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*

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

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

Objective:To establish a nomogram model based on elastic imaging parameters and ultrasound image features, and evaluate its predictive value in central lymph node metastasis (CLNM) in papillary thyroid carcinoma (PTC) .Methods:The clinical data of 168 patients (the research group) with papillary thyroid carcinoma who underwent thyroid surgery in our hospital from Jan. 2019 to Dec. 2021 were retrospectively collected, including gender, age, ultrasound elastography parameters (elasticity ratio, blue area ratio), and ultrasound examination indicators (nodule diameter, nodule number, internal echo, border, edge, aspect ratio, microcalcification, capsule invasion). Another 150 patients who underwent thyroid surgery in our hospital during the same period were selected as the validation group.According to the results of postoperative pathological examination, the the research group were divided into two groups: 64 cases (38.10%) of CLNM and 104 cases (61.90%) of non-CLNM. Binary logistic regression analysis was used to explore the influencing factors of CLNM in PTC patients, and a nomogram model based on elastic imaging parameters and ultrasound image features was established. The nomogram model was drawn to predict the receiver operating characteristic (ROC) curve of CLNM in PTC patients.Results:There were statistically significant differences in nodule diameter, edge, microcalcification, capsule invasion, blue area ratio, and elasticity ratio ( P<0.05). Most of the nodules in the CLNM group were ≥10 mm in diameter, with uneven margins, an aspect ratio of <1, microcalcifications and capsular invasion. Logistic regression analysis showed that nodule diameter, capsule invasion, blue area ratio and elastic ratio were risk factors for CLNM ( P<0.05). The AUC of the combined detection was 0.857 (0.777-0.937), and the sensitivity and specificity were 78.1% and 86.5%, respectively, and the AUC and sensitivity were significantly higher than the individual detection of each index ( P<0.05). In the research group, the sensitivity and specificity of the ultrasound parameter prediction model in predicting CLNM were 81.25% (52/64) and 84.62% (88/104), respectively. In the validation group, the sensitivity and specificity of the ultrasound parameter prediction model in predicting CLNM were 79.17% (38/48) and 85.29% (87/102), respectively. Conclusion:Elastography parameters (blue area ratio, elasticity ratio) and ultrasound image features (nodule diameter, capsular invasion) are the influencing factors of CLNM in PTC patients, and the combined prediction based on the above four indicators has good application value.

BACKGROUND:Intervertebral disc degeneration is clinically considered to be the main cause of low back pain,but due to the unclear pathogenesis of intervertebral disc degeneration,there is still a lack of effective means to delay the progression of the disease.Single-cell RNA sequencing technology can amplify and sequence mRNA at the single-cell level,reveal the gene expression intensity of a single cell,discover different cell subsets in tissues according to the heterogeneity of cells,study the pathogenesis of intervertebral disc degeneration at the molecular level,and provide a new theoretical basis for its early diagnosis and treatment. OBJECTIVE:To introduce the basic principles of single-cell RNA sequencing technology and review the research progress of single-cell RNA sequencing technology in intervertebral disc degeneration in recent years. METHODS:A computer was used to search PubMed,Web of Science,CNKI and WanFang databases for the literature published from 2012 to 2022.Key words were"single-cell RNA sequencing,intervertebral disc degeneration,sequencing Technology"in Chinese and English.Duplicate,poor-quality and irrelevant articles were excluded;a total of 70 articles were eventually included. RESULTS AND CONCLUSION:(1)We identified new cell subsets such as homeostatic chondrocytes,hypertrophy chondrocyte-like nucleus pulposus cells and fibrous nucleus pulposus cells,identified the marker genes and transcription factors of these cell subsets,and described the functions,differentiation paths and cell fate of these cell subsets during the development and progression of intervertebral disc degeneration,and proposed the concept of progenitor nucleus pulposus cells.A cell subpopulation with progenitor nucleus pulposus cells properties was identified and its effectiveness in treating intervertebral disc degeneration was verified in mice.(2)Fibro chondrocyte-like annulus fibrosus cells and annulus fibrosus stem cells with both cartilage and fiber properties were identified,and a new type of composite hydrogel was prepared by combining fibrous cartilage inducers silk fibroin and hyaluronic acid in vitro.Experiments in mice demonstrated that this hydrogel could repair both annulus fibrosus tissue and cartilage matrix,and was remarkably effective in the treatment of intervertebral disc degeneration.(3)Regulatory chondrocytes were found in endplate cartilage.Two distinct fates in the progression of intervertebral disc degeneration were analyzed and the differential genes in the two fates were identified.Intercellular communication analysis indicated that regulatory chondrocytes interact with endothelial cells to promote angiogenesis.(4)Immune cells such as macrophages,T cells,myeloid progenitor cells and neutrophils were identified in the degenerated intervertebral disc tissues,demonstrating the existence of immune response during intervertebral disc degeneration.It was found that apolipoprotein induced the polarization of macrophages M1 and M2 subtypes,and this polarization process affected the activity of progenitor nucleus pulposus cells by amplifying the inflammatory response through the MIF signaling pathway.

