OBJECTIVES: To investigate the efficacy of Lactobacillus plantarum ZG03 (L. plantarum ZG03) for ameliorating oxidative stress in zebrafish. METHODS: We evaluated the growth pattern of L. plantarum ZG03, observed its morphology using field emission scanning electron microscopy, and assessed its safety and potential efficacy with whole-genome sequencing for genetic analysis. FITC-labeled ZG03 was used to observe its intestinal colonization in zebrafish. In a zebrafish model of 2% glucose-induced oxidative stress, the effect of ZG03 was evaluated by assessing the changes in neutrophils in the caudal hematopoietic tissue (CHT), superoxide dismutase (SOD) activity, reactive oxygen species (ROS) levels, and malondialdehyde (MDA) content. Liquid chromatography-mass spectrometry-based targeted metabolomics was used for analyzing short-chain fatty acids (SCFAs) in the zebrafish, and the antioxidant effects of the key metabolites (acetate, propionate, and caproate) were tested. RESULTS: On MRS agar, L. plantarum ZG03 formed circular, smooth, moist, and milky-white colonies with a rod-shaped cell morphology. Genomic analysis revealed abundant sugar metabolism gene clusters. After inoculation of FITC-labeled L. plantarum ZG03 in zebrafish, green fluorescence was clearly observed in the intestinal bulb, mid-intestine, and hind intestine. In zebrafish with glucose-induced oxidative stress, L. plantarum ZG03 significantly reduced ROS levels and the number of neutrophils in the CHT with increased SOD activity. L.plantarum ZG03 significantly increased the content of SCFAs including acetic acid, propionic acid, and caproic acid in zebrafish metabolites. In addition, sodium acetate, sodium propionate, and sodium caproate in the SCFAs significantly increased SOD activity in the zebrafish models. CONCLUSIONS L. plantarum ZG03 ameliorates oxidative stress in a glucose-induced zebrafish model through its metabolites, particularly the SCFAs including acetic acid, propionic acid and caproic acid.
Animals ; Zebrafish/metabolism* ; Oxidative Stress ; Lactobacillus plantarum/metabolism* ; Fatty Acids, Volatile/metabolism* ; Probiotics ; Reactive Oxygen Species/metabolism* ; Superoxide Dismutase/metabolism*

Objective To investigate the efficacy of Lactobacillus plantarum ZG03(L.plantarum ZG03)for ameliorating oxidative stress in zebrafish.Methods We evaluated the growth pattern of L.plantarum ZG03,observed its morphology using field emission scanning electron microscopy,and assessed its safety and potential efficacy with whole-genome sequencing for genetic analysis.FITC-labeled ZG03 was used to observe its intestinal colonization in zebrafish.In a zebrafish model of 2%glucose-induced oxidative stress,the effect of ZG03 was evaluated by assessing the changes in neutrophils in the caudal hematopoietic tissue(CHT),superoxide dismutase(SOD)activity,reactive oxygen species(ROS)levels,and malondialdehyde(MDA)content.Liquid chromatography-mass spectrometry-based targeted metabolomics was used for analyzing short-chain fatty acids(SCFAs)in the zebrafish,and the antioxidant effects of the key metabolites(acetate,propionate,and caproate)were tested.Results On MRS agar,L.plantarum ZG03 formed circular,smooth,moist,and milky-white colonies with a rod-shaped cell morphology.Genomic analysis revealed abundant sugar metabolism gene clusters.After inoculation of FITC-labeled L.plantarum ZG03 in zebrafish,green fluorescence was clearly observed in the intestinal bulb,mid-intestine,and hind intestine.In zebrafish with glucose-induced oxidative stress,L.plantarum ZG03 significantly reduced ROS levels and the number of neutrophils in the CHT with increased SOD activity.L.plantarum ZG03 significantly increased the content of SCFAs including acetic acid,propionic acid,and caproic acid in zebrafish metabolites.In addition,sodium acetate,sodium propionate,and sodium caproate in the SCFAs significantly increased SOD activity in the zebrafish models.Conclusion L.plantarum ZG03 ameliorates oxidative stress in a glucose-induced zebrafish model through its metabolites,particularly the SCFAs including acetic acid,propionic acid and caproic acid.

