1.Consistency and risk factor analysis of 2 growth chart in the diagnosis of extrauterine growth restriction in preterm infants with a gestational age at birth of less than 32 weeks
Fan ZHANG ; Xiaohua ZHANG ; Shushu LI ; Shuping HAN
Chinese Journal of Pediatrics 2026;64(1):61-67
Objective:To investigate the diagnostic consistency of extrauterine growth restriction (EUGR) assessed by the Fenton 2013 preterm growth charts (Fenton 2013) and the growth charts by International Fetal and Newborn Growth Consortium for the 21st Century (IG-21).Methods:This multicenter retrospective cohort study included 5 591 preterm infants with a gestational age (GA) at birth of less than 32 weeks admitted to 19 member hospitals of the Neonatal Perinatal Collaborative Network of Suxinyun from January 1 st, 2019, to December 31 st, 2024. Clinical data including baseline characteristics, complications, feeding practices and anthropometrics were processed and analyzed. EUGR was assessed using both the Fenton 2013 and the IG-21. A decrease in weight Z-score at discharge compared to admission by more than 1 was defined as longitudinal EUGR, and discharge weight below the P10 for the corresponding corrected GA was defined as cross-sectional EUGR. Diagnostic consistency was assessed using the Kappa coefficient between the 2 standards, and diagnostic performance of the 2 standards was compared using the McNemar test. Risk factors for EUGR under different definitions were analyzed using univariate analysis and multivariate Logistic regression analysis. Results:A total of 5 591 preterm infants were included, with a GA at birth of (29.7±1.6) weeks and a birth weight of (1 360±315) g and at discharge with a corrected GA of (36.3±2.0) weeks and weight of (2 246±370) g. Detection rates of cross-sectional and longitudinal EUGR diagnosed by Fenton 2013 were both higher than those by IG-21 (37.0% (2 214/5 991) vs. 23.7% (1 324/5 591), 61.1% (3 662/5 991) vs. 30.7% (1 714/5 591), χ2=326.77 and 1 358.05, both P<0.001). Using Fenton 2013 as a reference, IG-21 demonstrated superior diagnostic value and consistency in identifying cross-sectional EUGR compared with longitudinal EUGR (sensitivity of 100.0% (3 377/3 377) vs. 99.6% (1 922/1 929), specificity of 59.8% (1 324/2 214) vs. 46.6% (1 707/3 662), positive predictive value of 79.1% (3 377/4 267) vs. 49.6% (1 922/3 877), negative predictive value of 100.0% (1 324/1 324) vs. 99.6% (1 707/1 714), accuracy of 84.1% (4 701/5 591) vs. 64.9% (3 629/5 591), and Kappa 0.64 vs. 0.37, all P<0.001). In multivariate Logistic regression models, risk factors common to EUGR across both standards included smaller GA at birth, lower birth weight, boy, early-onset sepsis, late-onset sepsis and the elder age at full enteral feeding (all P<0.05). Hemodynamically significant patent ductus arteriosus remained an independent risk factor for longitudinal EUGR regardless of whether by the Fenton 2013 or IG-21 standard (adjust odds ratio ( aOR) =1.25 and 1.27, 95% CI 1.09-1.42 and 1.11-1.45). In addition, under the IG-21 standard, severe bronchopulmonary dysplasia was an independent risk factor for cross-sectional EUGR ( aOR=1.54, 95% CI 1.00-2.38), while severe necrotizing enterocolitis was an independent risk factor for longitudinal EUGR ( aOR=2.18, 95% CI 1.01-4.73). Conclusions:IG-21 showed lower detection rates of both cross-sectional and longitudinal EUGR than Fenton 2013, suggesting greater clinical applicability of IG-21 by reducing overdiagnosis while maintaining sensitivity for predicting complications. Across both standards, cross-sectional EUGR facilitates early identification of growth restriction, whereas longitudinal EUGR better tracks dynamic growth patterns and complications of preterm infants.
