This study aims to reveal the effect and mechanism of Gentianella turkestanorum total extract(GTI) in ameliorating non-alcoholic steatohepatitis(NASH). UPLC-Q-TOF-MS was employed to identify the chemical components in GTI. SwissTarget-Prediction, GeneCards, OMIM, and TTD were utilized to screen the targets of GTI components and NASH. The common targets shared by GTI components and NASH were filtered through the STRING database and Cytoscape 3.9.0 to identify core targets, followed by GO and KEGG enrichment analysis. AutoDock was used for molecular docking of key components with core targets. A mouse model of NASH was established with a methionine-choline-deficient high-fat diet. A 4-week drug intervention was conducted, during which mouse weight was monitored, and the liver-to-brain ratio was measured at the end. Hematoxylin-eosin staining, Sirius red staining, and oil red O staining were employed to observe the pathological changes in the liver tissue. The levels of various biomarkers, including aspartate aminotransferase(AST), alanine aminotransferase(ALT), hydroxyproline(HYP), total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione(GSH), in the serum and liver tissue were determined. RT-qPCR was conducted to measure the mRNA levels of interleukin 1β(IL-1β), interleukin 6(IL-6), tumor necrosis factor α(TNF-α), collagen type I α1 chain(COL1A1), and α-smooth muscle actin(α-SMA). Western blotting was conducted to determine the protein levels of IL-1β, IL-6, TNF-α, and potential drug targets identified through network pharmacology. UPLC-Q-TOF/MS identified 581 chemical components of GTI, and 534 targets of GTI and 1 157 targets of NASH were screened out. The topological analysis of the common targets shared by GTI and NASH identified core targets such as IL-1β, IL-6, protein kinase B(AKT), TNF, and peroxisome proliferator activated receptor gamma(PPARG). GO and KEGG analyses indicated that the ameliorating effect of GTI on NASH was related to inflammatory responses and the phosphoinositide 3-kinase(PI3K)/AKT pathway. The staining results demonstrated that GTI ameliorated hepatocyte vacuolation, swelling, ballooning, and lipid accumulation in NASH mice. Compared with the model group, high doses of GTI reduced the AST, ALT, HYP, TC, and TG levels(P<0.01) while increasing the HDL-C, SOD, and GSH levels(P<0.01). RT-qPCR results showed that GTI down-regulated the mRNA levels of IL-1β, IL-6, TNF-α, COL1A1, and α-SMA(P<0.01). Western blot results indicated that GTI down-regulated the protein levels of IL-1β, IL-6, TNF-α, phosphorylated PI3K(p-PI3K), phosphorylated AKT(p-AKT), phosphorylated inhibitor of nuclear factor kappa B alpha(p-IκBα), and nuclear factor kappa B(NF-κB)(P<0.01). In summary, GTI ameliorates inflammation, dyslipidemia, and oxidative stress associated with NASH by regulating the PI3K/AKT/NF-κB signaling pathway.
Animals ; Non-alcoholic Fatty Liver Disease/genetics* ; Mice ; Network Pharmacology ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Chromatography, High Pressure Liquid ; Liver/metabolism* ; Mice, Inbred C57BL ; Humans ; Mass Spectrometry ; Tumor Necrosis Factor-alpha/metabolism* ; Disease Models, Animal ; Molecular Docking Simulation

Accurate identification of the glomerular ultrastructure is critical for the diagnosis of chronic kidney diseases,but the high cost of acquiring high-quality annotated data limits the application of fully-supervised learning.Therefore,a multi-class semi-supervised semantic segmentation framework based on segment anything model(MC4S-SAM)is proposed.After improving the mask decoder of segment anything model to enable multi-class semantic segmentation without requiring prompt information,the improved model is used to generate and refine pseudo-labels through a self-training strategy,and multi-level consistency regularization constraints are incorporated to enhance the model's performance.Experimental results show that,in the task of segmenting the glomerular mesangial ultrastructure,MC4S-SAM outperformes the fully-supervised model by 11.72%in mean intersection over union(mIoU)and 11.45%in mean Dice similarity coefficient(mDSC)when the labeled data accountes for 1/16 of the total.When the labeled data proportion is 1/4,the mIoU and mDSC reach 68.91%and 78.73%,respectively,demonstrating its significant potential for aiding the diagnosis of chronic kidney diseases.

