OBJECTIVE To comprehensively evaluate the quality of Sanzi powder from different batches based on 12 components quantitative analysis combined with chemical pattern recognition and entropy weight-TOPSIS method. METHODS The contents of 12 components in 15 batches of Sanzi powder （No. S1-S15） were determined by HPLC-MS/MS， such as ethyl gallate， gallic acid， ferulic acid， corilagin， genipin-1-O-β-D-gentiobioside， toosendanin， geniposide， caffeic acid， methyl deacetylated coumarinate， tannic acid， rutin， quercetin. Cluster analysis （CA）， principal component analysis （PCA）， and orthogonal partial least squares-discriminant analysis （OPLS-DA） were conducted on the assay results. Using variable importance projection （VIP） value＞1 and P＜0.05 as the evaluation criteria， the quality differential markers in Sanzi powder were screened. The entropy weight method was used to calculate the weight value， and TOPSIS method was used to rank the quality of 15 batches of Sanzi powder from superior to inferior. RESULTS The contents of the 12 components were 13.494-24.292， 2 069.608-3 188.100， 1.410-3.616， 1 065.030-2 630.584， 1 404.704-1 838.078， 101.640-354.268， 9 193.720-14 777.854， 1.240-5.060， 148.028-5 541.990， 4 261.422-5 607.438， 107.560- 195.512， 2.226-4.192 μg/g， respectively. The results of CA， PCA and OPLS-DA indicated that 15 batches of Sanzi powder could be clustered into two groups. Specifically， batches S3， S7， S10 and S15 were grouped into one category， and remaining batches were grouped into one category. VIP values of geniposide， quercetin， caffeic acid， and methyl deacetylated coumarinate were all greater than 1， with corresponding P-values less than 0.05. The results of the entropy weight-TOPSIS analysis revealed that methyl deacetylate exhibited the smallest information entropy and the highest weight. The relative closeness degrees of samples S3， S7， S10 and S15 ranged from 0.789 to 0.973， while the remaining samples ranged from 0.054 to 0.172. CONCLUSIONS The contents of 12 components in Sanzi powder could be determined accurately by using HPLC-MS/MS technology. Methyl deacetylated coumarinate， geniposide， quercetin and caffeic acid were identified as the quality differential markers. It was found that the overall quality of samples S3， S7， S10 and S15 were superior to that of other batches. Notably， the quality of Gardeniae Fructus decoction pieces emerges as a critical factor in ensuring the consistency of the preparation’s quality.

Objective: To investigate the status of overweight and obesity among primary and middle school students in Yuyao City, Zhejiang Province, so as to provide the basis for formulating weight management strategies for students. Methods: A census was conducted to investigate 72 primary schools and 23 middle schools students in Yuyao City in 2024, and data such as gender and age were collected through questionnaire surveys. Height and weight were measured to calculate the body mass index (BMI). Based on the BMI thresholds for different gender and age groups, overweight and obesity were determined. Results: Totally 75 082 individuals were surveyed, including 40 435 boys and 34 647 girls. Among them, 55 172 were primary school students and 19 910 were middle school students. A total of 8 677 overweight and 7 042 obese individuals were identified, with detection rates of 11.56% and 9.38%, respectively. The detection rates of overweight and obesity in boys were higher than in girls (14.01% vs. 8.69%, 11.58% vs. 6.81%, both P<0.05). The detection rate of obesity among students in rural schools was higher than students in urban schools (10.12% vs. 8.18%, P<0.05). The detection rates of overweight and obesity of students with non-local household registration were higher than students with local household registration (12.04% vs. 10.77%, 10.44% vs. 7.64%, both P<0.05). The detection rate of overweight showed an upward trend with age (P<0.05), while no significant age-related trend was observed for obesity (P>0.05). Conclusions The detection rates of overweight and obesity were notably high among primary and middle school students in Yuyao City. Boys, students in rural schools, students with non-local household registration, and older students are the key population.

