Objectives:To explore the impact of Gli1(gioma-associated oncogene homolog)on nucleus pulposus cells(NPCs)and its role in the fibrotic changes of the nucleus within intervertebral discs.Methods:A rat tail intervertebral disc puncture model was established.Histological staining was used to evaluate the levels of intervertebral disc degeneration and fibrotic changes in the untreated(negative control,NC)group and the puncture(puncture-induced intervertebral disc degeneration,PIDD)group.Western blot was used to assess the expression levels of Gli1 protein in the tissues.Gli1 protein was engineered to overexpress by lentivirus in NPCs and verified.The experiment was divided into three groups:control(Blank),Gli1 overexpression(Lv-Gli1),and virus control(lv-Ctrl).Immunofluorescence and Western blot were used to detect the expression levels of fibrotic markers FAP and FSP-1.Adding Gli1 inhibitor GANT61,cellular experiment was grouped into control(blank),Gli1 overexpression(Lv-Gli1),Gli1 overexpression+GANT61(Lv-Gli1+GANT61),and Gli1 overexpression+DMSO(Lv-Gli1+DMSO).Western blot was used to detect the expression levels of FAP and FSP-1 proteins.In vivo animal experiments were conducted in groups of puncture(PIDD),puncture treatment(PIDD+GANT61),and treatment control(PIDD+DMSO).Histological staining was used to evaluate the levels of intervertebral disc degeneration and fibrotic changes.Results:Histological staining results showed the histological score of degenerated discs significantly increased(P＜0.01).Western blot results showed a significant increase in Gli1 protein expression in the degenerated group(P＜0.05),which was consistent with the conclusions of the cellular experiments.NPCs overexpressing Gli1 protein were constructed,and immunofluorescence staining results showed that the expression of FAP and FSP-1 in the Gli1 overexpression group was significantly upregulated compared to the control and virus control groups.Western blot results further showed a significant increase in the expression of fibrotic markers in the Gli1 overexpression group(P＜0.05).In the Gli1 overexpression+GANT61 group of the cellular experiment,compared to the control and Gli1 overexpression groups,Western blot indicated that the expressions of FAP and FSP-1 proteins were downregulated(P＜0.05).In the in vivo experiment,the PIDD+GANT61 group showed a significant improvement in histological scores and collagen composition(P＜0.05).Conclusions:The expression of Gli1 protein is significantly upregulated in degenerated nucleus pulposus tissue and NPCs;Inhibition of Gli1 gene expression downregulates the expressions of FAP and FSP-1 proteins.The mechanism of intervertebral disc fibrosis and degeneration may be associated with the activation of Gli1.

Nine novel compounds, comprising seven tetrahydroanthraquinones (auxarthrolones A-G, 1-7), a γ-butenolide glycoside (malfilamentoside E, 26), and a γ-butenolide (auxarthrolide A, 27), together with eighteen known compounds (8-25) were isolated from rice-based solid culture of Auxarthron umbrinum (A. umbrinum) DSM3193 using the one strain many compounds (OSMAC) approach. The structural elucidation of these compounds was accomplished through nuclear magnetic resonance (NMR), mass spectrometry (MS), and NMR calculation combined with DP4+ analysis or MAEΔΔδ parameter, while the absolute configurations of new compounds were established through single-crystal X-ray diffraction, electronic circular dichroism (ECD) spectroscopic data analysis and/or chemical derivatization. Austrocortilutein (10) and auxarthrol H (14) demonstrated moderate cytotoxicity against U87 and U251 [half maximal inhibitory concentration (IC₅₀) 3.5-12.1 μmol·L^-1]. Additionally, auxarthrolone A (1), auxarthrol H (14), eupolyphagin B (15), and 7-hydroxy-2-(2-hydroxypropyl)-5-methylchromone (17) exhibited torsional effects on fibroblast proliferation challenges induced by oleic acid, thus demonstrating fibroblast proliferation-promoting activity.
4-Butyrolactone/pharmacology* ; Molecular Structure ; Anthraquinones/pharmacology* ; Humans ; Animals ; Mice ; Cell Line, Tumor ; Magnetic Resonance Spectroscopy

