High mobility group box 1 (HMGB1), when released extracellularly, plays a pivotal role in the development of spinal cord synapses and exacerbates autoimmune diseases within the central nervous system. In experimental autoimmune encephalomyelitis (EAE), a condition that models multiple sclerosis, the levels of extracellular HMGB1 and interleukin-33 (IL-33) have been found to be inversely correlated. However, the mechanism by which IL-33 deficiency enhances HMGB1 release during EAE remains elusive. Our study elucidates a potential signaling pathway whereby the absence of IL-33 leads to increased binding of P300/CBP-associated factor with HMGB1 in the nuclei of astrocytes, upregulating HMGB1 acetylation and promoting its release from astrocyte nuclei in the spinal cord of EAE mice. Conversely, the addition of IL-33 counteracts the TNF-α-induced increase in HMGB1 and acetylated HMGB1 levels in primary astrocytes. These findings underscore the potential of IL-33-associated signaling pathways as a therapeutic target for EAE treatment.
Animals ; Encephalomyelitis, Autoimmune, Experimental/metabolism* ; Astrocytes/metabolism* ; Interleukin-33/metabolism* ; HMGB1 Protein/metabolism* ; Acetylation ; Mice, Knockout ; Mice, Inbred C57BL ; p300-CBP Transcription Factors/metabolism* ; Mice ; Spinal Cord/metabolism* ; Cells, Cultured ; Female ; Signal Transduction

Arginine methylation is a critical post-translational modification that plays multifaceted biological functions. However, the manipulation of protein arginine methylation largely depends on genetic or pharmaceutic inhibition of the regulatory enzymes, protein arginine methyltransferases (PRMTs), or non-methylation substitution of corresponding arginine residue to lysine or alanine of protein of interest (POI), which inevitably affects other substrates, or disrupts the structure of POI. Thus, it urges an approach to specifically modulate the arginine methylation of a POI under physiological conditions. To this end, we report the discovery of a methylation tagging system (MeTAG), that enables targeted modification of protein arginine methylation. Through bridging the methyltransferase PRMT5 proximity to a POI, MeTAG facilitates the arginine methylation of POIs, including known arginine methylated proteins, androgen receptor (AR) and protein kinase B (AKT), as well as a neo-substrate E1A binding protein (p300), in a reversible and PRMT5-dependent manner. Moreover, MeTAG can regulate downstream signaling in a methylation dependent manner, leading to downregulation of PSMA mRNA level and activation of AKT. Therefore, MeTAG represents a feasible approach to modulate protein methylation and thereby perturbs protein function in biological and therapeutic contexts.

OBJECTIVES: To elucidate the molecular mechanism by which villin-like protein VILL (VILL) inhibits proliferation of nasopharyngeal carcinoma (NPC) cells. METHODS: Co-immunoprecipitation (CO-IP) assay, mass spectrometry, Western blotting, immunofluorescence staining, and GST pull-down assay were employed to identify and confirm the protein interacting with VILL that had the highest abundance in NPC cell lines. Transgenic experiments were conducted in both NPC cell lines and nude mice to validate the regulatory role of VILL and its target protein in NPC proliferation. Immunohistochemistry was utilized to assess the correlation of the expression levels of VILL and its target protein in clinical tissue specimens of NPC with the clinical features of the patients. RESULTS: In NPC cell lines (HONE1 EBV and S18), VILL was found to interact most abundantly with the E3 ubiquitin ligase LMO7, and both proteins co-localized in the cytoplasm with direct interactions. Overexpression of LMO7 partially counteracted the inhibitory effect of VILL on NPC cell proliferation. The expression of VILL was significantly downregulated in 136 NPC tissue samples compared to 67 non-cancerous nasopharyngeal tissues (P<0.00001) with close correlation with clinical T stage (P=0.04), N stage (P=0.01), and M stage (P=0.013), whereas LMO7 was highly expressed in all the NPC tissues. CONCLUSIONS VILL overexpression inhibits NPC proliferation probably by suppressing the oncogenic function of LMO7.
Nasopharyngeal Neoplasms/metabolism* ; Humans ; LIM Domain Proteins/metabolism* ; Cell Proliferation ; Cell Line, Tumor ; Animals ; Mice ; Nasopharyngeal Carcinoma ; Mice, Nude ; Transcription Factors/metabolism* ; Carcinoma ; Female ; Microfilament Proteins/genetics* ; Male ; Middle Aged