2.Overview of Diagnosis,Treatment and Mechanism Research of Functional Dyspepsia by Integrated Traditional Chinese and Western Medicine
Shengsheng ZHANG ; Zhaohong SHI ; Xiaofang LU ; Luqing ZHAO ; Danyan LI ; Shu ZHANG ; Lu ZHAO ; Yudi ZHUO ; Nian WANG ; Fan LIU ; Shuangyi LI ; Xudong TANG
Journal of Traditional Chinese Medicine 2026;67(4):397-403
Functional dyspepsia (FD) is a prioritized disease category where traditional Chinese medicine (TCM) demonstrates distinct therapeutic advantages. The current western medicine treatment for FD is mainly based on proton pump inhibitors and prokinetic agents, with digestive enzymes, probiotics and antidepressants serving as adjuvant medication, yet such therapies still have certain limitations. TCM treatment for FD includes oral administration of Chinese herbal formulas and Chinese patent medicines, as well as external TCM therapies such as acupuncture and moxibustion, acupoint application, hot medicinal compress therapy, rubbing with ointment, medicinal iontophoresis, auricular acupoint therapy and tui na (Chinese medical massage). The combined treatment of FD with integrated TCM and western medicine can significantly improve clinical effectiveness and reduce adverse reactions. The common mechanisms underlying the therapeutic effects of both TCM and western medicine revolve around the core pathological processes of FD, mainly focusing on restoring gastrointestinal motility, regulating the levels of brain-gut peptides, modulating intestinal microecology, and ameliorating inflammatory status. The differential mechanisms lie in the precise targeting feature of western medicine versus the holistic-regulating and multi-target characteristics of TCM, and the two approaches exert a synergistic effect to enhance efficacy. This paper proposes to leverage the advantages of TCM in holistic regulation and the strengths of western medicine in targeted treatment, so as to provide personalized and comprehensive treatment regimens for FD patients.
3.Experience Discussion on Feeding, Breeding and Genetic Management of Common Marmosets (Callithrix jacchus)
Li LI ; Xuebo LI ; Yongheng FAN ; Donghua HE ; Jianhong LI ; Liangtang CHANG
Laboratory Animal and Comparative Medicine 2026;46(1):107-118
The common marmoset (Callithrix jacchus) is a species of the genus Callithrix in the primate family Callitrichidae. Common marmosets are an important scientific research resource for the study of neuroscience, medicine, and pharmacy. The feeding and breeding of common marmosets provide important support for scientific research. At present, common marmoset resources in China are at a critical stage of development. This paper takes the Center for Excellence in Brain Science and Intelligence Technology of the Chinese Academy of Sciences (hereafter referred to as CEBSIT) as an example to discuss the experience of common marmoset feeding, breeding and genetic management. In terms of breeding environment and facilities, on the basis of the national standard, more stringent environmental parameter standards are implemented for environmental temperature, relative humidity, the number of air exchanges, and the average concentration of sedimentation bacteria. Two kinds of cages for common marmoset breeding and experimentation are designed. According to the differences in facility conditions and staffing, a cleaning and maintenance method combining "dry rearing" and "wet rearing" is introduced. In terms of feeding management, in order to meet the high nutritional needs of common marmosets, CEBSIT prepares special formulated diets, and introduces the transition method of common marmosets from diets mixed with formulated rations and pellets to exclusive commercial diets, and strictly regulates the operation procedures of formulated diets for common marmosets, and has accumulated a series of methods for preventing nutritional and metabolic diseases in common marmosets. In terms of animal health management, the health of common marmosets is ensured through measures such as regular quarantine, isolation and treatment, and epidemic prevention, and the health inspection process of common marmosets is described in detail, and hierarchical and classified management is carried out according to the physical condition of common marmosets. This paper also briefly describes the treatment of diarrhea and pneumonia that are common in common marmosets. In terms of breeding management, it covers the selection of breeding common marmosets, combining pairs of common marmosets into the same cage for breeding, group splitting and hand-rearing of infant common marmosets, and introduces the method of reintroducing hand-reared infant common marmosets to parental care from the perspective of animal welfare. In terms of genetic management, the common marmoset population in CEBSIT is maintained as a closed colony and follows a non-inbreeding method. CEBSIT has developed an inbreeding screening system to check pedigree information before pairing to avoid inbreeding. Under conditions without introducing new individuals from outside, CEBSIT has continuously bred the colony for 5 generations while maintaining genetic heterozygosity. This paper only discusses the feeding, breeding and genetic management of the common marmoset and aims to provide a useful reference for other institutions engaged in common marmoset breeding.
