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.

Objective To investigate the effects and mechanisms of silica dust on the apoptosis of alveolar epithelial cell (AEC) through in vitro and animal experiments. Methods i) In vitro experiment. A549 cells were stimulated with 100 mg/L silica suspension for 0, 12, 24 and 48 hours. The cell apoptosis rate was detected by flow cytometry. ii) Animal experiment. Specific pathogen-free male C57BL/6 mice were randomly divided into control, 14-day, 28-day, and 56-day groups, with five mice in each group. The mice in the control group were sacrificed at 56 days after being treated with 40.0 μL 0.9% sodium chloride solution, and the mice in the last three groups were sacrificed at 14, 28 and 56 days after being treated with 40.0 μL silica suspension with a mass concentration of 125 g/L via tracheal exposure method. The lung tissues of mice were collected to measure lung organ coefficients. Masson staining was used to detect the degree of pulmonary fibrosis, and Ashcroft scores were evaluated. The apoptosis of AEC in mice was observed by TUNEL immunofluorescence assay. iii) The mRNA relative expression of apoptosis-related genes in A549 cells and mouse lung tissue was detected using reverse transcription and real-time fluorescence quantitative polymerase chain reaction. Results i) In vitro experiment. The apoptosis rate of A549 cells increased with longer silica exposure (all P<0.05). The relative expression of B cell lymphoma-2 (BCL-2) mRNA in A549 cells in 24 h group and 48 h group decreased (both P<0.05), and the relative expression of BCL-2 associated X protein (BAX) mRNA increased (both P<0.05), compared with 0 h group. The mRNA relative expression of caspase (CASP) -3 and CASP-9 in A549 cells increased with longer silica exposure (all P<0.05). ii) Animal experiment. The lung organ coefficients and Ashcroft score in mice progressively increased (all P<0.05), the degree of pulmonary fibrosis was gradually aggravated, and TUNEL positive cells in lung tissue were gradually increased, while Bax, Casp-3 and Casp-9 mRNA relative expression increased with longer silica exposure (all P<0.05). Conclusion Silica dust may cause pulmonary fibrosis by inducing apoptosis of AEC, with a time-dependent effect. The mechanism may be related to the effect of silica dust on mitochondrial apoptosis through Bcl-2/Bax/Caspase-3 signaling pathway.

The innate immune system detects cellular stressors and microbial infections, activating programmed cell death (PCD) pathways to eliminate intracellular pathogens and maintain homeostasis. Among these pathways, pyroptosis, apoptosis, and necroptosis represent the most characteristic forms of PCD. Although initially regarded as mechanistically distinct, emerging research has revealed significant crosstalk among their signaling cascades. Consequently, the concept of PANoptosis has been proposed—an inflammatory cell death pathway driven by caspases and receptor-interacting protein kinases (RIPKs), and regulated by the PANoptosome, which integrates key features of pyroptosis, apoptosis, and necroptosis. The core mechanism of PANoptosis involves the assembly and activation of the PANoptosome, a macromolecular complex composed of three structural components: sensor proteins, adaptor proteins, and effector proteins. Sensors detect upstream stimuli and transmit signals downstream, recruiting critical molecules via adaptors to form a molecular scaffold. This scaffold activates effectors, triggering intracellular signaling cascades that culminate in PANoptosis. The PANoptosome is regulated by upstream molecules such as interferon regulatory factor 1 (IRF1), transforming growth factor beta-activated kinase 1 (TAK1), and adenosine deaminase acting on RNA 1 (ADAR1), which function as molecular switches to control PANoptosis. Targeting these switches represents a promising therapeutic strategy. Furthermore, PANoptosis is influenced by organelle functions, including those of the mitochondria, endoplasmic reticulum, and lysosomes, highlighting organelle-targeted interventions as effective regulatory approaches. Cardiovascular diseases (CVDs), the leading global cause of morbidity and mortality, are profoundly impacted by PCD. Extensive crosstalk among multiple cell death pathways in CVDs suggests a complex regulatory network. As a novel cell death modality bridging pyroptosis, apoptosis, and necroptosis, PANoptosis offers fresh insights into the complexity of cell death and provides innovative strategies for CVD treatment. This review summarizes current evidence linking PANoptosis to various CVDs, including myocardial ischemia/reperfusion injury, myocardial infarction, heart failure, arrhythmogenic cardiomyopathy, sepsis-induced cardiomyopathy, cardiotoxic injury, atherosclerosis, abdominal aortic aneurysm, thoracic aortic aneurysm and dissection, and vascular toxic injury, thereby providing critical clinical insights into CVD pathophysiology. However, the current understanding of PANoptosis in CVDs remains incomplete. First, while PANoptosis in cardiomyocytes and vascular smooth muscle cells has been implicated in CVD pathogenesis, its role in other cell types—such as vascular endothelial cells and immune cells (e.g., macrophages)—warrants further investigation. Second, although pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs) are known to activate the PANoptosome in infectious diseases, the stimuli driving PANoptosis in CVDs remain poorly defined. Additionally, methodological challenges persist in identifying PANoptosome assembly in CVDs and in establishing reliable PANoptosis models. Beyond the diseases discussed, PANoptosis may also play a role in viral myocarditis and diabetic cardiomyopathy, necessitating further exploration. In conclusion, elucidating the role of PANoptosis in CVDs opens new avenues for drug development. Targeting this pathway could yield transformative therapies, addressing unmet clinical needs in cardiovascular medicine.

