ObjectiveTo investigate the mechanism of Guizhi Fulingwan （GFW） against hepatic fibrosis， focusing on elucidating the regulatory effect of GFW on the mitochondrial DNA （mtDNA）/NOD-like receptor protein 3 （NLRP3）/cysteinyl aspartate-specific proteinase-1 （Caspase-1）/gasdermin D （GSDMD） signaling pathway. MethodsForty-two male Sprague-Dawley （SD） rats were randomly allocated into six groups （n=7）： control， model， low/medium/high-dose （0.14， 0.28， 0.56 g·kg^-1·d^-1） GFW （GFW-L， GFW-M， GFW-H）， and Dahuang Zhechong pills （DZW， 1.8 g·kg^-1·d^-1）. The rat model of hepatic fibrosis was induced by intraperitoneal injection of carbon tetrachloride. General conditions of the rats were observed. Serum levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） levels were measured. Liver histopathology and collagen deposition were observed through hematoxylin and eosin （HE） staining and Masson's trichrome staining. Transmission electron microscopy （TEM） was employed to observe structural alterations and damage of cellular ultrastructures including mitochondria. Mitochondrial membrane potential （MMP， ΔΨ_m） was detected by flow cytometry. Serum levels of interleukin-1β （IL-1β） and interleukin-18 （IL-18） were measured by enzyme-linked immunosorbent assay （ELISA）. The mRNA levels of mtDNA and NLRP3 in the liver tissue were quantified by Real-time polymerase chain reaction （Real-time PCR）. The protein levels of key molecules in the NLRP3/Caspase-1/GSDMD signaling pathway in the liver tissue were determined by Western blot. ResultsCompared with the control group， the model group exhibited a decrease in body weight （P<0.01）， an increase in liver index （P<0.01）， elevations in serum ALT and AST levels （P<0.01）， and typical fibrotic features such as disorganized hepatocytes， inflammatory infiltration， and increased collagen deposition in the liver tissue. TEM revealed significant karyotheca degeneration， mitochondrial swelling， endoplasmic reticulum expansion， and organelle efflux in the model group. In addition， the model group showed decreased ΔΨ_m （P<0.01）， up-regulated mRNA levels of mtDNA and NLRP3 （P<0.01） and protein levels of NLRP3， Caspase-1， and GSDMD （P<0.01） in the liver tissue， and increased serum levels of IL-1β and IL-18 （P<0.01）. Compared with that in the model group， the body weight increased in GFW-L， GFW-M， and DZW groups （P<0.05） and markedly increased in the GFW-H group （P<0.01）. The liver index decreased in the GFW groups and DZW group （P<0.01）. The serum ALT level declined in the GFW-L group （P<0.05）， and the serum ALT and AST levels decreased in the GFW-M， GFW-H， and DZW groups （P<0.01）. Histopathological damage and fibrosis were alleviated to varying degrees， and TEM revealed mitigated ultrastructural injuries including mitophagy， mitochondrial swelling， and endoplasmic reticulum expansion in the drug intervention groups. The ΔΨ_m increased in GFW groups without statistical significance. The mRNA level of mtDNA in the liver tissue was down-regulated in the GFW-M （P<0.05）， GFW-H （P<0.01）， and DZW （P<0.01） groups. The mRNA level of NLRP3 was down-regulated in GFW-M， GFW-H， and DZW groups （P<0.01）. Western blot analysis showed significantly down-regulated protein level of NLRP3 in all the GFW groups and the DZW group （P<0.01）. The protein level of GSDMD-N was down-regulated in GFW-H and DZW groups （P<0.01）. The protein level of cleaved Caspase-1 was down-regulated in GFW-M （P<0.05）， GFW-H （P<0.01）， and DZW （P<0.01） groups. In addition， the serum levels of IL-1β and IL-18 declined in GFW-H and DZW groups （P<0.01）. ConclusionGFW can suppress pyroptosis to ameliorate CCl₄-induced hepatic fibrosis， potentially through mitigating mitochondrial damage， inhibiting inflammasome assembly and activation， and blocking pro-inflammatory cytokine release.

