Objectiveto evaluate the effects and underlying mechanisms of a glucose restriction （GR） regimen on acrolein-induced neurotoxicity and AD-related pathology. MethodsImmortalized mouse hippocampal HT22 cells were pretreated with culture media containing different glucose concentrations （25 mmol/L， 17.5 mmol/L， and 12.5 mmol/L）， followed by incubation with 25 μmol/L acrolein for specified durations. Cell viability and apoptosis were assessed using MTT assay and flow cytometry， respectively. Changes in glutathione （GSH）， superoxide dismutase （SOD）， and malondialdehyde （MDA） levels were measured using enzyme linked immunosorbent assay（ELISA） kits. Alterations in the expression of BDNF/TrkB signaling proteins and key enzymes involved in amyloid precursor protein （APP） metabolism were evaluated by Western blot. ResultsPretreatment with GR significantly attenuated acrolein-induced loss of cell viability in HT22 cells. GR also alleviated the depletion of GSH and SOD， reduced MDA levels， and thereby ameliorated acrolein-induced oxidative damage（P＜0.05）. Furthermore， GR inhibited acrolein-triggered early apoptosis. Notably， the GR regimen protectively modulated the expression of BDNF/TrkB signaling pathway components and key APP-metabolizing enzymes （ADAM10 and BACE1）（P＜0.05）. ConclusionGR exerts neuroprotective effects against acrolein-induced toxicity， suggesting its potential in preventing or delaying the development of AD.

Blood turbidity and collateral disease are closely related to each other as the important component parts of the theoretical system of modern traditional Chinese medicine (TCM). The former focuses on blood and the latter on blood vessels and collaterals. By integrating these two theories, a theoretical basis for TCM syndrome differentiation and treatment of modern diseases can be provided. This article summarizes the correlation of origin, concept, treatment method and representative drugs of two theories, and points out that both blood turbidity and collateral disease prospers and develops through the integration of TCM classical theory and modern medical achievements. Theoretically, blood turbidity is the cause of collateral disease, and collateral disease is the result of aggravated blood turbidity. In many metabolic diseases, blood turbidity and collateral disease actually correspond to the main features of the early and late stages of the same disease, respectively. In treatment, clearing blood turbidity is consistent with dredging collaterals. When clearing blood turbidity, it is necessary to dredge the collaterals, and when dredging the collaterals, it is necessary to clear the blood turbidity. In terms of prescription and medication, Huazhuo Xingxue Decoction is the representative prescription of blood turbidity, which can be combined with Ramulus Cinnamomi, Sichuan lovage rhizome, earthworm, and other dredging collateral drugs. The representative prescription for collateral disease is Tongxinluo Capsule, which can be combined with lotus leaf, Fructus Crataegi, cassia seed, and other turbid-clearing drugs to enhance the curative effect.

Microbial communities such as those residing in the human gut are highly diverse and complex, and many with important implications for health and diseases. The effects and functions of these microbial communities are determined not only by their species compositions and diversities but also by the dynamic intra- and inter-cellular states at the transcriptional level. Powerful and scalable technologies capable of acquiring single-microbe-resolution RNA sequencing information in order to achieve a comprehensive understanding of complex microbial communities together with their hosts are therefore utterly needed. Here we report the development and utilization of a droplet-based smRNA-seq (single-microbe RNA sequencing) method capable of identifying large species varieties in human samples, which we name smRandom-seq2. Together with a triple-module computational pipeline designed for the bacteria and bacteriophage sequencing data by smRandom-seq2 in four human gut samples, we established a single-cell level bacterial transcriptional landscape of human gut microbiome, which included 29,742 single microbes and 329 unique species. Distinct adaptive response states among species in Prevotella and Roseburia genera and intrinsic adaptive strategy heterogeneity in Phascolarctobacterium succinatutens were uncovered. Additionally, we identified hundreds of novel host-phage transcriptional activity associations in the human gut microbiome. Our results indicated that smRandom-seq2 is a high-throughput and high-resolution smRNA-seq technique that is highly adaptable to complex microbial communities in real-world situations and promises new perspectives in the understanding of human microbiomes.