Objective To explore the mechanism of the gut-lung axis in paraquat-induced lung injury in mice, with a focus on analyzing the changes in intestinal gene expression and their potential roles. Methods Specific pathogen-free C57BL/6 wild-type mice were randomly divided into control, low-dose, and high-dose groups, with 10 mice in each group. Mice in the three groups received a single intragastric administration of paraquat solution at doses of 0, 25, or 50 mg/kg body weight. The mice were euthanized on day 21. Lung histopathological changes were assessed, and the differentially expressed genes (DEGs) in the intestinal tissues of mice in these two groups were analyzed through transcriptomics. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted to explore potential mechanisms of the gut-lung axis in paraquat-induced lung injury and fibrosis. Results Paraquat exposure induced dose-dependent pulmonary injury and fibrosis in the mice. The Ashcroft score of lung tissue was higher in the mice of low-dose group than that in the control group (P<0.05). Both the lung organ coefficient and Ashcroft score of lung tissues in the mice of high-dose group were higher than those in the control group and the low-dose group (all P<0.05). The result of transcriptomic analysis showed 146 DEGs, including 91 upregulated and 55 downregulated genes, in intestinal tissues of mice in the low-dose group, and 57 DEGs, including 47 upregulated and 10 downregulated genes in the high-dose group, compared with the control group. Notably, 19 DEGs were commonly altered in both low- and high-dose groups. The result of GO enrichment analysis showed that the DEGs were primarily involved in biological processes including "immune response", "oxidative stress" and "cell differentiation". The result of KEGG enrichment analyses showed that DEGs were primarily involved in key processes including "oxidative stress response path way", "immune response path way" and "digestion and absorption path way". Conclusion Paraquat exposure alters intestinal gene expression, particularly in genes in biological processes related to immune responses and oxidative stress. These changes may mediate inflammatory signaling via the gut-lung axis and contribute to the development of paraquat-induced pulmonary fibrosis.

