BACKGROUND: Enzalutamide, a second-generation androgen receptor (AR) pathway inhibitor, is widely used in the treatment of castration-resistant prostate cancer. However, after a period of enzalutamide treatment, patients inevitably develop drug resistance. In this study, we characterized leucine-rich repeated G-protein-coupled receptor 5 (LGR5) and explored its potential therapeutic value in prostate cancer. METHODS: A total of 142 pairs of tumor and adjacent formalin-fixed paraf-fin-embedded tissue samples from patients with prostate cancer were collected from the Pathology Department at Sun Yat-sen Memorial Hos-pital. LGR5 was screened by sequencing data of enzalutamide-resistant cell lines combined with sequencing data of lesions with different Gleason scores from the same patients. The biological function of LGR5 and its effect on enzalutamide resistance were investigated in vitro and in vivo . Glutathione-S-transferase (GST) pull-down, coimmunoprecipitation, Western blotting, and immunofluorescence assays were used to explore the specific binding mechanism of LGR5 and related pathway changes. RESULTS: LGR5 was significantly upregulated in prostate cancer and negatively correlated with poor patient prognosis. Overexpression of LGR5 promoted the malignant progression of prostate cancer and reduced sensitivity to enzalutamide in vitro and in vivo . LGR5 promoted the phosphorylation of glycogen synthase kinase-3β (GSK-3β) by binding heat shock protein 90,000 alpha B1 (HSP90AB1) and mediated the activation of the Wingless/integrated (WNT)/β-catenin signaling pathway. The increased β-catenin in the cytoplasm entered the nucleus and bound to the nuclear AR, promoting the transcription level of AR, which led to the enhanced tolerance of prostate cancer to enzalutamide. Reducing HSP90AB1 binding to LGR5 significantly enhanced sensitivity to enzalutamide. CONCLUSIONS LGR5 directly binds to HSP90AB1 and mediates GSK-3β phosphorylation, promoting AR expression by regulating the WNT/β-catenin signaling pathway, thereby conferring resistance to enzalutamide treatment in prostate cancer.
Male ; Humans ; Phenylthiohydantoin/pharmacology* ; Benzamides ; Receptors, G-Protein-Coupled/genetics* ; Nitriles ; Cell Line, Tumor ; HSP90 Heat-Shock Proteins/metabolism* ; Drug Resistance, Neoplasm/genetics* ; Prostatic Neoplasms/drug therapy* ; beta Catenin/metabolism* ; Receptors, Androgen/genetics* ; Animals ; Mice ; Wnt Signaling Pathway/physiology*

Acute-on-chronic liver failure （ACLF） is an acute and critical illness with a high short-term mortality rate， and current therapies mainly focus on elimination of causes， organ support， and prevention of complications. Although liver transplantation is the most effective treatment modality， its clinical application is limited， and traditional Chinese medicine has shown significant advantages and characteristics in the treatment of ACLF. In traditional Chinese medicine， ACLF is classified into the same category as diseases such as “jaundice”， and unlike traditional jaundice which is mostly characterized by excess and heat syndromes， the syndrome of ACLF has gradually transformed from Yang jaundice to Yin jaundice due to the changing disease spectrum of ACLF. With reference to the pathogenesis of ACLF in Western medicine and traditional Chinese medicine theories， this article discusses the essential pathogenesis of ACLF in traditional Chinese medicine， explores the evolution of ACLF syndromes， and reviews the research advances in the clinical efficacy and mechanisms of traditional Chinese medicine based on the three-factor differentiation-based treatment of damp-heat， blood stasis-heat， and spleen deficiency， as well as the safety of spleen-strengthening and Yang-warming drugs in the clinical treatment of ACLF， in order to provide ideas， methods， and evidence for the application of traditional Chinese medicine in ACLF.

