OBJECTIVE To investigate the arsenic trioxide(As2O3)-induced oxidative damage to mitochondrial DNA (mtDNA) in mouse oocytes and possible mechanisms. METHODS ① For in vitro assay,the mouse oocytes were denuded from ovaries of normal mice and incubated in medium for 20 h in different treatment groups:control,As2O3 1 and 2 μmol · L- 1,N-acetylcysteine (NAC) 5 mmol · L-1, 2,2,6,6-tetramethyl-1-piperidinyloxy(Tempo)1 mmol · L-1, As2O3(1 and 2 μmol · L-1)+NAC 5 mmol · L-1,As2O3 (1 and 2 μmol · L-1)+Tempo 1 mmol · L-1. ② For in vivo assay,mice were subjected to ip injection with physiological saline (normal control),As2O3 1 and 2 mg · kg-1,or As2O3 (1 and 2 mg · kg-1)+NAC 200 mg · kg-1, respectively. After 60 d,all the mice were sacrificed and their ovaries were quickly excised. Intracellular reactive oxygen species(ROS) levels were determined by 2′,7′-dichlorofluorescein-diacetate (DCFH-DA). The oxidative damage to mtDNA was induced using enzyme-linked immunosorbent assay(ELISA)for 8-hydroxy-2′-deoxyguanosine(8-OHdG). The expression of DNA polymerase γ(Polγ)and mitochondrial transcription factor A(mtTFA)was detected by Western blotting and the vitality of lysosomes was monitored by β-galactosidase(β-Gal)Assay Kit. RESULTS ①In vitro experiments,As2O3 elevated 8-OHdG levels of mtDNA in mouse oocytes accom? panied by increased levels of ROS (P<0.05),but co-treatment with NAC or Tempo significantly reduced ROS and 8- OHdG levels (P<0.05). Meanwhile, the expression levels of Pol γ and mtTFA were down-regulated by As2O3(P<0.05),but were markedly elevated by the addition of NAC or Tempo (P<0.05). ②In vivo assay,As2O3 elevated ROS as well as 8-OHdG levels of mtDNA in mouse oocytes,while the expression levels of Pol γ and mtTFA were down-regulated by As2O3(P<0.05). Co-treatment with NAC significantly reduced ROS and 8-OHdG levels,but markedly elevated Pol γ and mtTFA levels(P<0.05). Besides,a notable increase in β-Gal activity was shown in As2O3-treated mouse oocytes in vitro (P<0.05),while antioxidants efficiently reduced the activity (P<0.05). However,no significant changes were observed in the in vivo study. CONCLUSION The oxidative damage to mtDNA induced by As2O3 in mouse oocytes may be mediated by ROS and associated with down-regulation of protein levels of Pol γ and mtTFA as well as increment of lysosomal activity.

Objective: To uncover the underlying mechanisms of action of the Yinlai decoction on high-calorie diet-induced pneumonia through proteomics analysis. Methods: Based on the Gene Expression Omnibus (GEO) database, lung tissue samples from normal and high-fat diet (HFD) fed mice in the GSE16377 dataset were selected as test cohorts to identify differentially expressed genes and conduct bioinformatics analyses. In the animal experiments, mice were randomly divided into the control (N), high-calorie diet pneumonia (M), and Yinlai decoction treatment (Y) groups. Mice in the M group received high-calorie feed and a 0.5 mg/mL lipopolysaccharide solution spray for 30 min for 3 d. The mice in the Y group were intragastrically administered 2 mL/10 g Yinlai decoction twice daily for 3 d. Pathological evaluation of the lung tissue was performed. Differentially expressed proteins (DEPs) in the lung tissue were identified using quantitative proteomics and bioinformatics analyses. The drug-target relationships between Yinlai decoction and core DEPs in the lung tissue were verified using AutoDock Vina and Molecular Graphics Laboratory (MGL) Tools. DEPs were verified by western blot. Results: GEO data mining showed that an HFD altered oxidative phosphorylation in mouse lung tissue. The Yinlai decoction alleviated pathological damage to lung tissue and pneumonia in mice that were fed a high-calorie diet. A total of 47 DEPs were identified between the Y and M groups. Enrichment analysis revealed their association with energy metabolism pathways such as the tricarboxylic acid cycle (TCA) and oxidative phosphorylation. The protein-protein interaction network revealed that Atp5a1, Pdha1, and Sdha were the target proteins mediating the therapeutic effects of Yinlai decoction. Molecular docking results suggested that the mechanism of the therapeutic effect of Yinlai decoction involves the binding of brassinolide, praeruptorin B, chrysoeriol, and other components in Yinlai decoction to Atp5a1. Conclusion The Yinlai decoction alleviated lung tissue damage and pneumonia in mice that were fed a high-calorie diet by regulating the TCA and oxidative phosphorylation. Our study highlights the importance of a healthy diet for patients with pneumonia and provides a scientific basis for the prevention and treatment of pneumonia through dietary adjustments.

