OBJECTIVES: The risk factors and role of mother‒child gut microbiota in pediatric inflammatory bowel disease (PIBD) remain unclear. We aimed to explore the clinical risk factors associated with PIBD, analyze the characteristics of gut microbiota of children and their mothers, and examine the correlation of the microbial composition in mother‒child pairs. METHODS: We conducted a case-control study including children with PIBD and their mothers as the case group, as well as healthy children and their mothers as the control group. Questionnaires were used to collect information such as family illness history and maternal and early-life events. Fecal samples were collected from the children and mothers for microbiota 16S ribosomal RNA (rRNA) sequencing to analyze the composition and its potential association with PIBD. RESULTS: A total of 54 pairs of cases and 122 pairs of controls were recruited. A family history of autoimmune disease and antibiotic use during pregnancy were associated with an increased risk of PIBD, and a higher education level of the father was associated with a decreased risk of PIBD. Children with PIBD and mothers exhibited different gut microbiota compared to healthy children and mothers. Similarities were observed in the gut microbiota of mothers and children in the same groups. Some bacterial biomarkers of mothers discovered in this study had the power to predict PIBD in their offspring. CONCLUSIONS PIBD is influenced by maternal risk factors and has unique gut microbiota characteristics. The mother‒child gut microbiota is closely related, suggesting the transmission and influence of the gut microbiota between mothers and children. This study highlights the potential pathogenesis of PIBD and provides a basis for developing targeted interventions.
Humans ; Gastrointestinal Microbiome ; Female ; Risk Factors ; Case-Control Studies ; Male ; Child ; Inflammatory Bowel Diseases/etiology* ; Adult ; RNA, Ribosomal, 16S/genetics* ; Feces/microbiology* ; Mothers ; Pregnancy ; Child, Preschool

Functional neuroendocrine neoplasm, which secrete hormones and lead to diverse clinical symptoms, pose significant challenges in diagnosis and treatment. Early identification and standardized management are crucial for improving patient prognosis. This article summarized three clinically relatively common cases of Functional pancreatic neuroendocrine neoplasm (F-pNENs), focusing on analyzing their diagnostic key points and standardized therapeutic pathways. It aimed to provide reference for early clinical identification of rare tumors and improvement of standardized multidisciplinary diagnosis and treatment as well as personalized treatment strategies.

