Regulatory T cells (Tregs) are renowned for maintaining homeostasis and self-tolerance through their ability to suppress immune responses. For over two decades, Tregs have been the subject of intensive research. The immunosuppressive and migratory potentials of Tregs have been exploited, especially in the areas of cancer, autoimmunity and vaccine development, and many assay protocols have since been developed. However, variations in assay conditions in different studies, as well as covert experimental factors, pose a great challenge to the reproducibility of results. Here, we focus on human Tregs derived from clinical samples and highlighted caveats that should be heeded when conducting Tregs suppression and migration assays. We particularly delineated how factors such as sample processing, choice of reagents and equipment, optimization and other experimental conditions could introduce bias into the assay, and we subsequently proffer recommendations to enhance reliability and reproducibility of results. It is hoped that prioritizing these factors will reduce the tendencies of generating false and misleading results, and thus, help improve our understanding and interpretation of Tregs functional studies.Betulinic acid (BA) is a star member of the pentacyclic triterpenoid family, which exhibits great prospects for antitumor drug development. In an attempt to develop novel antitumor candidates, 21 BA-nitrogen heterocyclic derivatives were synthetized, in addition to four intermediates, 23 of which were first reported. Moreover, they were screened for in-vitro cytotoxicity against four tumor cell lines (Hela, HepG-2, BGC-823 and SK-SY5Y) by a standard methylthiazol tetrazolium (MTT) assay. The majority of these derivatives showed much stronger cytotoxic activity than BA. Remarkably, the most potent compound 7e (the half maximal inhibitory concentration (IC50) of which was 2.05 ± 0.66 μM) was 12-fold more toxic in vitro than BA-treated Hela. Furthermore, multiple fluorescent staining techniques and flow cytometry collectively revealed that compound 7e could induce the early apoptosis of Hela cells. Structure-activity relationships were also briefly discussed. The present study highlighted the importance of introducing nitrogen heterocyclic rings into betulinic acid in the discovery and development of novel antitumor agents.Biochars were produced with magnesium chloride as an additive for the sorption of hexavalent chromium dissolved in water using five types of straw (from taro, corn, cassava, Chinese fir, and banana) and one type of shell (Camellia oleifera) as the raw materials. The removal of hexavalent chromium by the six biochars mainly occurred within 60 min and then gradually stabilized. The kinetics of the adsorption process were second order, the Langmuir model was followed, and the adsorption of Cr(VI) by the six biochars was characterized by Langmuir monolayer chemisorption on a heterogeneous surface. Banana straw biochar (BSB) had the best performance, which perhaps benefitted from its special structure and best adsorption effect on Cr(VI), and the theoretical adsorption capacity was calculated as 125.00 mg/g. For the mechanism analysis, Mg-loaded biochars were characterized before and after adsorption by Fourier transform infrared spectroscopy (FTIR), X-ray diffractometry (XRD), and scanning electron microscopy/energy dispersive spectroscopy (SEM-EDS). The adsorption mechanism differed from the adsorption process of conventional magnetic biochar, and biochar interactions with Cr(VI) were controlled mainly by electrostatic attraction, complexation, and functional group bonding. In summary, the six Mg-loaded biochars exhibit great potential advantages in removing Cr(VI) from wastewater and have promising potential for practical use, especially BSB, which shows super-high adsorption performance.The study was conducted in vitro to investigate the effects of different ratios of Urtica cannabina and Leymus chinensis on fiber microstructure and digestibility in ruminal fluid. The experiment was divided into five groups based on the U. cannabina/L. chinensis ratios A (0100), B (3070), C (5050), D (7030), and E (1000). The culture medium was collected at 0, 1, 3, 6, 12, and 24 h. The results showed that (1) in vitro crude protein degradability (IVCPD) was higher in group A, whereas in vitro neutral detergent fiber degradability (IVNDFD) was higher in group C (p less then 0.05); (2) protozoa count was increased from 1 h to 3 h and decreased afterwards, with significant differences observed in several genera (p less then 0.05); (3) microbial crude protein (MCP) contents at 1, 3, 6, and 24 h were higher in groups A and C (p less then 0.05); (4) the basic tissue of U. cannabina was gradually degraded. At 24h, the secondary xylem vessel structure was observed in groups B and C, but not in groups D and E. In summary, there was higher neutral detergent fiber (NDF) digestibility, higher rumen MCP contents, and lower protozoa count, showing the significance of the 5050 ratio for microbial growth and fiber digestibility.Breakfast is one of the most important meals of the day. A good quality breakfast must include dairy products, cereals, and fruits. The aim of this study is to determine breakfast dietary patterns and their nutritional quality among Chilean university students. A cross-sectional non-probabilistic study was conducted in 200 university students between 18 and 27 years in Santiago, Chile. To identify dietary patterns and breakfast quality, a breakfast food survey was conducted. Patterns were identified by factor analysis. Most of the subjects (53%) ate breakfast daily, with a higher prevalence among females (60.2% vs. 43.7%, p less then 0.05); 68% did not consume fruits and 17.5% had good breakfast quality, with no differences by sex. Four breakfast dietary patterns were identified "dairy & cereals", "healthy", "traditional salty" and "traditional sweet" that together explained 35.6% of the total variance. There was no sex difference in predominant dietary patterns. The "dairy & cereals" and "traditional sweet" patterns were associated with regularly eating breakfast (β -0.47, p = 0.001; β -0.32, p = 0.020) and the "healthy" pattern with BMI ≥25 kg/m2 (β 0.35, p = 0.024). In conclusion, breakfast quality was inadequate due to low fruit consumption and energy intake. The four identified patterns included cereals, bread, dairy, fats and sugars. JTE 013 Results may be usual in the planning of future interventions aimed at improving breakfast consumption and quality in university students.