ObjectiveObesity, a global chronic metabolic disease, is closely associated with disruptions in lipid metabolism and gut microbiota. Current intervention strategies still have limitations in terms of safety and microecological regulation, necessitating the exploration of novel natural regulatory approaches. Based on the early pathological characteristics of obesity, this study innovatively employs a rectal delivery method alongside a high-fat diet (HFD)-induced obesity model to systematically evaluate the inhibitory effects, safety, and gut microbiota regulation mechanisms of leek-derived and konjac-derived extracellular vesicles on obesity development. By simulating early clinical intervention scenarios, this study aims to explore the preventive potential of plant-derived extracellular vesicles during the initial stages of obesity onset. MethodsExtracellular vesicles from leek and konjac were isolated using ultracentrifugation combined with density gradient centrifugation. Their nanoscale properties were characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM), and nanoparticle tracking analysis (NTA). Male C57BL/6J mice were randomly divided into four groups: normal control (NC), high-fat diet (HFD), leek-derived extracellular vesicles (LEVs), and konjac-derived extracellular vesicles (KEVs). Beginning simultaneously with HFD feeding, mice in the intervention groups received 20 g/L vesicles rectally every 3 d for 4 weeks. Body mass and body composition were monitored throughout. At endpoint, mouse serum, adipose tissue, and colonic contents were collected. Serum biochemical indices (lipid profile, liver and kidney function, cardiac markers) were assessed to evaluate safety and metabolic efficacy, while 16S rRNA sequencing was employed to analyze gut microbial structure and diversity. ResultsDLS, NTA, and TEM confirmed that both LEVs and KEVs exhibited typical cup-shaped nanostructures with average particle sizes of approximately 284 nm and 223 nm, respectively. LEVs and KEVs treatment significantly suppressed HFD-induced weight gain and elevation of body-fat percentage (P<0.05), and reduced accumulation of abdominal white and epididymal adipose tissue. Serological analyses showed that both vesicles lowered total cholesterol, triglycerides and LDL-cholesterol, and ameliorated liver enzyme profiles (ALT, AST), demonstrating lipid-metabolic regulation and hepatoprotective effects. No hepatic, renal or cardiac dysfunction was observed, indicating favorable safety. Gut microbiota analyses revealed that vesicle intervention partially restored HFD-depleted microbial diversity and reshaped community structure. Notably, LEVs markedly increased the relative abundance of the beneficial taxon Lachnospiraceae at the family level, which is known for producing short-chain fatty acids and enhancing intestinal barrier function. Furthermore, Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) functional prediction suggested that LEVs and KEVs modulated gut microbial functions through distinct mechanisms: LEVs downregulated pathways related to ribosomes and DNA replication while enhancing xenobiotic degradation, whereas KEVs tended to upregulate energy metabolism and protein synthesis toward healthy levels. ConclusionRectally administered LEVs and KEVs exhibit excellent safety and pronounced metabolic benefits during the early phase of obesity, suppressing weight gain, correcting lipid dysregulation, and exerting effects via modulation of gut microbial composition and function. This study provides systematic experimental evidence supporting plant-derived exosome-like vesicles as an early intervention strategy against obesity.

