ObjectiveThis study aims to systematically analyze the blood-absorbed components and metabolic profiles of Xiebaisan（XBS） in normal and chronic bronchitis （CB） mice using ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS）， while comparing differences between the two states. MethodsThirty female BABL/c mice were randomly divided into the normal group， the normal drug administration group， the CB group， the CB drug administration group and the dexamethasone group， with 6 mice in each group. The CB mouse model was established by inducing with ovalbumin （OVA）. The mice in the normal drug administration group and the CB drug administration group started to be gavaged with XBS（13.2 g·kg^-1） from the 21^st day， and the dexamethasone group mice were simultaneously gavaged with dexamethasone （0.5 mg·kg^-1） until the end of the 35^th day of the experiment. Subsequently， serum samples were collected and evaluated for their efficacy， based on the pharmacological evaluation indicators， to determine the efficacy of XBS in treating CB. Then the UPLC-Q-Exactive Orbitrap MS was employed to identify and analyze the chemical constituents， blood-absorbed components， and metabolites of XBS. Chemometric analysis was conducted to reveal metabolic profile differences under "dual states". Concurrently， Real-time PCR technology was utilized to detect the expression levels of key liver metabolic enzymes CYP2E1， CYP3A1， UGT1A1， and UGT1A6. ResultsA total of 28 prototype components and 158 metabolites （including 48 phase Ⅰ metabolites and 110 phase Ⅱ metabolites） of XBS were unambiguously identified in the serum of normal mice. Additionally， a comprehensive characterization was performed on a total of 32 prototype components and 178 metabolites （including 50 phase Ⅰ metabolites and 128 phase Ⅱ metabolites） of XBS in the serum of CB mice. Among them， 27 prototype components were detected in both states， including 12 flavonoids， 2 alkaloids， 3 triterpenes， 4 organic acids， 3 amides， 1 stilbene and 2 other compounds. The chemometrics analysis revealed no significant difference in the prototype components and metabolites of XBS between normal and CB mice； however， there was a significant increase in the in-vivo exposure of XBS in CB mice. Compared to normal mice， the levels of phase Ⅰ metabolites such as oxidation， reduction and methylation of blood components of XBS as well as phase Ⅱ metabolites of glucuronidation showed significant changes in CB mice. Real-time PCR further confirmed that these alterations were attributed to the upregulation of CYP2E1 （P<0.05）， CYP3A1 （P>0.05）， UGT1A1 （P<0.01） and UGT1A6 （P<0.01） enzymes expression in the liver of CB mice. ConclusionThis study elucidated the disparities in the levels of the blood-absorbed components and metabolic profiles of XBS in normal and CB mice， especially in oxidation， reduction， methylation in phase Ⅰ metabolism and glucoaldehyde acidification in phase Ⅱ metabolism. And there are related to the differences in the expression levels of phase Ⅰ and phase Ⅱ metabolic enzymes CYP2E1， CYP3A1， UGT1A1 and UGT1A6 in the liver.