Ulcerative colitis （UC） is a chronic， non-specific intestinal inflammatory disease. Its pathogenesis is complex， involving interactions among genetic， immune， and environmental factors， and remains incompletely elucidated. Mitochondrial damage can trigger the abnormal release of mitochondrial damage-associated molecular patterns （mtDAMPs）， activating inflammatory pathways and thereby exacerbating the inflammatory response in UC. Mitophagy， a core mitochondrial quality control mechanism， can clear damaged mitochondria and effectively reduce the abnormal release of mtDAMPs and the accumulation of harmful substances， thereby mitigating inflammatory damage resulting from mitochondrial dysfunction. Mitophagy plays a crucial role in maintaining the integrity of the intestinal epithelial barrier function and in the prevention and intervention of UC. Notably， the function of the spleen governing transportation and transformation in traditional Chinese medicine shares similarities with the role of mitochondria in energy transformation and substance metabolism. Furthermore， the pathological state of the spleen failing in transportation and transformation may be related to mitochondrial dysfunction， while the pathogenic characteristics of "endogenous turbid pathogen" align with the inflammatory cascade responses triggered by abnormal release of mtDAMPs. Based on this theoretical correlation， this paper aims to explore the correlation between the traditional Chinese medicine theory of "spleen dysfunction in essence distribution leading to endogenous turbid pathogens" and mitophagy. Using the correlation as an entry point， the potential role of mitophagy in the pathogenesis of UC was elucidated. Additionally， by considering the epidemiological characteristics of UC in the Southwest region of China， a new thinking for the treatment of UC from the perspective of turbid pathogens was proposed based on the "fortifying the spleen and resolving turbidity" method to provide theoretical support and research enlightenment for further exploring the prevention and treatment of UC with traditional Chinese medicine.

ObjectiveTo explore the mechanism by which Qifu Lizhong decoction restores intestinal mucosal barrier function in ulcerative colitis （UC） by regulating the myosin light chain kinase （MLCK）/phosphorylated myosin light chain （p-MLC） signaling pathway. MethodsA rat model of UC with spleen-kidney Yang deficiency syndrome and an in vitro intestinal barrier injury model were established. In vivo experiments were conducted to evaluate the body weight， disease activity index （DAI） scores， and changes in colonic pathological morphology in each group of rats. Enzyme-linked immunosorbent assay （ELISA） was used to measure the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and myeloperoxidase （MPO）. Transmission electron microscopy （TEM） was employed to observe the morphology and structure of tight junctions （TJ） in rat colonic epithelium. In vitro experiments were conducted to observe the morphology of F-actin in human colon adenocarcinoma cell line-2（Caco-2） cells by phalloidin staining. Combining both in vivo and in vitro approaches， immunohistochemistry （IHC）， immunofluorescence （IF）， and Western blot were applied to detect the expression and phosphorylation levels of TJ proteins ［zonula occludens-1 （ZO-1）， Occludin， Claudin-1］ as well as key proteins in the MLCK/p-MLC signaling pathway. ResultsIn vivo experiments demonstrated that， compared with the normal group， the model group exhibited significant body weight loss， elevated DAI scores and histopathological scores （P<0.01）， increased colonic TNF-α， IL-1β， and MPO levels （P<0.01）. TJ structures were disrupted， and the expression of ZO-1， Occludin， and Claudin-1 was significantly reduced （P<0.05， P<0.01）. The protein expression levels of myosin light chain （MLC）， MLCK， phosphorylated （p）-MLC， and p-MLCK significantly increased （P<0.01）. Compared with the model group， all Qifu Lizhong decoction dose groups exhibited increased body weight， as well as significantly decreased DAI scores and histopathological scores （P<0.05， P<0.01）. The colonic levels of MPO， TNF-α， and IL-1β were significantly reduced （P<0.05， P<0.01）. In the Qifu Lizhong decoction + inhibitor group， the expression of TJ proteins ZO-1， Occludin， and Claudin-1 significantly increased （P<0.01）， while the protein expression levels of MLC， MLCK， p-MLC， and p-MLCK were significantly reduced （P<0.01）. In vitro experiment results showed that， compared with the normal group， the TNF-α group exhibited disordered cytoskeletal architecture， weakened fluorescence signals of ZO-1， Occludin， and Claudin-1， and significantly elevated protein expression levels of MLC， MLCK， p-MLC， and p-MLCK （P<0.01）. Compared with the TNF-α group， the Qifu Lizhong decoction + inhibitor group restored the ordered arrangement of the F-actin cytoskeleton， normalized the expression and localization of TJ proteins ZO-1， Occludin， and Claudin-1， and significantly decreased the protein expression levels of MLC， MLCK， p-MLC， and p-MLCK （P<0.01）. ConclusionQifu Lizhong decoction may exert its therapeutic effects on UC by inhibiting the MLCK/p-MLC signaling pathway， thereby stabilizing TJ structure and repairing intestinal barrier integrity.

