ObjectiveTo investigate the mechanism by which Anmeidan improves learning and memory in insomnia rats by regulating the cyclic guanosine monophosphate-adenosine monophosphate synthase （cGAS）/stimulator of interferon genes （STING） signaling pathway to influence immunoinflammation. MethodsSixty SD rats were randomly divided into a blank group， a model group， a suvorexant group （30 mg·kg^-1）， and Anmeidan low-， medium-， and high-dose groups （4.55， 9.09， and 18.18 g·kg^-1）， with 10 rats in each group. The insomnia rat model was induced by intraperitoneal injection of p-chlorophenylalanine （PCPA）. Anmeidan decoction and normal saline were administered by gavage for 28 days at the corresponding doses. Morris water maze and new object recognition tests were used to assess learning and memory functions. Hematoxylin-eosin （HE） staining and Nissl staining were performed to observe hippocampal cell morphology. Enzyme-linked immunosorbent assay （ELISA） was used to measure the serum levels of interleukin-1 （IL-1）， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， interleukin-12 （IL-12）， interleukin-18 （IL-18）， and tumor necrosis factor-α （TNF-α）. Western blot and Real-time quantitative polymerase chain reaction（Real-time PCR） were used to detect the relative protein and mRNA expression levels of hippocampal cGAS and STING. ResultsCompared with the blank group， the 5-HT content in the model group was significantly reduced （P<0.01）. The latency to the upper platform and total distance were significantly increased （P<0.05， P<0.01）， while the residence time in the target quadrant and the number of platform crossings were significantly reduced （P<0.01）， and the relative recognition index for new objects was significantly lower （P<0.01）. The morphology and arrangement of hippocampal neurons were loose and disordered， with a decreased number of intracellular Nissl bodies. The relative expression levels of IL-1， IL-1β， IL-6， IL-8， IL-12， IL-18， TNF-α， cGAS， and STING pathway proteins and mRNA were significantly upregulated （P<0.01）. Compared with the model group， the latency to the upper platform in the high-dose Anmeidan group was significantly shortened （P<0.05）. In the medium- and high-dose Anmeidan groups and the suvorexant group， the residence time in the target quadrant and the number of platform crossings were significantly increased （P<0.01）. The total distance traveled was significantly reduced （P<0.01）， and the relative recognition index for new objects was significantly increased （P<0.01）. The hippocampal neurons were more neatly arranged， and the number of intracellular Nissl bodies increased. The expression of IL-1， IL-1β， IL-6， IL-8， IL-12， IL-18， TNF-α， and cGAS proteins and mRNA in the medium- and high-dose Anmeidan groups was significantly downregulated （P<0.05， P<0.01）. ConclusionAnmeidan improves learning and memory in insomnia rats， possibly by suppressing immunoinflammation through inhibition of the cGAS/STING signaling pathway.

ObjectiveTo elucidate the potential mechanisms by which the classic prescription Anmeidan alleviates cognitive impairment in insomnia model rats through metabolic profiling. MethodsA total of 60 SD rats were randomly divided into six groups： blank group， model group， low-， medium-， and high-dose Anmeidan groups， and the Suvorexant group， with 10 rats in each group. Except for the blank group， the insomnia model was established in all other groups via intraperitoneal injection of para-chlorophenylalanine. The Suvorexant group was administered Suvorexant solution （30 mg·kg^-1·d^-1） by gavage， while the low-， medium-， and high-dose Anmeidan groups received Anmeidan decoction （4.55， 9.09， 18.18 g·kg^-1·d^-1） by gavage. The blank group received an equivalent volume of normal saline. The open field test was used to assess spatial exploration and anxiety/depressive-like behaviors in rats. Serum levels of epidermal growth factor （EGF）， brain-derived neurotrophic factor （BDNF）， and vasoactive intestinal peptide （VIP） were measured using enzyme-linked immunosorbent assay （ELISA）. Untargeted metabolomics was employed to identify differential metabolites in rat serum， and systematic biological methods were applied to analyze the potential targets and pathways of Anmeidan. ResultsCompared to the blank group， the model group exhibited significant reductions in total distance traveled， average speed， number of entries into the central area， time spent in the central area， and frequency of upright events （P<0.01）， along with significant decreases in VIP， EGF， and BDNF levels （P<0.05，P<0.01）. A total of 100 differential metabolites were identified between the model and blank groups. Compared to the model group， the low-， medium-， and high-dose Anmeidan groups showed significant increases in total distance traveled， average speed， number of entries into the central area， time spent in the central area， and frequency of upright events （P<0.05，P<0.01）， as well as a significant increase in VIP levels （P<0.05，P<0.01）. Anmeidan significantly reversed abnormal changes in 67 metabolites compared to the model group. A combined analysis identified 134 potential targets of Anmeidan， with network topology analysis suggesting that Caspase-3， B-cell lymphoma 2 （Bcl-2）， nuclear transcription factor-κB （NF-κB）， interleukin-1β （IL-1β）， interleukin-2 （IL-2）， matrix metalloproteinase-9 （MMP-9）， and Toll-like receptor 4 （TLR4）， among others， may serve as key targets of Anmeidan. Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis revealed major enriched pathways， including the cyclic adenosine monophosphate （cAMP） signaling pathway， hypoxia inducible factor-1 （HIF-1） signaling pathway， and IL-17 signaling pathway. ConclusionThis study demonstrates that Anmeidan can recalibrate abnormal metabolic profiles in insomnia model rats to mitigate cognitive impairment， with its mechanisms of action potentially involving the regulation of immune-inflammatory responses， energy metabolism， and apoptosis-related pathways.

