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ObjectiveTo establish a real-time quantitative polymerase chain reaction （PCR） method for rapid and accurate identification of Cronobacter spp. in food samples and artificially contaminated samples.MethodsPrimers and probes were designed based on the conserved region of gyrB of Cronobacter spp. DNA. The method was verified using a specificity test， absolute sensitivity test， relative sensitivity test， and anti-interference test. Detection sensitivity was determined using artificially contaminated samples.ResultsThe method established in this study could specifically amplify seven kinds of Cronobacter spp.， but not the other Enterobacter species closely related to it and other pathogens common in food， suggesting that this method has good anti-interference ability. The absolute sensitivity was 1-10 pg and the relative sensitivity was 103 CFU/mL using Cronobacter sakazakii. It had good anti-interference ability at the genome and culture level. The sensitivity of artificially contaminated samples could reach 100 CFU/mL after incubation at 36 ℃ for 24 h.ConclusionThe real-time PCR method developed in this study is rapid， specific， sensitive， and stable for the detection of Cronobacter spp. in infant formula food samples and can provide technical reference for the detection of Cronobacter spp. in infant formula food.

Stem strength plays a crucial role in the growth and development of plants, as well as in their flowering and fruiting. It not only impacts the lodging resistance of crops, but also influences the ornamental value of ornamental plants. Stem development is closely linked to stem strength; however, the roles of the SPL transcription factors in the stem development of herbaceous peony (Paeonia lactiflora Pall.) are not yet fully elucidated. In this study, we obtained and cloned the full-length sequence of PlSPL14, encoding 1085 amino acids. Quantitative real-time PCR (qRT-PCR) analysis revealed that the expression level of PlSPL14 gradually increased with the stem development of P. lactiflora and was significantly expressed in vascular bundles. Subsequently, utilizing the techniques of virus-induced gene silencing (VIGS) and heterologous overexpression in tobacco (Nicotiana tabacum L.), it was determined that PlSPL14-silenced P. lactiflora had a thinner xylem thickness, a decreased stem diameter, and weakened stem strength, while PlSPL14-overexpressing tobacco resulted in a thicker xylem thickness, an increased stem diameter, and enhanced stem strength. Further screening of the interacting proteins of PlSPL14 using a yeast two-hybrid (Y2H) assay revealed an interactive relationship between PlSPL14 and PlSLR1 protein, which acts as a negative regulator of gibberellin (GA). Additionally, the expression level of PlSLR1 gradually decreased during the stem development of P. lactiflora. The above results suggest that PlSPL14 may play a positive regulatory role in stem development and act in the xylem, making it a potential candidate gene for enhancing stem straightness in plants. [ABSTRACT FROM AUTHOR]

Background: Serum vitamin D is associated with hyperuricemia. However, previous studies have been controversial, with limited focus on children and adolescents. Objective: This study aimed to examine the cross-sectional and longitudinal associations between serum vitamin D and serum uric acid (SUA) levels in children and adolescents. Methods: The cross-sectional survey comprised 4777 participants aged 6 to 18 years, while the longitudinal survey involved 1641 participants aged 6 to 12 years, all derived from an ongoing cohort study in Shenzhen, China. Restricted cubic splines were used to visualize the dose–response relationship between vitamin D and SUA and the risk of higher SUA status. Two-segment generalized linear models (GLM) and logistic models were used to assess the association between vitamin D and SUA and higher SUA status, respectively. The longitudinal analysis used GLM. Results: We observed an inverted U-shaped relationship between vitamin D and SUA (p-overall < 0.0001, p-nonlinear = 0.0002), as well as the risk of higher SUA status (p-overall = 0.0054, p-nonlinear = 0.0015), with the vitamin D inflection point at 24.31 and 21.29 ng/mL, respectively. A 10 ng/mL increment in 25(OH)D3 levels, when below 20.92 ng/mL, was associated with a 68% rise in the risk of higher SUA status (OR: 1.68, 95%CI: 1.07–2.66). Conversely, when 25(OH)D3 levels were above or equal to 20.92 ng/mL, a 10 ng/mL increment was associated with a 45% reduction risk of higher SUA status (OR: 0.55, 95%CI: 0.36–0.84). Longitudinal analysis indicated that the annual change of SUA was from −4.80 (β, 95%CI: −10.74, 1.13) to −9.00 (β, 95%CI: −15.03, −2.99) and then to −6.77 (β, 95%CI: −12.83, −0.71, p for trend = 0.0212) μmol/L when increasing the quartile of vitamin D3. Conclusions: An inverse U-shaped relationship was observed between vitamin D and SUA as well as the risk of higher SUA status. Sufficient vitamin D levels appear to play a preventative role against the age-related increase in SUA. Ensuring adequate vitamin D levels may be beneficial in improving uric acid metabolism. [ABSTRACT FROM AUTHOR]

Summary: Background: The relationship between sugar‐free beverage (SFB) intake and childhood obesity among Chinese children is unknown. Objectives: To describe the status of SFB consumption among children and adolescents in China and assess the association between SFB intake and different types of obesity. Methods: The study was based on the baseline data of an ongoing cohort project named Evaluation and Monitoring on School‐based Nutrition and Growth in Shenzhen (EMSNGS). Food frequency questionnaires were used to collect information on SFB consumption in 3227 students aged 9–17. Physical and clinical examinations were conducted by trained investigators and clinicians. Multivariable binary logistic regression models were performed to assess the association between SFB intake and general obesity, overweight/obesity, abdominal obesity, metabolically unhealthy overweight (MUOW)/metabolically unhealthy obesity (MUO). Results: The median age of the participants was 13.28 years. Among the participants, 55.2% were boys, and 66.1% were adolescents. The median SFB consumption was 16.67 mL/d. After adjusting for potential confounding factors, each 100 mL increase in daily SFB intake was associated with an increased risk of overweight/obesity (OR = 1.14; 95%CI: 1.06–1.23), abdominal obesity (OR = 1.12; 95%CI: 1.03–1.23), and MUOW/MUO (OR = 1.12; 95%CI: 1.02–1.21), respectively. Stratified analyses showed that family income may have an impact on the association between SFB intake and overweight/obesity (P for interaction = 0.021) and abdominal obesity (P for interaction = 0.031). Conclusion: SFB intake was positively associated with childhood obesity in Chinese children, particularly among individuals with high‐income families. [ABSTRACT FROM AUTHOR]