Objective: To explore the relationship between gastrointestinal heat retention syndrome and the inci-dence of pneumonia and recurrent respiratory tract infections (RRTIs) in children. Methods: A prospective cohort study was conducted in the pediatric outpatient department of Beijing Dongfang Hospital. Children without respiratory tract infections (RTIs) were consecutively recruited according to the selection criteria. A semi-structured questionnaire was used to record traditional Chi-nese medicine (TCM) symptoms and demographic and physiological characteristics. Gastrointestinal heat retention syndrome was considered to be a predisposing factor and was diagnosed according to a scale with reliability and validity. The participants were followed up for 12 months. Participants and their parents or guardians were contacted via clinical interviews and telephone every 6 months. Episodes of pneumonia and RTIs were recorded in detail. Results: A total of 420 children were included. Of participants, 370 (88.10％) were followed up for 12 months. The mean number of RTI episodes per participant was 5.37 (95％ CI: 5.14 to 5.60). In total, 186 participants in the gastrointestinal heat retention syndrome group and 184 participants in the non-gastrointestinal heat retention syndrome group completed the 12-month follow-up period. The base-line of both groups was comparable. The incidence of RRTIs in children with gastrointestinal heat retention syndrome was 1.27 (95％ CI: 1.01 to 1.59) times that in children without gastrointestinal heat retention syndrome. Logistic regression analysis revealed that abnormally increased appetite with frequent hunger, foul breath, dry stools, and dark red or purple fingerprints were positively correlated with the incidence of pneumonia. Irascibility and feverish feelings in the palms and soles were positively correlated with the occurrence of RRTI. Conclusions: Gastrointestinal heat retention syndrome is a risk factor for RRTIs in children. Studies with larger sample sizes and longer follow-up time are warranted to confirm the degree of causal risk associated with RTIs.

Objective:Gastrointestinal heat retention syndrome (GHRS) is associated with lung-heat syndrome and is related to recurrent respiratory infection.Upper respiratory tract infection (URTI) lung heat syndrome is common in children.The study will explore the effect of GHRS on the structure and function of gut microbiota in children with URTI lung-heat syndrome.Methods:Participants were divided into both groups using the self-developed URTI scale and the "GHRS Diagnostic Scale · Pediatric Part":GHRS-positive children (LS group) and GHRS-negative children (L group).General information,clinical symptoms,and stool were collected.We used 16S rRNA amplicon sequencing technology to determine the gene sequence of the V3-V4 region in feces and measure the gut microbiota of the both groups at the genus level.Results:A total of 23 children were included in the both groups.There were 12 cases in the LS group and 11 cases in the L group.There was no statistical difference between the both groups in age,gender,height,weight,and body mass index.The effective sequences shared by the both groups accounted for 85.66％ of the total.In the gut microbiota,there was no difference in the α diversity and the β diversity between the both groups.Compared with the L group,the LS group had a significant increase in the relative abundance of the Rurninococcus gnavus group,Prevotella-9,Staphylococcus,and Actinomyces(P ＜.05).The functions of the both groups of microbiota primarily concentrate on metabolism,genetic information processing,and environmental information processing.The relative abundance of signaling molecules and interactions in the LS group were higher than that in the L group (P ＜.05).The redun-dancy analysis (RDA) showed that the URTI score had the greatest impact on the distribution of microbiota.Conclusion:GHRS may affect the development of URTI lung-heat syndrome by changing the relative abundances of gut microbiota.

