Yiqi Kaimi prescription regulates protein phosphorylation to promote intestinal motility in slow transit constipation.

ETHNOPHARMACOLOGICAL RELEVANCE: The clinical efficacy of the Yiqi Kaimi prescription has been confirmed in slow transit constipation. However, the effects and biological mechanism of Yiqi Kaimi prescription are still unclear. AIMS OF THE STUDY: To identify the effects of Yiqi Kaimi prescription on intestinal motility; To reveal the potential key targets and pathways of Yiqi Kaimi prescription for the treatment of slow transit constipation. MATERIALS AND METHODS: The effects of Yiqi Kaimi prescription on slow transit constipation were investigated in a mouse model. The terminal ink propulsion experiment and fecal indocyanine green imaging was used to measure the intestinal transit time. Protein phosphorylation changes in colon tissues treated with Yiqi Kaimi prescription were detected using a Phospho Explorer antibody microarray. Bioinformatic analyses were performed using the Database for Annotation Visualization and Integrated Discovery (DAVID) and the Search Tool for the Retrieval of Interacting Genes (STRING). Western blot analysis and immunohistochemistry confirmed the observed changes in phosphorylation. RESULT: s: Yiqi Kaimi prescription significantly increased the intestinal transit rate (P < 0.05 vs. model) and reduced the time to first discharge of feces containing fecal indocyanine green imaging in mice (P < 0.05 vs. model). The administration of Yiqi Kaimi prescription induced phosphorylation changes in 41 proteins, with 9 upregulated proteins and 32 downregulated proteins. Functional classification of the phosphorylated proteins with DAVID revealed that the critical biological processes included tyrosine protein kinases, positive regulation of calcium-mediated signaling and response to muscle stretch. The phosphorylation of the spleen tyrosine kinase (SYK) at Tyr348 increased 2.19-fold, which was the most significant change. The phosphorylation level of the transcription factor p65 (RELA) at Thr505 was decreased 0.57-fold. SYK was a hub protein in the protein-protein interaction network and SYK and RELA formed the core of the secondary subnetwork. The key protein phosphorylation after treatment with Yiqi Kaimi prescription were verified by Western blot analysis and immunohistochemistry. CONCLUSION: Yiqi Kaimi prescription significantly enhanced intestinal motility. This effect was attributed to alterations in the phosphorylation levels of various target proteins. The observed changes in protein phosphorylation, including SYK and RELA, may serve as crucial factors in the treatment of slow transit constipation.

Differential expression of zinc finger CCHC-type superfamily proteins in thyroid carcinoma and their associations with tumor immunity.

BACKGROUND: The zinc-finger CCHC-type (ZCCHC) superfamily proteins are characterized by the shared sequence CX2-CX4-HX4-C and thought to own high affinity to single-stranded nucleic acids, particularly RNAs. In humans, a total of 24 ZCCHC proteins have been annotated in the HUGO Gene Nomenclature Committee (HGNC, https://www.genenames.org/ ) database with most of these members involved in multiple steps of RNA metabolism. Many studies have indicated that the ZCCHC genes play a regulatory role in the development and progression of solid tumors. To date, the expression pattern and prognostic value of ZCCHC factors in thyroid carcinomas have not been reported. METHODS: Bioinformatics analyses on the functions of ZCCHC factors in thyroid carcinoma (THCA) patients were performed based on various databases, i.e., TCGA, GEPIA, Kaplan-Meier Plotter, and TIMER. RESULTS: Compared with normal tissues, the expression of ZCCHC12 mRNA was significantly increased in THCA tissues. And it was associated with the overall survival of THCA patients, based on the Kaplan-Meier Plotter database. Furthermore, the expression levels of all ZCCHCs were correlated with tumor stages, implying its high relevance to THCA, specifically its immunity. CONCLUSION: The ZCCHC genes, represented by ZCCHC12, are differentially expressed in THCA staging. These genes are associated with immune infiltration of THCA and identified as the potential therapeutic targets for immunotherapy in THCA patients, which are possible novel biomarkers for the treatment of THCA.