Objective Chronic atrophic gastritis(CAG)is often accompanied by intestinal flora disorder and intestinal symptoms,forming the phenomenon of"stomach disease involving intestine".This study explored the dynamic correlation between intestinal symptoms and qi-stagnation degree in patients with CAG qi-stagnation syndrome and analyzed the characteristics of gut microbiota from the perspective of"spleen-stomach system serving as the pivotal hub of qi movement"in TCM.Methods According to the syndrome element differentiation method,410 patients with CAG were divided into four groups:non-qi-stagnation group,mild qi-stagnation group,moderate qi-stagnation group and severe qi-stagnation group.Correlation analysis and 16S intestinal flora sequencing technology were used to analyze the correlation and differential flora between the degree of CAG qi-stagnation and intestinal symptoms.Results Patients with CAG qi-stagnation syndrome were often accompanied by intestinal symptoms such as frequent flatulence,poor defecation and alternating loose-constipated stools.The frequency of cases was significantly positively correlated with the degree of qi-stagnation"non-mild-moderate-severe"(P<0.05).There was a difference in the abundance of gut microbiota between the four groups of CAG qi-stagnation none,mild,moderate and severe.The relative abundance of Streptococcus,Subdoligranulum,Eubacterium_coprostanoligenes_group and Haemophilus was positively correlated with the degree of qi-stagnation.The relative abundance of Ruminococcus_torques_group and Butyricicoccus showed a negative correlation,and Haemophilus was statistically significant among the four groups(P<0.05).Conclusion This study can provide clinical evidence and micro-mechanism for the connotation of"gastrointestinal co-morbidities"and"different diseases with the same syndrome",which may open up new ideas for clinical diagnosis and treatment.

Objective Chronic atrophic gastritis(CAG)is often accompanied by intestinal flora disorder and intestinal symptoms,forming the phenomenon of"stomach disease involving intestine".This study explored the dynamic correlation between intestinal symptoms and qi-stagnation degree in patients with CAG qi-stagnation syndrome and analyzed the characteristics of gut microbiota from the perspective of"spleen-stomach system serving as the pivotal hub of qi movement"in TCM.Methods According to the syndrome element differentiation method,410 patients with CAG were divided into four groups:non-qi-stagnation group,mild qi-stagnation group,moderate qi-stagnation group and severe qi-stagnation group.Correlation analysis and 16S intestinal flora sequencing technology were used to analyze the correlation and differential flora between the degree of CAG qi-stagnation and intestinal symptoms.Results Patients with CAG qi-stagnation syndrome were often accompanied by intestinal symptoms such as frequent flatulence,poor defecation and alternating loose-constipated stools.The frequency of cases was significantly positively correlated with the degree of qi-stagnation"non-mild-moderate-severe"(P<0.05).There was a difference in the abundance of gut microbiota between the four groups of CAG qi-stagnation none,mild,moderate and severe.The relative abundance of Streptococcus,Subdoligranulum,Eubacterium_coprostanoligenes_group and Haemophilus was positively correlated with the degree of qi-stagnation.The relative abundance of Ruminococcus_torques_group and Butyricicoccus showed a negative correlation,and Haemophilus was statistically significant among the four groups(P<0.05).Conclusion This study can provide clinical evidence and micro-mechanism for the connotation of"gastrointestinal co-morbidities"and"different diseases with the same syndrome",which may open up new ideas for clinical diagnosis and treatment.

