present a facile method for the preparation of biodegradable and robust PLA/organomodified montmorillonite (OMMT) composite films with excellent gas barrier performance. When the design of PLA/OMMT composite films, in addition to making OMMT have good intercalation effect in the matrix, the compatibility of intercalating polymer and matrix should also be considered. In this work, two polymers with high gas barrier properties, namely poly(vinyl alcohol) (PVA) and ethylene vinyl alcohol copolymer (EVOH), were selected to intercalate OMMT. The morphology and microstructures of the prepared PLA/PVA/OMMT and PLA/EVOH/OMMT composites were characterized by the X-ray diffraction measurement, scanning electron microscopy, and differential scanning calorimetry. It was shown that the good dispersibility of PVA in the PLA matrix, rather than the intercalation effect, was responsible for the improved gas barrier and mechanical properties of PLA/PVA/OMMT composite. The elongation at break increases from 4.5% to 22.7% when 1 wt % PVA is added to PLA/OMMT. Moreover, gas barrier of PLA/PVA1/OMMT measured as O2 permeability is 52.8% higher than that of neat PLA. This work provides a route to intercalate OMMT interlayer with high gas barrier polymers and thus can be a useful reference to fabricate PLA/OMMT composites with improved gas barrier and mechanical properties. A comparison of oxygen permeabilities with existing commercial packaging films indicates that the biodegradable PLA/PVA/OMMT may serve as a viable substitute for packaging film applications

Correlation of Gravity and Magnetic Field Changes Preceding Strong Earthquakes in Yunnan Province

The annual variation trend of the gravity and lithospheric magnetic field for adjacent periods are analyzed by using the observation of rover gravity and geomagnetic fields in Yunnan from 2011 to 2021, which tend to be consistent every year during the seismogenic process of a strong earthquake. Thus, this study normalizes the annual value of the adjacent periods for the gravity and lithospheric magnetic field. The normalized values are converted into two classifications that can be compared within [−1,1]. In Yunnan Province, a grid of 0.1° × 0.1° was used to compare the data correlation between the variation of gravity and the variation in the lithospheric magnetic field at the same location. The results are as follows. First, the variation trend of the gravity field and total magnetic field tend to be synchronous year to year in strong earthquake years. The range of consistency increases gradually with the approach of the earthquake year reaching its maximum one year before the earthquake. Throughout the region, the overlap number of normalized annual variations in gravity and magnetic field reaches its maximum, and the peak difference of kernel density curve reaches its minimum. Second, the correlation coefficient of the annual variation in the gravity and magnetic field increases year to year during the development of a strong earthquake within a smaller region surrounding the event. The maximum appears one year before the earthquake, and after the earthquake, the correlation decreases. The analysis of gravity and magnetic fusion characteristics can be employed for the prediction of strong earthquakes