Objective To analyze the correlation of cholecystectomy and changes in intestinal microbiota composition and function by observing functional characteristics of differential microbial communities.Methods A cross-sectional study was conducted on the patients(PC group,n=73)undergoing cholecystectomy in our hospital from 2020 to 2021.Another 56 healthy age-and gender-matched individuals(HC group)without a history of cholecystectomy were subjected and served as the control group.Fecal specimens were collected from the 2 groups.16S rRNA sequencing analysis was performed to examine the changes in composition and function of intestinal microbiota.Results There were no statistical differences between the 2 groups in baseline indicators,such as gender,age,BMI,smoking and drinking history,blood pressure,heart rate,and comorbidities,but significant difference was observed in total bilirubin(TBIL)between them(P<0.01).Alpha diversity analysis showed no significant difference in Chao1,Shannon,and Simpson indices between the 2 groups.Beta diversity analysis using the Bray-Curtis distance algorithm revealed a significant difference between the 2 groups at the class and genus levels(P<0.05).The analysis of microbiota relative abundance using LEFSE showed that Enterobacteriaceae,Lactobacillales,Citrobacter,Megasphaera,Lactobacillus,Enterococcus,Akkermansia,Streptococcus,Klebsiella,and Ruminococcus_gnavus were up-regulated in the PC group,and Lachnospiraceae,Sutterellaceae,Lachnospirales,Lachnospira,and Sutterella were down-regulated.Kyoto Encyclopedia of Genes and Genomes(KEGG)functional prediction analysis indicated that significant differences were seen between the 2 groups in metabolic pathways,including ascorbic acid(vitamin C)metabolism and aldonic acid metabolism(P<0.05),tricarboxylic acid cycle(TCA cycle)(P<0.05),glutathione metabolism(P<0.05),glutamic acid metabolism(P<0.05),secondary bile acid metabolism(P<0.05),and pentose phosphate pathway(P<0.01).Conclusion Cholecystectomy is closely associated with the structural alterations in the composition of intestinal microbiota.Variations in microbiota composition and function may induce perturbations in TCA cycle and glutathione metabolism,glutamate metabolism,secondary bile acid metabolism,and pentose phosphate pathways.

Objective To investigate the diversity and composition of gut fungi microbiota in patients with polycystic kidney disease(PKD)and its correlation with clinical indicators.Methods A total of 44 PKD patients,44 patients with non-polycystic chronic kidney disease(NPCKD)and 22 healthy controls(HC)admitted to our hospital from February 2023 to February 2024 were recruited.ITS1 DNA sequencing was applied to analyze the gut fungal composition.Bioinformatics analysis was used to compare the diversity and structural differences of fungi among the 3 groups.Pearson correlation analysis was performed to analyze the relationship between gut fungi and clinical indicators.Results There were no significant differences in baseline characteristics(gender,age,body mass index,etc.)among the 3 groups,but statistical differences were seen in terms of serum indicators(such as serum creatinine,blood urea nitrogen,uric acid,estimated glomerular filtration rate,etc.)(P<0.01).Alpha diversity analysis showed no significant difference was seen between the PKD and HC groups,but the PKD group had significant differences to the NPCKD group(P<0.01).Beta diversity analysis revealed significant differences among the 3 groups and in pairwise comparisons(P=0.001).Fungi composition analysis found that the abundance of Candida was significantly higher in the PKD group than the other 2 groups(P<0.01),while the abundances of Aspergillus and Cladosporium were significantly lower in the PKD group than the HC group(P<0.05).Linear discriminant analysis(LEFSe)indicated that Candida was significantly enriched,while Aspergillus and Cladosporium were significantly reduced in the PKD group.Correlation analysis revealed that the abundance of Cladosporium was negatively correlated with cyst diameter and immunoglobulin light chain Kappa/Lambda ratio in the PKD group(P<0.05),while the abundance of Candida was positively correlated with liver/kidney cyst diameter(P<0.01).Conclusion PKD patients exhibit characteristic changes in gut fungi diversity and composition.The abundances of Cladosporium and Candida are closely associated with clinical indicators of PKD patients.