ObjectiveObesity, a global chronic metabolic disease, is closely associated with disruptions in lipid metabolism and gut microbiota. Current intervention strategies still have limitations in terms of safety and microecological regulation, necessitating the exploration of novel natural regulatory approaches. Based on the early pathological characteristics of obesity, this study innovatively employs a rectal delivery method alongside a high-fat diet (HFD)-induced obesity model to systematically evaluate the inhibitory effects, safety, and gut microbiota regulation mechanisms of leek-derived and konjac-derived extracellular vesicles on obesity development. By simulating early clinical intervention scenarios, this study aims to explore the preventive potential of plant-derived extracellular vesicles during the initial stages of obesity onset. MethodsExtracellular vesicles from leek and konjac were isolated using ultracentrifugation combined with density gradient centrifugation. Their nanoscale properties were characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM), and nanoparticle tracking analysis (NTA). Male C57BL/6J mice were randomly divided into four groups: normal control (NC), high-fat diet (HFD), leek-derived extracellular vesicles (LEVs), and konjac-derived extracellular vesicles (KEVs). Beginning simultaneously with HFD feeding, mice in the intervention groups received 20 g/L vesicles rectally every 3 d for 4 weeks. Body mass and body composition were monitored throughout. At endpoint, mouse serum, adipose tissue, and colonic contents were collected. Serum biochemical indices (lipid profile, liver and kidney function, cardiac markers) were assessed to evaluate safety and metabolic efficacy, while 16S rRNA sequencing was employed to analyze gut microbial structure and diversity. ResultsDLS, NTA, and TEM confirmed that both LEVs and KEVs exhibited typical cup-shaped nanostructures with average particle sizes of approximately 284 nm and 223 nm, respectively. LEVs and KEVs treatment significantly suppressed HFD-induced weight gain and elevation of body-fat percentage (P<0.05), and reduced accumulation of abdominal white and epididymal adipose tissue. Serological analyses showed that both vesicles lowered total cholesterol, triglycerides and LDL-cholesterol, and ameliorated liver enzyme profiles (ALT, AST), demonstrating lipid-metabolic regulation and hepatoprotective effects. No hepatic, renal or cardiac dysfunction was observed, indicating favorable safety. Gut microbiota analyses revealed that vesicle intervention partially restored HFD-depleted microbial diversity and reshaped community structure. Notably, LEVs markedly increased the relative abundance of the beneficial taxon Lachnospiraceae at the family level, which is known for producing short-chain fatty acids and enhancing intestinal barrier function. Furthermore, Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) functional prediction suggested that LEVs and KEVs modulated gut microbial functions through distinct mechanisms: LEVs downregulated pathways related to ribosomes and DNA replication while enhancing xenobiotic degradation, whereas KEVs tended to upregulate energy metabolism and protein synthesis toward healthy levels. ConclusionRectally administered LEVs and KEVs exhibit excellent safety and pronounced metabolic benefits during the early phase of obesity, suppressing weight gain, correcting lipid dysregulation, and exerting effects via modulation of gut microbial composition and function. This study provides systematic experimental evidence supporting plant-derived exosome-like vesicles as an early intervention strategy against obesity.

Gastric cancer is a common malignant tumor of the digestive tract and can be classified as “fullness of the stomach”， “epigastric pain”， “noise” and other categories in the field of traditional Chinese medicine. Sijunzi decoction is composed of Panax ginseng， Poria cocos， Atractylodes macrocephala， and honey-fried Glycyrrhiza uralensis， and it has the effect of tonifying qi and strengthening the spleen. This article summarizes the active ingredients， mechanism of action， and clinical application research progress of Sijunzi decoction in treating gastric cancer. The results show that the main active ingredients of Sijunzi decoction include ginsenosides， atractylenolide， pachymic acid， glycyrrhizic acid， etc.； Sijunzi decoction and its effective ingredients can play an anti-gastric cancer role by inhibiting the proliferation of gastric cancer cell， inducing apoptosis of gastric cancer cell， enhancing gastric cancer cell chemotherapy sensitivity， and inhibiting invasion and metastasis of gastric cancer cell. In addition， Sijunzi decoction can enhance the efficacy of chemotherapy drugs， strengthen the immune function of the body and lower serum cancer marker levels during the clinical treatment of gastric cancer.

BACKGROUND:Angiogenesis is the main treatment target of cardiovascular diseases.Bone morphogenetic protein 2 can modulate angiogenesis,but the regulatory effect of endothelial cell-specific bone morphogenetic protein 2 on angiogenesis is unclear. OBJECTIVE:To investigate the effect of endothelial-specific bone morphogenetic protein 2 on angiogenesis. METHODS:(1)Bioinformatics analysis:Cellular expression specificity and abundance of bone morphogenetic protein 2 were meta-analyzed by the PanglaoDB single-cell transcriptome database.The endothelial cell transcriptome sequencing dataset of the mouse hindlimb model and endocardial transcriptome dataset of mice overexpressing bone morphogenetic protein 2 were reanalyzed to evaluate the effect of endothelial cell bone morphogenetic protein 2 on the angiogenesis pathway.(2)Validation in vivo:After establishing the mouse hindlimb model,we compared the blood perfusion between the affected and sham limb at 7,14,and 21 days.The expression of the colocation of bone morphogenetic protein 2 and CD31 was explored by immunofluorescence and immunohistochemical staining.(3)Validation in vitro:The cultured human umbilical vein endothelial cells in vitro were divided into a control group,a hypoxia group,and a bone morphogenetic protein 2 inhibitor Noggin intervention group.After being cultured for 24 hours,the angiogenesis of endothelial cells in each group was observed. RESULTS AND CONCLUSION:(1)Endothelial cells are an important cell subgroup expressing bone morphogenetic protein 2.Both in the mouse hindlimb ischemia model and endocardial cells overexpressing bone morphogenetic protein 2,bone morphogenetic protein 2 was significantly up-regulated,and the angiogenesis pathway was significantly activated.(2)In the mouse hindlimb model,bone morphogenetic protein 2-positive blood vessels around neoangiogenesis increased significantly at 7 days of ischemia(P<0.05),and decreased significantly after 2 weeks of ischemia(P<0.001).(3)In umbilical vein endothelial cells cultured in vitro,after hypoxic intervention,the migration and sprouting of endothelial cells increased significantly,and the expression of angiogenesis factors vascular endothelial growth factor and platelet-derived growth factor was significantly increased.Noggin significantly reduced hypoxia-induced endothelial cell angiogenesis(P<0.001)and down-regulated the expression of vascular endothelial growth factor and platelet-derived growth factor(P<0.01).(4)These findings verify that endothelial cell-specific bone morphogenetic protein 2 can regulate angiogenesis,and targeting endothelial cell bone morphogenetic protein 2 is a promising way to improve angiogenesis.