OBJECTIVES: To analyze the differentially expressed exosomal miRNAs in patients with chronic heart failure (CHF) complicated by hyperuricemia (HUA) and explore their potential as novel diagnostic molecular markers and their target genes. METHODS: This study was conducted among 30 CHF patients with HUA (observation group) and 30 healthy volunteers (control group) enrolled between September, 2020 and September, 2023. Peripheral blood samples were collected from 6 CHF patients with HUA for analyzing exosomal miRNAs by high-throughput sequencing, and the results were validated in the remaining 24 patients using qRT-PCR. GO and KEGG enrichment analyses were performed to predict the the target genes of the identified differential miRNAs. We also validated the differentially expressed miRNAs by animal experiment. RESULTS: A total of 42 differentially expressed exosomal miRNAs were detected in observation group by high-throughput sequencing; among them, miR-27a-5p was significantly upregulated (P=0.000179), and miR-139-3p was significantly downregulated (P=0.000058). In the 24 patients with both CHF and PUA, qRT-PCR validated significant upregulation of miR-27a-5p (P=0.004) and downregulation of miR-139-3p (P=0.005) in serum exosomes. When combined, miR-27a-5p and miR-139-3p had a maximum area under the curve (AUC) of 0.899 (95% CI: 0812-0.987) for predicting CHF complicated by HUA. GO and KEGG enrichment analyses suggested that the differential expressions of miR-27a-5p and miR-139-3p was associated with the activation of the AMPK-mTOR signaling pathway to activate the autophagic response. We obtained the same conclusion from animal experiment. CONCLUSIONS Upregulated exosomal miR-27a-5p combined with downregulated exosomal miR-139-3p expression can serve as a novel molecular marker for diagnosis of CHF complicated by HUA, and their differential expression may promote autophagy in cardiomyocytes by activating the AMPK-mTOR signaling pathway.
Humans ; Hyperuricemia/diagnosis* ; Heart Failure/genetics* ; MicroRNAs/metabolism* ; Exosomes/metabolism* ; Chronic Disease ; Male ; Female ; Middle Aged ; Animals

The aim of this study is to identify the cause of death of a South China tiger cub at the Meihuashan breeding institute of Fujian Province.Pathogenic bacteria were isolated and cultured from liver,spleen,lung and other tissue samples of the dead South China tiger aseptically.The iso-lated bacteria were identified through morphological observation,biochemical characterization,sequence analysis of housekeeping gene gyrB,virulence gene detection,animal pathogenicity test and drug sensitivity test.A pathogenic Aeromonas hydrophila strain,designated FJ/Tiger-201809 was successfully isolated from the trachea of dead South China tiger.The nucleotide sequence ho-mology between the isolate and 11 strains of Aeromonas gyrB ranged from 91.2％to 99.1％,with the highest homology of 99.1％observed with Aeromonas hydrophila(AF208251.1).Genetic evo-lution analysis showed that the isolated strain FJ/Tiger-201809 was in the same evolutionary branch as other reference strains of Aeromonas hydrophila and was closely related.The pathoge-nicity test in mice showed artificial infection of mice with the strain resulted in varying degrees of lesions in several organs of the mice,and the median lethal dose(LD50)was 1 × 107.8 CFU/mL.Virulence gene test results showed that the isolate FJ/Tiger-201809 carried two virulence genes,aer and act.The results of drug sensitivity test showed that FJ/Tiger-201809 was highly sensitive to enrofloxacin and ampicillin among 18 commonly used antibiotics,relatively sensitive to penicil-lin G and doxycycline,and resistant to the other 14 antibiotics.In conclusion,this study isolated and identified a strain of Aeromonas hydrophila from a dead South China tiger with multiple drug resistance and strong pathogenicity,which provided an important reference for the prevention and control of bacterial diseases in South China tiger.

The aim of this study is to identify the cause of death of a South China tiger cub at the Meihuashan breeding institute of Fujian Province.Pathogenic bacteria were isolated and cultured from liver,spleen,lung and other tissue samples of the dead South China tiger aseptically.The iso-lated bacteria were identified through morphological observation,biochemical characterization,sequence analysis of housekeeping gene gyrB,virulence gene detection,animal pathogenicity test and drug sensitivity test.A pathogenic Aeromonas hydrophila strain,designated FJ/Tiger-201809 was successfully isolated from the trachea of dead South China tiger.The nucleotide sequence ho-mology between the isolate and 11 strains of Aeromonas gyrB ranged from 91.2％to 99.1％,with the highest homology of 99.1％observed with Aeromonas hydrophila(AF208251.1).Genetic evo-lution analysis showed that the isolated strain FJ/Tiger-201809 was in the same evolutionary branch as other reference strains of Aeromonas hydrophila and was closely related.The pathoge-nicity test in mice showed artificial infection of mice with the strain resulted in varying degrees of lesions in several organs of the mice,and the median lethal dose(LD50)was 1 × 107.8 CFU/mL.Virulence gene test results showed that the isolate FJ/Tiger-201809 carried two virulence genes,aer and act.The results of drug sensitivity test showed that FJ/Tiger-201809 was highly sensitive to enrofloxacin and ampicillin among 18 commonly used antibiotics,relatively sensitive to penicil-lin G and doxycycline,and resistant to the other 14 antibiotics.In conclusion,this study isolated and identified a strain of Aeromonas hydrophila from a dead South China tiger with multiple drug resistance and strong pathogenicity,which provided an important reference for the prevention and control of bacterial diseases in South China tiger.