Objective:To investigate the effects of area of the posterior malleolus fracture and injury to the distal tibiofibular syndesmosis on functional recovery of the ankle joint in Bartoní?ek type 2 ankle fractures.Methods:A retrospective analysis was conducted of the clinical data of 47 patients with Bartoní?ek type 2 ankle fracture who had been treated at Department of Orthopedics, Huashan Hospital Affiliated to Fudan University from January 2016 to January 2022. There were 22 males and 25 females with an age of (46.0±15.6) years. All patients were treated by open reduction and closed reduction. Pearson correlation analysis and multiple regression analysis were used to investigate the relationships respectively between the American Association of Foot and Ankle Surgeons (AOFAS) ankle-hindfoot scores at the last follow-up and the preoperative proportion of posterior ankle fracture area, and the anterior and posterior tibiofibular distances.Results:All patients were followed up for (17.2±0.6) months after surgery. The Pearson correlation analysis showed that the proportion of posterior ankle fracture area ( P=0.160) and the posterior tibiofibular distance ( P=0.078) were significantly correlated with the AOFAS ankle-hindfoot score at the last follow-up. There was no significant correlation between the anterior tibiofibular distance and the AOFAS ankle-hindfoot score at the last follow-up ( P=0.689). The multiple regression analysis showed that the proportion of posterior ankle fracture area ( P=0.043) and the posterior tibiofibular distance ( P=0.022) had significant negative effects on the AOFAS ankle-hindfoot score at the last follow-up. Conclusions:In Bartoní?ek type 2 ankle fractures, the proportion of posterior ankle fracture area and the posterior tibiofibular distance are important predictors for postoperative functional recovery of the ankle. Therefore, in Bartoní?ek type 2 ankle fractures, surgical indications for the posterior malleolar fracture depend not only on the size of the fracture, but also on whether the fracture involves the lower tibiofibular syndesmosis.

Accurate identification of the glomerular ultrastructure is critical for the diagnosis of chronic kidney diseases,but the high cost of acquiring high-quality annotated data limits the application of fully-supervised learning.Therefore,a multi-class semi-supervised semantic segmentation framework based on segment anything model(MC4S-SAM)is proposed.After improving the mask decoder of segment anything model to enable multi-class semantic segmentation without requiring prompt information,the improved model is used to generate and refine pseudo-labels through a self-training strategy,and multi-level consistency regularization constraints are incorporated to enhance the model's performance.Experimental results show that,in the task of segmenting the glomerular mesangial ultrastructure,MC4S-SAM outperformes the fully-supervised model by 11.72%in mean intersection over union(mIoU)and 11.45%in mean Dice similarity coefficient(mDSC)when the labeled data accountes for 1/16 of the total.When the labeled data proportion is 1/4,the mIoU and mDSC reach 68.91%and 78.73%,respectively,demonstrating its significant potential for aiding the diagnosis of chronic kidney diseases.

Objective:To investigate the effects of area of the posterior malleolus fracture and injury to the distal tibiofibular syndesmosis on functional recovery of the ankle joint in Bartoní?ek type 2 ankle fractures.Methods:A retrospective analysis was conducted of the clinical data of 47 patients with Bartoní?ek type 2 ankle fracture who had been treated at Department of Orthopedics, Huashan Hospital Affiliated to Fudan University from January 2016 to January 2022. There were 22 males and 25 females with an age of (46.0±15.6) years. All patients were treated by open reduction and closed reduction. Pearson correlation analysis and multiple regression analysis were used to investigate the relationships respectively between the American Association of Foot and Ankle Surgeons (AOFAS) ankle-hindfoot scores at the last follow-up and the preoperative proportion of posterior ankle fracture area, and the anterior and posterior tibiofibular distances.Results:All patients were followed up for (17.2±0.6) months after surgery. The Pearson correlation analysis showed that the proportion of posterior ankle fracture area ( P=0.160) and the posterior tibiofibular distance ( P=0.078) were significantly correlated with the AOFAS ankle-hindfoot score at the last follow-up. There was no significant correlation between the anterior tibiofibular distance and the AOFAS ankle-hindfoot score at the last follow-up ( P=0.689). The multiple regression analysis showed that the proportion of posterior ankle fracture area ( P=0.043) and the posterior tibiofibular distance ( P=0.022) had significant negative effects on the AOFAS ankle-hindfoot score at the last follow-up. Conclusions:In Bartoní?ek type 2 ankle fractures, the proportion of posterior ankle fracture area and the posterior tibiofibular distance are important predictors for postoperative functional recovery of the ankle. Therefore, in Bartoní?ek type 2 ankle fractures, surgical indications for the posterior malleolar fracture depend not only on the size of the fracture, but also on whether the fracture involves the lower tibiofibular syndesmosis.