Purpose To investigate the expression of ubiquitin-specific protease 9X(USP9X)and programmed cell death ligand 1(PD-L1)in pulmonary sarcomatoid carcinoma(PSC),and to assess their clinicopathological signif-icance.Methods Immunohistochemical staining was performed on 48 PSC specimens using the EnVision two-step method to detect USP9X and PD-L1 expression.The relationships between their expressions levels and clinicopathologi-cal parameters were analyzed.Results USP9X was positively expressed in 81.3％(39/48)of PSC cases,and the USP9X positivity rate was significantly higher in lymph node-negative tumors compared with lymph node-positive tumors(P＜0.05).PD-L1 expression was observed in 70.8％(34/48)of cases,with a significantly higher positivity rate in male patients than in female patients(P＜0.01).Correlation analysis showed a significant positive association between USP9X and PD-L1 expression in PSC(P＜0.05).Conclusion Both USP9X and PD-L1 are highly expressed in PSC,and their expression levels are significantly positively correlated.Combined detection of USP9X and PD-L1 may aid in predicting the efficacy of immunotherapy in PSC.

Objective:To investigate the iodine nutrition status of children and pregnant women in different water iodine areas of Shandong Province.Methods:From February to September 2023, Leling City (iodine deficient), Gaotang County (moderate iodine), and Liangshan County (high iodine) in different water iodine areas of Shandong Province were selected as survey sites. One village was selected from each county (city) in five directions: east, west, south, north, and center. Forty children aged 8 to 10 years (balanced in age, half male and half female) and 20 pregnant women were selected as survey subjects in each village. Random urine samples of children and pregnant women were collected to test for urinary iodine. Meanwhile, thyroid examinations were conducted on children to calculate the goiter rate.Results:A total of 600 urine samples of children were tested, with a median urinary iodine level of 246.0 μg/L. The median urinary iodine levels of children in iodine deficient, moderate iodine and high iodine areas were 219.6, 208.0 and 446.0 μg/L, respectively ( n = 200, 200, 200). The median urinary iodine level of children in high iodine area was significantly higher than that in iodine deficient and moderate iodine areas ( P < 0.05). A total of 600 children underwent thyroid examinations, with a goiter rate of 5.8% (35/600). The goiter rate of children in iodine deficient, moderate iodine, and high iodine areas were 4.0% (8/200), 1.0% (2/200), and 12.5% (25/200), respectively. The goiter rate of children in high iodine area was significantly higher than that in iodine deficient and moderate iodine areas ( P < 0.05). A total of 247 urine samples of pregnant women were tested, with a median urinary iodine level of 158.2 μg/L. The median urinary iodine levels of pregnant women in iodine deficient, moderate iodine, and high iodine areas were 75.3, 175.2 and 321.2 μg/L, respectively ( n = 98, 84, 65). The median urinary iodine level of pregnant women in high iodine area was significantly higher than that in iodine deficient and moderate iodine areas, and the median urinary iodine level of pregnant women in moderate iodine area was significantly higher than that in iodine deficient area ( P < 0.05). Conclusion:The urinary iodine levels of children and pregnant women and the goiter rate of children in high iodine area of Shandong Province are significantly increased, and water iodine may be a key factor affecting the iodine nutrition status of the population.

