BACKGROUND:Rheumatoid arthritis is an inflammatory disease.Many studies have shown that wogonin has a good anti-inflammatory effect on rheumatoid arthritis,but its exact efficacy and specific mechanism of action remain to be clarified. OBJECTIVE:To investigate the mechanism of wogonin ameliorating joint inflammation by regulating endoplasmic reticulum stress pathway in rats with collagen-induced arthritis. METHODS:(1)At the animal level:Female Wistar rats were divided into healthy control group,arthritis model group and wogonin treatment group.Rat models of arthritis in the latter two groups were established by subcutaneous injection of bovine type Ⅱ collagen and adjuvant.In the wogonin group,wogonin was given by gavage for 28 consecutive days after modeling.During this period,the rats in each group were weighed,and arthritis score and ankle swelling were measured every 7 days.After the experiment,the pathological changes of the joint were observed,the mRNA and protein levels of endoplasmic reticulum stress pathway GRP78 and CHOP were detected by qRT-PCR,western blot,and immunohistochemistry.(2)At the cellular level,cell counting kit-8 was used to detect the cytotoxic effect of wogonin on fibroblast-like synoviocytes from rats with collagen-induced arthritis.The fibroblast-like synoviocytes induced by thapsigargin were treated with different concentrations of wogonin.The levels of interleukin-1β and tumor necrosis factor-α in the cell supernatant were detected by ELISA,and the intracellular reactive oxygen species in each group were determined by DCFH-DA probe method.The mRNA and protein levels of GRP78,IRE1α,XBP1s and CHOP were detected by qRT-PCR and western blot,respectively. RESULTS AND CONCLUSION:Compared with the healthy control group,arthritis index score and ankle swelling degree in the arthritis model group were increased(P<0.01),synovial hyperplasia,inflammatory cell infiltration,cartilage destruction and bone erosion were observed in pathological sections,and the mRNA and protein expressions of GRP78 and CHOP in the ankle were significantly increased(P<0.01),which were mainly located in synovial tissue and articular surface.Compared with the arthritis model group,the arthritis index score and ankle swelling degree in the wogonin treatment group were decreased(P<0.05),synovial hyperplasia and the number of inflammatory cells were decreased,cartilage destruction and bone erosion were alleviated,the mRNA and protein expression levels of GRP78 and CHOP in the ankle were decreased(P<0.05),particularly in synovial tissue and on the articular surface.There was no significant difference in body mass among the three groups(P>0.05).In the cell experiment,200 μmol/L wogonin significantly reduced the survival rate of fibroblast-like synoviocytes(P<0.01).Compared with the blank control group,the levels of interleukin-1β,tumor necrosis factor-α,content of reactive oxygen species,and mRNA and protein expression of GRP78,IRE1α,XBP1s,and CHOP in the thapsigargin group were significantly increased(P<0.05);compared with the thapsigargin group,50 and 100 μmol/L wogonin significantly reduced the levels of interleukin-1β and tumor necrosis factor-α in the cell supernatant(P<0.05,P<0.01),and 100 μmol/L wogonin significantly reduced the content of reactive oxygen species(P<0.01)and down-regulated the mRNA and protein expression levels of GRP78,IRE1α,XBP1s and CHOP(all P<0.05).These results suggest that wogonin can effectively alleviate joint inflammatory responses in rats with collagen-induced arthritis,and the endoplasmic reticulum stress pathway may be the key target of its intervention.