AIM: To investigate the expression changes of MMP-12 during the long-term axon regeneration induced by the lens injury after the optic nerve clamp trauma in sprague-dawley(SD)rats.METHODS: The optic nerve injury model and lens injury model of SD rats were established, and the 24 experimental animals were divided into control group; lens injury group; optic nerve injury group; lens injury combined with optic nerve injury group, with 6 rats in each group. Reference transcriptome sequencing was used to analyze the expression changes of differentially expressed genes in the injured optic nerve region, and relevant differentially expressed genes with high expression were screened. Quantitative real-time polymerase chain reaction(qRT-PCR)and enzyme-linked immunosorbent assay(ELISA)were used to quantify the expression changes of matrix metalloproteinase-12(MMP-12)in the injured optic nerve region.RESULTS: The Principal Component Analysis of transcriptome sequencing indicated that lens injury combined with optic nerve injury was the principal component of gene expression change. Analysis of gene expression differences showed that the expression of MMP-12 gene was up-regulated in the lens injury combined with optic nerve injury group. The mRNA expression level of MMP-12 in the lens injury combined optic nerve injury group was up-regulated compared with the control group, the optic nerve injury group and the lens injury group at 14d and 21d after successful modeling(P<0.05). At 7, 28d, there was no difference in expression among all groups. The protein expression level of MMP-12 in the lens injury combined with optic nerve injury group was up-regulated compared with the control group and optic nerve injury group at 7, 14 and 21d after successful modeling(P<0.05), and it was up-regulated in the lens injury group combined with optic nerve injury group compared with optic nerve injury group at 21d(P<0.05). At 28d, there was no difference in expression among all groups.CONCLUSION: The up-regulated expression of MMP-12 may be involved in the long-term regeneration of the optic nerve after lens injury.

Morphine is a frequently used analgesic that activates the mu-opioid receptor(MOR),which has prominent side effects of tolerance.Although the inefficiency of morphine in inducing the endocytosis of MOR underlies the development of morphine tolerance,currently,there is no effective therapy to treat morphine tolerance.In the current study,we aimed to develop a monoclonal antibody(mAb)precisely targeting MOR and to determine its therapeutic efficacy on morphine tolerance and the underlying molecular mechanisms.We successfully prepared a mAb targeting MOR,named 3A5C7,by hybridoma technique using a strategy of deoxyribonucleic acid immunization combined with cell immunization,and identified it as an immunoglobulin G mAb with high specificity and affinity for MOR and binding ability to antigens with spatial conformation.Treatment of two cell lines,HEK293T and SH-SY5Y,with 3A5C7 enhanced morphine-induced MOR endocytosis via a G protein-coupled receptor kinase 2(GRK2)/β-arrestin2-dependent mechanism,as demonstrated by immunofluorescence staining,flow cytometry,Western blotting,coimmunoprecipitation,and small interfering ribonucleic acid(siRNA)-based knock-down.This mAb also allowed MOR recycling from cytoplasm to plasma membrane and attenuated morphine-induced phosphorylation of MOR.We established an in vitro morphine tolerance model using differentiated SH-SY5Y cells induced by retinoic acid.Western blot,enzyme-linked immunosorbent assays,and siRNA-based knockdown revealed that 3A5C7 mAb diminished hyperactivation of adenylate cyclase,the in vitro biomarker of morphine tolerance,via the GRK2/β-arrestin2 pathway.Furthermore,in vivo hotplate test demonstrated that chronic intrathecal administration of 3A5C7 significantly alle-viated morphine tolerance in mice,and withdrawal jumping test revealed that both chronic and acute 3A5C7 intrathecal administration attenuated morphine dependence.Finally,intrathecal electroporation of silencing short hairpin RNA illustrated that the in vivo anti-tolerance and anti-dependence efficacy of 3A5C7 was mediated by enhanced morphine-induced MOR endocytosis via GRK2/β-arrestin2 pathway.Collectively,our study provided a therapeutic mAb,3A5C7,targeting MOR to treat morphine tolerance,mediated by enhancing morphine-induced MOR endocytosis.The mAb 3A5C7 demonstrates promising translational value to treat clinical morphine tolerance.