4.Biological Risk Control for Infectious Experiments in Cats in Animal Biosafety Level 2 Laboratory
He ZHAO ; Tao ZHANG ; Yuzhou XIAO ; Li LI ; Xuefang AN ; Fan ZHANG
Laboratory Animal and Comparative Medicine 2026;46(2):242-250
Cats, owing to their physiological and immunological similarities with humans, have become increasingly valuable as model animals in virology research, drug development, and vaccine evaluation. They are irreplaceable in studies of feline immunodeficiency virus, feline coronavirus, and other related pathogens. However, cats are temperamentally sensitive, exhibit strong stress responses, and possess well-developed nervous systems as well as sharp claws and teeth. Consequently, the biosafety risks associated with infectious experiments using cats in animal biosafety level 2 laboratory (ABSL-2) are significantly higher than those encountered with conventional rodents. Drawing on long-term ABSL-2 operational experience, this article systematically reviews the entire workflow of infectious experiments in laboratory cats — from animal selection, pre-entry preparation, reception and quarantine, housing management, to infectious experimental procedures and incident response — identifying and addressing critical risk points at each stage. For strain selection, SPF-grade shorthair cats with defined genetic backgrounds and docile temperaments are recommended; sex and age should be scientifically matched to experimental objectives. During pre-entry preparation, emphasis is placed on dual-credential personnel management, health surveillance, standardized disinfection of environments and cages, feed and water standards, and robust record-keeping. During reception and quarantine, standardized protocols are established for transport control, appearance inspection, isolation quarantine, pathogen exclusion, and positive-reinforcement training. During infectious experimentation, a "three-fixed" husbandry principle is clearly implemented: dedicated caretakers, fixed feeding/cleaning times, and fixed cage positions. Disinfectant selection, autoclaving of waste, and daily veterinary rounds are rigorously enforced. Operational risk control includes detailed measures for graded personal protection, animal anesthesia and restraint, zoned operation within biosafety cabinets, and disposal of experimental waste. Contingency plans are formulated to address animal death, escape, personnel exposure, and spills of infectious materials. This study provides a reproducible and scalable technical pathway and operational standard for conducting infectious experiments in laboratory cats in ABSL-2 laboratories, offering a reference for other facilities undertaking similar work.
5.Enhancement Effect of Porcine Inhibin Polyclonal Antibody on Superovulation in C57BL/6J Mice
He ZHAO ; Tao ZHANG ; Li LI ; Yuzhou XIAO ; Xuefang AN ; Fan ZHANG
Laboratory Animal and Comparative Medicine 2026;46(2):271-278
ObjectiveTo prepare rabbit anti-porcine inhibin polypeptide-keyhole limpet hemocyanin(KLH) conjugated polyclonal antibody and evaluate its effect on superovulation in C57BL/6J mice. MethodsNew Zealand white rabbits were immunized with a synthesized porcine inhibin polypeptide conjugated with KLH to produce anti-inhibin serum (AIS, i.e., inhibin polyclonal antibody). Female C57BL/6J mice received intraperitoneal injections of purified AIS in combination with pregnant mare serum gonadotropin (PMSG), followed by human chorionic gonadotropin (hCG) after 48 hours to induce superovulation. Oocytes obtained from superovulation were collected and counted 15 hours post-hCG administration, and the number of 2-cell embryos was assessed 24 hours after in vitro fertilization. ResultsAIS prepared by immunizing New Zealand White rabbits with KLH-conjugated porcine inhibin polypeptide was subjected to titer determination by indirect ELISA, showing titers reaching 1∶ 512 000. SDS-polyacrylamide gel electrophoresis analysis of ammonium sulfate-purified AIS revealed distinct 50 kDa and 25 kDa bands corresponding to the theoretical molecular weights of IgG antibody heavy and light chains, confirming successful production of porcine inhibin polyclonal antibody. Compared with conventional superovulation methods, AIS diluted 10-fold combined with PMSG significantly increased the number of oocytes obtained from superovulation in mice (P<0.05) by approximately 1.5-fold. ConclusionPorcine inhibin polyclonal antibody, as an improved superovulation reagent, can improve superovulation efficiency in C57BL/6J mice, and shows promising prospects for future applications.
