Study of Bitespiramycin Distribution in Rats and Cerebrospinal Fluid of Patients by a Sensitive LC-MS/MS Method with Rapid Sample Preparation.

Bitespiramycin, has been shown to have a therapeutic effect against respiratory tract inflammation, including a potential effect against COVID-19. A current clinical trial in China showed that bitespiramycin was an effective treatment for severe pneumonia and intracranial infection. However, there is lack of an analytical method to elucidate the distribution of bitespiramycin. In this study, a highly sensitive, rapid and reliable UPLC-MS/MS method was developed to comprehensively characterize the bitespiramycin distribution in various bio-samples, which is significantly improved upon the published work. A rapid sample preparation method was developed by using n-butanol as the solvent to extract bitespiramycin from different bio-samples. The extract was then directly analyzed by UPLC-MS/MS coupled with an alkaline-resistant column after centrifugation which avoids the time-consuming concentration process under nitrogen and redissolution. The method was employed to accurately quantify bitespiramycin and its metabolites in rat plasma, tissues, and human cerebrospinal fluid. Notably, the presence of bitespiramycin and its metabolites was identified for the first time in various rat organs including brain, testis, bladder and prostate as well as in human cerebrospinal fluid. This newly developed approach shows great promise for drug distribution assays including other antibiotics and can help elucidate the ADME of bitespiramycin.

CsRVE1 promotes seasonal greening of albino Camellia sinensis cv. Huangkui by activating chlorophyll biosynthesis.

Seasonal greening is a crucial survival strategy for albino tea cultivars, during which dysfunctional chloroplasts recover and chlorophyll biosynthesis increases in albino leaves. However, the regulatory mechanisms of seasonal greening in albino tea plants remain unclear. Here, we report that CsRVE1, a nuclear-located Myb-like transcription factor, can positively modulate the seasonal greening of albino Camellia sinensis cv. Huangkui leaves by activating the expression of genes involved in light harvesting and chlorophyll biosynthesis. The transcriptional expression of CsRVE1 increased during seasonal greening and was tightly correlated with increases in the expression of genes involved in light harvesting (CsLhcb) and chlorophyll biosynthesis (CsCHLH, CsHEMA1 and CsCAO). In vivo and in vitro molecular analyses showed that CsRVE1 can directly bind to the promoters of CsLhcb, CsCHLH and CsPORA, eventually leading to chlorophyll accumulation in tea leaves. Furthermore, transient suppression of CsRVE1 in tea leaves led to a decrease in target gene expression. In contrast, the overexpression of CsRVE1 in Arabidopsis led to chlorophyll increases and the activation of AtLhcb, AtPORA, AtCHLH, etc. These results identify CsRVE1 as an important promoter of seasonal greening that functions by regulating genes involved in chlorophyll biosynthesis in albino tea plants and shed new light on the regulatory mechanisms of leaf phenotypes in plants.

UV-B regulates seasonal greening of albino leaves by modulating CsHY5-inhibiting chlorophyll biosynthesis in Camellia sinensis cv. Huangkui.

Seasonal greening is crucial for albino plants but the underlying regulatory mechanism is unclear, especially concerning light regulation as one of the most important environmental factors for light-sensitive albino tea plants. Here, we report that the UV-B signal regulates the seasonal greening process of albino leaves by modulating CsHY5-inhibiting chlorophyll biosynthesis in Camellia sinensis cv. Huangkui. Reduction of solar UV-B in plantation promoted the seasonal greening of albino 'HK' leaves by inhibiting CsHY5 transcription and activating genes involved in light-harvesting CsLhlb and the chlorophyll biosynthetic pathway (CsCHLH, CsHEMA1, and CsPORA), leading to enrichment of chlorophyll accumulation and recovery of dysfunctional chloroplasts. In contrast, indoor supplementary UV-B exposure reduced chlorophylls by activating CsHY5 but inhibiting chlorophyll biosynthetic genes. In vivo and in vitro molecular analyses showed that CsHY5 can directly bind to the promoters of CsLhlb, CsCHLH, CsHEMA1, and CsPORA. These results indicate that CsHY5 acts as a repressor for the seasonal greening of the albino tea plants in response to the UV-B signal. This is the first study that investigates the regulatory role of the CsHY5-mediated UV-B signal in regulating the seasonal greening of the albino tea plant, which improves our understanding of light regulation in leaf phenotypes of higher plants.

