[Developmental toxicity of Cry1Ab protein in the embryonic stem-cell model].

OBJECTIVE: To evaluate the developmental toxicity of Cry1Ab protein by studying its effects on cell proliferation and differentiation ability using a developmental toxicity assessment model based on embryonic stem-cell. METHODS: Cry1Ab protein was tested in seven dose groups (31.25, 62.50, 125.00, 250.00, 320.00, 1 000.00, and 2 000.00 µg/L) on mouse embryonic stem cells D3 (ES-D3) and 3T3 mouse fibroblast cells, with 5-fluorouracil (5-FU) used as the positive control and phosphate buffer saline (PBS) as the solvent control. Cell viability was detected by CCK-8 assay to calculate the 50% inhibitory concentration (IC50) of the test substance for different cells. Additionally, Cry1Ab protein was tested in five dose groups (125.00, 250.00, 320.00, 1 000.00, and 2 000.00 µg/L) on ES-D3 cells, with PBS as the solvent control and 5-FU used for model validation. After cell treatment, cardiac differentiation was induced using the embryonic bodies (EBs) culture method. The growth of EBs was observed under a microscope, and their diameters on the third and fifth days were measured. The proportion of EBs differentiating into beating cardiomyocytes was recorded, and the 50% inhibition concentration of differentiation (ID50) was calculated. Based on a developmental toxicity discrimination function, the developmental toxicity of the test substances was classified. Furthermore, at the end of the culture period, mRNA expression levels of cardiac differentiation-related markers (Oct3/4, GATA-4, Nkx2.5, and ß-MHC) were quantitatively detected using real-time quantitative polymerase chain reaction (qPCR) in the collected EBs samples. RESULTS: The IC50 of 5-FU was determined as 46.37 µg/L in 3T3 cells and 32.67 µg/L in ES-D3 cells, while the ID50 in ES-D3 cells was 21.28 µg/L. According to the discrimination function results, 5-FU was classified as a strong embryotoxic substance. There were no statistically significant differences in cell viability between different concentrations of Cry1Ab protein treatment groups and the control group in both 3T3 cells and ES-D3 cells (P>0.05). Moreover, there were no statistically significant differences in the diameter of EBs on the third and fifth days, as well as their morphology, between the Cry1Ab protein treatment groups and the control group (P>0.05). The cardiac differentiation rate showed no statistically significant differences between different concentrations of Cry1Ab protein treatment groups and the control group (P>0.05). 5-FU significantly reduced the mRNA expression levels of ß-MHC, Nkx2.5, and GATA-4 (P < 0.05), showing a dose-dependent trend (P < 0.05), while the mRNA expression levels of the pluripotency-associated marker Oct3/4 exhibited an increasing trend (P < 0.05). However, there were no statistically significant differences in the mRNA expression levels of mature cardiac marker ß-MHC, early cardiac differentiation marker Nkx2.5 and GATA-4, and pluripotency-associated marker Oct3/4 between the Cry1Ab protein treatment groups and the control group (P>0.05). CONCLUSION: No developmental toxicity of Cry1Ab protein at concentrations ranging from 31.25 to 2 000.00 µg/L was observed in this experimental model.

Hypoglycemic Effects and Mechanisms of Buckwheat-Oat-Pea Composite Flour in Diabetic Rats.

Nutritional intervention is a basic way to prevent and treat diabetes mellitus. Appropriate whole grain intake daily is recommended. The study aimed to explore the feasibility of a kind of buckwheat-oat-pea composite flour (BOP, quality ratio of buckwheat:oats:peas = 6:1:1) as a stable food substitution and its underlying mechanisms. High-fat food (HFD) and streptozotocin injection were used to induce diabetes in rats, and buckwheat, oats, and three different doses of BOP were added to the HFD separately for diet intervention. The whole study lasted for 10 weeks, and the glucose tolerance test, lipids, liver injury, and gut microbiota were evaluated in the last week. The diabetic rat model was successfully induced. The BOP significantly changed the glucose and lipids metabolism, decreased liver injury, and changed the composition of the gut microbiota of diabetic rats. The outcomes of the current study revealed that BOP is a potential stable food substitution.

Self-Reported Sedentary Behavior and Metabolic Syndrome among Children Aged 6-14 Years in Beijing, China.

(1) Objective: This study aimed to examine the prevalence of metabolic syndrome (MetS) in children aged 6−14 years in Beijing, and to determine whether sedentary behavior is a risk factor. (2) Methods: Using a multistage stratified cluster random sampling method, 3460 students were selected for the Nutrition and Health Surveillance in Schoolchildren of Beijing (NHSSB). Data on children's sedentary behavior time and MetS indicators were collected using the questionnaires, physical measurements, and laboratory tests. MetS was defined according to the CHN2012 criteria, and logistic regression analysis was used to compare the effects of different sedentary time on MetS and its components. (3) Results: The overall prevalence of MetS among children aged 6−14 in Beijing was 2.4%, and boys, suburban children, and older age were associated with a higher prevalence (χ2 values were 3.947, 9.982, and 27.463, respectively; p < 0.05). In boys, the prevalence rates of abdominal obesity, hyperglycemia, high triglycerides (TG), and low high-density lipoprotein cholesterol (HDL-C) were higher in the high-level sedentary behavior group than those in the low-level sedentary behavior group (p < 0.05); and in girls, the prevalence rates of high TG, low HDL-C, and MetS were higher in the high-level sedentary behavior group than those in the low-level sedentary behavior group (p < 0.05). After adjusting for confounding factors, the multivariate logistic regression results showed that compared with children with low-level sedentary behavior, the risks of abdominal obesity and low HDL-C were higher in boys with high-level sedentary behavior (odds ratio (OR) 1.51, 95% confidence interval (CI) 1.10−2.07, p = 0.011; OR 2.25, 95% CI 1.06−4.76, p = 0.034, respectively); while the risk of abdominal obesity was higher in girls with medium and high-level sedentary behavior (OR 1.52, 95% CI 1.01−2.27, p = 0.043; OR 1.59, 95% CI 1.04−2.43, p = 0.032, respectively). (4) Conclusions: Higher sedentary behavior time was related to the higher risk of MetS components among children aged 6−14 in Beijing. Reducing sedentary behavior may be an important method for preventing metabolic diseases.

Effects of transgenic Bacillus Thuringiensis maize (2A-7) on the growth and development in rats.

This study aimed to determine the effects of genetically modified insect-resistant maize (2A-7) on the growth and development in developing rats. Rats were fed a diet formulated with 2A-7 maize and were compared with rats fed a diet formulated with non-transgenic maize (CK group) and rats fed AIN-93G diet (BC group). 2A-7 maize was formulated into diets at ratios of 82.4% (H group) and 20.6% (L group); non-transgenic maize was formulated into diets at a ratio of 82.4%. From the first day of pregnancy, adult rats were divided into four groups and fed with the above four diets, respectively. Weaning on postnatal day 21, the diets of offspring were consistent with their parents. The results showed that body weight, hematology, serum biochemistry, organ weight, organ coefficients and allergenicity of offspring fed with 2A-7 maize were comparable with those in the CK and BC groups. In physiological and behavioral development experiments, there was no statistically significant difference among groups. Although mCry1Ab proteins were detected in organs and serum, no histopathological changes were observed among groups. In conclusion, A-7 maize cause no treatment-related adverse effects on offspring, indicating that 2A-7 maize is safe for developing rats.