Glioma represents the most prevalent malignant tumor of the central nervous system, with chemotherapy serving as an essential adjunctive treatment. However, most chemotherapeutic agents exhibit limited ability to penetrate the blood-brain barrier (BBB). This study introduced a novel dual-targeting strategy for glioma therapy by modulating the formation of nanobody-driven protein coronas to enhance the brain and tumor-targeting efficiency of hydrophobic cisplatin prodrug-loaded lipid nanoparticles (C8Pt-Ls). Specifically, nanobodies (Nbs) with fibrinogen-binding capabilities were conjugated to the surface of C8Pt-Ls, resulting in the generation of Nb-C8Pt-Ls. Within the bloodstream, Nb-C8Pt-Ls could bound more fibrinogen, forming the protein corona that specifically interacted with LRP-1, a receptor highly expressed on the BBB. This interaction enabled a "Hitchhiking Effect" mechanism, facilitating efficient trans-BBB transport and promoting effective brain targeting. Additionally, the protein corona interacted with LRP-1, which is also overexpressed in glioma cells, achieving precise tumor targeting. Computational simulations and SPR detection clarified the molecular interaction mechanism of the Nb-fibrinogen-(LRP-1) complex, confirming its binding specificity and stability. Our results demonstrated that this strategy significantly enhanced C8Pt accumulation in brain tissues and tumors, induced apoptosis in glioma cells, and improved therapeutic efficacy. This study provides a novel framework for glioma therapy and underscores the potential of protein corona modulation-based dual-targeting strategies in advancing treatments for brain tumors.

Objective:To explore the effect of behavioral activation therapy on subthreshold depression and quality of life in patients with diabetes foot amputation, and to provide reference for clinical psychotherapy and quality of life improvement in patients with diabetes foot amputation.Methods:A randomized controlled trial was conducted to select 50 patients with diabetes foot amputation from September to December 2023 in the People′s Hospital Affiliated to Shandong First Medical University by convenient sampling. The participants were divided into an control group and a observation group, with 25 cases in each group, using a random number table method. The patients in the control group received routine nursing care of diabetes feet, and the patients in the observation group were given behavior activation therapy on the basis of the control group. The Centre for Epidomiological Studies Depression Scale (CES-D), Behavioral Activation for Depression Scale-Short Form (BADS-SF), and Short Form Health Measure (SF-12) were used to assess patients′ subthreshold depression, behavior, and quality of life before the intervention, at the end of the intervention, and 3 months after the intervention.Results:The final number of patients who completed the study in the control group and observation group were 23 and 24, respectively. There were 11 males and 12 females in the control group, with an age of (69.70 ± 7.01) years. There were 12 males and 12 females in the observation group, with an age of (69.17 ± 6.83) years. At the end of the intervention and 3 months after the intervention, the CES-D scores of the observation group were (21.13 ± 2.19) and (18.83 ± 1.86) points, respectively, which were significantly lower than those of the control group (25.70 ± 1.72) and (25.91 ± 1.65) points, and the differences were statistically significant ( t=-7.93, -13.80, both P<0.05). At the end of the intervention and 3 months after the intervention, the BADS-SF behavioral activation dimension scores in the observation group were (17.67 ± 2.41) and (18.50 ± 2.19) points, respectively, higher than the control group's (11.83 ± 1.78) and (11.57 ± 2.02) points, and the differences were statistically significant ( t=31.65, 34.45, both P<0.05). The behavioral avoidance dimension scores were (9.48 ± 1.50) and (9.65 ± 1.99) points, respectively, lower than the control group's (10.75 ± 1.42) and (11.88 ± 1.57) points, and the differences were statistically significant ( t=6.53, 13.17, both P<0.05). Three months after the intervention, the total score of SF-12 in the observation group was (65.96 ± 2.94) points, and the psychological dimension score was (37.50 ± 4.14) points, both of which were higher than those in the control group (60.30 ± 2.42) and (30.13± 5.15) points, and the differences were statistically significant ( t=51.54, 5.42, both P<0.05). There were statistically significant differences in time, between groups, and interaction effects in CES-D score, scores of two dimensions of BADS-SF and SF-12 total score ( F values were 10.04-190.71). Conclusions:Behavioral activation therapy can improve the symptoms of subthreshold depression in patients with diabetes foot amputation, enhance their behavioral activation level, and improve the quality of life to a certain extent.