Objective To analyze the diversity and composition of gut microbiota in individuals with circadian rhythm disruption and their correlation with cognition.Methods Night shift workers and regular shift workers were subjected from our hospital during August 2022 and October 2024.The participants with circadian rhythm disorders were assigned into an experimental group(n=24),and those with normal circadian rhythms were into a control group(n=24).Their height,weight,age,gender,body mass index(BMI)and fresh fecal samples were collected,and Montreal Cognitive Assessment(MoCA)and Mini-Mental State Examination(MMSE)were used to evaluate their mental status.Metagenomics,Alpha and Beta diversity analyses,Linear Discriminant Analysis Effect Size(LEfSe),and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were employed to investigate the diversity and function characteristics of gut microbiota in the participants.Results There were no statistical differences between the 2 groups in baseline data,such as height,weight,gender,age,and BMI(P>0.05).Alpha diversity analysis indicated that no statistical differences were observed in the ACE,Chao1,Shannon,or Simpson indices between the 2 groups,while beta diversity analysis revealed significant differences(P<0.01),suggesting different structure of gut microbiota between them.In the experimental group,the abundance of Faecalibacterium prausnitzii and Agathobacter rectalis was decreased,while that of Escherichia coli and Phocaeicola vulgaratus was increased,with significant differences when compared with the control group(P<0.05).Additionally,KEGG functional analysis showed that the experimental group had obviously higher expression levels in Th17 cell differentiation and the IL-17 signaling pathway than the control group(P<0.05).Agathobacter rectalis and Faecalibacterium prausnitzii were positively correlated with MoCA score and MMSE score(P<0.05,P<0.01).Agathobacter rectalis was negatively correlated with the IL-17 signaling pathway and Th17 cell differentiation.Conclusion Individuals with circadian rhythm disorders have significant changes in the structure and function of gut microbiota when compared to those with normal circadian rhythms.Agathobacter rectalis may be involved in the regulation of the IL-17 signaling pathway and differentiation of Th17 cells,thereby possibly impacting the increases of cognitive score related to circadian rhythm disorders.

Objective To study the expression and clinical significance of neural Wiskott-Alrdich syndrome protein(N-WASP)in pla-centas with preeclampsia.Methods This study included a total of 65 pregnant women:15 in the early-onset preeclampsia group,15 in the early-onset control group,15 in the late-onset preeclampsia group,and 20 in the late-onset control group.Real-time fluorescence quan-titative PCR(RT-qPCR)was used to detect the relative expression of N-WASP mRNA in placental tissues.Western blotting and immu-nohistochemistry were used to detect the expression and position of N-WASP protein in placental tissues from each group.Results RT-qPCR revealed significantly lower N-WASP mRNA expression levels in the placental tissue of the early-onset preeclampsia group compared to those in the early-onset control group(0.50±0.19 vs.0.93±0.73,P＜0.05).The N-WASP mRNA expression levels in late-onset preeclampsia placenta were significantly lower than those in the late-onset control group(0.83±0.34 vs.1.15±0.34,P＜0.05).Western blotting revealed significantly lower N-WASP protein expression in the placental tissue of early-onset preeclampsia compared to that in the early-onset control group(0.35±0.17 vs.0.72±0.21,P＜0.05).The N-WASP protein expression in late-onset preeclampsia placenta was significantly lower than that in the late-onset control group(0.39±0.16 vs.0.76±0.20,P＜0.05).The N-WASP mRNA expression in the placenta negatively correlated with the occurrence of early-onset(r =-0.37,P = 0.042)and late-onset preeclampsia(r =-0.39,P = 0.019).Immunohistochemistry revealed that N-WASP protein was localized in the cytoplasm of syncytiotrophoblasts,cytotrophoblasts,villous stromal cells,and vascular endothelial cells.Conclusion The low expression of N-WASP may be closely associated with preeclampsia.