Background Studies on the relationships of environmental noise exposure with negative emotions and sleep quality have long been reported. Self-perceived noise intensity is not only related to environmental noise exposure, but also reflects an individual's susceptibility to noise; however, few studies on self-perceived noise intensity, negative emotions, and sleep quality have been reported, and it is not clear whether negative emotions play a mediating role in the relationship between self-perceived noise intensity and sleep quality. Objective To analyze the mediating role of negative emotions (anxiety, depression, and stress) in the relationship between self-perceived noise intensity and sleep quality in noise-exposed workers, and to provide a scientific basis for addressing psychological problems and sleep quality induced by noise exposure. Methods Stratified cluster sampling was used to select noise-exposed workers from a large equipment manufacturing plant in Chengdu from May to June 2023, and demographic characteristics were investigated using a self-designed general information questionnaire; self-perceived noise intensity was reported by the study subjects as the noise intensity of their workplaces; sleep quality was synthesized from three indicators: night sleep duration, self-perceived sleep quality, and sleep-related symptoms; the Anxiety-Depression-Stress Scale (DASS-21 scale) was used to evaluate anxiety, depression, and stress of a worker. Structural equation modeling was used to analyze the pathways of negative emotions (anxiety, depression, and stress) between self-perceived noise intensity and sleep quality. Results The positive rates of negative emotions were 33.1% (anxiety symptoms, 215/649), 26.0% (depression symptoms, 169/649), and 14.0% (stress symptoms, 91/649), respectively in a total of 649 noise-exposed workers. Self-perceived noise intensity was positively correlated with sleep quality score (r=0.218, P<0.001) and negative emotions (anxiety, depression, and stress) (r=0.167, 0.145, 0.167, P<0.001); sleep quality score was positively correlated with negative emotions (anxiety, depression, and stress) (r=0.512, 0.447, 0.513, P<0.001). The results of path analysis showed that the negative emotions (anxiety, depression, and stress) partially mediated between self-perceived noise intensity and sleep quality (β=0.123，P<0.001). Self-perceived noise intensity was positively correlated with negative emotions and sleep quality, and there was a path of self-perceived noise intensity → negative emotion → sleep quality (P<0.001), and the mediating effect accounted for 42.71%. Conclusion The positive rates of anxiety, depression, and stress are high among the noise-receiving workers in this manufacturing industry, and negative emotions characterized by anxiety, depression, and stress partially mediate the relationship between self-perceived noise intensity and sleep quality.