Objectives:To explore the impact of Gli1(gioma-associated oncogene homolog)on nucleus pulposus cells(NPCs)and its role in the fibrotic changes of the nucleus within intervertebral discs.Methods:A rat tail intervertebral disc puncture model was established.Histological staining was used to evaluate the levels of intervertebral disc degeneration and fibrotic changes in the untreated(negative control,NC)group and the puncture(puncture-induced intervertebral disc degeneration,PIDD)group.Western blot was used to assess the expression levels of Gli1 protein in the tissues.Gli1 protein was engineered to overexpress by lentivirus in NPCs and verified.The experiment was divided into three groups:control(Blank),Gli1 overexpression(Lv-Gli1),and virus control(lv-Ctrl).Immunofluorescence and Western blot were used to detect the expression levels of fibrotic markers FAP and FSP-1.Adding Gli1 inhibitor GANT61,cellular experiment was grouped into control(blank),Gli1 overexpression(Lv-Gli1),Gli1 overexpression+GANT61(Lv-Gli1+GANT61),and Gli1 overexpression+DMSO(Lv-Gli1+DMSO).Western blot was used to detect the expression levels of FAP and FSP-1 proteins.In vivo animal experiments were conducted in groups of puncture(PIDD),puncture treatment(PIDD+GANT61),and treatment control(PIDD+DMSO).Histological staining was used to evaluate the levels of intervertebral disc degeneration and fibrotic changes.Results:Histological staining results showed the histological score of degenerated discs significantly increased(P＜0.01).Western blot results showed a significant increase in Gli1 protein expression in the degenerated group(P＜0.05),which was consistent with the conclusions of the cellular experiments.NPCs overexpressing Gli1 protein were constructed,and immunofluorescence staining results showed that the expression of FAP and FSP-1 in the Gli1 overexpression group was significantly upregulated compared to the control and virus control groups.Western blot results further showed a significant increase in the expression of fibrotic markers in the Gli1 overexpression group(P＜0.05).In the Gli1 overexpression+GANT61 group of the cellular experiment,compared to the control and Gli1 overexpression groups,Western blot indicated that the expressions of FAP and FSP-1 proteins were downregulated(P＜0.05).In the in vivo experiment,the PIDD+GANT61 group showed a significant improvement in histological scores and collagen composition(P＜0.05).Conclusions:The expression of Gli1 protein is significantly upregulated in degenerated nucleus pulposus tissue and NPCs;Inhibition of Gli1 gene expression downregulates the expressions of FAP and FSP-1 proteins.The mechanism of intervertebral disc fibrosis and degeneration may be associated with the activation of Gli1.

Objective:To summarize experience of managing adult Langerhans cell histiocytosis(LCH) in hypothalamic-pituitary region(HPR) from Shanghai Huashan Hospital.Methods:Adult HPR-LCH patients diagnosed at oar endocrinology department from January 2013 to February 2022 were included. Clinical characteristics and treatment response were retrospectively analyzed.Results:A total of 27 adult HPR-LCH patients were included, with 14 cases involving the hypothalamus(H group) and 13 cases without(group NH). The common radiological findings included thickening of the pituitary stalk(25/27, 92.6%). At the time of diagnosis, 14 cases(51.9%) presented with panhypopituitarism, and 19 cases(70.4%) exhibited metabolic abnormalities. The group H had higher proportions of adrenal insufficiency, central hypothyroidism, panhypopituitarism, and diabetes compared to group NH(78.6% vs 23.1%; 78.6% vs 23.1%; 92.9% vs 30.8%, 35.7% vs 0%, respectively, all P<0.05). Hypothalamus syndrome was identified in 71.4%(10/14) of group H. The inital diagnosis rate was 79.2%(19/24), with 48.1% and 51.9% through biopsy of sellar and extrasellar lesions, respectively. Repeated biopsies confirmed the diagnosis in 25.9%(7/27) of cases. The peripheral lesions included bone, thyroid, lung, lymph node, thymus and liver. Out of 20 cases treated with chemotherapy, the objective response rate was 85% at 12 weeks. Four cases received local therapy, one case received traditional Chinese medicine treatment, one case abandoned treatment, and one case was lost to follow-up. The median follow-up time was 28(range 15 to 54) months. During this period, there were 3 deaths in group H and 1 death in group NH. Conclusion:Adult HPR-LCH patients presented with diabetes insipidus and high prevalences of hypopituitarism, hypothalamus syndrome and metabolic abnormalities. Typical imaging features were pituitary stalk thickening. A solitary mass in the HPR was usually very small, posing a great challenge for early diagnosis. Systemic evaluation would help to clarify the diagnosis. Patients with hypothalamus involvement had a higher mortality rate, suggesting the hypothalamus as a risk organ with poor prognosis.