Adipose tissue is a critical energy reservoir in animals and humans, with multifaceted roles in endocrine regulation, immune response, and providing mechanical protection. Based on anatomical location and functional characteristics, adipose tissue can be categorized into distinct types, including white adipose tissue (WAT), brown adipose tissue (BAT), beige adipose tissue, and pink adipose tissue. Traditionally, adipose tissue research has centered on its morphological and functional properties as a whole. However, with the advent of single-cell transcriptomics, a new level of complexity in adipose tissue has been unveiled, showing that even under identical conditions, cells of the same type may exhibit significant variation in morphology, structure, function, and gene expression——phenomena collectively referred to as cellular heterogeneity. Single-cell transcriptomics, including techniques like single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA sequencing (snRNA-seq), enables in-depth analysis of the diversity and heterogeneity of adipocytes at the single-cell level. This high-resolution approach has not only deepened our understanding of adipocyte functionality but also facilitated the discovery of previously unidentified cell types and gene expression patterns that may play key roles in adipose tissue function. This review delves into the latest advances in the application of single-cell transcriptomics in elucidating the heterogeneity and diversity within adipose tissue, highlighting how these findings have redefined the understanding of cell subpopulations within different adipose depots. Moreover, the review explores how single-cell transcriptomic technologies have enabled the study of cellular communication pathways and differentiation trajectories among adipose cell subgroups. By mapping these interactions and differentiation processes, researchers gain insights into how distinct cellular subpopulations coordinate within adipose tissues, which is crucial for maintaining tissue homeostasis and function. Understanding these mechanisms is essential, as dysregulation in adipose cell interactions and differentiation underlies a range of metabolic disorders, including obesity and diabetes mellitus type 2. Furthermore, single-cell transcriptomics holds promising implications for identifying therapeutic targets; by pinpointing specific cell types and gene pathways involved in adipose tissue dysfunction, these technologies pave the way for developing targeted interventions aimed at modulating specific adipose subpopulations. In summary, this review provides a comprehensive analysis of the role of single-cell transcriptomic technologies in uncovering the heterogeneity and functional diversity of adipose tissues.