ObjectiveTo investigate the effects of modified Xiaoyaosan （JJXYS） on behavioral abnormalities and hippocampal mitochondrial quality control （MQC） in the rat model of post-myocardial infarction depression （PMD） and preliminarily explore its potential mechanism. MethodsA rat model of PMD was established by left anterior descending coronary artery ligation combined with chronic unpredictable mild stress （CUMS）. Rats were randomized into a control group， a model group， a fluoxetine （FLX， 10 mg·kg^-1） group， and low-， medium-， and high-dose JJXYS （JJXYS-L/M/H， 1.12， 2.24， 4.48 g·kg^-1， respectively） groups. Depressive-like behaviors were evaluated by body weight monitoring， sucrose preference test， open field test， and forced swimming test. Hematoxylin-eosin staining and Nissl staining were used to observe hippocampal histomorphology and neuronal changes. Enzyme-linked immunosorbent assay was conducted to determine the serum levels of 5-hydroxytryptamine （5-HT）， dopamine （DA）， interleukin-1 beta （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-alpha （TNF-α）. The mRNA levels of MQC-related genes including peroxisome proliferator-activated receptor-gamma coactivator-1 alpha （PGC-1α）， nuclear respiratory factor 1 （Nrf1）， and transcription factor A， mitochondrial （TFAM） in the hippocampal tissue were measured by real-time PCR. The expression of proteins related to the dynamin-related protein 1 （Drp1）/PTEN-induced putative kinase 1 （PINK1）/Parkin signaling pathway was determined by Western blot. ResultsCompared with the control group， the model group showed restricted body weight gain， aggravated depressive-like behaviors， declined serum 5-HT and DA levels， evident hippocampal neuronal damage and reduced Nissl bodies， as well as downregulated expression of MQC-related genes and proteins （P<0.05）. Compared with the model group， both FLX and JJXYS alleviated the above changes to varying degrees. Moreover， the JJXYS-M and JJXYS-H groups showed more pronounced effects， improving behavioral performance， restoring 5-HT and DA levels， alleviating hippocampal pathological injury， and upregulating the expression of PGC-1α/Nrf1/TFAM mRNA and Drp1/PINK1/Parkin signaling pathway-related proteins （P<0.05）. ConclusionJJXYS can significantly alleviate depressive-like behaviors and neurotransmitter imbalance in the rat model of PMD by regulating hippocampal MQC and upregulating the Drp1/PINK1/Parkin-related pathway. This study provides experimental evidence for the intervention of PMD with JJXYS.

Objective: To investigate the assocation of sedentary behavior among college students on psychological health issues, such as depressive, anxiety, and stress symptoms, and to analyze the moderating role of takeaway consumption behavior in the context, in order to provide a scientific basis for reducing emotional symptoms among college students. Methods: A stratified cluster sampling method was employed to conduct a questionnaire on 3 427 first year students of a higher education institution in Hefei of Anhui Province from May to June 2021. The study variables included demographic characteristics, sedentary time, takeaway consumption behavior, and emotional (symptoms depressive, anxiety, and stress symptoms). The Spearman correlation analysis was used to analyze the association between variables, and linear regression analysis was used to analyze the association between takeaway consumption behavior and depressive, anxiety and stress symptoms among college students with sedentary time. Results: Both sedentary time and takeaway consumption behavior were positively correlated with depressive, anxiety and stress symptoms among college students ( r =0.10, 0.10, 0.10; 0.10, 0.11, 0.11, all P <0.05). The results of linear regression analysis showed that the interaction term between takeaway consumption behavior and sedentary time was positively correlated with symptoms of depressive, anxiety, and stress symptoms among college students (depression: β =0.04, anxiety: β =0.04, stress: β =0.04, all P <0.05). The results of the simple slope test demonstrated that regardless of the level of takeaway consumption behavior, sedentary time was positively correlated with the depressive symptoms of college students; compared with low takeaway consumption behavior, high takeaway consumption behavior ( β=0.77, P <0.01) enhanced the association between sedentary time and depressive symptoms among college students. In addition, under the condition of high takeaway consumption behavior, sedentary time was positively correlated with the anxiety and stress symptoms of college students (anxiety: β =0.64; stress: β =0.71, both P <0.01); while under the condition of low takeaway consumption behavior, sedentary time was not related to the anxiety and stress symptoms of college students ( β =0.17, 0.22, both P >0.05). Conclusions Sedentary behavior is related to a the emotional symptoms of depressive, anxiety, and stress among college students. Takeaway consumption behavior may exacerbate this impact.