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*

Obesity and metabolic dysfunction-associated steatotic liver disease (MASLD) are linked to numerous chronic conditions, including cardiovascular disease, atherosclerosis, chronic kidney disease, and type II diabetes. Previous research identified the natural flavonoid diosmin, derived from Chrysanthemum morifolium, as a regulator of glucose metabolism. However, its effects on lipid metabolism and underlying mechanisms remained unexplored. The AMP-activated protein kinase (AMPK) pathway serves a critical function in glucose and lipid metabolism. The relationship between diosmin and the AMPK pathway has not been previously documented. This investigation examined diosmin's capacity to reduce lipid content through AMPK pathway activation in hepatoblastoma cell line G2 (HepG2) and 3T3-L1 cells. The study revealed that diosmin inhibits lipogenesis, indicating its potential as an anti-obesity agent in obese mice. Moreover, diosmin demonstrated effective MASLD alleviation in vivo. These findings suggest that diosmin may represent a promising therapeutic candidate for treating obesity and MASLD.
Diosmin/administration & dosage* ; Animals ; AMP-Activated Protein Kinases/genetics* ; Humans ; Non-alcoholic Fatty Liver Disease/enzymology* ; Mice ; Obesity/enzymology* ; Hep G2 Cells ; Male ; 3T3-L1 Cells ; Mice, Inbred C57BL ; Signal Transduction/drug effects* ; Lipid Metabolism/drug effects* ; Chrysanthemum/chemistry* ; Lipogenesis/drug effects*

OBJECTIVE To explore the potential of Aconiti Kusnezoffii Radix in affecting diabetes mellitus type 2(T2DM) and the potential mechanism for T2DM related symptoms based on systematic pharmacology, bioinformatics, molecular docking and in vitro and in vivo experiments. METHODS The database was used to search the related chemical components of Aconiti Kusnezoffii Radix, predict the potential targets and intervene related diseases. The differential genes of T2DM relative healthy people were retrieved from GEO database, mapped with the action target of Aconiti Kusnezoffii Radix, and placed in DAVID database for biological function enrichment. The sensitivity of the target gene to T2DM was analyzed by one-way ANOVA, binary logistic regression analysis and ROC curve. The binding position and interaction force between chemical compounds of Aconiti Kusnezoffii Radix and target proteins were analyzed by molecular docking technology. The effect of Aconiti Kusnezoffii Radix and its chemical compounds on the expression of target protein was verified by T2DM model in vivo and in vitro. RESULTS Through database retrieval and analysis, 304 kinds of target related diseases(P value<0.05, FDR<0.05) were obtained, and T2DM with the highest degree value(Degree=59) was selected and analyzed. The 43 target genes were obtained from the intersection of differential genes in T2DM relatively healthy people and potential action targets of Aconiti Kusnezoffii Radix. A total of 9 genes with significant differences were obtained by one-way ANOVA, 5 meaningful genes were obtained by binary logistic regression analysis, and 3 genes with area under ROC curve AUC>0.5 were obtained. By molecular docking (+)-Isoboldine binds to proteins APEX1, CASP1 and CBFB, Napelline binds to proteins CBX1 and EHMT2 through different forces such as hydrogen bond interaction, ligand interaction, hydrophobic interaction, ionizability and electrostatic interaction, so as to increase the ability of ligands to target proteins. After 2 weeks of treatment with Aconiti Kusnezoffii Radix aqueous extract, Aconiti Kusnezoffii Radix may alleviated the symptoms of T2DM by improving peripheral neuropathy. Moreover, Aconiti Kusnezoffii Radix could affect the protein expression of APEX1, CASP1, CBFB, CBX1 and EHMT2 in rat liver tissue. The effect of (+)-Isoboldine and Napelline chemical compounds in Aconiti Kusnezoffii Radix on the target protein of the model in vitro was consistent with that in vivo. CONCLUSION It is preliminarily revealed that Aconiti Kusnezoffii Radix can be used as a potential therapeutic drug to improve T2DM peripheral neuropathy, which lays a theoretical foundation for the research and development of Chinese Mongolian medicine and the excavation of new target drugs for the treatment of T2DM.