Inactivation of Vibrio parahaemolyticus and retardation of quality loss in oyster (Crassostrea gigas) by ultrasound processing during storage.

The health problems caused by foodborne pathogens of raw oysters have been widely concerned. Traditional heating methods tend to lead the loss of the original nutrients and flavors, in this study, the nonthermal ultrasound technology was applied to inactivate Vibrio parahaemolyticus on raw oysters, and the retardation effects on microbial growth and quality loss of oysters stored at 4 ℃ after ultrasonic treatment were also investigated. After treated by 7.5 W/mL ultrasound for 12.5 min, the Vibrio parahaemolyticus in oysters was reduced by 3.13 log CFU/g. By measuring total aerobic bacteria and total volatile base nitrogen, the growth trend after ultrasonic treatment was delayed compared with heat treatment, and the shelf life of oysters was prolonged. At the same time, ultrasonic treatment delayed the changes of color difference and lipid oxidation of oysters during cold storage. Texture analysis showed that ultrasonic treatment helped maintain the good textural structure of oysters. Histological section analysis also demonstrated that muscle fibers were still tightly packed after ultrasonic treatment. Low-field nuclear magnetic resonance (LF-NMR) illustrated that the water in the oysters was well maintained after ultrasonic treatment. In addition, gas chromatograph - ion mobility spectrometer (GC-IMS) showed that ultrasound treatment could better preserve the flavor of oysters during cold storage. Therefore, it is believed that ultrasound can inactivate foodborne pathogens of raw oysters and keep its freshness and original taste better during storage.

Stresses make microbe undergo programmed cell death: Mechanisms and opportunities.

In recent years, programmed cell death (PCD) has attracted the attention of many scholars as an active death process. PCD refers to an active extinction process that occurs after cells receive a certain signal or are stimulated by certain factors in order to maintain the stability of the internal environment. It is orchestrated by precise molecular circuitry. Initially, PCD was thought to exhibit altruistic behavior of "active devotion", a physiological characteristic of multicellular organisms. However, many evidences show that PCD also exists in microbe, and researchers in recent years have found that some stressors during modern food processing could cause microbe undergo different forms of programmed death, including ultraviolet, gamma radiation, cold plasma, ultrasound, weak carboxylic acids and H2O2. The attention to these situations will help researchers to explore novel mechanisms of sterilization. At present, microbial PCD can be further divided into three types, including apoptosis-like death, autolytic death, and toxin-antitoxin systems based on the differences in their intrinsic mechanisms. In order to better utilize these PCD mechanisms, four directions are provided from searching for new antimicrobial targets, development of new types of preservatives, engineering design in the field of synthetic biology and modification of fermentation strains. We believe that this review can provide useful help for scholars to carry out further research on microbial PCD.

The microbial stress responses of Escherichia coli and Staphylococcus aureus induced by chitooligosaccharide.

Intermediate-state bacteria produced in the process of bacteriostasis have always been a potential threat to public health, but they are very easy to be overlooked. As a natural and non-toxic biological antibacterial agent, chitooligosaccharide (COS) has attracted the public's attention. However, little is known about the microbial stress response during the antibacterial process of COS. In this study, the antibacterial mechanisms of COS were expounded, and the formation of sublethal and viable but nonculturable (VBNC) state were further investigated. The COS was shown to bind to the cell envelopes, and the permeability and integrity of bacterial cell membrane were damaged severely, accompanied by the increase of intracellular reactive oxygen species and decrease of adenosine triphosphate content. Flow cytometry analysis indicated that COS finally inactivated Escherichia coli through the sublethal injury process. While for Staphylococcus aureus, some cells were induced into VBNC state by COS, causing incomplete inactivation.

Continuous rise of the tropopause in the Northern Hemisphere over 1980-2020.