The study aims to investigate the effects of adding different proportions of Codonopsis radix compound crude extracts to the rabbit diet on growth performance,immune status,intesti-nal enzyme activity,structure,and microbial composition.A total of 96 5-week-old New Zealand White rabbits were randomly divided into 4 groups,with 6 replicates per group.The control group(BC)was fed a basal diet,while the experimental groups(CM-H and CM-L)were fed a basal diet supplemented with 1 000 mg/kg and 500 mg/kg of Codonopsis radix compound crude extracts,re-spectively.The antibiotic group(CK)was fed a basal diet supplemented with 300 mg/kg of keto-tifen.The experimental period was 42 days.Blood samples were collected at days 21 and 42,and se-rum biochemical and immune markers were determined.Intestinal segments and contents were col-lected at day 42 for analysis of intestinal health.The results showed that compared with the BC group,the average daily gain,feed-to-gain ratio,and diarrhea rate were significantly higher(P＜0.05)in the CM-H and CM-L groups.The total cholesterol(Tchol)content in the serum was sig-nificantly lower in the CM-H group at day 21 and the CM-L group at day 42(P＜0.05).The high-density lipoprotein(HDL)was significantly higher in the CM-H and CM-L groups than in the CK group at day 42(P＜0.05),and the total protein(TP)in the serum was significantly higher in the CM-H and CM-L groups than in the BC group(P＜0.05).The IgG and IgM levels in the serum were significantly higher in the CM-H and CM-L groups than in the BC group(P＜0.05).In the CM-H and CM-L groups,the content of acetic acid in the colon was significantly higher than that in the BC group(P＜0.05).The content of propionic acid in the colon of the CM-L group was also significantly higher than that in the BC group(P＜0.05).The content of α-amylase in the duode-num,the content of trypsin in the duodenum,the pancreas,and the ileum of the CM-H group were significantly higher than those in the BC group(P＜0.05),and the content of trypsin in the duode-num of the CM-H group was significantly higher than those in the BC group and the CM-L group(P＜0.05).Compared with the BC group,the content of GPX1 in the ileum and jejunum of the CM-L group and the ileum of the CM-H group was significantly increased(P＜0.05),and the length of the villi in the duodenum of the CM-H group was significantly increased(P＜0.05).Compared with the BC group,the expression level of ZO-1 in the ileum of the CM-H group was significantly upregulated(P＜0.05),and the expression level of Claudin in the jejunum of the CM-H group and the CM-L group was significantly higher than that in the CK group(P＜0.05).The high-throughput sequencing results showed that the Sob index was significantly higher in the CM-L group compared to the BC group(P＜0.05).At the phylum level,the Firmicutes and Bacteroid-ota phyla were the main phyla.At the genus level,Akkermansia and Ruminococcus were the main genera.The relative abundance of Papillibacter and Eubacterium_ruminantium_group in the CM-L group was significantly higher than that in the CK group(P＜0.05).In summary,adding a Codonopsis radix compound crude extract to the diet can improve the growth performance,immu-nity,antioxidant capacity,integrity of intestinal mucosal structure,enzyme activity in the intestine,and increase the diversity of microorganisms in the blind intestine when the diet is supplemented with 500 mg/kg of Codonopsis radix compound crude extract.

To explore the immunomodulatory effects of a new vegetable oil adjuvant(named E515)containing vitamin E(VE)and ginsenosides(GS)on rabbit peripheral blood lymphocytes and Bordetella of rabbit inactivated vaccine.E515,Bordetella bronchiseptica(Bb)and LPS were co-cultured with rabbit peripheral blood lymphocytes in vitro,and the lymphocyte conversion rate was detected by CCK8 method,and the content of lymphocyte supernatant cytokines was detected by ELISA method.After rabbits were immunized with E515-Bb vaccine,the antibody level was detec-ted by indirect ELISA,the serum cytokine content was detected by ELISA,and the protective effect of E515-Bb vaccine on rabbits was observed by challenge test.In vitro cell experiments showed that E515 could significantly increase lymphocyte proliferation and TH1/TH2 cytokine se-cretion in rabbit peripheral blood.In vivo animal experiments showed that E515 adjuvant could sig-nificantly enhance the level of Bb specific antibody induced by Bordetella vaccine in rabbits.In-crease the secretion level of TH1/TH2 cytokines and decrease the secretion level of TNF-α;It can effectively protect rabbits against Bordetella infection with a protection rate of 91.67％.Therefore,E515 as a new vegetable oil adjuvant deserves further study.