Humans ; Gastrointestinal Microbiome/genetics* ; Bacteriophages/physiology* ; High-Throughput Nucleotide Sequencing ; Sequence Analysis, RNA/methods* ; Bacteria/virology*

Objective:To investigate the effects of repetitive transcranial magnetic stimulation(rTMS) on the core symptoms, brain functional connectivity and activation in children with attention deficit hyperactivity disorder (ADHD) using functional near-infrared spectroscopy (fNIRS).Methods:From September 2022 to March 2024, a total of 35 children with ADHD were selected as research subjects and they were randomly divided into observation group ( n=17) and control group ( n=18). The control group received conventional rehabilitation therapy, while the observation group received rTMS therapy in addition to the conventional therapy. Both groups were treated every other day, with each course of treatment lasting four weeks, and a total of three courses of treatment were administered consecutively. The clinical symptoms of the children with ADHD were assessed using Swanson, Nolan, and Pelham-Ⅳ(SNAP-Ⅳ) before and after treatment. fNIRS was used to detect the relative concentration changes of oxyhemoglobin (HbO 2) and deoxyhemoglobin (HbR) in the prefrontal cortex under resting-state and Go/Nogo task conditions before and after treatment. Statistical analysis was performed using SPSS 26.0 software. Paired sample t-test were used for within-group comparisons, and independent sample t-test were used for between-group comparisons. Results:(1) After treatment, the scores for inattention, hyperactivity/impulsivity and oppositional defiant behavior in the two groups were significantly lower than before treatment ( t=3.51-18.86, all P<0.05). The scores for inattention, hyperactivity/impulsivity and oppositional defiant behavior in the observation group were significantly lower than those in the control group ( t=2.21, 2.03, 2.39, all P<0.05). (2) After treatment, the functional connectivity strength between all regions of interest in both groups was significantly higher than before treatment ( t=3.53-37.90, all P<0.05). The functional connectivity strength of the left dorsolateral prefrontal cortex (0.25±0.03, 0.21±0.03), right dorsolateral prefrontal cortex (0.12±0.02, 0.09±0.02), left medial prefrontal cortex (0.13±0.02, 0.10±0.01) and right medial prefrontal cortex (0.31±0.04, 0.24±0.06) in the observation group was significantly higher than those in the control group (all P<0.05). (3) In the Go/Nogo task, after treatment, the average HbO 2 concentrations in the left dorsolateral prefrontal cortex, right dorsolateral prefrontal cortex, left medial prefrontal cortex, right medial prefrontal cortex, left temporal lobe, and right temporal lobe in both groups were all higher than before treatment (all P<0.05). After treatment, the average HbO 2 concentrations in the left dorsolateral prefrontal cortex, right dorsolateral prefrontal cortex, left medial prefrontal cortex and right medial prefrontal cortex of the observation group were significantly higher than those of the control group (all P<0.05). Conclusion:rTMS therapy can improve the core symptoms of children with ADHD, which may be related to the strength of brain functional connectivity and activation of ADHD brain function by rTMS.