Objective:To investigate the experience and results of the modified lateral prostate capsule sparing robot-assisted radical cystectomy-orthotopic ileal neobladder (LPCS-RARC-OIN).Methods:From December 2018 to November 2020, 19 patients received LPCS-RARC-OIN by a single surgeon in Sun Yat-sen Memorial Hospital, Sun Yat-sen University. LPCS-RARC-OIN was performed on male patients with high-risk non-muscle-invasive bladder cancer or muscle-invasive bladder cancer cT 2N 0M 0 without tumour in the bladder neck or urethra, and prostate cancer was ruled out by MRI and serum PSA<2.5ng/ml. The average age was 57.6 years, the average IIEF-5 score was 20.4. Separating the prostatic adenoma and the lateral prostate capsule from the base to the apex of the prostate, and retaining the lateral prostate surgical capsule or lateral prostate capsule about 1-2mm thickness. Patients were followed up and urinary function, sexual function and oncological outcomes were recorded. Results:All 19 operations were finished successfully. The average operation time was 279.9 (225-345) min and average estimated blood loss was 88.9 (30-200) ml. The average postoperative hospital stays was 15.8 (9 -23) days. The average lymph node yields was 23.3 (11-42). All surgical margins were negative and no incidental prostate cancer was found. 2 weeks, 1 month, 3 months and 6 months after catheter removal, the daytime and nighttime continence were 42.1% (8/19)and 36.8% (7/19), 63.2% (12/19)and 63.2% (12/19), 78.9% (15/19) and 73.7% (14/19), 94.7% (18/19) and 89.5% (17/19), respectively. 3 months and 6 months after operation, the average IIEF-5 score was 7.2 and 10.1 points respectively. The average follow-up was 10.6 months (5.4-26.1 months)and no recurrence or distant metastasis was found in this study.Conclusions:LPCS-RARC-OIN could improve the urinary and sexual function in selected patients. However, the long-term follow up is needed for functional and oncological outcomes.

Objective:To investigate the safety and efficacy of low molecular weight heparin (LMWH) bridging program in the perioperative period of mini-percutaneous nephrolithotomy (mPCNL) for long-term antithrombotic patients.Methods:The clinical data of 50 patients who received long-term antithrombotic therapy with mPCNL in Sun Yat-sen Memorial Hospital from January 2013 to December 2017 were retrospectively analyzed. Perioperative anticoagulation plans were drawn up after discussion with an internist. Patients with high thrombosis risk were bridged with LMWH during the perioperative period. Resumed LMWH anticoagulation within 48 hours after surgery. Patients with low or medium thrombosis risk directly discontinued anticoagulation one week before surgery. Preoperative anticoagulation was resumed within 48 hours after removing the nephrostomy tube in all patients. We analyzed the general information before surgery, data during surgery, postoperative hemoglobin changes and stone-free rate (SRF) of all cases. 21 patients were treated with LMWH bridging (bridging group), and 29 patients were directly discontinued with anticoagulant drugs (non-bridging group). There was no statistical difference between the two groups in age [(59.7±7.1) vs. (52.4±10.4)years] , gender [(male/female), 14/7 vs. 19/10], BMI [ (24.3±3.9) kg/m 2 vs. (24.7±5.1) kg/m 2], S. T.O.N.E. score (7.4±1.1 vs. 6.9±1.0), stone surface area [ 314.0(31.4-1 130.4) mm 2 vs. 282.5(64.7-866.0) mm 2], the number of calculi involved in calyces (6/15 vs. 13/16) and stone-related surgical history [ 34% (7/21) vs. 24% (7/29) ]. Results:In the bridging group, 18 patients (86%) performed single-channel mPCNL, 3 patients (14%) underwent dual-channel mPCNL, and the operation time was 80 (35-180) min. In the non-bridging group, 27 patients (93%) underwent single-channel mPCNL, 2 patients (7%) performed dual-channel mPCNL, and the operation time was 80 (30-60) min. The mean changes in hemoglobin in the bridging group and the non-bridging group was 18 (-2 -66) g/L and 14 (-25-64) g/L, respectively ( P = 0.073). The average postoperative hospital stay in the bridging group was (8.6 ± 3.5) days, and the non-bridging group was (7.1 ± 2.3) days ( P= 0.057). Two patients in each group received blood transfusion, and no patients received interventional embolization. The SRF of bridging group and non-bridging group was 81.0% (17/21) vs. 75.9% (22/29) ( P = 0.67) 1 month after the operation. During the perioperative period, no patients had thrombotic complications. Conclusions:When mPCNL was required for long-term antithrombotic treatment patients, the use of LWMH alternatives during the perioperative period did not increase bleeding related complications.

Anoctamin 1 (ANO1) is a kind of calcium-activated chloride channel involved in nerve depolarization. ANO1 inhibitors display significant analgesic activity by the local peripheral and intrathecal administration. In this study, several thiophenecarboxylic acid and benzoic acid derivatives were identified as novel ANO1 inhibitors through the shape-based virtual screening, among which the 4-arylthiophene-3-carboxylic acid analogues with the best ANO1 inhibitory activity were designed, synthesized and compound