Objective: To establish anin vitromodel of ferroptosis induced by Erastin and RAS-selective lethal 3(RSL3) in hepatoma cells, and to provide theoretical basis for the development of novel therapeutic strategies for HCC. Methods: Hepatoma cells(HCCLM3, HepG2, Hep3B, Huh7 and PLC/PRF/5) in logarithmic growth phase were treated with Erastin(0-40 μmol/L) and RSL3(0-10 μmol/L) at double concentrations respectively. After 24 h, CCK-8 method was used to detect cell viability, draw growth curve, calculate IC50, and HCC cells sensitive to inducers were selected for follow-up experiments. The effect of inducer on the state of hepatoma cells was observed under light microscope, and immunoblotting and flow cytometry were used to verify whether the ferroptotic modelin vitrowas successfully constructed. Results: Huh7, Hep3B and HepG2 cells were sensitive to Erastin and RSL3, but HCCLM3 and PLC/PRF/5 were insensitive to Erastin and RSL3. When the concentration of Erastin and RSL3 reached the maximum, the survival rate was still above 65%. Huh7, Hep3B and HepG2 cells were selected for subsequent experiments. Compared with the control group, the expression of Glutathione peroxidase 4(GPX4), a ferroptotic marker, was down-regulated in a concentration-dependent manner. In Huh7, Hep3B and HepG2 cells, lipid reactive oxygen species(ROS) levels significantly increased after 24 h treatment with 10 μmol/L and 20 μmol/L Erastin, respectively; in Huh7 cells, lipid ROS levels significantly increased after 24 h treatment with 0.5 μmol/L and 1 μmol/L RSL3, respectively; in Hep3B and HepG2 cells, lipid ROS levels significantly increased after 24 h treatment with 1 μmol/L and 2 μmol/L RSL3, respectively, compared with control group. Conclusion Huh7, Hep3B and HepG2 cells are highly sensitive to Erastin and RSL3. Huh7, Hep3B and HepG2 cells treated with 10 μmol/L Erastin for 24 h are good models for simulating ferroptosis induced by Erastinin vitro, Huh7 cells treated with 0.5 μmol/L RSL3 for 24 h and Hep3B and HepG2 cells treated with 1 μmol/L RSL3 for 24 h are good models for simulating ferroptosis induced by RSL3in vitro.

OBJECTIVES: To explore the effects of exogenous trigger (hCG/GnRHa) versus endogenous LH surge in natural cycle IVF/ICSI (NC-IVF/ICSI) for patients with diminished ovarian reserve (DOR). METHODS: A retrospective analysis was conducted on 1,118 NC-IVF/ICSI cycles from two reproductive centers between 2013 and 2024. Propensity score matching (PSM) and multivariate logistic regression were used to adjust for confounding factors. The trigger-day hormone threshold was determined using receiver operating characteristic (ROC) curve analysis. Outcome measures included oocyte retrieval rate, 2PN fertilization rate, clinical available embryo rate, high-quality embryo rate, fresh cycle clinical pregnancy rate (CPR), and live birth rate (LBR). RESULTS: After adjusting for confounders via PSM and logistic regression, the exogenous trigger group demonstrated significantly better outcomes across all the evaluated parameters (oocyte retrieval rate, 2PN fertilization rate, transferable embryo rate, high-quality embryo rate, fresh cycle CPR, and LBR) than the endogenous LH surge group (P<0.05). Age-stratified analysis revealed that for the entire cohort, exogenous triggering significantly increased the number of transferable embryos and high-quality embryos (P<0.001). In the 35-39 years old subgroup, exogenous triggering showed significant advantages in oocyte yield, high-quality embryo rate, CPR, and LBR (P<0.05) and resulted in the most pronounced improvement in LBR (OR=6.25, 95% CI: 1.34-29.23). ROC analysis established a decision-day LH threshold of 19.055 mIU/mL (AUC=0.945, specificity=93.3%) for precise stratification of the clinical pathways. CONCLUSIONS For DOR patients undergoing NC-IVF/ICSI, exogenous triggering comprehensively improves the treatment outcomes, particularly providing significant live birth benefits for women aged 35-40 years. An individualized protocol incorporating the LH threshold (19.055 mIU/mL) effectively enhances embryonic developmental potential and live birth rates.