BACKGROUND:In recent years,numerous studies have shown that autophagy and mitophagy play an important role in the regulation of bone metabolism.Under non-physiological conditions,mitophagy breaks the balance of bone metabolism and triggers metabolism disorders,which affect osteoblasts,osteoclasts,osteocytes,chondrocytes,bone marrow mesenchymal stem cells,etc. OBJECTIVE:To summarize the mechanism of mitophagy in regulating bone metabolic diseases and its application in clinical treatment. METHODS:PubMed,Web of Science,CNKI,WanFang and VIP databases were searched by computer using the keywords of"mitophagy,bone metabolism,osteoblasts,osteoclasts,osteocytes,chondrocytes,bone marrow mesenchymal stem cells"in English and Chinese.The search time was from 2008 to 2023.According to the inclusion criteria,90 articles were finally included for review and analysis. RESULTS AND CONCLUSION:Mitophagy promotes the generation of osteoblasts through SIRT1,PINK1/Parkin,FOXO3 and PI3K signaling pathways,while inhibiting osteoclast function through PINK1/Parkin and SIRT1 signaling pathways.Mitophagy leads to bone loss by increasing calcium phosphate particles and tissue protein kinase K in bone tissue.Mitophagy improves the function of chondrocytes through PINK1/Parkin,PI3K/AKT/mTOR and AMPK signaling pathways.Modulation of mitophagy shows great potential in the treatment of bone diseases,but there are still some issues to be further explored,such as different stages of drug-activated mitophagy,and the regulatory mechanisms of different signaling pathways.

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.

ObjectiveIn the realm of forensic science, dust is a valuable type of trace evidence with immense potential for intricate investigations. With the development of DNA sequencing technologies, there is a heightened interest among researchers in unraveling the complex tapestry of microbial communities found within dust samples. Furthermore, striking disparities in the microbial community composition have been noted among dust samples from diverse geographical regions, heralding new possibilities for geographical inference based on microbial DNA analysis. The pivotal role of microbial community data from dust in geographical inference is significant, underscoring its critical importance within the field of forensic science. This study aims to delve deeply into the nuances of fungal community composition across the urban landscapes of Beijing, Fuzhou, Kunming, and Urumqi in China. It evaluates the accuracy of biogeographic inference facilitated by the internal transcribed spacer 2 (ITS2) fungal sequencing while concurrently laying a robust foundation for the operational integration of environmental DNA into geographical inference mechanisms. MethodsITS2 region of the fungal genomes was amplified using universal primers known as 5.8S-Fun/ITS4-Fun, and the resulting DNA fragments were sequenced on the Illumina MiSeq FGx platform. Non-metric multidimensional scaling analysis (NMDS) was employed to visually represent the differences between samples, while analysis of similarities (ANOSIM) and permutational multivariate analysis of variance (PERMANOVA) were utilized to statistically evaluate the dissimilarities in community composition across samples. Furthermore, using Linear Discriminant Analysis Effect Size (LEfSe) analysis to identify and filter out species that exhibit significant differences between various cities. In addition, we leveraged SourceTracker to predict the geographic origins of the dust samples. ResultsAmong the four cities of Beijing, Fuzhou, Kunming and Urumqi, Beijing has the highest species richness. The results of species annotation showed that there were significant differences in the species composition and relative abundance of fungal communities in the four cities. NMDS analysis revealed distinct clustering patterns of samples based on their biogeographic origins in multidimensional space. Samples from the same city exhibited clear clustering, while samples from different cities showed separation along the first axis. The results from ANOSIM and PERMANOVA confirmed the significant differences in fungal community composition between the four cities, with the most pronounced distinctions observed between Fuzhou and Urumqi. Notably, the biogeographic origins of all known dust samples were successfully predicted. ConclusionSignificant differences are observed in the fungal species composition and relative abundance among the cities of Beijing, Fuzhou, Kunming, and Urumqi. Employing fungal ITS2 sequencing on dust samples from these urban areas enables accurate inference of biogeographical locations. The high feasibility of utilizing fungal community data in dust for biogeographical inferences holds particular promise in the field of forensic science.