Aim To investigate the targeting mechanism of miR-23b on PINKl/Parkin pathway in transdifferentiation of NRK-52E cellsinduced by TGF-β1, and to elucidate the intervention mechanism of Qingshen granules drug-containing serum on NRK-52E cell transdifferentiation. Methods Ultra-high performance liquid chromatography ( UPLC ) fingerprinting method was used to analyze Qingshen granules. The NRK-52E transdifferentiation model induced by TGF-β1 was constructed. The NRK-52E cells were divided into simulated no-load control group, miR-23b-5p simulated group, inhibitor no-load control group, and miR-23b-5p inhibitor group, after transfection with siRNA, and the effect of miR-23b-5p on PINK1 expression was ob-served. The NRK-52E cells were then divided into normal group, TGF-(31 group, Qingshen granule group, miR-23 b-mimic group, miR-23 b-mimic group, and miR-23b-mimic + Qingshen granule group. Western blot was used to detect the expression of Pinkl, Parkin, LC3 n, Beclin-1, P62 and a-SMA proteins, and RT- PCR was used to detect the expression of miR-23 b-5p, Pinkl, Parkin, Beclin-1 and a-SMA mRNA in NRK- 52E cells. Dual-Luciferase Reporter gene experiment was used to detect the targeting relationship between miR-23b-5p and PINKL Results UPLC fingerprinting method found 11 active components in Qingshen granules. After overexpression of miR-23b-5p, the expression of PINkl mRNA significantly increased (P < 0. 05). And after silencing of miR-23 b-5 p expression, the expression of PINkl mRNA also significantly decreased (P < 0. 05 ). Dual-Luciferase Reporter Assay showed that Rno-miR-23b-5p could significantly down- regulate the luciferase activity of Rno-PINKl-WT (P < 0. 05 ), but could not down-regulate the luciferase activity of mutant Rno-PINKl -mut ( P > 0. 05 ). The experimental results showed that the expressions of miR- 23b-5p, Pinkl, Parkin, Beclin-1, LC3 II and LC3 II/ I ratio in TGF-β1 group were significantly lower than those in normal group, but the expressions of P62 and a-SMA were significantly higher than those in normal group ( P <0.05). The expressions of miR-23 b-5 p, Pinkl, Parkin, Beclin-1, LC3 II and LC3 11/ I ratio in Qingshen granule group and miR-23 b-mimic group were significantly higher than those in TGF-β1 group, and the expressions of P62 and a-SMA were significantly lower than those in TGF-β1 group (P < 0. 05 ). The performance of miR-23 b-mimic + Qingshen granule group was better than that of miR-23 b-mimic group (P < 0. 05 ). Conclusions Qingshen granules can up- regulate the expression of miR-23b-5p in NRK-52E cellsand inhibit the transdifferentiation process of NRK- 52E cells by enhancing the mitochondrial autophagy activity mediated by PINKl/Parkin pathway.

Abstract: To explore the mechanism of the intestinal microecology regulation by polysaccharide prebiotics, ELISA, histopathologic analysis, immunohistochemical analysis, 16S rRNA high-throughput sequencing, and gas chromatography-mass spectrometry were applied to investigate the effects of fermented polysaccharides on changes in the intestinal microbiota and short-chain fatty acids (SCFAs) in mice with dextran sulfate sodium (DSS)-induced colitis model and their relationship with the level of intestinal inflammation and barrier protein expression. It was found that fermented Lycium barbarum polysaccharides (FLBP) significantly reduced intestinal inflammation level, improved colonic tissue structure, up-regulated the expression of tight junction proteins Claudin-1 and ZO-1, and significantly increased the content of intestinal SCFAs in mice. Gut bacteria analyses showed that FLBP enriched intestinal Dubosiella and Akkermansia in mice and decreased the abundance of Turicibacter, Faecalibaculum, and Escherichia-Shigella. Results showed that remodeled Dubosiella activated by FLBP played a dominant role in ameliorating colitis by significantly increasing SCFAs content, improving intestinal barrier and reducing intestinal inflammation. The study aimed to provide a safer and better option for the amelioration of colitis and to provide a theoretical basis for the development of functional foods with FLBP.

Objective To explore the distribution of traditional Chinese medicine(TCM)syndrome elements in patients with multi-drug resistant bacteria-infected pneumonia.Methods Clinical data of 126 patients with multi-drug resistant bacteria-infected pneumonia admitted to the intensive care unit of Lung Disease Centre of Qingdao Hospital of Traditional Chinese Medicine from May 2020 to July 2022 were retrospectively collected.The clinical data included the patients'gender,age,underlying diseases,history of bad additions of smoking and alcohol,multi-drug resistant bacteria,and the information of four diagnostic methods of TCM,etc.The disease-nature syndrome elements in patients with drug-resistance to various strains of drug-resistant bacteria were extracted,and then deficiency-excess syndrome differentiation was carried out.Results(1)A total of 201 strains of multi-drug resistant bacteria were detected in 126 patients with multi-drug resistant bacterial pneumonia.The main pathogenic species were Gram-negative bacteria,and the proportion accounted for 95.52%(192/201),which was significantly higher than that of Gram-positive bacteria[4.48%(9/201)],with a statistically significant difference(χ2 = 166.612,P＜0.001).Klebsiella pneumoniae accounted for the highest percentage of 23.38%in the gram-negative bacterium.(2)A total of 12 syndrome elements were extracted from the 126 patients.The excess syndrome elements were predominated by phlegm and heat,and the deficiency syndrome elements were predominated by yin deficiency.There was no statistically significant difference in the distribution of yin deficiency,blood deficiency,heat,phlegm,fluid-retention and damp syndrome elements among patients with different strains of drug-resistant bacterial infection(P＞0.05).(3)Of the 126 patients,62 cases(49.21%)had simple excess syndrome,one case(0.79%)had simple deficiency syndrome,and 63 cases(50.00%)had concurrent deficiency-excess syndrome.Among the 126 patients,there were 19 cases of single syndrome element,41 cases of concurrent two-syndrome element,49 cases of concurrent three-syndrome element,16 cases of concurrent four-syndrome element,and one case of concurrent five-syndrome element.And the combined syndrome element of phlegm-heat-yin deficiency occurred most frequently for 26 times.Conclusion Gram-negative bacteria are the primary infectious pathogens for the patients with multi-drug resistant bacterial infections,and the TCM syndrome elements of the patients are characterized by the concurrence of deficiency and excess and simple excess syndrome,mainly manifesting as phlegm,heat,and yin deficiency.