Melanoma is an aggressive malignant tumor. The crucial role of circular RNAs has been documented in many types of cancer, including melanoma. The objective of this study was to uncover the function of circ_0084043 in the biological process of melanoma and associated mechanism of action. The expression of circ_0084043, miR-31, and Krüppel-like factor 3 (KLF3) was determined by qRT-PCR. Cell proliferation and apoptosis were monitored by the MTT assay and flow cytometry assay, respectively. The progression of glycolysis was evaluated according to the levels of glucose consumption, lactate production, and ATP concentration using appropriate detection kits. The relationship between miR-31 and circ_0084043 or KLF3 was predicted by the bioinformatics tool and ascertained by the dual-luciferase reporter assay. The protein levels of KLF3 and glucose transporter 1 (Glut1) were quantified by western blot. A xenograft model was established to ascertain the role of circ_0084043 in vivo. As a result, circ_0084043 expression was reinforced in melanoma tissues and cells. Circ_0084043 knockdown inhibited cell proliferation, induced cell apoptosis, and restrained glycolysis. MiR-31 was a target of circ_0084043, and miR-31 deficiency reversed the role of circ_0084043 knockdown. KLF3 was targeted by miR-31, and KLF3 upregulation abolished the effects of miR-31 enrichment. Moreover, circ_0084043 knockdown impeded tumor growth in vivo and suppressed the level of Glut1 by modulating miR-31 and KLF3. Circ_0084043 promoted cell proliferation and glycolysis, and blocked apoptosis through the circ_0084043–miR-31–KLF3 regulatory axis in melanoma.

Background and aims: Childhood and adolescence are critical periods for linear growth and preventing stunting. Current evidence indicates that dietary protein intake in children and adolescents is often two to three times higher than the recommendations in many regions worldwide. However, few studies have focused on the association between high protein intake and linear growth and stunting in this population. We aim to investigate this association in children and adolescents aged 6 to 18 years in a population with relatively high protein consumption. Methods: We conducted a large cross-sectional study involving 3299 participants from Shenzhen, a modern metropolis of China. Protein intake, including total protein, animal protein, and plant protein, was evaluated by a food-frequency questionnaire and expressed as grams per kilogram of body weight per day (g·kg−1·d−1) and as a percentage of total energy intake (%E). The primary outcomes were body height and height-for-age Z score (HAZ). Generalized linear models and logistic regression analyses were employed to examine the associations between protein intake and outcomes. We also conducted stratified analyses across different genders and pubertal stages in the aforementioned associations. Results: The mean protein intake was 1.81 g·kg−1·d−1 (17% E). After adjusting for serum calcium, zinc, vitamin D3, vitamin A levels, birth outcomes, lifestyle, and parental characteristics, each standard deviation increase of 1 in protein intake (0.64 kg−1·d−1) is found to be associated with a −5.78 cm change in body height (95% CI: −6.12, −5.45) and a −0.79 change in HAZ (95% CI: −0.84, −0.74). Consistent results were observed when protein intake was expressed as %E or specifically as animal or plant protein. Moreover, the relationship between protein intake and linear growth remained consistent across genders in different pubertal stages, similar to that of the overall participants. Conclusions: Our findings highlight the potential hazards of high protein intake on linear growth in children and adolescents. Caution should be exercised when promoting increased protein consumption in children and adolescents who already have a high intake of protein. [ABSTRACT FROM AUTHOR]

Metabolic dysfunction-associated fatty liver disease (MAFLD) is the crucial pathogenesis for intra-hepatic and extra-hepatic diseases, especially in elderly adults. Lifestyle management may be a modifiable cost-effective measure for MAFLD prevention, but the evidence is limited. A total of 23,408 middle-aged and elderly individuals were included in a longitudinal study from 2008 to 2018. Combined lifestyle scores (range 0–6) were evaluated by BMI, smoking, drinking, diet, physical activity, and sleep. Logistic regression models were used to calculate ORs for the risks of MAFLD and specific subtypes. The mean age of participants was 61.7 years, and 44.5% were men. Compared with poor lifestyle (scores 0–2), ORs (95% CIs) of the ideal lifestyle (scores 5–6) were 0.62 (0.57–0.68) for MAFLD, 0.31 (0.28–0.34) for MAFLD with excess weight and obesity, 0.97 (0.75–1.26) for MAFLD with diabetes, and 0.56 (0.51–0.62) for MAFLD with metabolic dysregulation. Additionally, lifestyle improvement was associated with lower risks of MAFLD (OR, 0.76; 95% CI, 0.68–0.86), MAFLD with excess weight and obesity (OR, 0.72; 95% CI, 0.63–0.81), MAFLD with diabetes (OR, 0.74; 95% CI, 0.54–1.02) and MAFLD with metabolic dysregulation (OR, 0.49; 95% CI, 0.43–0.55), respectively. Our findings suggest that adherence to a combined healthy lifestyle was associated with lower risks of MAFLD, particularly in excess weight/obese individuals or those with metabolic dysregulation. [ABSTRACT FROM AUTHOR]