6.Effect and Mechanisms of Bushen Tongluo Prescription on Pulmonary Fibrosis via Inhibiting Macrophage Polarization Through Wnt3a/β-catenin Signaling Pathway
Yanxia LIANG ; Xuelian YU ; Wenwen WANG ; Guangsen LI ; Hongfei XING ; Maorong FAN ; Bin YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):112-123
ObjectiveThis study aimed to investigate whether Bushen Tongluo prescription inhibits macrophage polarization by regulating the Wnt3a/β-catenin signaling pathway, thereby reducing epithelial-mesenchymal transition and excessive extracellular matrix deposition, in order to elucidate the anti-pulmonary fibrosis mechanisms of Bushen Tongluo prescription and provide a new theoretical basis for the clinical treatment of pulmonary fibrosis. MethodsFifty male Sprague-Dawley (SD) rats were randomly divided into a blank group, model group, pirfenidone group, and high- and low-dose Bushen Tongluo prescription groups. Except for the blank group, the pulmonary fibrosis model was established by intratracheal instillation of bleomycin. Intervention was initiated on day 28 after modeling. The high- and low-dose Bushen Tongluo prescription groups were administered Bushen Tongluo prescription at doses of 30.88, 15.44 g·kg-1, respectively, by intragastric gavage. The pirfenidone group was administered pirfenidone capsules at 110 mg·kg-1 by intragastric gavage. The blank and model groups were given an equal volume of normal saline by gavage, once daily for 90 days. After treatment, the level of transforming growth factor-β1 (TGF-β1) in bronchoalveolar lavage fluid (BALF) was detected by enzyme-linked immunosorbent assay (ELISA). Morphological changes in lung tissue and the collagen volume fraction were compared. The protein distribution and expression of E-cadherin, cytokeratin 19, α-smooth muscle actin (α-SMA), vimentin, collagen type Ⅰ (Col Ⅰ), and collagen type Ⅲ (Col Ⅲ) in lung tissue were detected by immunohistochemistry. The protein distribution and expression of CD68, arginase-1 (Arg-1), inducible nitric oxide synthase (iNOS), Wnt3a, and β-catenin in lung tissue were detected by immunofluorescence. The protein expression of Wnt3a and β-catenin in lung tissue was detected by Western blot, and the mRNA expression of Wnt3a and β-catenin was detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultsCompared with the blank group, a large number of inflammatory cells infiltrated the airway walls, alveolar spaces, and interstitial tissue in the model group, with obvious fibrous tissue hyperplasia. The level of TGF-β1 in BALF was significantly increased. The protein expression of E-cadherin and cytokeratin 19 in lung tissue was decreased, whereas the protein expression of α-SMA, Vimentin, Wnt3a, β-catenin, Col Ⅰ, and Col Ⅲ was increased. The fluorescence-positive area ratios of CD68, Arg-1, iNOS, Wnt3a, and β-catenin in lung tissue were increased. The protein and mRNA expression levels of Wnt3a and β-catenin in lung tissue were significantly increased (P<0.01). Compared with the model group, all treatment groups showed varying degrees of improvement in inflammatory cell infiltration and fibrous tissue hyperplasia in the airway walls, alveolar spaces, and interstitial tissue, decreased TGF-β1 levels in BALF, increased protein expression of E-cadherin and cytokeratin 19 in lung tissue, decreased protein expression of α-SMA, Vimentin, Col Ⅰ, and Col Ⅲ, decreased fluorescence-positive area ratios of CD68, Arg-1, iNOS, Wnt3a, and β-catenin in lung tissue, and decreased protein and mRNA expression levels of Wnt3a and β-catenin in lung tissue (P<0.05, P<0.01). ConclusionBushen Tongluo prescription can improve bleomycin-induced pulmonary fibrosis in rats by inhibiting epithelial-mesenchymal transition and reducing excessive extracellular matrix deposition. The mechanism may be related to inhibition of the Wnt3a/β-catenin signaling pathway and the macrophage polarization mediated by this pathway.
7.Association between screen behaviors with overweight and obesity among children and adolescents
Chinese Journal of School Health 2026;47(4):486-489
Objective:
To investigate the prevalence of overweight and obesity among children and adolescents in Yangzhou City, and its association with screen behaviors, so as to provide scientific evidence for weight management among students.