Humans ; Female ; Ovarian Reserve ; Pregnancy ; Propensity Score ; Retrospective Studies ; Fertilization in Vitro ; Sperm Injections, Intracytoplasmic ; Chorionic Gonadotropin ; Pregnancy Rate ; Logistic Models ; Ovulation Induction/methods* ; Gonadotropin-Releasing Hormone ; Adult ; Oocyte Retrieval

Objective: To investigate the role of butyric acid-producing bacteria in long bone homeostasis in mice with periodontitis under a high-fat/high-sugar diet and to provide new insights for the prevention and treatment of periodontitis and related bone metabolic diseases. Methods: This study has been approved by the Animal Welfare and Ethics Committee of the Experimental Animal Center. Initially, 14 mice were randomly divided into the CON group (the control group) and the LIG group (the periodontitis group). Mice in the LIG group had experimental periodontitis induced by ligating the second maxillary molars bilaterally and were fed a high-fat and high-sugar diet. After 8 weeks, samples were collected. Micro-computed tomography (Micro-CT) was used to analyze alveolar bone resorption and various parameters of the proximal tibia trabecular bone, including bone mineral density (BMD), bone volume per tissue volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp). After decalcification, hematoxylin and eosin (HE) staining was performed on maxillary bone sections to assess periodontal tissue inflammation and connective tissue destruction. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect related genes in the distal femur and proximal tibia bone tissues, including osteocalcin (OCN), osteogenic transcription factor (Osterix), osteoprotegerin (OPG), tartrate resistant acid phosphatase (TRAP), osteoclast-associated receptor (OSCAR), receptor activator of nuclear factor kappa-B (RANK), and receptor activator of nuclear factor kappa-B ligand (RANK-L). Subsequently, the other 28 mice were randomly divided into the CON group (the control group), LIG group (the periodontitis group), CON + butyric acid-producing bacteria (BP) group, and LIG + BP group. The breeding, sampling, and sample detection methods remained the same. Finally, the other 28 mice were randomly divided into the CON group (the control group), LIG group (the periodontitis group), CON + sodium butyrate (SB) group, and LIG + SB group. The breeding, sampling, and sample detection methods remained the same. Results: ①Periodontitis modeling was successful. Compared with the CON group, the LIG group exhibited significant alveolar bone resorption of the maxillary second molar, aggravated periodontal tissue inflammation, and connective tissue destruction. ②Periodontitis exacerbated long bone resorption in mice fed a high-fat high-sugar diet. Compared with the CON group, the LIG group had significantly lower BMD, BV/TV, Tb.N, and Tb.Th (P<0.05), and significantly higher Tb.Sp (P<0.05). HE staining of the proximal tibia showed that the trabeculae in the LIG group were sparse and disordered, with some areas showing fractures or dissolution. The expression of osteoblast markers (OCN, Osterix, OPG) was significantly lower in the LIG group (P<0.05), while the expression of the osteoclast marker TRAP showed an increasing trend (P>0.05). The ratio of RANK-L/OPG was significantly higher in the LIG group compared with the CON group (P<0.05). ③ Supplementation with butyric acid-producing bacteria alleviates periodontitis-induced disruption of long bone homeostasis in mice fed a high-fat/high-sugar diet. Compared with the LIG group, BMD and Tb.Th were significantly higher in the LIG + BP group. HE staining of the proximal tibia showed that bone resorption was mitigated in the LIG + BP group compared with the LIG group. The expression of OCN and Osterix was significantly higher in the LIG + BP group, while the expression of osteoclast-specific genes (OSCAR, RANK, RANK-L) was significantly lower (P<0.05). ④ Supplementation with butyrate alleviates periodontitis-induced disruption of long bone homeostasis in mice fed a high-fat/high-sugar diet. Compared with the LIG group, BV/TV and Tb.N were significantly higher in the LIG + SB group, and Tb.Sp was significantly lower (P<0.05). HE staining of the proximal tibia showed that bone resorption was mitigated in the LIG + SB group compared with the LIG group. The expression of Osterix, OPG, OSCAR, TRAP, and RANK was significantly lower in the LIG + SB group compared with the LIG group (P<0.05). Conclusion Periodontitis disrupts the long bone homeostasis of mice fed a high-fat high-sugar diet, aggravating long bone resorption. Supplementation with butyric acid-producing bacteria or butyrate can effectively alleviate the disruption of long bone homeostasis caused by periodontitis.