Objective To observe the clinical efficacy of Xingnao Kaiqiao Acupuncture(with the functions of awakening the brain and opening the orifices)combined with acupuncture at pericardium meridian points in the treatment of post-stroke sleep reversal(PSSR).Methods Sixty patients with PSSR were randomly divided into observation group and control group,30 patients in each group.Both groups were given conventional treatment,the control group was given oral use of Alprazolam,and the observation group was given the combination of acupuncture a at pericardial meridian points,and 10 days of treatment was one course of treatment.After 10 days of treatment,the clinical efficacy of the two groups was evaluated.The changes in the Pittsburgh Sleep Quality Index(PSQI)and Ascens Insomnia Scale(AIS)scores,as well as the Hamilton Depression Scale(HAMD)scores were observed before and after treatment in the two groups.The changes in cortisol levels at 0,8,and 16 o'clock were compared before and after treatment between the two groups.Results(1)After treatment,the PSQI scores of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving PSQI scores,and the difference was statistically significant(P＜0.05).(2)After treatment,the AIS and HAMD scores of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the AIS and HAMD scores,and the difference was statistically significant(P＜0.05).(3)After treatment,the cortisol level of patients in the two groups at 0,8,and 16 o'clock was significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the cortisol level at 0,8,and 16 o'clock was significantly superior to the control group,and the difference was statistically significant(P＜0.05).(4)The total effective rate was 86.67%(26/30)in the observation group and 80.00%(24/30)in the control group.The efficacy of the observation group was slightly superior to that of the control group,but the difference was not statistically significant(P＞0.05).Conclusion Xingnao Kaiqiao Acupuncture combined with acupuncture at pericardium meridian points for the treatment of PSSR can significantly improve the clinical symptoms of the patients,so as to improve the quality of life of the patients,and the therapeutic efficacy is remarkable.

Objective To explore the role and possible molecular mechanism of Isocitrate dehydrogenase 1(IDH1)gene in proliferation and migration of intrahepatic cholangiocarcinoma(iCCA)cell HuCCT1.Methods HuCCT1 cells with IDH1 gene knockout(HuCCT1IDH1-/-)were constructed by CRISPR/Cas9 gene editing technology.To investigate the capacities of proliferation,migration and invasion of HuCCT1WT(HuCCT1 cells with wild-type IDH1 gene)and HuCCT1IDH1-/-cells,assays of CCK-8,clone formation,scratch and transwell were performed.Western blotting was used to detect the expression levels of epithelial-mesenchymal transition(EMT)associated proteins E-cadherin,N-cadherin,Vimentin,MMP-9,Wnt3a and β-catenin in two groups of cells.The transcriptome sequencing data of HuCCT1WT and HuCCT1IDH1-/-cells were analyzed by bioinformatics methods,Western blotting was used to verify the expression of signaling pathway-related proteins.Results Compared with HuCCT1WT cells,HuCCT1IDH1-/-cells showed the number of proliferation and clone formation significantly reduced(P<0.05),the proportion of cells blocked in G2/M phase was significantly increased(P<0.01),the rate of scratch healing was significantly decreased(P<0.01),and the number of migrated cells(P<0.001)and invaded cells(P<0.05)was significantly reduced.qRT-PCR assay showed that the expression levels of IDH1,Vimentin,MMP-9 and genes related to the regulation of G2/M cycle proliferation,Cyclin A2,Cyclin B1 and CDK1 mRNA were down-regulated in HuCCT1IDH1-/-cells(P<0.05),and the expression of CDH1 mRNA encoding E-cadherin was up-regulated(P<0.01);Western blotting assay showed that the expression level of E-cadherin in HuCCT1IDH1-/-cells was significantly increased(P<0.05),and the expression level of N-cadherin,Vimentin and MMP-9 protein was significantly decreased(P<0.05)than that in HuCCT1WT cells.Data of transcriptome sequencing revealed 1476 differentially expressed genes(DEGs)between two groups of HuCCT1 cells.Go enrichment analysis showed the DEGs were significantly enriched in cell biological processes associated with inflammatory response,cell signaling and cell metabolism.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis suggested that the DEGs may be involved in some signaling pathways such as Wnt,MAPK,Rap1,Hippo and TNF,which are closely related to the regulation of proliferation and invasion of tumor cells.Western blotting verification results showed that compared with HuCCT1WT cells,the relative expression of Wnt3a and β-catenin proteins of HuCCT1IDH1-/-cells was significantly decreased(P<0.05).Conclusions IDH1 gene may participate in the control of biological functions of HuCCT1 cells,including cell proliferation,migration,invasion and epithelial mesenchymal transition.The mechanism may be related to the activation of the Wnt/β-catenin signaling pathway.