Objective To analyze the symptomatic improvement effects of vitamin D in children with autism spectrum disorder(ASD)based on the microbiota-gut-brain axis.Methods Seventy-two children with ASD were randomly divided into an observation group and a control group,with 36 cases in each group.Three cases dropped out in the control group.The observation group received 1200 IU/day of vitamin D supplementation in addition to conventional rehabilitation training,while the control group received only conventional rehabilitation training.The intervention lasted for 12 weeks.Assessments were conducted before and after the intervention using the childhood autism rating scale(CARS),autism behavior checklist(ABC),and repetitive behavior scale-revised(RBS-R).Resting-state functional connectivity of the brain was measured using near-infrared functional imaging,and serum levels of 25(OH)D3,inflammatory cytokines,and gut microbiota were analyzed.The differences in these indicators before and after the intervention were compared between the two groups to evaluate clinical efficacy.Results The between-group differences in pre-and post-intervention changes showed that the observation group had significantly greater improvements than the control group in the following measures:CARS scores(-5.92±1.40 vs.-2.55±1.43),RBS-R scores(-5.99±1.01 vs.-3.10±1.47),resting-state brain functional connectivity(0.19±0.15 vs.0.10±0.18),serum 25(OH)D3 levels[(34.89±8.18)ng/mL vs.(0.68±6.73)ng/mL],serum interleukin-6(IL-6)levels[(-6.60±6.07)pg/mL vs.(-0.74±9.45)pg/mL],IL-1β levels[(-2.56±1.33)pg/mL vs.(-0.04±2.13)pg/mL],and tumor necrosis factor-α(TNF-α)levels[(-4.09±3.85)pg/mL vs.(0.21±4.05)pg/mL](P<0.05).Post-intervention,significant differences in gut microbial β-diversity were observed between the two groups(R2=0.030,P=0.040,Adonis).LEfSe analysis revealed that the observation group exhibited enrichment in Clostridia(LDA=4.747,P=0.003),Clostridiales(LDA=4.747,P=0.003),Clostridiaceae(LDA=3.476,P=0.001),Lachnospiraceae(LDA=4.709,P=0.004),Odoribacteraceae(LDA=3.458,P=0.027),Odoribacter(LDA=3.458,P=0.027),Burkholderiales(LDA=3.339,P=0.038),Firmicutes(LDA=4.764,P=0.003),and Betaproteobacteria(LDA=3.338,P=0.037).Conclusion Vitamin D supplementation can modulate gut microbial diversity in children with ASD,significantly influence the abundance of specific gut microbiota,reduce systemic inflammatory cytokines,enhance brain functional connectivity,and alleviate clinical symptoms of ASD.

Objective To investigate the effect of total paeony glycoside(TPG)on cerebral ischemia-reperfusion injury(CI/RI)of rats.Methods The rats were randomly divided into sham surgery(sham)group,CI/RI model group(simple CI/RI group),positive control group(nimodipine group,5 mg/kg),low-dose TPG group(TPG-L group,27 mg/kg),a high-dose TPG group(TPG-H group,54 mg/kg)and a high-dose TPG+NOD-like receptor thermal protein domain associated protein 3(NLRP3)activator diethyl dithiocarbamate(DDC)group(TPG-H+DDC group,54 mg/kg TPG and 30 mg/kg DDC),with 18 rats in each,administered once a day for 7 consecutive days.After the administration,the neurological deficit score of the rats was evaluated.Nissl staining microscopy was applied to observe neuronal activity in brain tissue.2,3,5-triphenyltetrazolium chloride(TTC)staining microscopy was applied to detect the area of cerebral infarction in rats.The level of interleukin-1β and IL-18 in brain tissue was measured by ELISA method.Western blot was applied to detect the expression of purinergic receptor P2X ligand-gated ion channel 7(P2X7R)/NLRP3 signaling pathway related proteins and pyroptosis related proteins such as apoptosis associated speck like protein containing a CARD(ASC)and cysteine protease 1(caspase-1)proteins in brain tissue.Results Compared with the sham group,the neurological deficit score,infarct area,level of IL-1β and IL-18 in brain tissue and protein expression of P2X7R,NLRP3,ASC,and caspase-1 in brain tissue of rats in the simple CI/RI group were significantly increased(P<0.05),while the proportion of Nissl body positive cells in brain tissue was significantly reduced(P<0.05).The change in corresponding indicators of rats in the nimodipine group,TPG-L group,and TPG-H group was opposite to those in the simple CI/RI group(P<0.05).NLRP3 acti-vator DDC antagonized the inhibitory effect of TPG on cell pyroptosis in CI/RI rats.Conclusions TPG may inhibit brain injury in CI/RI rats by down-regulating the P2X7R/NLRP3 pathway.