The study aims to investigate the effects of adding different proportions of Codonopsis radix compound crude extracts to the rabbit diet on growth performance,immune status,intesti-nal enzyme activity,structure,and microbial composition.A total of 96 5-week-old New Zealand White rabbits were randomly divided into 4 groups,with 6 replicates per group.The control group(BC)was fed a basal diet,while the experimental groups(CM-H and CM-L)were fed a basal diet supplemented with 1 000 mg/kg and 500 mg/kg of Codonopsis radix compound crude extracts,re-spectively.The antibiotic group(CK)was fed a basal diet supplemented with 300 mg/kg of keto-tifen.The experimental period was 42 days.Blood samples were collected at days 21 and 42,and se-rum biochemical and immune markers were determined.Intestinal segments and contents were col-lected at day 42 for analysis of intestinal health.The results showed that compared with the BC group,the average daily gain,feed-to-gain ratio,and diarrhea rate were significantly higher(P＜0.05)in the CM-H and CM-L groups.The total cholesterol(Tchol)content in the serum was sig-nificantly lower in the CM-H group at day 21 and the CM-L group at day 42(P＜0.05).The high-density lipoprotein(HDL)was significantly higher in the CM-H and CM-L groups than in the CK group at day 42(P＜0.05),and the total protein(TP)in the serum was significantly higher in the CM-H and CM-L groups than in the BC group(P＜0.05).The IgG and IgM levels in the serum were significantly higher in the CM-H and CM-L groups than in the BC group(P＜0.05).In the CM-H and CM-L groups,the content of acetic acid in the colon was significantly higher than that in the BC group(P＜0.05).The content of propionic acid in the colon of the CM-L group was also significantly higher than that in the BC group(P＜0.05).The content of α-amylase in the duode-num,the content of trypsin in the duodenum,the pancreas,and the ileum of the CM-H group were significantly higher than those in the BC group(P＜0.05),and the content of trypsin in the duode-num of the CM-H group was significantly higher than those in the BC group and the CM-L group(P＜0.05).Compared with the BC group,the content of GPX1 in the ileum and jejunum of the CM-L group and the ileum of the CM-H group was significantly increased(P＜0.05),and the length of the villi in the duodenum of the CM-H group was significantly increased(P＜0.05).Compared with the BC group,the expression level of ZO-1 in the ileum of the CM-H group was significantly upregulated(P＜0.05),and the expression level of Claudin in the jejunum of the CM-H group and the CM-L group was significantly higher than that in the CK group(P＜0.05).The high-throughput sequencing results showed that the Sob index was significantly higher in the CM-L group compared to the BC group(P＜0.05).At the phylum level,the Firmicutes and Bacteroid-ota phyla were the main phyla.At the genus level,Akkermansia and Ruminococcus were the main genera.The relative abundance of Papillibacter and Eubacterium_ruminantium_group in the CM-L group was significantly higher than that in the CK group(P＜0.05).In summary,adding a Codonopsis radix compound crude extract to the diet can improve the growth performance,immu-nity,antioxidant capacity,integrity of intestinal mucosal structure,enzyme activity in the intestine,and increase the diversity of microorganisms in the blind intestine when the diet is supplemented with 500 mg/kg of Codonopsis radix compound crude extract.

To explore the immunomodulatory effects of a new vegetable oil adjuvant(named E515)containing vitamin E(VE)and ginsenosides(GS)on rabbit peripheral blood lymphocytes and Bordetella of rabbit inactivated vaccine.E515,Bordetella bronchiseptica(Bb)and LPS were co-cultured with rabbit peripheral blood lymphocytes in vitro,and the lymphocyte conversion rate was detected by CCK8 method,and the content of lymphocyte supernatant cytokines was detected by ELISA method.After rabbits were immunized with E515-Bb vaccine,the antibody level was detec-ted by indirect ELISA,the serum cytokine content was detected by ELISA,and the protective effect of E515-Bb vaccine on rabbits was observed by challenge test.In vitro cell experiments showed that E515 could significantly increase lymphocyte proliferation and TH1/TH2 cytokine se-cretion in rabbit peripheral blood.In vivo animal experiments showed that E515 adjuvant could sig-nificantly enhance the level of Bb specific antibody induced by Bordetella vaccine in rabbits.In-crease the secretion level of TH1/TH2 cytokines and decrease the secretion level of TNF-α;It can effectively protect rabbits against Bordetella infection with a protection rate of 91.67％.Therefore,E515 as a new vegetable oil adjuvant deserves further study.