【Objective】 To establish the internal quality control standard of leukocyte-depleted suspended red blood cell volume in our center, so as to guide the preparation of components, strengthen the internal quality control and improve the quality of blood preparations. 【Methods】 A total of 1 523 bags of whole blood collected using two manufacturers′ leukocyte-depleted blood bags from March to August 2023 at our center were extracted. The blood before and after filtration were weighed, and the volume of whole blood collected, the volume of filtration loss and the product volume based on the formula and measured specific gravity were calculated. According to the data distribution characteristics, the reference range of leukocyte-depleted suspended red blood cell volume was determined, and the differences of whole blood collection volume, filtration loss capacity and product capacity between the two manufacturers were analyzed. The quality control data of leukocyte-depleted red blood cells over the past year with the difference between another 100 bags of these cells and the reference interval were compared, and the effectiveness of reference interval was validated. 【Results】 The median whole blood collection volume in the sample size was 402.0 mL, with a median filtration loss of 42.4 mL, and an average volume of leukocyte-depleted suspended red blood cells at 322.5 mL. The whole blood collection volume (A: median 404.4 mL; B: median 397.7 mL, P<0.01) and the volume of leukocyte-depleted suspended red blood cell products (A: mean 331.4 mL; B: mean 312.0 mL, P<0.01) using manufacturer A′s leukocyte-depleted blood bag were both higher, with a lower filtration loss capacity (A: median 39.5 mL; B: median 46.6 mL, P<0.01). The standard deviation of the volume of leukocyte-depleted suspended red blood cell was 19.6, and the reference interval was 284.1-360.9 mL. The validation samples and quality control sampling data showed no difference from the interval samples (P>0.05). 【Conclusion】 According to the actual situation of our center, the volume standard of leukocyte-depleted suspended red blood cells in our center is determined to be 284.1-360.9 mL.

ObjectiveAngle-closure glaucoma (ACG) is one of the major eye-blinding diseases. To diagnose ACG, it is crucial to examine the anterior chamber angle. Current diagnostic tools include slit lamp gonioscopy, water gonioscopy, ultrasound biomicroscopy (UBM), and anterior segment optical coherence tomography (AS-OCT). Slit lamp and water gonioscopy allow convenient observation of the anterior chamber angle, but pose risks of invasive operation and eye infections. UBM can accurately measure the structure of the anterior chamber angle. However, it is complex to operate and unsuitable for patients, who have undergone trauma or ocular surgery. Although AS-OCT provides detailed images, it is costly. The aim of this study is to explore a non-invasive, non-destructive optical reflection tomography (ORT) technique. This technique can achieve low-cost three-dimensional imaging and full-field anterior chamber angle measurement of the porcine eye. MethodsThe experiment involved assembling an optical reflection tomography system, which included a complementary metal oxide semiconductor (CMOS) camera, a telecentric system, a stepper motor, and a white light source, achieving a spatial resolution of approximately 8.5 μm. The process required positioning the porcine eye at the center of the field of the imaging system and rotating it around its central axis using a stepper motor. Reflection projection images were captured at each angle with an exposure time of 1.0 ms and an interval of 2°. The collected reflection-projection data were processed using a filtered reflection tomography algorithm, generating a series of two-dimensional slice data. These slices essentially represented cross-sectional views of the three-dimensional structural image, and were reconstructed into a complete three-dimensional structural image. Based on the reconstructed three-dimensional structural image of the porcine eye, the anterior chamber angles at different positions were measured, and a distribution map of these angles was drawn. Simultaneously, the ORT measurements were compared with the standard results obtained from optical coherence tomography (OCT) to assess the accuracy of ORT measurements. ResultsIn this study, we successfully obtained the reflection projection data of a porcine eye using ORT technology, reconstructed its three-dimensional structural image, and measured the anterior chamber angle, generating the corresponding distribution map. To better distinguish the different structural parts of porcine eye, the three-dimensional structural image was marked with blue, green, and yellow dashed lines from the outer to the inner layers. The area between the blue and green dashed lines corresponded to the sclera. The area between the green and yellow dashed lines corresponded to the iris. The area inside the yellow dashed line corresponded to the pupil. The three-dimensional structural image clearly revealed the key anatomical features of the porcine eye. It was able to measure the anterior chamber angle at different positions. Additionally, the anterior chamber angle measurements of the porcine eye using ORT were compared with the measurements obtained using a TEL320C1 type OCT system, showing an average deviation of 0.51° and a mean square error MSE of 0.317. ConclusionORT is a non-invasive, non-destructive, low-cost, and high-resolution imaging technique capable of achieving three-dimensional structural imaging and full-field anterior chamber angle measurement of a porcine eye. This technology offers a new perspective for the diagnosis of angle-closure glaucoma and is significant for the screening, diagnosis, and monitoring of eye diseases, potentially benefiting clinics and small hospitals in remote areas in the future.