Objective To analyze the symptomatic improvement effects of vitamin D in children with autism spectrum disorder(ASD)based on the microbiota-gut-brain axis.Methods Seventy-two children with ASD were randomly divided into an observation group and a control group,with 36 cases in each group.Three cases dropped out in the control group.The observation group received 1200 IU/day of vitamin D supplementation in addition to conventional rehabilitation training,while the control group received only conventional rehabilitation training.The intervention lasted for 12 weeks.Assessments were conducted before and after the intervention using the childhood autism rating scale(CARS),autism behavior checklist(ABC),and repetitive behavior scale-revised(RBS-R).Resting-state functional connectivity of the brain was measured using near-infrared functional imaging,and serum levels of 25(OH)D3,inflammatory cytokines,and gut microbiota were analyzed.The differences in these indicators before and after the intervention were compared between the two groups to evaluate clinical efficacy.Results The between-group differences in pre-and post-intervention changes showed that the observation group had significantly greater improvements than the control group in the following measures:CARS scores(-5.92±1.40 vs.-2.55±1.43),RBS-R scores(-5.99±1.01 vs.-3.10±1.47),resting-state brain functional connectivity(0.19±0.15 vs.0.10±0.18),serum 25(OH)D3 levels[(34.89±8.18)ng/mL vs.(0.68±6.73)ng/mL],serum interleukin-6(IL-6)levels[(-6.60±6.07)pg/mL vs.(-0.74±9.45)pg/mL],IL-1β levels[(-2.56±1.33)pg/mL vs.(-0.04±2.13)pg/mL],and tumor necrosis factor-α(TNF-α)levels[(-4.09±3.85)pg/mL vs.(0.21±4.05)pg/mL](P<0.05).Post-intervention,significant differences in gut microbial β-diversity were observed between the two groups(R2=0.030,P=0.040,Adonis).LEfSe analysis revealed that the observation group exhibited enrichment in Clostridia(LDA=4.747,P=0.003),Clostridiales(LDA=4.747,P=0.003),Clostridiaceae(LDA=3.476,P=0.001),Lachnospiraceae(LDA=4.709,P=0.004),Odoribacteraceae(LDA=3.458,P=0.027),Odoribacter(LDA=3.458,P=0.027),Burkholderiales(LDA=3.339,P=0.038),Firmicutes(LDA=4.764,P=0.003),and Betaproteobacteria(LDA=3.338,P=0.037).Conclusion Vitamin D supplementation can modulate gut microbial diversity in children with ASD,significantly influence the abundance of specific gut microbiota,reduce systemic inflammatory cytokines,enhance brain functional connectivity,and alleviate clinical symptoms of ASD.