Objective To explore clinical application value of fetal heart quantification(HQ)technology in evaluating morphology and function of fetal heart in gestational hypertension pregnancy patients.Methods A total of 85 women with gestational hypertension(GH)during gestational age of 20-40 weeks from March 2020 to March 2024 had been selected as experimental group.According to blood pressure,urine protein,and symptoms of pregnant women,experimental group was divided into GH group(n=35),mild preeclampsia(MPE)group(n=28),and severe preeclampsia(SPE)group(n=22).Additionally,150 normal pregnant women with matched gestational age were randomly selected as control group.Fetal HQ technique was adopted to obtain cardiac morphological and functional indicators of groups,and statistical analysis was conducted.Results There was statistically significant difference in global spherical index(GSI)in each group(P＜0.05).Patients in SPE group had rounder fetal hearts.Bariance analysis was performed on fractional area change(FAC),Tei index,and global longitudinal strain(GLS)for each group of patients.There were statistically significant differences in right ventricular GLS and right ventricular Tei index(P＜0.05).The absolute value of right ventricular GLS in SPE group and MPE group was lower than that in control group,while right ventricular Tei index in SPE group was higher than that in control group.Conclusion Fetal HQ technique provides quick and easy quantitative assessment of fetal heart shape and function.Gestational hypertension not only changes the fetal heart morphology,but also affects the fetal heart function,and the fetal right heart system is more affected than the fetal left heart.

Objective: To explore the changes of mechanosensitive channels Piezos (Piezo 1 and Piezo 2) in neurogenic bladder (NB) of young rats and their effects，so as to provide reference for clinical search of new therapeutic targets. Methods: A total of 30 female young SD rats were divided into 5 groups based on random number table method：sham operation group (sham)，2-week nerve transection group (NB-2W)，6-week nerve transection group (NB-6W)，2-week nerve transection + Piezos inhibitor group (NB-P-2W) and 6-week nerve transection + Piezos inhibitor group (NB-P-6W)，with 6 rats in each group.The NB models were constructed by transecting the L6 and S1 spinal nerves of young rats.The NB-2W and NB-6W groups were not intervened after modeling，while the NB-P-2W and NB-P-6W groups were intraperitoneally injected with Piezos inhibitor GsMTx4 (10 mg/kg) every 2 days after modeling.Bladder cystometry and ultrasound were performed after 2 and 6 weeks of transection.The expressions of Piezos and fibrosis-related indexes (Collagen Ⅰ and α-smooth muscle actin) were detected in bladder tissues. Results: The results of bladder cystometry showed that the basal bladder pressure in NB-2W group was significantly increased，while it was slightly decreased but was still higher in NB-6W group than in the sham group (P<0.05).Basal bladder pressure was lower in NB-P-2W group than in NB-2W group，but was higher than that in the sham group; basal bladder pressure was lower in NB-P-6W group than in NB-6W group，but higher than that in the sham group (P<0.05).Compared with the sham group，the NB-2W and NB-6W groups had firstly increased and then decreased maximum cystometric capacity (MCC) (P<0.05).Compared with NB-2W group，NB-P-2W group had lower bladder leakage point pressure (BLPP)，but higher MCC and bladder compliance (BC) (P<0.05).Compared with NB-6W group，NB-P-6W group had significantly lower BLPP but higher MCC and BC (P<0.05).HE and MASSON staining and ultrasound results showed that，with the extension of nerve transection time，bladder fibrosis gradually worsened，the bladder wall became rough and thickened，calculi were visible inside，and hydronephrosis gradually appeared; the degree of fibrosis in NB-P-2W and NB-P-6W groups was less than that in NB-2W and NB-6W groups，and no hydronephrosis was observed in the upper urinary tract.In addition，Western blotting and immunohistochemical results showed that NB-2W and NB-6W groups had significantly higher relative expression levels of Piezos，Collagen Ⅰ and α-SMA than the sham group (P<0.01)，while NB-P-2W and NB-P-6W groups had lower relative expression levels of Piezos,Collagen Ⅰ and α-SMA than NB-2W and NB-6W groups (P<0.01). Conclusion: The increased expressions of mechanosensitive channels Piezos in NB young rats may be involved in the progression of bladder fibrosis，but its mechanism needs further study.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is a prevalent type of cancer with a high mortality rate in its late stages. One of the major challenges in OSCC treatment is the resistance to epidermal growth factor receptor (EGFR) inhibitors. Therefore, it is imperative to elucidate the mechanism underlying drug resistance and develop appropriate precision therapy strategies to enhance clinical efficacy. METHODS: To evaluate the efficacy of the combination of the Ca 2+ /calmodulin-dependent protein kinase II (CAMK2) inhibitor KN93 and EGFR inhibitors, we performed in vitro and in vivo experiments using two FAT atypical cadherin 1 ( FAT1 )-deficient (SCC9 and SCC25) and two FAT1 wild-type (SCC47 and HN12) OSCC cell lines. We assessed the effects of EGFR inhibitors (afatinib or cetuximab), KN93, or their combination on the malignant phenotype of OSCC in vivo and in vitro . The alterations in protein expression levels of members of the EGFR signaling pathway and SRY-box transcription factor 2 (SOX2) were analyzed. Changes in the yes-associated protein 1 (YAP1) protein were characterized. Moreover, we analyzed mitochondrial dysfunction. Besides, the effects of combination therapy on mitochondrial dynamics were also evaluated. RESULTS: OSCC with FAT1 mutations exhibited resistance to EGFR inhibitors treatment. The combination of KN93 and EGFR inhibitors significantly inhibited the proliferation, survival, and migration of FAT1 -mutated OSCC cells and suppressed tumor growth in vivo . Mechanistically, combination therapy enhanced the therapeutic sensitivity of FAT1 -mutated OSCC cells to EGFR inhibitors by modulating the EGFR pathway and downregulated tumor stemness-related proteins. Furthermore, combination therapy induced reactive oxygen species (ROS)-mediated mitochondrial dysfunction and disrupted mitochondrial dynamics, ultimately resulting in tumor suppression. CONCLUSION Combination therapy with EGFR inhibitors and KN93 could be a novel precision therapeutic strategy and a potential clinical solution for EGFR-resistant OSCC patients with FAT1 mutations.
Humans ; ErbB Receptors/metabolism* ; Mouth Neoplasms/metabolism* ; Cell Line, Tumor ; Animals ; Drug Resistance, Neoplasm/genetics* ; Cadherins/metabolism* ; Carcinoma, Squamous Cell/metabolism* ; Mice ; Mutation/genetics* ; Mice, Nude ; Protein Kinase Inhibitors/therapeutic use* ; Cetuximab/pharmacology* ; Afatinib/therapeutic use* ; Cell Proliferation/drug effects* ; Signal Transduction/drug effects*