Methods:
In May 2025, an electronic questionnaire survey was conducted among children and adolescents in Yangzhou City. A total of 3 722 participants were selected from grades 4 to 12 in 18 primary and secondary schools (108 classes) by using stratified cluster random sampling. The Chi square test was used to compare the differences in the detection rates of overweight and obesity among children and adolescents with 5 types of screen behaviors (watching TV, playing electronic games, scrolling short videos, screen based learning, electronic socializing) in different time groups each day (never, >0~<2 h, ≥2 h). Multivariate Logistic regression analysis was performed to examine the associations of five types of screen behaviors, presence of electronic devices in the bedroom, and screen use during meals on the weight status of children and adolescents.
Results:
The prevalence of overweight and obesity among children and adolescents was 37.3%. For all five types of screen behaviors, the differences in the distribution of overweight and obesity detection rates among children and adolescents across the three time spent categories were statistically significant ( χ 2=30.76- 70.78 , all P <0.01). After adjusting for confounding factors, multivariate Logistic regression analysis revealed that frequent or always using screens during meals( OR =1.63, 95% CI =1.14~2.31), playing video games ( OR =1.28, 95% CI =1.11-1.48), browsing short videos ( OR =1.29, 95% CI=1.09-1.54), and screen based learning ( OR =1.26, 95% CI =1.10-1.44) were significantly associated with overweight and obesity among children and adolescents (all P <0.05).
Conclusions
Excessive screen use is positively correlated with the incidence of overweight and obesity in children and adolescents. Targeted interventions on screen behaviors among children and adolescents are therefore warranted.
8.Prognostic significance of TRIM28 elevation in non-M3 acute myeloid leukemia
Siqi GONG ; Cong LI ; Mengmeng FAN ; Huiping WANG ; Wanqiu ZHANG ; Xue LIANG ; Qianshan TAO ; Qiang HONG ; Zhimin ZHAI
Acta Universitatis Medicinalis Anhui 2026;61(2):301-308
ObjectiveTo clarify the expression of TRIM28 in non-M3 acute myeloid leukemia (AML) and its correlation with clinical indicators and prognosis, and to further explore the effect of TRIM28 expression levels on the proliferation and apoptosis of AML cells using small interfering RNA. MethodsThe GSE34577 dataset was analyzed using R software to compare TRIM28 expression between healthy controls and non-M3 acute myeloid leukemia (AML) patients. Clinical samples from non-M3 AML patients were collected, with TRIM28 expression levels measured using real-time quantitative PCR (qPCR). The analysis focused on correlations between TRIM28 expression and various clinical indicators, treatment efficacy, and patient prognosis. Furthermore, small interfering RNA (siRNA) technology was employed to downregulate TRIM28 expression in human primary AML cells (HL60 cell line). The effects on cell proliferation and apoptosis were then assessed through CCK-8 assays and flow cytometry, respectively. ResultsThe results showed that TRIM28 was up-regulated in non-M3 AML of both online database GSE34577 and clinical samples (P<0.000 1), TRIM28 expression of new diagnosis group and relapsed refractory group was higher than iron deficiency anemia group (P<0.01), and there was no significance between different French-American-British classification systems subtype. TRIM28 expression was higher in non-M3 AML patients with a poor genetic prognosis stratified as moderate than in the good prognosis group, and TRIM28 expression was associated with NPM1 combined with the FLT3-ITD mutation, positively correlated with age, bone marrow blast, peripheral blood blast and white blood cell, negatively correlated with hemoglobin. In addition, interference TRIM28 greatly inhibited cell proliferation and promoted cell apoptosis. ConclusionThis study reveals that TRIM28 is highly expressed in non-M3 AML and associated with prognosis, and plays a key role in the proliferation and apoptosis of AML cells, suggesting that TRIM28 may serve as a novel therapeutic target for non-M3 AML.