AIM:To investigate the effect of Huangqi-Danggui decoction(HQDG)on the brain tissue of rats with cerebral ischemia/reperfusion(I/R)injury for 7 d by regulating macroautophagy and chaperone-mediated autophagy(CMA),and to explore its mechanism.METHODS:Male SD rats were randomly divided into sham group,model group,HQDG group and Xuesaitong(XST)group.Determination of main chemical components of HQDG by liquid chro-matography-mass spectrometry.The model of middle cerebral artery occlusion/reperfusion in rats was established by the left modified thread embolism method,and the changes of cerebral blood flow were observed by laser speckle blood flow imager.Zea Longa score was used to observe the neurological deficit.HE staining was used to observe the degree of nerve cell injury.The changes of neurovascular unit and autophagosomes in brain tissue were observed by transmission electron microscopy.Immunohistochemical method was used to detect the expression of LC3,P62,lysosome-associated membrane protein-2A(LAMP-2A),heat shock protein 70(HSP70)and myocyte enhancer factor 2D(MEF2D)proteins.Western blot was used to detect the expression of autophagy-related proteins P62 and LC3-Ⅱ/LC3-I.RESULTS:Compared with the sham group,the neurological deficit score in model group was significantly higher(P<0.01).A large number of nerve cells showed necrosis and nuclear dissolution,with the cell arrangement being disordered.The number of autophagosomes increased.The protein expression levels of LC3,LAMP-2A,HSP70 and MEF2D in brain tissue increased,while the ex-pression level of P62 protein decreased(P<0.05 or P<0.01).Compared with the model group,the scores of neurological deficit in brain tissue in HQDG and XST groups were significantly lower(P<0.01).Cell damage was significantly re-duced.The number of autophagosomes further increased.The expression levels of LAMP-2A,HSP70,MEF2D and P62 proteins in brain tissue decreased,while the expression levels of LC3-Ⅱ/LC3-I protein increased(P<0.05 or P<0.01).CONCLUSION:HQDG can alleviate cerebral ischemia/reperfusion injury in rats and exert neuroprotective effects by ac-tivating macroautophagy and reducing CMA.

Objectives:To explore the causal relationship between body mass index,uric acid and congestive heart failure(CHF),and provide genetic evidence to support the association between uric acid-mediated body mass index and the risk of CHF. Methods:Using the published data set of genome-wide association studies(GWAS)in East Asia,inverse variance weighting(IVW)method was used as the main analysis method,and MR-Egger method,weighted median model(WME),simple model and weighted model were used to analyze the causal relationship between body mass index,uric acid and CHF.MR-Egger regression was used to detect pleiotropy,Cochran Q test was used to detect heterogeneity,leave-one-out sensitivity was used to detect bias,funnel plot was drawn to detect bias,and MR-PRESSO package was used to remove outlier single nucleotide polymorphism(SNP).After Mendelian analysis,the mediating effect ratio was calculated,and the reverse Mendelian randomization study results between body mass index,uric acid and CHF were analyzed. Results:IVW method showed that body mass index(OR=1.685,95%CI:1.417-2.003,P<0.001)and uric acid(OR=1.225,95%CI:1.087-1.380,P<0.001)were risk factors of CHF in two-sample Mendelian analysis.Body mass index(OR=1.204,95%CI:1.139-1.273,P<0.001)was a risk factor for uric acid.The mediating effect of uric acid was 7.23%.The P values of MR-Egger regression intercept terms were all>0.05,that is,there was no pleiotropy of the selected SNP,and the causal inference method was valid.The Cochran Q test P values of body mass index and uric acid,and body mass index and CHF were<0.01,indicating heterogeneity.IVW analysis of CHF and body mass index in reverse Mendelian analysis was OR=0.977,95%CI:0.947-1.008,P>0.05 and CHF and uric acid was OR=1.000,95%CI:0.963-1.038,P>0.05,so the reverse causal inference was not valid.However,the analysis of uric acid and body mass index showed pleiotropy,so the causal inference method was invalid. Conclusions:There is a causal and positive correlation between body mass index and CHF.There is a causal relationship and positive correlation between uric acid and CHF.However,uric acid is an incomplete mediator between body mass index and CHF.