Objective To evaluate the diagnostic value of prostate health index(PHI)and prostate health index density(PHID)in identifying intermediate-to high-risk prostate cancer(PCa).Methods Clinical data of 160 treatment-na?ve patients with highly suspected PCa,who underwent initial prostate biopsy in our hospital during Jul.2022 and Feb.2024,were retrospectively analyzed.Data included age,body mass index(BMI),prostate volume(PV),total prostate-specific antigen(tPSA),free PSA(fPSA),[-2]proPSA(p2PSA),PHI and PHID.Biopsy-positive results were stratified according to the EAU D'Amico risk criteria.Receiver operating characteristic(ROC)curve and multivariate logistic regression analysis were employed to assess the diagnostic performance of PHI and PHID in predicting PCa and identifying intermediate-to high-risk PCa.Results There were statistically significant differences in tPSA,p2PSA,PHI and PHID between the negative and positive groups,as well as among the low-,medium-and high-risk groups(P＜0.01).Both PHI and PHID demonstrated good diagnostic performance in predicting PCa(AUC=0.820 8 and 0.875 7,respectively;all P＜0.001),and in identifying intermediate-to high-risk PCa(AUC=0.838 0 and 0.878 3,respectively;all P＜0.001).Compared to the baseline model,the incorporation of PHI and PHID individually into the multivariate model significantly improved the screening performance for PCa(AUC=0.910 and 0.898,respectively;all P＜0.001).Conclusion PHI and PHID exhibit high diagnostic efficacy in screening PCa,particularly in identifying intermediate-to high-risk disease.