BACKGROUND:Lumbar disc degeneration is a common disease that causes lower back pain and lower limb neurological symptoms.The balance of collagen metabolism plays an important role in maintaining the stability of the intervertebral discs.OBJECTIVE:To review the research progress in the imbalance of collagen metabolism in intervertebral disc degeneration.METHODS:The first author searched for relevant literature published before May 2024 in CNKI,PubMed and Web of Science databases.Search terms were"degenerative disc disease,""collagen metabolism,""collagenase family,""collagen synthesis related factors,"and"collagen breakdown related factors"in Chinese and English.Seventy-six articles were finally included for review.RESULTS AND CONCLUSION:In the process of intervertebral disc degeneration,the balance of collagen metabolism plays a crucial role in maintaining the stability of the normal intervertebral disc.When intervertebral disc degeneration occurs,a large amount of pro-inflammatory factors,collagenase,and oxidative stress reactions occur in the intervertebral disc,which increases the breakdown of collagen in the intervertebral disc.At the same time,it inhibits the generation of growth factors,collagen synthase,and collagen synthesis-related factors,resulting in a decrease in collagen synthesis in the intervertebral disc.The combined effect of the above two conditions disrupts the balance of collagen metabolism in the intervertebral disc,further exacerbating the process of intervertebral disc degeneration.

OBJECTIVE:Dry needling therapy is widely used in various myofascial pain syndromes.The purpose of this study was to systematically evaluate the clinical effect of dry needling in the treatment of knee joint diseases.METHODS:PubMed,EBSCO,ScienceDirect,Web of Science,CINAHL,Cochrane Library,CNKI and other databases were searched for relevant literature.Randomized controlled trials with dry needling as the main treatment method and patients diagnosed with knee joint disease were selected.Two evaluators independently screened the articles,scored the methodological quality,and extracted the data.The main indicators were visual analog scale score,and the secondary indicators were the Western Ontario McMaster Universities(WOMAC)score,pressure pain threshold,knee mobility and Kujala score(knee function score scale).RESULTS:A total of 15 randomized controlled trials involving 698 patients were included in the Meta-analysis.The results showed that compared with non-dry needling,dry needling at myofascial trigger points had a significant advantage in visual analog scale score,the WOMAC pain score and WOMAC stiffness score[MD=-0.63,95%confidence interval(CI):-1.06 to-0.19,P=0.005;MD=-0.74,95%CI:-1.32 to-0.17,P=0.01;MD=-0.43,95%CI:-0.77 to-0.09,P=0.01).However,there was no significant advantage in WOMAC total score,WOMAC functional score,pressure pain threshold,knee mobility,and Kujala score.CONCLUSION:Dry needling can effectively treat knee pain and stiffness;however,the clinical advantages of dry needling in improving other knee joint dysfunction and its follow-up effect are not well documented.Therefore,in some older patients with chronic knee pain or joint stiffness,the use of dry needling can be carefully considered.

Objective: To investigate the inhibitory effect of Elesclomol (ES) + Cu2 + on the proliferation of human hepatoma cell lines PLC/PRF/5 and Huh-7 and its potential to induce Cuproptosis. Methods: Human hepatoma cell lines PLC/PRF/5 and Huh_7 cells were Cultured in vitro. ES solution , Cu2 + solution and copper chelating agent ammonium tetrathiomolybdate VI (ATTM) solution was treated separately or in combination. The effect of ES + Cu2 + on the survival rate of human hepatoma cell lines PLC/PRF/5 and Huh_7 cells and the effect of ES + Cu2 + on the survival rate after pretreatment with copper chelating agent ATTM were evaluated using CCK_8 kit. The cell death induced by ES + Cu2 + was detected by flow cytometry and the changes of ES + Cu2 + after pretreatment with copper chelating agent ATTM. The expression of Cuproptosis related proteins ATPase copper transporting beta (ATP7B) ,ferredoxin 1 (FDX1) , dihydrolipoamide s_acetyltransferase(DLAT) and superoxide dismutase 1 (SOD1) were detected by Western blot. The effect of ES + Cu2 + on cell proliferation and the reverse effect after ATTM pretreatment was detected by cell scratch assay. Results: The toxicity of ES + Cu2 + to human hepatocellular carcinoma cell lines PLC/PRF/5 and Huh_7 was significantly dose_dependent (P < 0. 05) . Compared with the control group , the combined application of ES and Cu2 + had a more significant inhibitory effect on hepatocellular carcinoma cells than ES or Cu2 + alone (P < 0. 05) , and copper chelating agent ATTM could reverse the inhibitory effect of ES + Cu2 + on hepatocellular carcinoma cells (P < 0. 05) . Flow cytometry results showed that compared with the control group , the proportion of cell death in PLC/PRF/5 and Huh_7 cells treated with ES + Cu2 + increased , while the proportion of cell death decreased after ATTM intervention (P < 0. 05) . The results of cell scratch test showed that the migration ability of PLC/PRF/5 and Huh_7 cells was decreased after ES + Cu2 + treatment , however, the addition of ATTM reversed the inhibitory effect of ES + Cu2 + on cell migration (P < 0. 05) . Compared with the control group , the expression levels of copper death related proteins ATP7B , FDX1 , DLAT and SOD1 decreased after ES + Cu2 + treatment , but the addition of ATTM reversed the expression trend of these proteins (P < 0. 05) . Conclusion The combination of ES and Cu2 + can effectively inhibit the proliferation and migration of PLC/PRF/5 and Huh_7 of hepatocellular carcinoma cells , and induce Cuproptosis , which provides a new strategy for the treatment of hepatocellular carcinoma.