Aim To observe the effect of lipopolysac-charide(LPS)induced supernatant of BV-2 cells on the inflammatory response and apoptosis of HT22 neu-rons.Methods After the concentration and time of LPS were determined by CCK-8 method,BV-2 cells were cultured with medium without LPS and medium containing LPS,the morphological changes of BV-2 microglia were observed by inverted microscope,and the CD86/CD206 ratio of BV-2 microglia was detected by immunofluorescence.Subsequently,BV-2 cell cul-ture supernatants were isolated and added to HT22 neuronal culture to observe the effect on the inflamma-tory response of HT22 neurons.The proliferation of HT22 neurons was detected by CCK-8 method and EdU method.The structural changes of HT22 neurons were observed under the microscope and examined by urani-um-lead staining.The levels of cytokines interleukin-1β(IL-1β),interleukin-10(IL-10),nuclear factor kappa-B(NF-κB)and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent as-say(Elisa).Neuronal apoptosis was detected by the TUNEL method.The protein expressions of Bax,Bcl-2 and inflammatory factors were detected by Western blot.Results After induction with 1 mg·L-1 LPS,BV-2 cells exhibited increased cell body size,thicker protrusions on both side,and some cells showed de-formed protrusions,the CD86/CD206 ratio in BV-2 cells decreased,promoting the transformation of BV-2 cells from M2 type to M1 type.After treating with the culture supernatant of BV-2 cells,HT22 neuronal cell activity and proliferation were reduced,axons short-ened,and the number of cells decreased.Neuronal cell bodies were enlarged and some cells were de-formed,with damaged cell membranes,round cell nu-clei but displaced nucleoli from the normal position,swollen mitochondria with vacuoles,reduced internal ridge structures,and increased levels of inflammatory factors NF-κB,IL-1 β,and TNF-α(P＜0.05 or P＜0.01),while the anti-inflammatory factor IL-10 de-creased(P＜0.05),protein expression of the pro-apoptotic indicator Bax increased(P＜0.01),and the protein expression of the anti-apoptotic indicator Bcl-2 decreased(P＜0.05).Conclusion After induction of BV-2 cell polarization by LPS,the supernatant could inhibit HT22 neuronal cell viability,upregulate inflam-matory factor expression and promote apoptosis.

Objective:To compare the survival outcomes of different subsequent treatment regimens in patients with locally advanced hypopharyngeal squamous cell carcinoma (HPSCC) who achieved partial response (PR) after neoadjuvant chemotherapy based on the gross tumor volume regression rate (GTVRR).Methods:This retrospective study included patients with locally advanced HPSCC treated at the Department of Head and Neck Surgery, Beijing Tongren Hospital, from January 2011 to December 2023. The cohort included 135 males and 3 females, aged from 35 to 77 years. All patients received 2-3 cycles of TPF regimen (paclitaxel+cisplatin+5-fluorouracil) neoadjuvant chemotherapy. Subsequent treatments included concurrent chemoradiotherapy or surgery combined with postoperative adjuvant radiotherapy. The impacts of different subsequent treatment modalities on the survivals and prognoses of patients were compared based on GTVRR thresholds of 50% and 70%. The χ 2 test was used to analyze influencing factors; survival analysis and intergroup comparisons were performed using the Kaplan-Meier method and Log-rank test; prognostic factors were assessed using univariate and multivariate Cox regression analyses. Results:The 5-year OS and PFS rates were 56.5% and 47.9%, respectively, while, the 10-year OS and PFS rates were 25.8% and 21.2%, respectively. The median OS was 75 months, and the median PFS was 48 months. The laryngeal function preservation rate for the entire cohort was 83.3%. The patients who underwent surgery combined with postoperative radiotherapy had significantly better OS and PFS outcomes than those treated with concurrent chemoradiotherapy ( P<0.05). Stratification based on GTVRR revealed that the surgery plus postoperative radiotherapy regimen was particularly effective for PR patients with a GTVRR of 30%-70%, showing significantly better OS and PFS compared to the concurrent chemoradiotherapy group ( P<0.05). Conclusion:The optimal subsequent treatment for PR-HPSCC may be surgery-based comprehensive treatment, particularly for patients with a GTVRR of 30%-70%. This study offers valuable insights for the stratified treatment of HPSCC, which could contribute to improving overall patient prognosis.