Objective:To explore the effect of behavioral activation therapy on subthreshold depression and quality of life in patients with diabetes foot amputation, and to provide reference for clinical psychotherapy and quality of life improvement in patients with diabetes foot amputation.Methods:A randomized controlled trial was conducted to select 50 patients with diabetes foot amputation from September to December 2023 in the People′s Hospital Affiliated to Shandong First Medical University by convenient sampling. The participants were divided into an control group and a observation group, with 25 cases in each group, using a random number table method. The patients in the control group received routine nursing care of diabetes feet, and the patients in the observation group were given behavior activation therapy on the basis of the control group. The Centre for Epidomiological Studies Depression Scale (CES-D), Behavioral Activation for Depression Scale-Short Form (BADS-SF), and Short Form Health Measure (SF-12) were used to assess patients′ subthreshold depression, behavior, and quality of life before the intervention, at the end of the intervention, and 3 months after the intervention.Results:The final number of patients who completed the study in the control group and observation group were 23 and 24, respectively. There were 11 males and 12 females in the control group, with an age of (69.70 ± 7.01) years. There were 12 males and 12 females in the observation group, with an age of (69.17 ± 6.83) years. At the end of the intervention and 3 months after the intervention, the CES-D scores of the observation group were (21.13 ± 2.19) and (18.83 ± 1.86) points, respectively, which were significantly lower than those of the control group (25.70 ± 1.72) and (25.91 ± 1.65) points, and the differences were statistically significant ( t=-7.93, -13.80, both P<0.05). At the end of the intervention and 3 months after the intervention, the BADS-SF behavioral activation dimension scores in the observation group were (17.67 ± 2.41) and (18.50 ± 2.19) points, respectively, higher than the control group's (11.83 ± 1.78) and (11.57 ± 2.02) points, and the differences were statistically significant ( t=31.65, 34.45, both P<0.05). The behavioral avoidance dimension scores were (9.48 ± 1.50) and (9.65 ± 1.99) points, respectively, lower than the control group's (10.75 ± 1.42) and (11.88 ± 1.57) points, and the differences were statistically significant ( t=6.53, 13.17, both P<0.05). Three months after the intervention, the total score of SF-12 in the observation group was (65.96 ± 2.94) points, and the psychological dimension score was (37.50 ± 4.14) points, both of which were higher than those in the control group (60.30 ± 2.42) and (30.13± 5.15) points, and the differences were statistically significant ( t=51.54, 5.42, both P<0.05). There were statistically significant differences in time, between groups, and interaction effects in CES-D score, scores of two dimensions of BADS-SF and SF-12 total score ( F values were 10.04-190.71). Conclusions:Behavioral activation therapy can improve the symptoms of subthreshold depression in patients with diabetes foot amputation, enhance their behavioral activation level, and improve the quality of life to a certain extent.

Objective To analyze the characteristics of blood transfusion consultation cases and establish the consulta-tion route,so as to provide reference for blood transfusion doctors to participate in blood transfusion consultation practice.Methods The cases involved in clinical transfusion consultation in the blood transfusion department of our hospital from 2020 to 2023 were collected from the hospital information system(HIS),and then classified by department and consultation type to summarize the main points of transfusion consultation,formulate transfusion consultation routes,and conduct typical cases analysis.Results There were 315 clinical transfusion consultations from 2020 to 2023,with an increasing trend year by year(26 in 2020,67 in 2021,81 in 2022,141 in 2023).The consultations involved 24 departments,including cardio-vascular medicine 14.0%(44/315),orthopedics 12.7%(40/315),intensive care medicine 8.9%(28/315),general medi-cine 8.3%(28/315),cardiopulmonary disease 6.0%(19/315),etc.There were 8 categories of consultations,including 35.6%(112/315)autologous ozonized blood transfusion,23.8%(75/315)plasma exchange,14.9%(47/315)perioperative mass blood preparation(transfusion),11.4%(36/315)platelet-rich plasma therapy and 6.3%(20/315)autologous blood collection,etc.The clinical blood transfusion consultation route was formulated according to the consultation points.Six pa-tients with various diseases were treated by blood transfusion department.With effective treatment measures taken,all of them improved and were discharged.Conclusion The summary of key points of clinical blood transfusion consultation and formulation of the blood transfusion consultation route by department of blood transfusion are conducive to the implementation of blood transfusion consultation and guarantee the safety of patients.