Objective To propose a classification method for ultrasound images of pedicle screw canals based on radiomics analysis,and to evaluate the integrity of the screw canal.Methods With thoracolumbar spine specimens from 4 fresh cadavers,50 pedicle screw canals were pre-established and ultrasound images of the canals were acquired.A total of 2 000 images(1 000 intact and 1 000 damaged canal samples)were selected.The dataset was randomly divided in a 4∶1 ratio using 5-fold cross-validation to form training and testing sets(consisting of 1 600 and 400 samples,respectively).Firstly,the optimal radius of the region of interest was identified using the Otsu's thresholding method,followed by feature extraction using pyradiomics.Principal component analysis and the least absolute shrinkage and selection operator algorithm were employed for dimensionality reduction and feature selection,respectively.Subsequently,3 machine learning models(support vector machine[SVM],logistic regression,and random forest)and 3 deep learning models(visual geometry group[VGG],ResNet,and Transformer)were used to classify the ultrasound images.The performance of each model was evaluated using accuracy.Results With a region of interest radius of 230 pixels,the SVM model achieved the highest classification accuracy of 96.25%.The accuracy of the VGG model was only 51.29%,while the accuracies of the logistic regression,random forest,ResNet,and Transformer models were 85.50%,80.75%,80.17%,and 75.18%,respectively.Conclusion For ultrasound images of pedicle screw canals,the machine learning model performs better than the deep learning model as a whole,and the SVM model has the best classification performance,which can be used to assist physicians in diagnosis.

AIM:This study aims to explore the impact of Escherichia coli(E.coli)high-pathogenicity island(HPI)on pyroptosis and intestinal inflammation.METHODS:Kunming mice and IPEC-J2 cells(porcine small intesti-nal epithelial cells)were treated with HPI-containing E.coli strain(HPI+),HPI-deleting E.coli strain(ΔHPI),or lipo-polysaccharide(LPS).The intestinal lactate dehydrogenase(LDH)activity,superoxide dismutase(SOD)activity,IgA expression and secretory IgA(SIgA)content were assessed in mice.The expression of key regulatory factors in the nucleo-tide-binding oligomerization domain-like receptor protein 3(NLRP3)/caspase-1 signaling pathway-related proteins in mouse intestinal tract and IPEC-J2 cells was analyzed by RT-qPCR,immunohistochemical staining and Western blot.The levels of interleukin-1β(IL-1β)and IL-18 in mouse serum and IPEC-J2 cell culture supernatants were measured by ELISA.The pivotal role of NLRP3 in HPI+infection was confirmed by silencing NLRP3 in IPEC-J2 cells using siRNA.RE-SULTS:The HPI+infection markedly decreased SOD activity,increased IgA+B cell count,and induced the LDH release and SIgA secretion in the mouse intestine compared with ΔHPI infection.The results of ELISA,HE staining and TUNEL staining indicated that E.coli HPI triggered DNA damage,tissue injury and inflammation in mouse intestinal epithelial cells.Western blot revealed an increase in intestinal gasdermin D N-terminal fragment(GSDMD-N)protein level with HPI+infection compared with ΔHPI infection.E.coli HPI significantly up-regulated mRNA and protein expression of NLRP3,apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),caspase-1,GSDMD,IL-1β and IL-18 in mouse intestinal tissues and IPEC-J2 cells,accompanied by the elevated secretion of IL-1β and IL-18.The confo-cal microscopy demonstrated an enhanced assembly of the NLRP3 inflammasome with HPI+infection compared with ΔHPI infection,leading to colocalization of NLRP3 and caspase-1.Furthermore,NLRP3 silencing in IPEC-J2 cells attenuated E.coli HPI-induced cell inflammation,damage,and NLRP3/caspase-1 signaling pathway activation.CONCLUSION:The presence of HPI enhances the virulence of E.coli and exacerbates intestinal inflammation.Moreover,pyroptosis,regu-lated by the NLRP3/caspase-1 signaling pathway,plays a pivotal role in the intestinal injury induced by E.coli HPI.

  Objective  To analyze the clinical characteristics of thyroid LCH to enhance understanding of the disease.  Methods  We retrospectively studied the clinical data from six thyroid LCH patients who hospitalized in Huashan Hospital Affiliated to Fudan University from January 2015 to January 2022.We analyzed the ultrasound and ¹⁸F FDG-PET/CT imaging characteristics of thyroid LCH.  Results  The six patients diagnosed (2 males and 4 females) were between 18 and 58 years old.All patients had diabetes insipidus.MRI revealed thickened pituitary stalk.Two cases had central hypothyroidism, while four cases euthyroidism.Three cases tested positive for thyroid antibodies.Ultrasound showed thyroid nodules of TI-RADS 3 in three cases, TI-RADS 4 in two cases, and 1 with nodular goiter.Ultrasound showed that all sic cases indicated low echogenicity, 5 of which clear boundaries, 4 of which uneven echo distribution, 5 of which irregular shape, and noen has calcification.¹⁸F FDG-PET/CT indicated high uptake nodules with SUVmax values all above 10.4 cases were diagnosed by surgical excision and the other 2 by coarse-needle aspiration biopsy.When diagnosed, two cases had liver and thymus involvement respectively.One case had lung and bone involvement respectively.After treatment, 4 cases showed that nodular goiter shrank, while the other two with liver involvement progressed fast and no assessment made.  Conclusions  Thyroid LCH presented low echogenicity, clear boundaries, irregular shape, without calcification, and high uptake in ¹⁸F FDG-PET/CT.A definite diagnosis of pituitary stalk thicking accompanied by thyroid nodules, especially those with hypoechoic and irregular nodules, can be achieved by coarse-needle aspiration biopsy and langerin-specific pathological staining.