9.Association of mixed exposure to lithium, vanadium, uranium, and bismuth in early pregnancy with gestational weight gain
Jiao LI ; Qi LI ; Shuang CHENG ; Jiayi SONG ; Xiaohui GUO ; Xiang WANG ; Di CHENG ; Kefeng FAN ; Ju WANG
Journal of Environmental and Occupational Medicine 2026;43(4):475-484
Background Gestational weight gain is closely related to maternal and infant health outcomes. Pregnant women are simultaneously exposed to four metals—lithium (Li), vanadium (V), uranium (U), and bismuth (Bi)—through inhalation of fine particulate matter and consumption of contaminated food and water. Existing studies suggest that exposure to these metals may be associated with gestational weight gain. However, no study has yet explored the complex relationships between exposure to mixtures of these four metals and weight gain at different stages of pregnancy. Objective To investigate the associations between mixed exposure to Li, V, U, and Bi in early pregnancy and the average weekly gestational weight gain during both early pregnancy and mid-to-late pregnancy. Methods This prospective study recruited eligible women in early pregnancy from an obstetrics clinic of a tertiary hospital in Jinan, China, between September 2021 and July 2023. Pre-pregnancy weight, current weight (at 11+0 to 13+6 weeks of gestation), and spot urine samples (≥5.0 mL) were collected at enrollment. Urinary concentrations of Li, V, Bi, and U were determined using inductively coupled plasma mass spectrometry. Participants were followed up in late pregnancy (≥28 weeks of gestation) to collect information on physical activity via questionnaire; weight measurements at the last antenatal visit (35+0 to 37+6 weeks of gestation) were obtained from the hospital information system. After adjusting for covariates, multiple linear regression and generalized additive models were used to assess the associations of individual metals with weekly weight gain in early pregnancy and in mid-to-late pregnancy. Bayesian kernel machine regression (BKMR) and quantile-based g-computation (Qgcomp) were applied to evaluate the joint effects of the metal mixture exposure on weekly weight gain at the two gestational stages. Results A total of 313 pregnant women were included. The geometric means of urinary Li, V, U, and Bi concentrations were 37.07, 0.20, 0.06, and 0.04 μg·L−1, respectively; after creatinine adjustment, the corresponding values were 46.82, 0.25, 0.07, and 0.05 μg·g−1 (Cr). The mean weekly gestational weight gain was (0.19±0.25) kg in early pregnancy and (0.53 ± 0.18) kg in mid-to-late pregnancy. Both multiple linear regression and generalized additive models showed that urinary V concentration was positively associated with average weekly gestational weight gain in early pregnancy, while no significant associations were found for other metals or for gestational weight gain in mid-to-late pregnancy. In the BKMR model with early-pregnancy weight gain as the outcome, V had the strongest association [posterior inclusion probability (PIP)=0.773]. When other metals were fixed at their medians, V showed a positive non-linear association with the outcome. A significant single-metal effect of V and its interaction with Li were observed. Compared with the 50th percentile of the metal mixture, the average weekly weight gain in early pregnancy increased by 0.016 (95%CI: 0.003, 0.029) and 0.018 (95%CI: 0.001, 0.036) at the 60th and 65th percentiles, respectively; conversely, at the 25th percentile, it decreased by 0.026 (95%CI: 0.002, 0.050). Overall, the joint effect of the metal mixture on early- pregnancy weight gain showed an upward trend. In the BKMR model for mid-to-late pregnancy gestational weight gain, all PIPs were<0.5, and no significant single-metal effects, interactions, or joint effects were identified. Qgcomp results confirmed a positive association between the metal mixture and early-pregnancy weight gain (b=0.031, 95%CI: 0.010, 0.051; P<0.01), with V contributing the highest positive weight (0.71). No significant association was found for weight gain in mid-to-late pregnancy (b=0.007, P=0.339). Conclusion Higher levels of co-exposure to the Li, V, Bi, and U metal mixture during early pregnancy may be associated with increased average weekly weight gain in early pregnancy. Among these metals, V exhibits a predominant role and appears to interact with Li. No association is observed between early-pregnancy metal mixture exposure and average weekly gestational weight gain in mid-to-late pregnancy. These findings suggest that monitoring and managing metal exposure during early pregnancy may be crucial for the rational regulation of gestational weight gain.