ObjectiveTo clarify the pharmacodynamic characteristics and neuroinflammatory mechanisms of Ruyi Zhenbaowan （RYZBW） in treating nociceptive hypersensitivity and central sensitisation of spinal cord in the mouse model of central post-stroke pain （CPSP）. MethodSPF-grade male ICR mice of 8 weeks old were assigned into the sham operation （Sham）， model （CPSP）， low-， medium-， and high-dose （0.303， 0.607 1.214 g·kg^-1） RYZBW （RYZBW-L， RYZBW-M， and RYZBW-H， respectively）， and pregabalin （PGB， 0.046 g·kg^-1， positive control） groups. The rat model of CPSP was established by injection of type Ⅳ collagenase into the ventral posterior lateral nucleus of the thalamus on day 1. Rats were administrated with corresponding drugs or normal saline （Sham and CPSP groups） by gavage from day 14 to day 17. The mechanical pain sensitivity test was performed on days 0， 3， 4， 7， 10， 14， 17. On day 18， the L₅ segment of spinal cord was collected for the detection of inflammatory cytokines by immunoinflammatory microarray， CXC chemokine ligand 16 （CXCL16） by enzyme-linked immunosorbent assay， and calcitonin gene-related peptide （cGRP） by immunohistochemistry. In addition， fluorescence dual-labeling was employed to determine the expression levels of CXCL16， the dendritic cell marker CD11c， the macrophage marker CD68， the microglia marker TMEM119， the endothelial cell markers CD31 and CXCR6， and the T cell marker CD3. ResultCompared with the Sham group， the mechanical pain threshold of the CPSP group was significantly lower than that of the Sham group from day 3 to day 17， with stable hyperalgesia symptoms. On the 7^th day， the mechanical pain threshold of the PGB group was significantly higher than that of the CPSP group， with significant analgesic effect （P<0.01）. On days 10-17, the mechanical pain threshold of the RYZBW-H group was significantly higher than that of the CPSP group， showing a stable analgesic effect （P<0.05）. On the 17^th day， the analgesic effect of RYZBW was dose-effect correlated （R²=0.303 7）. From day 4 to day 17， the mechanical pain threshold of RYZBW-H group was positively correlated with time （R²=0.111 5）. The above results suggested that the analgesia of RYZBW was time-dependent. On the 17 th day， the expression of central sensitization marker cGRP in the spinal dorsal horn of CPSP mice was significantly increased compared with the Sham group （P<0.05）， and RYZBW down-regulated it in a dose-dependent manner （R²=0.500 8）， suggesting that RYZBW significantly inhibited the central sensitization of the spinal cord caused by CPSP. The results of spinal cord inflammation chip on the 17^th day showed that compared with CPSP group， RYZBW-H group inhibited CXCL16 expression （P<0.01）.The results of ELISA based on independent repeated samples showed that RYZBW inhibited the expression of CXCL16 protein in spinal cord in a dose-dependent manner （R²=0.250 4）. The results of immunofluorescence double labeling showed that compared with Sham group, the expression of CXCL16 in CD11c positive dendritic cells in CPSP group increased， and the number of CD68 positive cells increased （P<0.05）. Compared with CPSP group， RYZBW down-regulated it: the expression of CXCL16 in CD31 positive endothelial cells， CD68 positive macrophages and TMEM119 positive microglia increased， and the number and cell body area of TMEM119 positive microglia increased significantly （P<0.05）. The number of CD3 positive T cells （P<0.05） and the expression of CXCR6 in CD3 positive T cells were increased. RYZBW inhibited the activation of endothelial cells and macrophages in a dose-dependent manner， and reduced the infiltration of microglia and T cells （R²=0.691 4， R²=0.551 5， R²=0.653 2， R²=0.180 6， R²=0.287 5， R²=0.298 6，R²=0.511 6）. ConclusionRYZBW can effectively alleviate nociceptive hypersensitivity and central sensitisation of the spinal cord in CPSP mice by regulating CXCL16-CXCR-6， inhibiting the infiltration and activation of microglia and macrophages， and the activation of dendritic cells， endothelial cells， and T cells.