Objective To assess the risk factors for carbapenem-resistant Acinetobacter baumannii(CRAB)bloodstream infection(BSI)and 28-day short-term mortality in elderly patients,and provide reference for the pre-vention and treatment of CRAB BSI.Methods Clinical data of patients aged ≥60 years and diagnosed with AB BSI in a hospital in Yulin City from January 2013 to December 2022 were retrospectively analyzed,including demogra-phic and microbiological characteristics,as well as clinical outcomes of the patients.Variables which were significant in univariate analysis were selected for multivariate analysis using binary logistic regression model and Cox propor-tional hazards model.Independent risk factors for infection were further determined,and survival analysis was per-formed using Kaplan-Meier curve.Results A total of 150 patients were included in the study,out of which 16 pa-tients(10.7％)had CRAB BSI and 134 had carbapenem-sensitive AB(CSAB)BSI.The 28-day short-term mortali-ty of AB BSI in elderly patients was 15.3％(23/150,95％CI:9.6％-21.1％),and the short-term mortality of CRAB BSI was higher than that of CSAB([56.3％,9/16]vs[10.4％,14/134]).Deep venous catheterization(OR:15.598,95％CI:1.831-132.910)and combined infections of other sites(OR:15.449,95％CI:1.497-159.489)were related to CRAB BSI in elderly patients.The independent risk factors for 28-day mortality in elderly patients with AB BSI were hemodialysis(OR:11.856,95％CI:2.924-48.076),intensive care unit admission(OR:9.387,95％CI:1.941-45.385),and pulmonary infection being suspected source of bacteremia(OR:7.019,95％CI:1.345-36.635).Conclusion The occurrence of CRAB BSI in elderly patients is related to the combined infection of other sites and deep vein catheterization.Hemodialysis,admission to ICU,and pulmonary infection being suspected source of bacteremia are independent risk factors for the prognosis of AB BSI in elderly patients.

Objective To explore specific magnetic resonance imaging(MRI)features of somatic symptoms in depression by comparing the differences of brain gray matter volume in depression patients with and without somatic symptoms using voxel-based morphometry(VBM).Methods A total of 52 depression patients were recruited and divided into somatic and no somatic symptoms group according to the patient health questionnaire-15 score(＞9 and≤9,respectively).Forty gender-age-matched healthy volunteers were recruited as the control group.All subjects underwent MRI scanning.Imaging data were analyzed to explore the differences in brain gray matter between groups using VBM.Results Compared with control group,gray matter volume increased in the right superior temporal gyrus,and decreased in the right inferior orbitofrontal gyrus,right inferior temporal gyrus,left inferior orbitofrontal gyrus,and left superior temporal gyrus for depressive patients with somatic symptoms(P＜0.001);gray matter volume increased in the right middle temporal gyrus,and decreased in the right superior temporal gyrus and right inferior temporal gyrus for depressive patients without somatic symptoms(P＜0.001).Only the volume in the right tongue and left cingulate gyrus increased in depressive patients with somatic symptoms compared with that in patients without somatic symptoms(P＜0.01).Conclusion VBM-MRI has demonstrated increased volume in the tongue and cingulate gyrus in the somatic symptoms of depression.