Dysphagia is a common complication of stroke. Combining the principles of traditional Chinese medicine with modern research findings， it is proposed that "latent pathogen in the cerebral collaterals" constitutes the core pathogenesis of post-stroke dysphagia （PSD）. In clinical practice， treatment is tailored according to the location of PSD. During the oral stage， when the pathogen invades the face and mouth， resulting in excessive salivation， acupoints are primarily selected from the foot shaoyin （少阴） kidney channel， in combination with ren mai （任脉） ， du mai （督脉）， chong mai （冲脉） and the spleen channel， to replenish essence and fill the marrow， dispel dampness and unblock the channels. In the pharyngeal stage， as the pathogen obstructs the throat， disrupting normal swallowing， the therapy emphasizes dredging the shaoyang （少阳） channel and warming and tonifying the jueyin （厥阴） channel， by taking acupoints mainly from the hand and foot shaoyang channels， along with the jueyin channels， so as to soothe the liver and promote bile secretion， regulate and harmonize qi and blood. During the esophageal stage， where the pathogen damages the esophagus， impeding food passage， the treatment emphasizes activating the yangming （阳明） channels and regulating taiyin （太阴） channels； acupoints are mainly selected from the foot yangming stomach channel， along with the taiyin channels， aiming to warm yang， unblock the channels and dispel stasis.

Objective To investigate the protective effect and mechanisms of total alkaloids of Bulbus Fritillaria Pallidiflora (TA-BFP) on pulmonary fibrosis in silicosis mice. Methods i) Bulbus Fritillaria Pallidiflora (BFP) powder was collected by heating and refluxing twice with 70.00% ethanol, and the dried concentrated sample was purified using cation-exchange and macroporous adsorbent resins to obtain TA-BFP. The total alkaloids level was detected using ultraviolet spectrophotometry and the characteristic peaks in TA-BFP were identified using the high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD). ii) Specific pathogen-free male C57BL/6J mice were randomly divided into the control group, model group and intervention group, with 15 mice in each group. Mice in the model group and the intervention group were treated with 80 μL silica suspension with a mass concentration of 62.5 g/L, while the control group was treated with an equal volume of 0.9% sodium chloride solution using non-exposed tracheal instillation method. On the 28th day after modeling, mice in the intervention group were given TA-BFP at a dose of 60 mg/kg body mass, and the model group and control group were given 2.00% Tween-80 solution at the same volume by gastric gavage, once per day, for 28 days. The mouse lung tissue pathology was observed, lung organ coefficient was calculated, and inflammation and fibrosis scores were assessed after the intervention period. Serum levels of mouse interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay. Results i) The total alkaloid level of TA-BFP in the BFP powder was 50.36% after elution and purification. A total of nine alkaloid components in TA-BFP were identified by HPLC-ELSD. ii) The results of lung histopathology showed that no obvious changes of inflammation and fibrosis were identified in the lungs of mice in the control group, obvious changes of inflammation and fibrosis were identified in the lungs of mice in the model group, and the intervention group showed reduced inflammation and fibrosis in the lungs of mice compared with that of the model group. The lung organ coefficient, pulmonary inflammation score, pulmonary fibrosis score, and levels of serum IL-1β, IL-6, and TNF-α were significantly higher in the model group than those in the control group (all P<0.05). The lung organ coefficient, pulmonary inflammation score, pulmonary fibrosis score, and levels of serum IL-1β, and TNF-α were significantly higher in the intervention group than those in the control group (all P<0.05), while the pulmonary inflammation score, pulmonary fibrosis score, and levels of serum IL-1β, IL-6, and TNF-α were significantly lower in the intervention group than those in the model group (all P<0.05). Conclusion TA-BFP alleviates pulmonary inflammation in silicosis mice, thereby delaying the progression of pulmonary fibrosis. The mechanism may involve in inhibiting the expression of inflammatory factors IL-1β, IL-6, and TNF-α.