10.Effect of Oral Sodium Butyrate on Skeletal Muscle Atrophy via The Gut-muscle Axis in Antibiotic-pretreated CT26 Tumor-bearing Mice and Its Mechanism
Shu-Ling ZHANG ; Jun-Wei WANG ; Shi-Liang HU ; Tu-Tu WANG ; Shun-Chang LI ; Jia FAN ; Jun-Zhi SUN
Progress in Biochemistry and Biophysics 2026;53(3):724-739
ObjectiveTo explore the effect of oral sodium butyrate on skeletal muscle atrophy in CT26 tumor mice through the gut microbiota-skeletal muscle axis and its potential mechanism. MethodsSixty SPF BALB/c male mice aged 8 weeks were randomly divided into a normal control group (NC, n=18) and a ABX-depleted group (ABX, n=42). The ABX mice were pretreated with a quadruple antibiotic cocktail via oral gavage (0.2 ml per administration, once daily, 6 d per week, for 2 weeks), whereas NC received an equal volume of sterile water. The quadruple antibiotic cocktail consisted of metronidazole (1 g/L), vancomycin (0.5 g/L), ampicillin (1 g/L), and gentamicin (1 g/L). Following successful pretreatment, six mice from each group were randomly selected for gut microbiota sequencing analysis and designated as the Abx group and the NC0 group, respectively. Theremaining mice in ABX were subcutaneously inoculated in the dorsum with 0.2 ml of CT26 cell suspension (at a cell density of 1×107/ml). Then these mice were randomly allocated into three subgroups: a control tumor bearing model group (0_NaB, n=12), a tumor-bearing model group receiving low-dose oral sodium butyrate (L_NaB, n=12), a tumor-bearing model group receiving high-dose oral sodium butyrate (H_NaB, n=12). And mice in NC were inoculated at the same site with 0.2 ml of normal saline. The administration dose for L_NaB was 0.3 g/(kg·d), that for H_NaB was 0.5 g/(kg·d), while NC and 0_NaB were given the same volume of normal saline (0.2ml per time, once daily, 6 d per week, for 4 weeks). The general condition of mice was monitored, and forelimb grip strength gastrocnemius muscle mass and its muscle fiber cross-sectional area were measured for each group. The structural changes in gut microbiota were assessed by 16S rRNA sequencing of cecal contents. Pathological alterations in the intestinal wall were examined via HE staining. Serum and gastrocnemius muscle levels of TNF‑α, IL-6, IL-1β, and LPS were quantified using ELISA. The protein expression of ZO-1 and occludin in the small intestine, as well as proteins associated with the TLR4/MyD88/NF-κB signaling pathway in the gastrocnemius muscle, were detected by Western blot analysis. Results(1) The alpha-diversity in Abx was significantly lower than that in NC0 (P<0.01), a significant decrease of the mass and muscle fiber cross-sectional area of the gastrocnemius (P<0.01), with the majority of gut microbiota being effectively depleted. (2) Compared with NC, the subcutaneous tumors of mice in 0_NaB were prominent, a significant increase of the mass and muscle fiber cross-sectional area of the gastrocnemius, accompanied by a significant decrease in body weight at the end of the 3th and 4th week (P<0.05), and a significant weakening of the forelimb grasping strength at the 5th and 6th week (P<0.01). Compared with 0_NaB, the tumor mass of mice in L_NaB and H_NaB showed a significant decreasing trend, and the grip strength of the forelimbs significantly increased at the 5th and 6th week (P<0.05, P<0.01). (3) Compared with 0_NaB, the Shannon and Observed species indices in α diversity of L_NaB and H_NaB were significantly increased (P<0.05). At the genus level, compared with 0_NaB, L_NaB exhibited a significant decrease in the relative abundance of Parasutterella (P< 0.01), while H_NaB showed significant reductions in the relative abundances of both Escherichia-Shigella and Parasutterella (P < 0.01). (4) Compared with 0_NaB, the small intestinal tissue structure in L_NaB and H_NaB was more intact, the infiltration of inflammatory cells was significantly reduced, and the capillaries were slightly dilated. The expression levels of ZO-1 and occludin proteins in L_NaB were significantly increased (P<0.01). (5) The LPS concentration in the gastrocnemius muscle and the protein expression levels of TLR4, MyD88, p-IκBα, and p-NF‑κB p65 in L_NaB and H_NaB were significantly lower than those in 0_NaB (P<0.05). The serum TNF‑α concentration in H_NaB and TNF-α concentration in the gastrocnemius muscle of the L_NaB and H_NaB were significantly lower than those in 0_NaB (P<0.05, P<0.01, P<0.01). ConclusionOral administration of NaB can improve gut microbiota α diversity, adjusting its composition, improving intestinal mucosal barrier function, reducing the LPS-induced pro-inflammatory response, and delaying skeletal muscle atrophy. The underlying mechanism may involve down regulation of TLR4/MyD88/NF-κB signaling in skeletal muscle.


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