Therapeutic effect and underlying mechanism of Shenkang injection against cisplatin-induced acute kidney injury in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Shenkang injection (SKI), a Chinese patent medicine injection, has been approved for the treatment of chronic kidney disease (CKD) due to its definite clinical therapeutic efficacy. However, the effect and associated underlying mechanism of Shenkang injection against cisplatin (CDDP)-induced acute kidney injury (AKI) has not yet been well elucidated. AIM OF THE STUDY: This study aims to investigate the therapeutic effect and associated underlying mechanism of Shenkang injection against CDDP-induced AKI. MATERIALS AND METHODS: We established a CDDP-induced AKI mouse model to evaluate renal function by biochemical markers measurement and to observe histopathological alterations by haemotoxylin and eosin (HE)-staining sections of renal. In addition, the distribution of representative components of SKI in the kidneys of mice was evaluated by liquid chromatography tandem mass spectrometry (LC-MS/MS). Furthermore, the degree of oxidative stress and inflammation were assessed by detecting the levels of inflammatory cytokines and oxidants, while the related mechanisms were elucidated by network pharmacology. RESULTS: CDDP could induce excessive inflammation and severe injury to the kidneys of mice. However, SKI significantly ameliorated the kidney damages and improved the renal function by reducing the levels of renal function markers (SCr, BUN and urine protein), and inhibiting the production of inflammatory cytokines IL-34, IL-6 and TNF-α. SKI repaired oxidative balance through up-regulation of antioxidants SOD and GSH and down-regulated oxidants MDA. Moreover, 4 components from SKI were detected in the kidney by LC-MS/MS quantification. In addition, pharmacology network indicated the PI3K/AKT, TNF, MAPK, and p53 were the possible signaling pathways for the therapeutic effect of SKI against CDDP-induced AKI, which were related to inflammation, oxidative stress and apoptosis. CONCLUSION: In the present study, we for the first time demonstrated that SKI alleviates CDDP-induced nephrotoxicity by antioxidant and anti-inflammation via regulating PI3K/AKT, MAPK, TNF, and p53 signaling pathways. The study may provide a scientific rationale for the clinical indication of SKI.

A hybrid explicit implicit staggered grid finite-difference scheme for the first-order acoustic wave equation modeling.

Implicit staggered-grid finite-difference (SGFD) methods are widely used for the first-order acoustic wave-equation modeling. The identical implicit SGFD operator is commonly used for all of the first-order spatial derivatives in the first-order acoustic wave-equation. In this paper, we propose a hybrid explicit implicit SGFD (HEI-SGFD) scheme which could simultaneously preserve the wave-equation simulation accuracy and increase the wave-equation simulation speed. We use a second-order explicit SGFD operator for half of the first-order spatial derivatives in the first-order acoustic wave-equation. At the same time, we use the implicit SGFD operator with added points in the diagonal direction for the other first-order spatial derivatives in the first-order acoustic wave-equation. The proposed HEI-SGFD scheme nearly doubles the wave-equation simulation speed compared to the implicit SGFD schemes. In essence, the proposed HEI-SGFD scheme is equivalent to the second-order FD scheme with ordinary grid format. We then determine the HEI-SGFD coefficients in the time-space domain by minimizing the phase velocity error using the least-squares method. Finally, the effectiveness of the proposed method is demonstrated by dispersion analysis and numerical simulations.

Screening of the ubiquitin-proteasome system activators for anti-Alzheimer's disease by the high-content fluorescence imaging system.

Ubiquitin-proteasome system (UPS) plays an important role in neurodegenerative diseases, such as Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD). The discovery of UPS activators for anti-neurodegenerative diseases is becoming increasingly important. In this study, we aimed to identify potential UPS activators using the high-throughput screening method with the high-content fluorescence imaging system and validate the neuroprotective effect in the cell models of AD. At first, stable YFP-CL1 HT22 cells were successfully constructed by transfecting the YFP-CL1 plasmid into HT22 cells, together with G418 screening. The degradation activity of the test compounds via UPS was monitored by detecting the YFP fluorescence intensity reflected by the ubiquitin-proteasome degradation signal CL1. By employing the high-content fluorescence imaging system, together with stable YFP-CL1 HT22 cells, the UPS activators were successfully screened from our established TCM library. The representative images were captured and analyzed, and quantification of the YFP fluorescence intensity was performed by flow cytometry. Then, the neuroprotective effect of the UPS activators was investigated in pEGFP-N1-APP (APP), pRK5-EGFP-Tau P301L (Tau P301L), or pRK5-EGFP-Tau (Tau) transiently transfected HT22 cells using fluorescence imaging, flow cytometry, and Western blot. In conclusion, our study established a high-content fluorescence imaging system coupled with stable YFP-CL1 HT22 cells for the high-throughput screening of the UPS activators. Three compounds, namely salvianolic acid A (SAA), salvianolic acid B (SAB), and ellagic acid (EA), were identified to significantly decrease YFP fluorescence intensity, which suggested that these three compounds are UPS activators. The identified UPS activators were demonstrated to clear AD-related proteins, including APP, Tau, and Tau P301L. Therefore, these findings provide a novel insight into the discovery and development of anti-AD drugs.