Testicular radiation injury is a structural and functional abnormality of the testes caused directly or indirectly by radiation, which disrupts spermatogenesis and compromises male fertility. The development of effective preventive and therapeutic interventions is essential because of the high prevalence of this condition in clinical settings and its profound effect on patients′ reproductive health and overall well-being. The concept of "the struggle between vital qi and pathogen" is first seen in the Treatise on Cold Pathogenic Diseases. It denotes the dynamic struggle between vital and pathogenic qi. The occurrence, development, and sequelae of all diseases reflect this ongoing conflict. In this context, this study defines the "vital qi" of the testis as its capacity to generate and preserve the essence of reproduction and to resist damage. The pathogenic qi associated with testicular radiation injury is categorized into two types: ionizing poison and retaining evil. The pathogenesis of testicular radiation damage is delineated into three stages by integrating the characteristics of vital and pathogenic qi: the injury, adhesion, and recovery phases. Based on the theoretical framework advanced by this study, the therapeutic approach for testicular radiation injury should adhere to the fundamental principle of strengthening vital qi and eliminating pathogenic factors. Although the primary focus of treatment should be on strengthening vital qi, it should also be complemented by strategies to eliminate pathogenic influences. This paper aims to provide a novel perspective and strategic approach to the traditional Chinese medicine diagnosis, prevention, and treatment of testicular radiation injury. By elucidating the process of testicular radiation injury and its corresponding treatment principles, it seeks to offer valuable insights for clinical practice.

ObjectiveTo clarify the protective effect of Danlou tablet， a representative traditional Chinese medicine of the stagnation of phlegm and blood stasis co-treatment method， on vascular endothelial function in patients with atherosclerosis （AS）. MethodsA randomized controlled trial was conducted. From September 2023 to November 2023， a total of 72 patients who were diagnosed at Guang'anmen Hospital， China Academy of Chinese Medical Sciences and met the inclusion and exclusion criteria for carotid atherosclerosis （CAS） combined with coronary atherosclerotic heart disease （CHD） and stable angina pectoris （SAP） were enrolled. The patients were randomly divided into a control group （receiving conventional Western medicine treatment） and an observation group （receiving Danlou tablet combined with conventional Western medicine treatment）， with 36 cases in each group. The intervention lasted for 12 weeks. The frequency of angina pectoris attacks was recorded to evaluate the clinical efficacy of Danlou tablet. Peripheral blood samples were collected from patients， and the expression levels of serum endothelial injury markers before and after treatment were detected by enzyme-linked immunosorbent assay （ELISA）. The nitrate reductase method was employed to evaluate the protective effect of Danlou tablet on vascular function. The expression levels of serum inflammatory factors and lipoproteins were determined by ELISA and an automatic biochemical analyzer （dynamic timed scatter turbidimetry and enzymatic method） to assess the anti-inflammatory and lipid-regulating effects of Danlou tablet. ResultsIn terms of angina pectoris attacks， compared with that in the control group， the frequency of attacks in the observation group was reduced （P<0.05）. In terms of endothelial injury markers， compared with the levels before treatment within the same group， the levels of endothelin-1 （ET-1）， intercellular adhesion molecule-1 （ICAM-1）， and vascular cell adhesion molecule-1 （VCAM-1） in the peripheral blood of the observation group were decreased （P<0.05）， while the levels of nitric oxide （NO） and vascular endothelial growth factor （VEGF） were increased （P<0.05）. Compared with the control group after treatment， the differences in ET-1， NO， ICAM-1， and VCAM-1 were significant （P<0.05）. In terms of serum inflammatory factors， after treatment， the interleukin-6 （IL-6） level in the observation group was decreased significantly （P<0.05）. Compared with the control group after treatment， the IL-6 level in the observation group was decreased significantly （P<0.01）. In terms of serum lipoproteins， after treatment， the level of low-density lipoprotein cholesterol （LDL-C） in the observation group was decreased （P<0.05）. After treatment， the level of high-density lipoprotein cholesterol （HDL-C） in the observation group was significantly higher than that in the control group （P<0.05）. In terms of safety evaluation， no serious adverse events occurred in either group during the intervention period. ConclusionDanlou tablet applied to patients with CAS combined with CHD can improve endothelial function， reduce inflammatory indicators， alleviate symptoms， improve the quality of life of patients， and demonstrate good safety.