Objective:To analyze the treatment effects and side effects of non-surgical comprehensive treatment for locally advanced hypopharyngeal carcinoma invading cervical esophagus.Methods:A retrospective analysis was performed on sixty-six patients with locally advanced hypopharyngeal carcinoma invade the esophagus. These patients were treated in the Department of Otolaryngology, Head and Neck Surgery of Chinese People′s Liberation Army General Hospital between January 2011 and May 2022, including sixty-five males and one female, aged 43-71 years. Treatment regimen consisted of induction chemotherapy and concurrent chemoradiothrapy and epidermal growth factor receptor (EGFR)-targeted therapy, three of these cases were treated with programmed cell death 1 (PD-1) immunotherapy. The Kaplan-Meier method was used for survival analysis. Side effects were evaluated with the established CTCAE (Common Terminology Criteria for Adverse Events) 5.0 criteria. The factors affecting prognosis were analyzed by Cox multivariate regression analysis.Results:Sixty-four (97.0%, 64/66) patients completed the radiotherapy and chemotherapy plan. The most common grade three side effects were radioactive oropharyngeal mucositis (89.1%, 57/64) and leukopenia (23.4%, 15/64). Five (7.8%, 5/64) patients showed grade three hoarseness; two patients (3.1%, 2/64) suffered from grade three swallowing dysfunction and required feeding tube and intravenous nutrition; the remaining patients(89.1%) retained good vocal and swallowing functions. The overall survival (OS) of all patients was 81.5% after one year, 54.0% after three years, and 39.9% after five years; the progression-free survival (PFS) was 78.3% after one year, 54.9% after three years, and 42.6% after five years; local control rate (LCR) was 80.9% after one year, 62.5% after three years, and 52.0% after five years. T4a patients showed better OS, PFS and LCR than T4b patients, with statistically significant differences (χ 2=8.10, 8.27, and 6.64, respectively, all P<0.05). Cox multivariate regression analysis showed that lymph node metastasis was an independent factor affecting prognosis (χ 2=10.21, P<0.05). Conclusion:Non-surgical comprehensive treatment can provide with another option of radical treatment for locally advanced hypopharyngeal carcinoma with cervical esophagus invasion, offering the patients higher rate of larynx and esophageal preservation with tolerable side effects.

Sodium salicylate is an anti-inflammatory medication with a side-effect of tinnitus. Here, we used mouse cochlear cultures to explore the effects of salicylate treatment on cochlear inner hair cells (IHCs). We found that IHCs showed significant damage after exposure to a high concentration of salicylate. Whole-cell patch clamp recordings showed that 1-5 mmol/L salicylate did not affect the exocytosis of IHCs, indicating that IHCs are not involved in tinnitus generation by enhancing their neuronal input. Instead, salicylate induced a larger peak amplitude, a more negative half-activation voltage, and a steeper slope factor of Ca²⁺ current. Using noise analysis of Ca²⁺ tail currents and qRT-PCR, we further found that salicylate increased the number of Ca²⁺ channels along with Ca_V1.3 expression. All these changes could act synergistically to enhance the Ca²⁺ influx into IHCs. Inhibition of intracellular Ca²⁺ overload significantly attenuated IHC death after 10 mmol/L salicylate treatment. These results implicate a cellular mechanism for tinnitus generation in the peripheral auditory system.
Animals ; Calcium ; Exocytosis ; Hair Cells, Auditory, Inner ; Mice ; Sodium Salicylate/pharmacology* ; Tinnitus/chemically induced*

OBJECTIVE: To compare the curative effect on diarrhea-predominant irritable bowel syndrome (IBS-D) between acupuncture for regulating METHODS: A total of 231 patients with IBS-D were randomized into an acupuncture group (154 cases) and a western medication group (77 cases) at the ratio of 2 to 1. In the acupuncture group, acupuncture was applied to acupoint regimen for regulating RESULTS: After treatment and in follow-up, the total scores of IBS-SSS in the patients of the two groups were all reduced as compared with those before treatment ( CONCLUSION Acupuncture for regulating
Acupuncture Therapy ; Diarrhea/therapy* ; Humans ; Irritable Bowel Syndrome/therapy* ; Quality of Life ; Serotonin Plasma Membrane Transport Proteins/genetics* ; Spleen ; Treatment Outcome