OBJECTIVE To investigate the effects of medicated serum of Siwutang on autophagy of ovarian granulosa cells(KGN cells)in polycystic ovarian syndrome(PCOS)and its underlying mechanism.METHODS Blank serum and different-concentration medicated serum of Siwutang were prepared by intragastric administration of normal saline and different doses of Siwutang[0.52,1.04,2.08 g/(kg·d)]in 3-month-old female SD rats.After screening the intervention concentration of Siwutang medicated serum,KGN cells were divided into control group(without any treatment),dehydroepiandrosterone(DHEA)group(treated with 50 μmol/L DHEA for 48 h),blank serum group(treated with 50 μmol/L DHEA for 48 h and with 10％blank serum for 72 h)and medium-concentration of Siwutang medicated serum group(treated with 50 μmol/L DHEA for 48 h and with 10％medium-concentration Siwutang medicated serum for 72 h).The number of autophagosomes was observed in each group,and protein expressions of pathway-related proteins[fructose-1,6-bisphosphatase 1(FBP1),mammalian target of rapamycin(mTOR),phosphorylated mTOR(p-mTOR)],autophagy-related proteins[p62,microtubule-associated protein 1 light chain 3(LC3)]and mRNA expression of FBP1 were also detected.The(transfected)cells were further divided into Siwutang group(treated with 10％medium dose of Siwutang medicated serum for 72 h after 48 h intervention with 50 μmol/L DHEA),Siwutang+si-NC group[negative control small interfering RNA(siRNA)transfected cells treated with 50 μmol/L DHEA for 48 h,and then with 10％medium-concentration of Siwutang medicated serum for 72 h]and Siwutang+si-FBP1 group(FBP1 siRNA transfected cells treated with 50 μmol/L DHEA for 48 h,and then with 10％medium-concentration Siwutang medicated serum for 72 h).The effects of knocking down FBP1 on the above-mentioned effects of Siwutang were detected.RESULTS Compared with control group,DHEA group exhibited an increase in the number of autophagosomes,an elevated LC3-Ⅱ/LC3-Ⅰ and p-mTOR/mTOR,as well as increases in protein and mRNA expressions of FBP1,and decreased protein expression of p62(P＜0.05).Compared to both DHEA group and blank serum group,the medium-concentration of Siwutang medicated serum group showed a decrease in the number of autophagosomes,a decrease in LC3-Ⅱ/LC3-Ⅰ,and increases in p-mTOR/mTOR,protein expression of p62,protein and mRNA expressions of FBP1(P＜0.05).After knocking down FBP1,compared with Siwutang+si-NC group,Siwutang+si-FBP1 group showed a significant decrease in cell viability,protein expression of p62,protein and mRNA expressions of FBP1 as well as p-mTOR/mTOR,and an increase in LC3-Ⅱ/LC3-Ⅰ(P＜0.05).CONCLUSIONS Siwutang can promote the phosphorylation of mTOR protein by up-regulating the protein and mRNA expressions of FBP1 in KGN cells,thus inhibiting autophagy of KGN cells.

Objective:In this study,three methods of removing microglial cells from primary astrocyte culture were compared and analyzed to explore the characteristics of different purification methods.It provides the basis for research-ers to select the appropriate primary astrocyte culture method.Methods:The cerebral cortex cells of SD neonatal rats were isolated and platted and divided into an unpurified group(control group),a traditional oscillation purification group(TO group),a cytosine β-D-arabinofuranoside(AraC)+TO group,and an AraC+L-leucine methyl ester(LME)group according to different purification methods.Cell counts were used to calculate the cell production after different purification treatments.The cell morphology of each group was observed under an optical microscope,and the proportion of glial fibrillary acidic protein(GFAP)positive cells was calculated by immunofluorescence staining.AlamarBlue HS was used to detect cell viability after passage.Results:Compared with the control group,the cell purity(P＜0.0001)and cell viability(P＜0.0001)of purified cells in the TO group were significantly increased after pas-sage,while the total number of purified cells was significantly decreased(P＜0.001).These trends were more obvious in the AraC+TO and AraC+LME group,and the cell purity(P＜0.0001 or P＜0.0001)and cell viability(P＜0.01 or P＜0.001)of purified cells in the AraC+TO and AraC+LME group after passage were significantly higher than those in the TO group.The total amount of cells obtained was significantly decreased compared with the TO group(P＜0.01 orP＜0.01).There was no significant difference between the AraC+TO and AraC+LME group.Conclusion:The culture rich in astrocytes(cell purity＞90％)was obtained by the TO method,and the cell yield was high,which could be applied to routine cell research.The highly enriched astrocyte cultures(cell purity＞99％)obtained by the AraC+TO and AraC+LME purification methods can be applied to study the precise role of astrocytes in the context of neuroinflammation.