This study compared the effects on bone metabolism and morphology of pathological obesity induced by excessive fat intake in a non-hibernator (mice) versus healthy obesity due to pre-hibernation fattening in a hibernator (ground squirrels). Kunming mice were fed a high-fat diet to provide a model of pathological obesity (OB group). Daurian ground squirrels fattened naturally in their pre-hibernation season (PRE group) were used as a healthy obesity model. Micro-computed tomography (micro-CT) and three-point bending tests were used to determine the microstructure and mechanical properties of bone. Western blots were used to analyze protein expression levels related to bone metabolism (Runt-related transcription factor 2 (RunX2), osteocalcin (OCN), alkaline phosphatase (ALP), osteoprotegerin (OPG), receptor activator of nuclear factor-‍κB ligand (RANKL), cathepsin K, matrix metallopeptidase 9 (MMP9), patched protein homolog 1 (Ptch1), phosphorylated β‍-‍catenin (P‍-‍β‍-‍catenin), and glycogen synthase kinase-3β (GSK-3β)). Compared with controls, there was no obvious bone loss in the OB mice, and the stiffness of the femur was increased significantly. Compared with summer active squirrels, bone formation was enhanced but the mechanical properties did not change in the PRE group squirrels. In OB mice, western blots showed significantly increased expression levels of all proteins except RunX2, OPG, and Ptch1. PRE ground squirrels showed significantly increased expression of most proteins except OCN and Ptch1, which decreased significantly, and P‍-‍β‍-‍catenin and OPG, which did not change. In conclusion, for non-hibernating mice, moderate obesity had a certain protective effect on bones, demonstrating two-way regulation, increasing both bone loss and bone formation. For pre-hibernating ground squirrels, the healthy obesity acquired before hibernation had a positive effect on the microstructure of bones, and also enhanced the expression levels of proteins related to bone formation, bone resorption, and Wnt signaling.
Mice ; Animals ; Hibernation ; Core Binding Factor Alpha 1 Subunit/metabolism* ; Glycogen Synthase Kinase 3 beta/metabolism* ; Diet, High-Fat ; X-Ray Microtomography ; Sciuridae/metabolism* ; Obesity

Glycyrrhizae Radix et Rhizoma,a traditional Chinese medicine also known as Gan Cao(GC),is frequently included in clinical prescriptions for the treatment of pneumonia.However,the pharmacological com-ponents of GC for pneumonia treatment are rarely explored.Gan An He Ji oral liquid(GAHJ)has a simple composition and contains GC liquid extracts and paregoric,and has been used clinically for many years.Therefore,GAHJ was selected as a compound preparation for the study of GC in the treatment of pneumonia.We conducted an in vivo study of patients with pneumonia undergoing GAHJ treatments for three days.Using the intelligent mass spectrometry data-processing technologies to analyze the meta-bolism of GC in vivo,we obtained 168 related components of GC in humans,consisting of 24 prototype components and 144 metabolites,with 135 compounds screened in plasma and 82 in urine.After analysis of the metabolic transformation relationship and relative exposure,six components(liquiritin,liquiritigenin,glycyrrhizin,glycyrrhetinic acid,daidzin,and formononetin)were selected as potential effective components.The experimental results based on two animal pneumonia models and the in-flammatory cell model showed that the mixture of these six components was effective in the treatment of pneumonia and lung injury and could effectively downregulate the level of inducible nitric oxide synthase(iNOS).Interestingly,glycyrrhetinic acid exhibited the strongest inhibition on iNOS and the highest exposure in vivo.The following molecular dynamic simulations indicated a strong bond between glycyrrhetinic acid and iNOS.Thus,the current study provides a pharmaceutical basis for GC and reveals the possible corresponding mechanisms in pneumonia treatment.