Hepatic hydrothorax (HH) is a challenging complication of liver cirrhosis associated with portal hypertension, and its pathogenesis and therapeutic measures remain unknown. This article summarizes and reviews the advances and challenges in the research on the pathogenesis, clinical manifestations, diagnosis, and treatment of HH and proposes a multidisciplinary treatment strategy, including reducing the production of ascites, preventing effusion from entering the thoracic cavity, removing pleural effusion, occluding the pleural cavity, and performing liver transplantation, so as to provide a reference for more clinicians.

Objective:To compare the clinical efficacy of percutaneous transhepatic choledochoscope lithotomy (PTCSL) with laparoscopic choledocholithotomy (LD) in treatment of choledocholithiasis.Methods:Data of 132 patients with choledocholithiasis treated at the First Affiliated Hospital of Guangzhou Medical University from July 2012 to December 2018 were retrospectively analyzed. There were 75 males and 57 females, with an average age of 62.7 years. For 76 patients underwent PTCSL (the PTCSL group) and 56 underwent LD (the LD group). The data of the patients the success rate of lithotomy, stone residual rate, operation time, postoperative complications and stone recurrence, chronic cholangitis, and acute cholangitis 1 month after operation were compared between the two groups.Results:The ratio of upper abdominal operation history and biliary tract infection in the PTCSL group was higher than that in the LD group, and the difference was statistically significant (both P<0.05). In the PTCSL group, the calculi were successfully removed in 64 patients in one treatment session, while residual calculi were removed through subsequent sinus choledochoscopy in 9 patients. In the remaining 3 patients, the residual calculi were removed with LD or laparotomy operations. Postoperative complications occurred in 14 patients (19.2%, 14/73). In the LD group, the calculi were successfully removed in one session in 46 patients while in 8 patients the residual calculi were removed by choledochoscopy (1 patient still had residual calculi after choledochoscopy). The remaining 2 patients underwent open surgery due to anatomical difficulties. Postoperative complications occurred in 11 patients (20.4%, 11/54). There were no significant differences between the two groups in the one-off stone removal rate, postoperative stone residual rate, final stone removal rate and postoperative complication rate (all P>0.05). The operation time of the PTCSL group was (156±60) min, which was significantly shorter than the LD group (203±59) min ( P<0.05). There was no significant difference between the two groups in the incidence of postoperative chronic cholangitis and recurrence rate of calculi (both P>0.05). The incidence of acute cholangitis in the PTSCL group was significantly higher than that in the LD group ( P<0.05). Conclusion:PTCSL was as safe and effective as LD, with fewer complications and faster recovery. It is especially suitable for patients with previous upper abdominal surgery, recurrence of calculi and repeated biliary tract infection.

Objective: To analyze the clinical characteristics of patients with relapsed/refractory primary central nervous system lym-phoma (PCNSL) and to explore the factors that influence the prognosis, in order to provide evidence for the clinical diagnosis and treat-ment. Methods: Sixty-four patients with relapsed/refractory PCNSL diagnosed from October 2006 to August 2015 were selected. The clinical features, treatment plans, and laboratory examination data were retrospectively analyzed. Cox regression was used for multi-variate analysis. Results: Univariate and multivariate analyses showed that progression-free survival of first time (PFS1)≤1 year and Kar-nofsky performance status (KPS) score<70 points were independent prognostic factors in patients with first relapsed/refractory PCNSL. The median PFS2 and overall survival of second time (OS2) were 19 and 21 months, respectively, in patients with PFS1≥1 year, where-as the median progression free survival of second time (mPFS2) and OS2 were 10 and 14 months, respectively, in patients with PFS1<1 year. The median PFS2 (mPFS2) in patients with first relapse/refractory KPS score≥70 points and those with KPS score<70 points were 40 and 10 months, respectively, and the median OS2 were 43 and 12 months, respectively. The median PFS for the methotrexate (MTX) and non-MTX groups was 18 and 10 months, respectively. Multivariate analysis showed that the salvage therapy was a relevant factor influencing the patient's PFS. However, univariate analysis showed that the median OS2 in the MTX and non-MTX groups was 23 and 12 months, respectively, with significant difference but without any correlation with prognosis. Conclusions: progression-free sur-vival (PFS)≤1 year and KPS score<70 were independent prognostic factors in patients with first relapsed/refractory PCNSL. Patients with relapsed/refractory PCNSL who continuously received high-dose MTX-based treatment may have improved long-term treatment outcomes.