Objective:To investigate the clinical efficacy of probiotics combined with omeprazole in the treatment of acute gastroenteritis in children.Methods:This study used a prospective design. A total of 110 children with acute gastroenteritis admitted to The First Hospital of Yulin from January to December 2023 were selected for this study. The children were divided into two groups: a control group and an observation group ( n = 55 per group) using a random numbering method. The control group received omeprazole enteric-coated capsules in addition to conventional treatment, while the observation group was treated with butyric acid-producing bifidobacterium and omeprazole enteric-coated capsules. The treatment course for both groups lasted 2 weeks. Clinical symptom scores were compared between the two groups before and after treatment. Additionally, the time to relief of major symptoms, the proportions of T cell subpopulations, changes in serum inflammatory markers, and the incidence of adverse reactions were also compared between the two groups. Results:After treatment, the scores of vomiting, loss of appetite, and diarrhea in the observation group were (1.04 ± 0.14), (0.84 ± 0.09), and (1.55 ± 0.17), respectively, all of which were significantly lower than those in the control group [(1.67 ± 0.18), (1.05 ± 0.12), (2.31 ± 0.25); t = 20.49, 10.38, 18.64, all P<0.001]. The time to relief from vomiting, fever, and diarrhea in the observation group were (1.53 ± 0.52) days, (1.39 ± 0.32) days, and (1.67 ± 0.42) days, which were significantly shorter than those in the control group [(2.88 ± 0.82) days, (2.59 ± 0.31) days, (2.69 ± 0.71) days; t = 10.31, 19.98, 9.17, all P<0.001]. The proportions of CD 3+, CD 4+, and CD 8+ T cells in the observation group were (69.74 ± 7.67)%, (45.32 ± 6.80)%, and (27.61 ± 3.31)%, respectively. They were (62.48 ± 9.37)%, (38.66 ± 5.80)%, and (33.42 ± 4.01)% respectively in the control group. There were significant differences in the proportions of CD 3+, CD 4+, and CD 8+ T cells between the two groups ( t = -4.45, -5.53, 8.29, all P<0.001). The levels of high-sensitivity C-reactive protein, procalcitonin, and interleukin-6 in the observation group were (4.22 ± 0.51) mg/L, (2.55 ± 0.38) ng/L, and (10.25 ± 1.44) ng/L, respectively, all of which were significantly lower than those in the control group [(7.68 ± 1.08) mg/L, (4.78 ± 0.57) ng/L, (18.53 ± 2.22) ng/L; t = 21.48, 24.14, 23.21, all P<0.001]. The level of interleukin-2 in the observation group was significantly higher than that in the control group [(69.74 ± 7.67) pg/L vs. (56.87 ± 8.53) pg/L; t = -8.32, P<0.001]. There was no significant difference in the incidence of adverse reactions between the observation and control groups [7.27% (4/55) vs. 3.64% (2/55); χ2 = 0.18, P>0.05). Conclusions:The combination of probiotics and omeprazole can effectively improve the clinical manifestations of acute gastroenteritis in children, accelerate symptom recovery, enhance immune function, and demonstrate good safety.

Objective:To investigate the effects of repetitive transcranial magnetic stimulation(rTMS) on the core symptoms, brain functional connectivity and activation in children with attention deficit hyperactivity disorder (ADHD) using functional near-infrared spectroscopy (fNIRS).Methods:From September 2022 to March 2024, a total of 35 children with ADHD were selected as research subjects and they were randomly divided into observation group ( n=17) and control group ( n=18). The control group received conventional rehabilitation therapy, while the observation group received rTMS therapy in addition to the conventional therapy. Both groups were treated every other day, with each course of treatment lasting four weeks, and a total of three courses of treatment were administered consecutively. The clinical symptoms of the children with ADHD were assessed using Swanson, Nolan, and Pelham-Ⅳ(SNAP-Ⅳ) before and after treatment. fNIRS was used to detect the relative concentration changes of oxyhemoglobin (HbO 2) and deoxyhemoglobin (HbR) in the prefrontal cortex under resting-state and Go/Nogo task conditions before and after treatment. Statistical analysis was performed using SPSS 26.0 software. Paired sample t-test were used for within-group comparisons, and independent sample t-test were used for between-group comparisons. Results:(1) After treatment, the scores for inattention, hyperactivity/impulsivity and oppositional defiant behavior in the two groups were significantly lower than before treatment ( t=3.51-18.86, all P<0.05). The scores for inattention, hyperactivity/impulsivity and oppositional defiant behavior in the observation group were significantly lower than those in the control group ( t=2.21, 2.03, 2.39, all P<0.05). (2) After treatment, the functional connectivity strength between all regions of interest in both groups was significantly higher than before treatment ( t=3.53-37.90, all P<0.05). The functional connectivity strength of the left dorsolateral prefrontal cortex (0.25±0.03, 0.21±0.03), right dorsolateral prefrontal cortex (0.12±0.02, 0.09±0.02), left medial prefrontal cortex (0.13±0.02, 0.10±0.01) and right medial prefrontal cortex (0.31±0.04, 0.24±0.06) in the observation group was significantly higher than those in the control group (all P<0.05). (3) In the Go/Nogo task, after treatment, the average HbO 2 concentrations in the left dorsolateral prefrontal cortex, right dorsolateral prefrontal cortex, left medial prefrontal cortex, right medial prefrontal cortex, left temporal lobe, and right temporal lobe in both groups were all higher than before treatment (all P<0.05). After treatment, the average HbO 2 concentrations in the left dorsolateral prefrontal cortex, right dorsolateral prefrontal cortex, left medial prefrontal cortex and right medial prefrontal cortex of the observation group were significantly higher than those of the control group (all P<0.05). Conclusion:rTMS therapy can improve the core symptoms of children with ADHD, which may be related to the strength of brain functional connectivity and activation of ADHD brain function by rTMS.