Objectives:To analyze the consistency of evaluating left ventricular hemodynamics (HDF) based on single plane and multi plane cine sequences of magnetic resonance mitral valve orifice.Methods:A prospective study was conducted on 48 healthy adults, and two methods were used to measure the mitral valve diameter and calculate HDF parameters. The first method was to measure the diameter of the mitral valve opening in the left ventricular three chamber cine sequence; The second method is to measure the mitral valve diameter using cine sequences of two chamber, three chamber, and four chamber hearts, and then take the average value. Paired t-tests were used to compare the differences in HDF measured by two methods, and Pearson correlation coefficient ( r), intra group correlation coefficient ( ICC), and Bland-Altman analysis were used to test the consistency and reproducibility of the two methods. Results:The root mean square (RMS) of longitudinal HDF calculated using single plane and multi plane mitral valve diameters were [(17.28±4.41)% vs (17.21±4.61)%] ( P=0.379) for the entire cardiac cycle, [(21.45±5.54)% vs (21.49±5.68)%] ( P=0.646) for systolic phase, and [(12.78±4.10)% vs (12.54±4.24)%] ( P=0.106) for diastolic phase, respectively. The difference in the calculation results of HDF parameters related to ventricular function was not statistically significant (all P>0.05), and there was good consistency ( r=0.924-0.996, ICC=0.924-0.995). The two HDF parameters related to atrial function were sensitive to the measurement method of mitral valve orifice diameter [RMS of longitudinal HDF during active atrial emptying: (3.26±1.51)% vs (3.32±1.55)%, P=0.006; longitudinal HDF pulse during active atrial emptying: (-2.60±1.28)% vs (-2.76±1.30)%, P<0.001]. Conclusions:The ventricular function related HDF parameters obtained from the analysis of mitral valve orifice diameter using single plane and multi plane methods have good consistency, and can be evaluated using relatively simple single plane methods for left ventricular HDF.

Objective To investigate the effect of esketamine on depression and improvement of postoperative cognitive function in patients with major depressive disorder treated by electroconvulsive therapy(ECT).Methods A total of 115 patients with major depressive disorder,37 males and 78 fe-males,aged≥12 years,BMI 18-30 kg/m2,ASA physical status Ⅰ or Ⅱ were selected to receive ECT.The patients were randomly divided into two groups using a random number table:the esketamine group(group E,n=56)and the control group(group C,n=59).Group E received esketamine,propofol,and succincholine during anesthesia induction,and group C received propofol and succincholine during an-esthesia induction.ECT indicators(including EEG twitch time,twitch time,stimulus intensity,and twitch energy index)were recorded during each ECT session.Montreal cognitive assessment(MoCA)and Mini-mental state examination(MMSE)were used to evaluate cognitive function at baseline and after ECT treat-ment.Hamilton depression(HAMD)scale was used to score depression.The occurrence of headache,nau-sea,vomiting,dizziness,delirium,and other adverse reactions were recorded.Results Compared with group C,the EEG convulsion time in group E was significantly prolonged(P＜0.05),the incidence of Mo-CA score decreased by≥1 point and MMSE score decreased by≥1 point in group E after the end of ECT treatment was significantly reduced(P＜0.05),the incidence of nausea,vomiting,and dizziness were sig-nificantly decreased(P＜0.05).Conclusion Using esketamine in ECT for the treatment of major depres-sive disorder can improve cognitive function after ECT.