Purpose To investigate the relationship be-tween the expressions of CD3,CD4,CD8,CD20,CD68,PD-1 and PD-L1 and the clinical prognosis of peritoneal mesothelioma(PM).Methods Clinical data of 69 PM patients were collect-ed.EnVision two-step immunohistochemical method was used to detect the expression of CD3,CD4,CD8,CD20,CD68,PD-1 and PD-L1 in PM.Associations between expression levels and survival were estimated by univariate and multivariate Cox pro-portional-hazards models.Results There were no significant differences in the expressions of CD3,CD4,CD8,CD20,CD68,and PD-1 in tumor infiltrating lymphocytes(TILs)of ep-ithelioid and non-epithelioid PM.The expression of PD-L1 in non-epithelioid PM TILs was higher than that in epithelioid PM TILs,but the difference was not statistically significant.The median overall survival(mOS)time of PM was 19.1 months.Multivariate models identified asbestos exposure(P=0.002),PCI score(P=0.034),histological type(P=0.036),and CD4 expression(P=0.043)was independent prognostic factors for PM.Conclusion Asbestos exposure,PCI score,histologi-cal type and CD4 expression in TILs may exert significant im-pacts on survival of PM patients.

OBJECTIVE: This study aimed to investigate the therapeutic effects of Xiaoyao San (XYS), a herbal medicine formula, on exercise capacity and liver mitochondrial metabolomics in a rat model of depression induced by chronic unpredictable mild stress (CUMS). METHODS: A total of 24 male SD rats were randomly divided into four groups: control group (C), CUMS control group (M), Venlafaxine positive treatment group (V), and XYS treatment group (X). Depressive behaviour and exercise capacity of rats were assessed by body weight, sugar-water preference test, open field test, pole test, and rotarod test. The liver mitochondria metabolomics were analyzed by using liquid chromatography-mass spectrometry (LC-MS) method. TCMSP database and GeneCards database were used to screen XYS for potential targets for depression, and GO and KEGG enrichment analyses were performed. RESULTS: Compared with C group, rats in M group showed significantly lower body weight, sugar water preference rate, number of crossing and rearing in the open field test, climbing down time in the pole test, and retention time on the rotarod test (P < 0.01). The above behaviors and exercise capacity indices were significantly modulated in rats in V and X groups compared with M group (P < 0.05, 0.01). Compared with C group, a total of 18 different metabolites were changed in the liver mitochondria of rats in M group. Nine different metabolites and six metabolic pathways were regulated in the liver mitochondria of rats in X group compared with M group. The results of network pharmacology showed that 88 intersecting targets for depression and XYS were obtained, among which 15 key targets such as IL-1β, IL-6, and TNF were predicted to be the main differential targets for the treatment of depression. Additionally, a total of 1 553 GO signaling pathways and 181 KEGG signaling pathways were identified, and the main biological pathways were AGE-RAGE signaling pathway, HIF-1 signaling pathway, and calcium signaling pathway. CONCLUSION XYS treatment could improve depressive symptoms, enhance exercise capacity, positively regulate the changes of mitochondrial metabolites and improve energy metabolism in the liver of depressed rats. These findings suggest that XYS exerts antidepressant effects through multi-target and multi-pathway.