Objective To retrospectively analyze of factors influencing early preterm birth(EPB)and late preterm birth(LPB)in pregnancy women with placenta previa complicated by placenta accreta spectrum disorders(PAS),and assess maternal and infant outcomes.Methods We included 590 cases of pregnancy women with placenta previa complicated by PAS who underwent cesarean sections at five hospitals in Wuhan and Xianning cities between January 2018 and June 2021.These patients were divided into three groups based on delivery gesta-tional age:EPB,LPB,and term birth(TB).A multiple logistic regression model was employed to analyze the risk factors associated with EPB and LPB.Additionally,differences in early maternal and infant outcomes among these groups were examined.Results Among 590 pregnancy women with placenta previa complicated by PAS,the proportions of EPB and LPB were 9.7%and 54.4%.The use of uterine contraction inhibitors prior to cesarean section,vaginal bleeding,and previous cesarean sections history were identified as risk factors for both EPB and LPB.The proportion of severe postpartum hemorrhage was comparable between the EPB group and the LPB group;however,the incidence of neonatal asphyxia,low birth weight infants,and the rate of newborns transferred to the Neonatal Intensive Care Unit(NICU)within 24 hours after cesarean delivery were significantly higher in the EPB group compared to the LPB group.Conclusions Placenta previa complicated by PAS predominantly leads to LPB.The history of prior cesarean sections,uterine contractions,and vaginal bleeding prior to cesarean section,are sig-nificantly associated with both EPB and LPB.During the perinatal period,efforts should be made to extend gesta-tional weeks under close monitoring to minimize the incidence of premature births and thereby improve early mater-nal and infant outcomes.

Objective:To compare intact dissection and segmented dissection of cochlear surface preparation in adult mice.Methods:From February to March, 2019, Six adult C57BL/6 mice were randomly divided into 2 groups: one group (3 mice) for the intact dissection while the other group (3 mice) for the segmented dissection. Cochlear hair cells were labeled with phalloidin for evaluation of the integrity of the basilar membrane.Results:The basilar membranes can be completely dissected from the cochlea by two approaches. The average dissection time is (16.33±1.86)min with the intact dissection approach while (23.66±3.88) min with the segmented dissection( t=-4.173, P=0.002). Immunofluorescence analysis showed all cochlear hair cells werevisible and intact in two groups. Conclusion:Cochlear basilar membrane can be dissected intact in a short time through both approaches. The approaches selection is dependent on the purpose of experiment and operators′ experience.

Objective: To understand the prevention and control of coal-burning-borne endemic fluorosis in Hubei Province, and to evaluate the effects of prevention and control measures on coal-burning-borne endemic fluorosis. Methods: According to the requirements of national "Coal-burning-borne Endemic Fluorosis Monitoring Program", 9 fixed monitoring villages and 15 non-fixed villages in Jianshi, Badong and Zhuxi were selected as the monitoring sites every year from 2012 to 2017, 10 households were selected in each village to carry out the dynamic monitoring of the usage of stove and health behavior related to the corn and pepper for human consumption. At the same time, the monitoring survey of children's dental fluorosis and urinary fluoride were carried out. Results: From 2012 to 2017, the residents of the monitoring villages mainly used the improved stoves, the usage rates of the improved stove were all > 96%. There were statistically significant differences in the qualified rate and correct usage rate of the improved stove between each year (fixed monitoring villages: χ²= 48.71, 63.72, P < 0.05; non-fixed monitoring villages: χ²= 74.02, 84.80, P < 0.05). The correct usage rates of the improved stove were all 100.00% from 2015 to 2017. There were statistically significant differences in the correct rate of drying, keeping and washing eating corn and pepper between each year (fixed monitoring villages: χ²= 10.18, 10.41, 4.99, 15.71, 4.67, 6.80, P < 0.05; non-fixed monitoring villages: χ²= 6.22, 6.23, 18.86, 49.90, 23.88, 6.42, P < 0.05). From 2015 to 2017, all the rest rates were 100.00% except for the washing rate of pepper before eating. It showed a significant decline of dental fluorosis detection rate in 8 to 12 years old children annually (χ²=251.89, P < 0.05), from 33.57% (330/983) of 2012 to 10.26% (111/1 082) of 2017. The geometric means of urinary fluoride of children were 0.62, 0.60, 0.54, 0.53, 0.54 and 0.47 mg/L, respectively. There was statistically significant difference between each year (H=56.91, P < 0.05). Conclusions The prevention and control measures on coal-burning-borne endemic fluorosis of the 3 monitoring counties are implemented well in Hubei Province. Health behavior and life style are gradually formed, and the general condition gets better obviously.