Tropopause height (H) is a sensitive diagnostic for anthropogenic climate change. Previous studies showed increases in H over 1980­2000 but were inconsistent in projecting H trends after 2000. While H generally responds to temperature changes in the troposphere and stratosphere, the relative importance of these two contributions is uncertain. Here, we use radiosonde balloon observations in the Northern Hemisphere (NH) over 20°N to 80°N to reveal a continuous rise of H over 1980­2020. Over 2001­2020, H increases at 50 to 60 m/decade, which is comparable to the trend over 1980­2000. The GPS radio occultation measurements from satellites and homogenized radiosonde records are in good agreement with those results. The continuous rise of the tropopause in the NH after 2000 results primarily from tropospheric warming. A large trend in H remains after major natural forcings for H are removed, providing further observational evidence for anthropogenic climate change.

Deeper insight into hydrolysis mechanisms of polyester/cotton blended fabrics for separation by explicit solvent models.

Aiming to get a deeper and accurate understanding on separation of polyester/cotton blended fabrics in subcritical water, the hydrolysis mechanisms of cellulose and polyester were studied using dispersion-corrected density functional theory (DFT-D) method with and without explicit H2O under the conductor-like screening model (COSMO) set. The number and locations of explicit H2O were determined by their likely functions including being dissociation and solvent and catalyst. The calculations disclosed that explicit H2O provide inductive activation on glycosidic bond of cellulose and ester groups at the center of polyester and the assistance on the transfer of proton as proton-carrier and as catalyst of proton shuttle, affecting the reaction and activation energies in a realistic manner. In addition, the number of explicit H2O molecules functioning as catalyst of proton shuttle may also has a strong influence on catalytic activity. Based on the improved explicit solvation models, the overall activation energies of proposed hydrolysis mechanisms for cellulose and polyester are 14.81 and 21.46 kcal/mol respectively, which explains the preferential hydrolysis of cellulose from experimental results.

Quercetin-loaded mixed micelles exhibit enhanced cytotoxic efficacy in non-small cell lung cancer in vitro.

In the present study, quercetin (QUR)-loaded mixed micelles (QUR-M) were prepared with the aim of improving the physicochemical and anticancer efficacy of QUR in lung cancer cells. The mixed micelles comprised tocopheryl polyethylene glycol 1000 succinate (TPGS) and a 1,2-distearoyl-sn-glycero-3-phosphatidylethanolamine derivative of polyethylene glycol. The nanosized QUR-M exhibited a pH-responsive and controlled release of QUR that is likely to be beneficial in cancer treatment. The results of an MTT assay clearly demonstrated that the anticancer effect of QUR-M in A549 cancer cells was stronger compared with that of free QUR at 24 and 48 h time points. The half-maximal inhibitory concentrations of QUR and QUR-M were observed to be 12.45 and 6.42 µg/ml, respectively. When stained with Hoechst 33342 and observed using a confocal laser scanning microscope, A549 cells treated with QUR-M exhibited severe chromatin condensation and apoptotic body formation of the nuclei. Overall, high intracellular uptake, sustained drug release and the presence of TPGS in the mixed micelles may result in an increased inhibitory effect against cell proliferation and improved therapeutic efficacy in lung cancers.

Optimizing a desirable fare structure for a bus-subway corridor.

This paper aims to optimize a desirable fare structure for the public transit service along a bus-subway corridor with the consideration of those factors related to equity in trip, including travel distance and comfort level. The travel distance factor is represented by the distance-based fare strategy, which is an existing differential strategy. The comfort level one is considered in the area-based fare strategy which is a new differential strategy defined in this paper. Both factors are referred to by the combined fare strategy which is composed of distance-based and area-based fare strategies. The flat fare strategy is applied to determine a reference level of social welfare and obtain the general passenger flow along transit lines, which is used to divide areas or zones along the corridor. This problem is formulated as a bi-level program, of which the upper level maximizes the social welfare and the lower level capturing traveler choice behavior is a variable-demand stochastic user equilibrium assignment model. A genetic algorithm is applied to solve the bi-level program while the method of successive averages is adopted to solve the lower-level model. A series of numerical experiments are carried out to illustrate the performance of the models and solution methods. Numerical results indicate that all three differential fare strategies play a better role in enhancing the social welfare than the flat fare strategy and that the fare structure under the combined fare strategy generates the highest social welfare and the largest resulting passenger demand, which implies that the more equity factors a differential fare strategy involves the more desirable fare structure the strategy has.