AIM To prepare the soluble microneedles of Aconitum brachypodum Diels alkaloids.METHODS Centrifugal molding method was adopted in the preparation of soluble microneedles.With chondroitin sulfate consumption,PVP K120 consumption and 40%ethanol consumption as influencing factors,piercing rate as an evaluation index,the formulation was optimized by Box-Behnken response surface method,after which the morphology,piercing performance,drug content and in vitro transdermal performance were investigated.RESULTS The optimal formulation was determined to be 123 mg for chondroitin sulfate consumption,298 mg for PVP K120 consumption,and 2.4 mL for 40%ethanol consumption,the piercing rate was 98.3%.The soluble microneedles were yellow and square patch with conoid needle,which could pierce aluminum foil and rat skin,along with the drug content of(0.94±0.025)mg.The soluble microneedle group demonstrated the accumulative permeability rate of 91.4%within 24 h,which was higher than that in the gel ointment group,and the permeability accorded with Higuchi equation.CONCLUSION The soluble microneedles of A.brachypodum alkaloids exhibit good mechanical strength,which can achieve effective transdermal delivery of drugs.

Objective To understand the spectrum and changes of pathogens causing healthcare-associated infec-tion(HAI)in neonatal intensive care unit(NICU).Methods Clinical medical records of neonates with HAI in a hospital from January 2018 to December 2022 were collected,spectrum of pathogens causing HAI were and analyzed retrospectively.Results A total of 7 597 hospitalized neonates were investigated,and 240 of whom had 263 cases of HAI,with an HAI incidence of 3.16％and healthcare-associated case infection incidence of 3.46％.96 cases(36.50％)were bloodstream infection,70(26.62％)were respiratory system infection,and 57(21.67％)were in-fection without clear sites.A total of 170 pathogens were detected from specimens,78(45.88％)of which were Gram-positive bacteria,with Staphylococcus spp.accounting for the highest proportion,78(45.88％)were Gram-negative bacteria,mainly Klebsiella pneumoniae,and 14(8.24％)were fungi.The detection rate of Gram-negative bacteria showed an upward trend from 2018 to 2022(P＜0.01).Conclusion The majority of HAI in NICU is bloodstream infection.In recent years,the detection rate of Gram-negative bacteria has been increasing year by year,and it is necessary to streng-then the prevention and control of HAI in clinical practice.

OBJECTIVE To rapidly analyze the chemical constituents of Rubus sachalinensis Leveille by HPLC-Q-Exactive-MS/MS. METHODS Chromatographic separation was carried out on CAPCELL PAK MGII C18(4.6 mm×250 mm, 5 μm) column at the temperature of 30 ℃. The mobile phase was acetonitrile-0.1% formic acid by gradient elution, with a flow rate of 1.0 mL∙min−1, and the injection volume of 20 µL. The MS spectrum was acquired in negative ion modes using HESI ion source. RESULTS The molecular and structural formulae of the compounds were determined based on the exact mass number and ChemSpider and PubChem databases. By comparing the retention time of the corresponding reference standards and those reported in the literature, primary mass spectra, and secondary mass spectrometry pyrolysis fragments, combined with fragmentation regularity of such compounds, a total of 71 compounds were identified from Rubus sachalinensis Leveille, including 30 organic acids, 22 flavonoids, 7 triterpenoid saponins, 5 coumarins, 1 lignan, 1 gallotannin and 2 aromatic compounds. CONCLUSION This method can quickly and accurately identify the complex chemical constituents in Rubus sachalinensis Leveille, and provide scientific basis for the basic research on the medicinal substances of Rubus sachalinensis Leveille.