Objective To systematically review the correlation between fluoride exposure through drinking water and dental fluorosis of school-age children,and to provide a theoretical basis for further development of prevention strategies against dental fluorosis.Methods Such databases as China National Knowledge Infrastructure (CNKI),WanFang Data,VIP Database and China Biology Medicine Disc (CBM) were searched through computer to find out the relationship between fluoride exposure through dringking water and occurrence of dental fluorosis.The retrieval time was from January 1,2000 to January 1,2018.Meta-analysis was performed using RevMan 5.3.Funnel plot and fail-safe method were used to evaluate publication bias,and these data were analyzed sensitivity with random and fixed effect models.Results Totally 20 literatures entered into this study,79 814 people in fluoride exposure group,and 181 876 people in control group.The meta-analysis showed that,20 literatures were inhomogeneous through the heterogeneity test,which was analyzed in the random effect model,the pooled odds ratio (OR) value and 95％ confidence interval (95％CI) was 4.25 (3.66-4.94),which suggested that the risk of dental fluorosis in the fluoride exposure group was 4.25 times higher than that in control group.Funnel plot was asymmetrical,the fail-safe number was 47 791.56,which was 2 389.6 (47 792/20) times higher than included literatures.Literatures publication bias was small,sensitivity analysis revealed that the results were basically reliable.Conclusion Excessive fluoride exposure through dringking water could be one of the main risk factors leading to dental fluorosis.

Objective To analyze the status of iodine nutrition after implementing new standard of iodized salt in Hubei Province,and to evaluate the effectiveness of prevention and control measures.Methods In 2016,in 103 counties (cities,districts) of Hubei Province,the townships (streets) under the jurisdiction of each county (city,district) were divided into five regions:east,south,west,north,and middle.One township (street) was selected from each region,and one primary school was selected from each township (street).From each primary school 40 children aged 8-10 (half males half females) were selected to collect salt samples from their households.Twenty pregnant women from townships (streets) near the selected schools were chosen to collect edible salt samples from their households.According to the method of population proportionate sampling (PPS),35 counties (cities,districts) were chosen from 103 counties (cities,districts).Thyroid volume of children aged 8-10 was examined,and urinary iodine of children and pregnant women were tested.Iodine nutrition criteria:the median urinary iodine of children ＜ 100 μg/L is iodine deficiency,100-＜ 200 μg/L is iodine appropriate,200-＜ 300 μg/L is iodine overdose,≥300 μg/L is iodine excess;pregnant women urinary iodine median ＜ 150 μg/L is iodine deficiency,150-＜ 250 μg/L is iodine appropriate,250-＜ 500 μg/L is iodine overdose,and ≥ 500 μg/L is iodine excess.Results Totally 30 967 edible salt samples from children's and pregnant women's households were examined,and the median of salt iodine was 23.85 mg/kg.The coverage rate of iodized salt was 99.46％ (30 799/30 967),the consumption rate of qualified iodized salt was 94.00％ (29 108/30 967).Totally 6 789 children aged 8-10 were examined thyroid volume and detected urinary iodine,the goiter rate was 0.85％ (58/6 789) and the median urinary iodine was 263.54 μg/L.Totally 3 348 urine samples of pregnant women were examined,and the median urinary iodine was 166.71 μg/L.Conclusions The salt iodine content of residents and the goiter rate of children in Hubei Province meet the national standard for eliminating iodine deficiency disorders.The iodine nutrition level of children and pregnant women is iodine appropriate or iodine overdose.The iodine nutrition level monitoring of key populations should be continuously strengthened.