Objective:To investigate the clinical efficacy of probiotics combined with omeprazole in the treatment of acute gastroenteritis in children.Methods:This study used a prospective design. A total of 110 children with acute gastroenteritis admitted to The First Hospital of Yulin from January to December 2023 were selected for this study. The children were divided into two groups: a control group and an observation group ( n = 55 per group) using a random numbering method. The control group received omeprazole enteric-coated capsules in addition to conventional treatment, while the observation group was treated with butyric acid-producing bifidobacterium and omeprazole enteric-coated capsules. The treatment course for both groups lasted 2 weeks. Clinical symptom scores were compared between the two groups before and after treatment. Additionally, the time to relief of major symptoms, the proportions of T cell subpopulations, changes in serum inflammatory markers, and the incidence of adverse reactions were also compared between the two groups. Results:After treatment, the scores of vomiting, loss of appetite, and diarrhea in the observation group were (1.04 ± 0.14), (0.84 ± 0.09), and (1.55 ± 0.17), respectively, all of which were significantly lower than those in the control group [(1.67 ± 0.18), (1.05 ± 0.12), (2.31 ± 0.25); t = 20.49, 10.38, 18.64, all P<0.001]. The time to relief from vomiting, fever, and diarrhea in the observation group were (1.53 ± 0.52) days, (1.39 ± 0.32) days, and (1.67 ± 0.42) days, which were significantly shorter than those in the control group [(2.88 ± 0.82) days, (2.59 ± 0.31) days, (2.69 ± 0.71) days; t = 10.31, 19.98, 9.17, all P<0.001]. The proportions of CD 3+, CD 4+, and CD 8+ T cells in the observation group were (69.74 ± 7.67)%, (45.32 ± 6.80)%, and (27.61 ± 3.31)%, respectively. They were (62.48 ± 9.37)%, (38.66 ± 5.80)%, and (33.42 ± 4.01)% respectively in the control group. There were significant differences in the proportions of CD 3+, CD 4+, and CD 8+ T cells between the two groups ( t = -4.45, -5.53, 8.29, all P<0.001). The levels of high-sensitivity C-reactive protein, procalcitonin, and interleukin-6 in the observation group were (4.22 ± 0.51) mg/L, (2.55 ± 0.38) ng/L, and (10.25 ± 1.44) ng/L, respectively, all of which were significantly lower than those in the control group [(7.68 ± 1.08) mg/L, (4.78 ± 0.57) ng/L, (18.53 ± 2.22) ng/L; t = 21.48, 24.14, 23.21, all P<0.001]. The level of interleukin-2 in the observation group was significantly higher than that in the control group [(69.74 ± 7.67) pg/L vs. (56.87 ± 8.53) pg/L; t = -8.32, P<0.001]. There was no significant difference in the incidence of adverse reactions between the observation and control groups [7.27% (4/55) vs. 3.64% (2/55); χ2 = 0.18, P>0.05). Conclusions:The combination of probiotics and omeprazole can effectively improve the clinical manifestations of acute gastroenteritis in children, accelerate symptom recovery, enhance immune function, and demonstrate good safety.