Objective To evaluate the efficacy and safety of Qingpeng ointment for the treatment of localized eczema in children.Methods A multicenter,randomized,open-labeled,tacrolimus 0.03％ ointment-controlled clinical trial was conducted.A total of 442 children with localized eczema were enrolled into this study,and divided into 2 groups to topically apply Qingpeng ointment (Qingpeng group) and tacrolimus 0.03％ ointment (tacrolimus group) respectively twice a day for 2 weeks.The children were followed up before the treatment and 1,2 weeks after the treatment,and the efficacy and safety were evaluated at the same time.Results Clinical data from 426 children were included in the efficacy analysis.At 1,2 weeks after the treatment,there were no significant differences in the response rate between the Qingpeng group and tacrolimus group (1 week after the treatment:78.92％ vs.81.77％,x2 =0.545,P =0.460;2 weeks after the treatment:98.65％ vs.97.54％,x2 =0.721,P =0.396).However,the pruritus scores at 1,2 weeks after the treatment were both significantly lower in the Qingpeng group than in the tacrolimus group (1 week:F =14.001,P =0.000;2 weeks:F =11.252,P =0.001).At 1 week after the treatment,the incidence rate of adverse reactions was significantly lower in the Qingpeng group than in the tacrolimus group (P ＜ 0.05).Mild itching,burning sensation and other local irritant sensations were the most common adverse reactions.Conclusion Qingpeng ointment is safe and effective for the treatment of localized eczema in children with good tolerability.

Objective To identify factors associated with gastrointestinal heat retention in preschool children,and to provide a foundational understanding for future clinical investigations. Methods A case-control study was performed,which involved children from kindergartens in the Longgang District of Shenzhen City,Guangdong Province,from May to July 2021. Using the Children's Gastrointestinal Heat Retention Diagnostic Self-assessment Scale,subjects were allocated into a case group (children diagnosed with gastrointestinal heat retention) and a control group (children without this condition). An online survey was used to collect data on dietary behaviors,caregivers' feeding behaviors,early antibiotic use,daily routines,and birth conditions. SPSS 27.0 software was used to facilitate precise sociodemographic matching and paired logistic regression analysis to explore the association between gastrointestinal heat retention and the above factors. Results From the analysis of 51,252 matched cases,the study found that several factors contributed to an increased risk of gastrointestinal heat retention. These factors included reduced food intake compared to peers,reports of picky eating by caregivers,distractions during meals,pronounced dietary preferences,disinterest in food,meal durations ≥ 25 min,reluctance to sample new foods,consistent refusal of specific food types for over one month,irregular meal locations,coercive feeding practices,use of micronutrient supplements,allowing children too much freedom in food choice,persuading children to eat,infrequent encouragement to experiment with new foods,early antibiotic introduction,inadequate sleep,and premature birth (P<0.05). In contrast,exclusive breastfeeding in the first six months,engagement in moderate to massive physical activity,and regular napping patterns were associated with a reduced risk of gastrointestinal heat retention (P<0.05). Conclusion The suboptimal dietary habits,improper feeding practices,insufficient physical activity,inadequate sleep,and premature antibiotic exposure may be significant risk factors for gastrointestinal heat retention. Future research dedicated to unraveling the cause of gastrointestinal heat retention should prioritize these elements.

Objective: To search for specific metabolites in the lungs of pneumonia rats fed with a high-calorie diet, as well as explore the changes in the lung metabolites of young rats treated with Yinlai Decoction (YD) and its effects on inflammation-related metabolic pathways.Methods: Lipopolysaccharides (LPS) and a special high-calorie diet were used to induce Sprague Dawley (SD) rats to simulate the intestinal state of infant pneumonia. Liquid chromatography-mass spectrometry technology (LC-MS/MS) was used to detect metabolites in each group. Supervised orthogonal partial least squares discriminant analysis (OPLS-DA) model values were used for the detection results to find the differential metabolites. The metabolic pathways that are involved with the differential metabolites were clarified through enrichment analysis and topological analysis. Finally, the T cell receptor signaling pathway (TCR) signal conversion was analyzed by the network pharmacology method. Results: In the high-calorie diet combined with pneumonia group (M3), a total of 55 metabolites were determined to be different from the normal group (N). A total of 36 metabolites were determined to be different from those in the lung metabolites of the YD treatment group (T1). YD had a regulatory effect on glutathione metabolism, arginine and proline metabolism, ascorbic acid and aldehyde metabolism and phenylalanine metabolism. And the small molecule metabolites could act on the FYN and lymphocyte-specific protein tyrosine kinase (LCK) target proteins in the TCR signaling pathway, thereby affecting the immune function of the lungs. Conclusion: A high-calorie diet can cause abnormal sphingolipid metabolism in the lungs of young rats, thereby creating chronic lung inflammation in young rats. YD has a beneficial effect when used to treat young rats with LPS-induced pneumonia fed on high-calorie diets. Its mechanisms of action may affect the body's immune pathways by regulating the oxidative stress pathway affected by glutathione metabolism.