Objective:To investigate the efficacy and safety of intravesical injections of botulinum toxin A (BTX-A) for the treatment of pediatric refractory urinary frequency and incontinence (UFI), and to analyze the factors predicting the therapeutic effect of BTX-A injections.Methods:A retrospective case series study was made on 35 children with UFI [(12.3±4.2) years old], including 13 males (37.1%) and 22 females (62.9%), treated in the First Affiliated Hospital of Zhengzhou University from January 2021 to March 2024.Urotherapy and drug treatments were ineffective in all children, who then received intravesical injections of BTX-A.The urodynamic study (UDS) was performed 1 week before treatment and 3 months after treatment, and the Overactive Bladder Symptom Score (OABSS) and Incontinence Quality of Life (I-QOL) were collected.Perioperative and postoperative adverse events were also recorded.A global response assessment (GRA) score of ≥2 at 3 months postoperatively was defined as effective treatment.According to the postoperative GRA score, the patients were divided into effective and ineffective groups to explore the predictive factors affecting the outcome of BTX-A treatment, such as age, gender, preoperative urodynamic parameters, and the types of urinary incontinence. t-test was used to compare quantitative data such as age, UDS parameters, OABSS, and I-QOL scores.The comparison of count data such as gender and urinary incontinence types was conducted using Fisher′s exact test. Results:The bladder compliance, bladder capacity ratio, OABSS and I-QOL scores of patients significantly improved after BTX-A injection (all P<0.05).The 77.1% (27/35) of the patients were satisfied with BTX-A treatment effects.The main adverse event during the follow-up was a temporary increase in postvoid residual urine after injecting BTX-A (7 cases), which was alleviated by the Creade action or clean intermittent catheterization.There was significant difference in age [(13.00±4.32) years vs.(10.00±2.67) years] and preoperative bladder compliance [(11.21±5.74) mL/cmH 2O vs.(5.13±2.42) mL/cmH 2O] between effective (27 cases) and ineffective groups (8 cases) ( t=2.383, 2.899, all P<0.05).The differences in preoperative bladder capacity ratio, maximum filling detrusor pressure, postvoid residual urine, voiding efficiency, gender, and the types of urinary incontinence were not statistically significant between the two groups (all P>0.05). Conclusions:Intravesical injection of BTX-A is a safe and effective treatment for children with refractory UFI.A young age and poor bladder compliance at treatment are associated with poor prognosis of BTX-A treatment.

Lower urinary tract dysfunction (LUTD) is common in children, but treatment is difficult in some cases, and it might causes upper urinary tract damage in the late stage. Recently, botulinum toxin A(BTX-A) has been recommended as an effective procedure for refractory LUTD, which has significantly improved symptoms and reduced the occurrence of upper urinary tract damage in children with LUTD. At present, there is a lack of international consensus on the application indication and dose selection of BTX-A in LUTD children, and the safety and long-term efficacy of repeated BTX-A injection are also clinical concerns. This article reviews the current application perspectives and progress of BTX-A in the treatment of children with LUTD, especially the clinical application indications and methods.BTX-A provides a minimally invasive, safe and effective treatment option for children with LUTD, which can significantly improve clinical symptoms and urodynamic parameters. With the further clarification of indications and the improvement of administration methods, BTX-A will be able to assist more children with LUTD in enhancing their quality of life and lessening their physical and mental burden.

Objective To evaluate the diagnostic value of prostate health index(PHI)and prostate health index density(PHID)in identifying intermediate-to high-risk prostate cancer(PCa).Methods Clinical data of 160 treatment-na?ve patients with highly suspected PCa,who underwent initial prostate biopsy in our hospital during Jul.2022 and Feb.2024,were retrospectively analyzed.Data included age,body mass index(BMI),prostate volume(PV),total prostate-specific antigen(tPSA),free PSA(fPSA),[-2]proPSA(p2PSA),PHI and PHID.Biopsy-positive results were stratified according to the EAU D'Amico risk criteria.Receiver operating characteristic(ROC)curve and multivariate logistic regression analysis were employed to assess the diagnostic performance of PHI and PHID in predicting PCa and identifying intermediate-to high-risk PCa.Results There were statistically significant differences in tPSA,p2PSA,PHI and PHID between the negative and positive groups,as well as among the low-,medium-and high-risk groups(P＜0.01).Both PHI and PHID demonstrated good diagnostic performance in predicting PCa(AUC=0.820 8 and 0.875 7,respectively;all P＜0.001),and in identifying intermediate-to high-risk PCa(AUC=0.838 0 and 0.878 3,respectively;all P＜0.001).Compared to the baseline model,the incorporation of PHI and PHID individually into the multivariate model significantly improved the screening performance for PCa(AUC=0.910 and 0.898,respectively;all P＜0.001).Conclusion PHI and PHID exhibit high diagnostic efficacy in screening PCa,particularly in identifying intermediate-to high-risk disease.