The study was designed to investigate the peripheral anti-inflammatory effect of nicotinamide riboside(NR)in experimental autoimmune encephalomyelitis(EAE)mice and its mechanisms.Female C57BL/6 mice were induced by MOG35-55 to prepare EAE model,which then randomly divided into EAE model group and NR treatment group.Mice in EAE model group were given normal saline at a dose of 200 μl/d and mice in NR treatment group were given NR at a dose of 500 mg/kg(200 μl/d)by intragastric administration.Clinical score and body weight of mice in each group were observed and recorded.After mice were sacrificed on the 28th day after immunization,frozen sections of spleen and spinal cord were prepared and proteins of spinal cord were extracted.HE staining was used to detect peripheral inflammatory cells infiltrating spinal cord;immunofluorescence staining was used to detect the number of CD4+T cells and CD68+macrophages in spinal cord of mice;Western blot was used to detect the expression of IFN-γ and IL-1β in spinal cord of mice;immunofluorescence staining was used to detect the number of ROCK-Ⅰ+cells,TLR4+cells,p-NF-κB+cells,TNF-α+cells,IL-1β+cells,IFN-γ+cells,IL-6+cells,IL-10+cells,IL-17+cells,iNOS+cells and Arg-1+cells in spleen of mice.Data showed that compared with EAE model group,NR significantly delayed the onset time of EAE mice(P<0.05),decreased clinical score(P<0.05 or P<0.01),alleviated weight loss,prevented peripheral inflammatory cells from infiltrating spinal cord,decreased the number of CD4+T cells and CD68+macrophages in spinal cord(P<0.01),down-regulated the expression of IFN-γ and IL-1β of spinal cord(P<0.05),inhibited the expression of ROCK-Ⅰ,TLR4 and p-NF-κB in spleen of mice(P<0.01),reduced the secretion of IFN-γ,iNOS,IL-6 and other pro-inflammatory factors in spleen(P<0.05 or P<0.01),and increased the secretion of anti-inflammatory factors Arg-1 and IL-10 in spleen(P<0.05 or P<0.01).In conclusion,NR can effectively alleviate the clinical symptoms of EAE mice and significantly reduce inflammatory response of peripheral and central nervous system,and its mechanism may be related to the inhibition of Rho/ROCK signaling pathway and TLR4/NF-κB signaling pathway in spleen of EAE mice.

Objective:To standardize the origin of Rubi Fructus by using ITS2 and matK molecular markers to identify Rubi Fructus and its analogues. Methods:The ITS and matK sequences of Rubus chingii Hu, Rubus corchorifolius L.f., Rubus hirsutus Thunb., Rubus parvifolius L., Rubus buergeri Miq. and Rubus lambertianus Ser. were amplified and sequenced. The hidden Markov model was used to remove 5.8S and 28S from ITS sequences. A total of 25 ITS2 sequences were obtained. And a total of 22 matK sequences were obtained by proofreading by Glustal software. MEGA software was used for matK and ITS2 sequences analysis, intraspecific and interspecific genetic distances caculation, and neighbor joining (NJ) phylogenetic tree construction. ITS2 secondary structure was predicted by ITS2 database and aligned by 4Sale software. Profile neighbor-joining (PNJ) phylogenetic tree was constructed based on combined ITS2 sequence and its secondary structure by ProfDistS software. Results:An obvious barcoding gap between Rubi Fructus and its analogues was showed. The topological relationship between NJ tree and PNJ tree was consistent, and each taxon exhibits monophyly. The ITS2 secondary structure of Rubi Fructus was significant different from its analogues.Conclusions:It is recommended that both ITS2 and matK markers can serve as DNA barcodes for identifying Rubi Fructus and its analogues. The addition of ITS2 secondary structure information can enrich the identification results and provide theoretical support for resource research and variety selection of Rubi Fructus.

Objective To investigate the role and mechanism of lysine acetyltransferase 8(KAT8)on the prolifera-tion of colorectal cancer cells.Methods The expression of KAT8 in cancerous tissues and adjacent tissues of color-ectal cancer patients was analyzed by RNA-seq data of TCGA database.Cell proliferation was detected by colony-forming unit assays and CCK8.The GEO database was used to analyze the differential genes of KAT8 knockdown cells and control cells and perform functional pathway enrichment analysis.In the colorectal cancer database hosted on cBioPortal,that conducted an analysis examining the correlation between KAT8 and genes in the regulation of N6-methyladenosine(m6A)modification.Western blot technique was employed to assess the protein expression lev-els of KAT8 and METTL3.Results Compared to human normal colorectal tissue,KAT8 was highly expressed in colorectal cancer(P＜0.05).Knockdown or selective inactivation of KAT8 inhibited colorectal cancer cells prolifera-tion(P＜0.05).In colorectal cancer cell lines,knocking down KAT8 reduced m6A modification levels(P＜0.05).Knocking down KAT8 inhibited METTL3 expression(P＜0.05).Over-expression of METTL3 reversed cell prolifera-tion which was inhibited by knockdown KAT8(P＜0.05).Conclusions KAT8 facilitates the proliferation of color-ectal cancer cell lines through regulation of METTL3-mediated m6A modifications.