Objective:To explore the structural and functional characteristics of microbiota in oropharynx of sub-healthy children with gastrointestinal heat retention syndrome(GHRS)differentiated by traditional Chinese medicine,and screen the biometric operational taxonomic units(OTUs)to assist the clinical diagnosis.Methods:We recruited children according to the"GHRS diagnostic scale",collected their oropharyngeal swabs,and sequenced the 16S rDNA V4 region.We described the bacterial structure with alpha-indexes,beta-distances,and relative abundances;moreover,we screened the differential genera/OTUs with Wilcoxon rank-sum test,MetagenomeSeq analysis,and linear discriminant analysis effect size(LEfSe)analysis,in which biometric OTUs were selected to construct the receiver operating characteristic curve to verify the diagnostic value.The bacterial function was predicted with Kyoto Encyclopedia of Genes and Genomes pathways according to 16S rDNA gene by using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States.Results:The study population was composed of 10 children with GHRS and 10 healthy control children.GHRS children were more likely to overeat(gluttony,P=.033).Alpha-indexes,such as Sobs,abundance-based coverage estimator,Bootstrap,and Qstat,were significantly higher in the GHRS group,while beta-distances did not exhibit any significant intergroup differences.There were 9 differently distributed non-predominant genera between the groups in Wilcoxon rank-sum test,as well as 13 non-predominant genera in MetagenomeSeq analysis and 3 non-predominant OTUs in LEfSe analysis.OTU44 and OTU196 were used to construct the receiver operating characteristic curve,and the area under curve was 0.92.Predicted functions showed that pathways related to oxidative phosphorylation and carbon metabolism were enriched in healthy control samples,while the pathway related to renin secretion was remarkably enriched in GHRS samples.Conclusion:Unique oropharyngeal microbial structure and function were identified in GHRS children.OTU44 and OTU196 were specific OTUs,which could be used as biomarkers of GHRS to assist clinical diagnosis.

Objective:To investigate the association between metabolic syndrome and the risk of early renal function injury in chronic kidney disease(CKD) in the elderly.Methods:A retrospective cohort was established based on health check-up data of 4 495 elderly residents in Mengzi City, Yunnan Province from January 2016 to December 2018. The medial history, living habits, and related physical examination information were collected. Cox hazard regression model was used to explore the association between metabolic syndrome, along with its components, and the early renal function injury in CKD. Results:The median age of the elderly was 71.00(67.00, 75.00) years, with metabolic syndrome detection rate of 21.98%. Early renal function injury of CKD developed in 1 300(28.92%) subjects during the follow-up. Univariate Cox regression showed that the number of metabolic syndrome components was associated with the risk of early kidney development in CKD. The HRs were 1.23 (95% CI 1.03-1.47, P=0.022) with 1 component, 1.54 (95% CI 1.28-1.84, P<0.001) with 2, and 1.38 (95% CI 1.14-1.67, P<0.001) with 3 or more. Multivariate Cox regression showed that elevated fasting triglycerides( HR=1.20, 95% CI 1.07-1.36, P=0.003) and lower high density lipoprotein-cholesterol(HDL-C; HR=1.25, 95% CI 1.09-1.43, P=0.002) were risk factors for early kidney injury in CKD, while doing some physical activity( HR=0.57, 95% CI 0.33-0.98, P=0.042), or on daily basis( HR=0.57, 95% CI 0.49-0.66, P<0.001) was a protective factor for early kidney injury in CKD. Conclusion:The abnormality of one or more metabolic components can significantly increase the risk of early kidney injury in the elderly with CKD. Elevated triglyceride and decreased HDL-C may be the risk factors.

OBJECTIVE： To establish a method for simultaneous determination of isoquercitrin， astragalin and salvianolic acid B in Moringa oleifera leaves granules. METHODS： HPLC method was adopted. The determination was performed on Cosmosil-C18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution （gradient elution） at flow rate of 1.3 mL/min. The column temperature was 40 ℃ and detection wavelength was set at 260 nm. The sample size was 10 μL. RESULTS： The linear ranges of isoquercitrin， astragalin and salvianolic acid B were 0.017-0.341， 0.010-0.194， 0.010-0.195 mg/mL， respectively （all r＞0.999 0）. The detection limits were 0.085， 0.143， 0.117 μg/mL， and the limits of quantitation were 0.283， 0.476， 0.392 μg/mL， respectively. RSDs of precision， stability （24 h） and reproducibility tests were all lower than 2.0％ （n＝6）. Average recoveries were 101.22％， 98.76％ and 98.72％， and RSDs were 0.66％，0.30％，0.30％ （n＝6）， respectively. CONCLUSIONS： The established method is simple， accurate and reproducible. It can be used for simultaneous determination of isoquercitrin， astragalin and salvianolic acid B in M. oleifera leaves granules.