Objective:To develop a deep learning-based system for real-time recognition and classification of portal hypertensive gastropathy (PHG) and evaluate its ability to assist junior endoscopists.Methods:A total of 2 848 gastroscopy images from 832 patients with liver cirrhosis were selected from Digestive Endoscopy Center databases of Renmin Hospital of Wuhan University, Wuhan Hospital of Traditional Chinese and Western Medicine, and the Second Hospital of Jingzhou from January 2015 to October 2023. This system referred to 3 endoscopic features of Baveno Ⅱ scoring system. Three models were developed respectively for gastric antral vascular ectasia (GAVE), mosaic-like pattern (MLP), and red marks (RM). The specific classification references were as follows: (1) GAVE model: 0 no, 1 yes; (2) MLP model: 0 no, 1 mild, 2 severe; (3) RM model: 0 no, 1 isolated, 2 fused. The classification results for endoscopic characteristics of PHG of 3 endoscopy experts were taken as the gold standard. The yolov8-m model was used for training. The training dataset, validation dataset, and test dataset were allocated at a ratio of 8∶1∶1. The test dataset was used to evaluate the performance of models and their auxiliary effects on endoscopists. The accuracy, recall, precision, specificity and Kappa coefficient were calculated. Results:The accuracy, recall, specificity of GAVE model were 96.0% (48/50), 87.5% (7/8) and 97.6% (41/42). There was no significant difference between its accuracy and the gold standard ( χ2=316.226, P=1.000). The precision of GAVE1 and GAVE0 were 87.5% (7/8) and 97.6% (41/42) respectively. The accuracy of MLP model was 84.1% (132/157), and there was no significant difference compared with the gold standard ( χ2=3.286, P=0.193). The precision and recall of MLP2 were 88.2% (15/17) and 75.0% (15/20). The precision and recall of MLP1 were 77.9% (60/77) and 88.2% (60/68). The precision and recall of MLP0 were 90.5% (57/63) and 82.6% (57/69). The accuracy of RM model was 87.9% (123/140), and there was no significant difference compared with the gold standard ( χ2=2.891, P=0.409). The precision and recall of RM2 were 94.7% (18/19) and 78.3% (18/23). The precision and recall of RM1 were 72.2% (26/36) and 81.3% (26/32). The precision and recall of RM0 were 92.9% (79/85) and 92.9% (79/85). The mean accuracy of the three junior endoscopists, with and without the assistance of the GAVE model, MLP model, and RM model, respectively increased from 95.3% to 99.3%, from 83.9% to 91.9%, and from 81.9% to 83.1%. The overall consistency analysis of the 3 junior endoscopists with the gold standard indicated that the consistency of the GAVE model before and after assistance was extremely strong (both an overall Kappa of 1.000); the consistency before assistance of the MLP model was moderate (with an overall Kappa of 0.601), which increased to extremely strong after assistance (with an overall Kappa of 0.964); and the consistency of the RM model before and after assistance was also relatively strong (with an overall Kappa of 0.792 before and 0.798 after). Conclusion:The deep learning system accurately identifies and classifies PHG features and significantly enhances diagnostic performance of junior endoscopists.