Objective:To investigate the impact of circular RanGTPase activating protein 1 (circRANGAP1) on the biological behavior of trophoblast cells in preeclampsia and its potential mechanisms.Methods:Placental tissues were collected from preeclampsia patients and age- and gestational age- matched control pregnant women admitted to Shengjing Hospital of China Medical University from August 2020 to December 2022 (eight cases each in the early-onset preeclampsia group and early-onset control group, and 24 cases each in the late-onset preeclampsia group and late-onset control group). The expression levels of circRANGAP1 and eukaryotic translation initiation factor 4A3 (EIF4A3) mRNA in placental tissues were detected by real-time quantitative polymerase chain reaction (RT-qPCR), and EIF4A3 protein expression was assessed by Western blotting. In HTR-8/Svneo cells, the proliferation, migration, and invasion abilities were evaluated by cell counting assay, scratch assay, Transwell invasion assay, and the regulatory effect of EIF4A3 on circRANGAP1 was examined by RNA binding protein immunoprecipitation (RIP). Changes of circRANGAP1 expression in HTR-8/Svneo cells were detected by RT-qPCR after EIF4A3 knockdown. Statistical analysis was performed using independent sample t-test, non-parametric Chi-square test, or Pearson correlation analysis. Results:(1) There was no significant difference in circRANGAP1 expression between the early-onset preeclampsia group and the early-onset control group. However, circRANGAP1 expression was higher in the late-onset preeclampsia group compared to the late-onset control group [(3.764±3.297) vs. (0.960±0.720), t=4.07, P<0.001]. In late-onset preeclampsia patients, circRANGAP1 expression was positively correlated with both systolic and diastolic blood pressure (systolic: r=0.639, P<0.01; diastolic: r=0.800, P<0.001). There was no significant difference in EIF4A3 mRNA and protein expression between the early-onset preeclampsia group and the early-onset control group, but EIF4A3 mRNA and protein expression were higher in the late-onset preeclampsia group compared to the late-onset control group [mRNA: (2.963±3.081) vs. (1.149±0.667), t=2.30, P=0.028; protein: (2.504±1.008) vs. (0.258±0.180), t=4.39, P=0.005]. (2) After small interfering (si) RNA knockdown, there was no significant difference in mRANGAP1 expression, but circRANGAP1 expression decreased [(1.000±0.004), (0.465±0.031), and (0.621±0.030)], with si-1 showing the highest knockdown efficiency ( t=23.59, P=0.002). Specific knockdown of circRANGAP1 resulted in increased proliferation [(1.297±0.058) vs. (1.456±0.030), t=－5.97, P<0.001], invasion [(94.400± 6.504) vs. (219.000±19.870), t=－13.32, P<0.001], and migration [(25.493±3.498)% vs. (58.456±3.277)%, t=－15.38, P<0.001] abilities of trophoblast cells. (3) There are six binding sites for EIF4A3 in the upstream region of circRANGAP1 pre-mRNA. EIF4A3 can bind through regions a and b, but not region c. After siRNA knockdown, EIF4A3 expression decreased [(1.003±0.101), (0.276±0.060), (0.398±0.074), and (0.184±0.017)], with si-3 showing the highest knockdown efficiency. After EIF4A3 knockdown, circRANGAP1 expression in trophoblast cells decreased [(1.004±0.118) vs. (0.480±0.039), t=5.96, P=0.027]. Conclusion:circRANGAP1, regulated by EIF4A3, inhibits the proliferation, migration, and invasion abilities of trophoblast cells, thereby participating in the pathogenesis of preeclampsia.

Objective:To investigate the clinicopathological features, diagnosis and prognosis of polymorphous low-grade neuroepithelial tumor of the young (PLNTY).Methods:Two cases of PLNTY diagnosed during January 2016 to December 2019 were collected from Ningbo Diagnostic Pathology Center, Zhejiang, China. The clinical features, histopathological characteristics, immunohistochemical and molecular genetic findings were analyzed and the relevant literature was reviewed.Results:The two patients were both female, at the ages of 14 and 25 years, respectively. Both patients presented with seizure attacks. The imaging study showed a mixed signal in the cerebral cortex, located in the occipital and temporal lobes, respectively. Microscopically, the tumors were characterized by the invariable presence of oligodendroglioma-like appearance, often with calcification. Immunohistochemically, the tumors were diffusely and intensely CD34 positive with ramified, CD34-expressing neural elements in regional cortex. The tumors were positive for GFAP, Olig2 and ATRX, and negative for IDH1, Neu N, nestin and EMA. The Ki-67 labeling index was less than 2%. The case number 2 harbored the BRAF V600E mutation, while the case number 1 was negative for both the BRAF V600E mutation and 1p/19q codeletion. Both patients recovered very well and were free of seizures after the following-up of 2 and 24 months, respectively.Conclusions:PLNTY is an uncommon neuroepithelial tumor. Histopathologic and immunohistochemical examinations are necessary for establishing the diagnosis and for excluding oligodendroglioma. PLNTY should be considered as a benign tumor corresponding to WHO Grade I. The prognosis is overall good after complete tumor-resection.

Traumatic brain injury (TBI) is one of the diseases with high morbidity,mortality,and disability,which seriously endangers human health.Primary and secondary injuries caused by TBI are cascade reaction of various pathophysiological interactions.Because of its many injury factors and complex mechanisms,the treatment and therapeutic effect of TBI are limited at present.In recent years,animal experiments and clinical studies have shown that stem cell therapy could alleviate TBI-mediated neurological damage to a certain extent.Therefore,activation of endogenous neural cells response and transplantation of exogenous stem cells may be new strategies for TBI treatment.This article reviews the research progress of activation of endogenous neural cells response and transplantation of exogenous stem cells after TBI,and focuses on the therapeutic effects and possible mechanisms of stem cell transplantation in TBI treatment.