Objective To explore the mechanism of Luzhongjiangu Decoction in treating Osteonecrosis of the femoral head(ONFH).Methods Animal experiments were conducted to validate the therapeutic effect of Luzhongjiangu Decoction on ONFH.Network pharmacology was employed to screen potential targets,and molecular docking was used to verify the affinity between ligands and receptors.Fifty rats were randomly divided into blank group,model group,high-dose Luzhongjiangu Decoction groups,medium-dose Luzhongjiangu Decoction groups and low-dose Luzhongjiangu Decoction groups.After treatment,the therapeutic effect was evaluated by bone histomorphometry,bone histopathology and the expression of osteogenic related proteins.The serum metabolic differences were detected by UHPLC-QE-MS non-targeted metabonomics technique,followed by metabolic pathway enrichment analysis.Network pharmacology and metabolomics were integrated,and molecular docking was employed to further investigate the underlying mechanisms..Results The experimental results showed that Luzhongjiangu Decoction can significantly increase the expression of OCN,OPN and other osteogenic related factors,increase bone mineral density,alleviate the destruction of bone trabeculae,compared with that in the model group(P<0.01).The metabolomics test showed that compared with model group,Luzhongjiangu Decoction could callback 34 kinds of serum metabolic differences,involving 21 metabolic pathways,the main metabolic pathways were arachidonic acid metabolism,steroid hormone biosynthesis,primary bile acid biosynthesis,lysine degradation and so on.Network pharmacological results suggested that 185 active components and 1040 targets,551 disease targets and 76 intersection targets were obtained.The key pathways of Luzhongjiangu Decoction in the treatment of ONFH,which were analyzed by KEGG,involved HIF-1 signaling pathway,thyroid hormone signaling pathway,Calcium signaling pathway and so on.Integrated analysis showed that Luzhongjiangu Decoction may treat ONFH through seven metabolic pathways including arginine and proline metabolism,arachidonic acid metabolism,and steroid hormone biosynthesis;the molecular docking results also revealed correlation between potential pharmacodynamic substances and core target genes.Conclusion Luzhongjiangu Decoction can regulate the balance between proliferation and apoptosis of bone cells in a hypoxic environment by affecting the metabolic pathways such as arachidonic acid metabolism and HIF-1 signaling pathway,thereby exerting a therapeutic effect on osteonecrosis of the femoral head.

Aim To explore the effect of salvianolic acid B(SalB)on improving non-alcoholic fatty liver by regulating Lcn2.Methods In vivo experiments,8-week-old male C57BL/6J mice were fed with regular maintenance diet as the control group,while 8-week-old ApoE-/-mice were fed with high-fat diet and random-ly divided into the model group and SalB group.After eight weeks of feeding,mice in the SalB group were ga-vaged with SalB at a low dose of 15 mg·kg-1·d-1 and a high dose of 30 mg·kg-1·d-1,while mice in the control and model groups were gavaged with equal doses of normal saline.The results of liver RNA-seq revealed differentially expressed genes between the model group and the SalB group.HE staining and Oil Red O staining were used to observe pathological chan-ges in liver tissue.The kit was used to detect lipid,ox-idative stress,and inflammation in serum and liver tis-sue.RT-qPCR was employed to detect the mRNA lev-els of Lcn2,SREBP-1C,and enzymes related to lipid synthesis.In vitro experiments established a NAFLD model by inducing L02 and LX-2 cells with palmitic acid(PA)for 24 hours,and the effect of SalB on non-alcoholic fatty liver in vitro was detected by co treat-ment of SalB(30 pmol·L-1)and PA(0.2 mmol·L-1).The assay kit was used to detect the content of TC and TG in L02 cells,the cell oil red O staining to detect the accumulation of lipids in L02 cells,and RT-qPCR to detect the mRNA levels of Lcn2,SREBP-1 C,and genes related to lipid metabolism in LX-2 cells.Results The biochemical indicators of serum and liver in the model group were abnormal,with elevated levels of lipid,inflammation,and oxidative stress in liver tis-sue.Large areas of lipid vacuoles and deposition were observed,and PA induced L02 cells also exhibited sig-nificant lipid accumulation.These liver lesions were significantly improved after intervention with SalB.The effect of SalB on regulating lipid metabolism and in-flammation was found from RNA-seq.The mRNA lev-els of liver and LX-2 cells showed significant upregula-tion of Lcn2,SREBP-1 C,and enzymes related to lipid metabolism in the model group,and markedly downreg-ulation in the SalB group.Conclusions SalB can im-prove non-alcoholic fatty liver,and its mechanism may be related to down-regulating the expression of Lcn2,SREBP-1 C,and lipid synthase.