Humins with Efficient Electromagnetic Wave Absorption: A By-Product of Furfural Conversion to Isopropyl Levulinate via a Tandem Catalytic Reaction in One-Pot.

Both one-pot catalytic conversion of furfural (FAL) to isopropyl levulinate (PL) and carbonization of by-product (humins) for electromagnetic wave absorption are discussed, which provides inspiration that humins can be applied to electromagnetic wave absorption. In the former, phosphotungstic acid (PW) is employed as a homogeneous catalyst to convert FAL to PL via a tandem reaction in one pot, with the formation of a vast amount of humins. With FAL and various intermediates as substrates, it was found that humins was a polymerization product of FAL, furfuryl alcohol (FOL) and furfuryl ester (FE) with furan rings. In addition, the in situ attenuated total reflection infrared (ATR-IR) spectra also provided a basis for the proposed reaction route. In the latter, with the humins as raw material, P species and WO3 doped nano-porous carbon (Humins-700) platform formed after high-temperature annealing is used for electromagnetic wave absorption and manifests desirable absorption performance. The minimum reflection loss (RLmin ) value is -47.3 dB at 13.0 GHz with a thickness of 2.0 mm and the effective absorption bandwidth reaches 4.5 GHz (11.2-5.7 GHz).

MiR-1260b inhibitor enhances the chemosensitivity of colorectal cancer cells to fluorouracil by targeting PDCD4/IGF1.

Colorectal cancer (CRC) is the most common malignant tumor type and has become resistant to 5-fluorouracil (5-FU) in recent decades, which is one of the most popular therapies. Recently, microRNA (miRNA or miR) has been investigated as a potential therapeutic strategy for CRC. However, there has been little investigation of the underlying mechanism of the association between expression of miRNA and chemosensitivity. The present study aimed to investigate the effect of miR-1260b inhibitor on CRC cells, and their chemosensitivity to 5-FU, by treating them with the miR-1260b inhibitor. miR-1260b inhibitor was demonstrated to significantly promote the proliferation and invasion of the CRC cell line, HCT116, and to increase the apoptotic rate. Furthermore, it was validated that programmed cell death 4 (PDCD4) was a direct target of miR-1260b inhibitor in CRC with bioinformatics tools and a luciferase assay. Western blot analysis revealed that miR-1260b inhibitor could significantly decrease PDCD4 expression, and downregulate the expression of phosphorylated-Akt (p-Akt) and phosphorylated-extracellular-signal-regulated kinase (p-ERK). In conclusion, it was confirmed that the anti-tumor effect of the miR-1260b inhibitor was conducted by blocking the phosphorylated 3-kinase/Akt pathway as dysregulated protein expression induced by miR-1260b inhibitor was rescued by insulin-like growth factor. Notably, miR-1260b inhibitor could significantly enhanced the chemoresponse of HCT116 cells to 5-FU via reduced proliferation, increased apoptosis, and downregulation of PDCD4, p-Akt and p-ERK protein expression. In summary, the present study may provide a novel direction for future clinical therapy to enhance the chemosensitivity of tumor cells.

Regularizing active set method for nonnegatively constrained ill-posed multichannel image restoration problem.

In this paper, we consider the nonnegatively constrained multichannel image deblurring problem and propose regularizing active set methods for numerical restoration. For image deblurring problems, it is reasonable to solve a regularizing model with nonnegativity constraints because of the physical meaning of the image. We consider a general regularizing l(p)-l(q) model with nonnegativity constraints. For p and q equaling 2, the model is in a convex quadratic form, therefore, the active set method is proposed since the nonnegativity constraints are imposed naturally. For p and q not equaling 2, we present an active set method with a feasible Newton-conjugate gradient solution technique. Numerical experiments are presented for ill-posed three-channel blurred image restoration problems.