Prompt gamma rays are a kind of secondary radiation produced in particle therapy, and prompt gamma information largely reflects the incidence of particles. Consequently, use of prompt gamma information to verify the range of particles is a promising verification method. In this article, the concept of prompt gamma ray in vivo range verification and the advantages of prompt gamma verification over existing methods were introduced. Secondly, the progress in developing a method for range verification using prompt gamma in recent years was reviewed, and the advantages and disadvantages of three methods including prompt gamma imaging (PGI), prompt gamma timing (PGT) and prompt gamma spectroscopy (PGS) were discussed. Finally, these three methods were summarized, and the development trend of prompt gamma rays for in vivo range monitoring was prospected.

Objective:To investigate the correlation between long-term use of low-dose aspirin and Helicobacter pylori(HP)infection in elderly people and its effectiveness on HP eradication and recurrence.Methods:A retrospective analysis was conducted on 2 834 elderly people aged 60 and above who underwent the C 13-or C 14-urea breath test(UBT)for the first time in the Physical Examination Center of the First Hospital of Lanzhou University between March 2010 and December 2019.According to the results of UBT, people were divided into an HP infection group with 1 510 patients and a non-HP infection group with 1 324 patients.Univariate and multivariate Logistic regression analysis were used to investigate the relationship between aspirin use and HP infection.Additionally, in a prospective case-control analysis, 544 elderly hypertensive patients diagnosed with HP infection between March 2015 and December 2020 were selected and divided into an aspirin group(402 cases)and an observation group(142 cases), based on whether aspirin was used.The aspirin group was further divided into a 1 to <2 years group(134 cases), a ≥2 to <5 years group(142 cases)and a ≥5 years group(126 cases)based on the duration of aspirin treatment.The rates of HP eradication, safety and one-year post-treatment HP recurrence with bismuth-containing quadruple therapy were compared. Results:The overall HP infection rate was 53.28%(1 510/2 834).Univariate analysis showed that the infection rate in women was higher than in men[56.86%(584/1 027) vs.51.25%(926/1 807), χ2=8.307, P=0.004].The infection rate in aspirin users was higher than in non-aspirin users[57.29%(920/1 606) vs.48.05%(590/1 228), χ2=23.866, P<0.001], with no significant difference between aspirin use for 1-<2 years, ≥2-<5 years and ≥5 years[60.22%(162/269) vs.56.4%(273/484) vs.56.86%(485/853), χ2=1.166, P=0.558].Fasting blood glucose levels in the HP infection group were higher than in the non-HP infection group[(5.92±1.78)mmol/L vs.(5.77±1.40)mmol/L, t=2.317, P=0.021].Multivariate Logistic regression analysis showed the risk of HP infection in women was higher than in men( OR=1.254, 95% CI: 1.075-1.464, P=0.004).Long-term aspirin use increased the risk of HP infection( OR=1.450, 95% CI: 1.249-1.684, P<0.001).Among the 544 cases selected for eradication therapy, 522 completed the treatment protocol, with 479 achieving successful eradication.The overall eradication rate was 91.76%(479/522)according to per-protocol(PP)analysis and 88.05%(479/544)according to intention-to-treat(ITT).After 1 year, 472 cases underwent reexamination, with an overall recurrence rate of 3.6%(17/472).There was no statistical significance in the eradication rate and recurrence rate among the groups with different durations of aspirin treatment and the observation group. Conclusions:Long-term use of low-dose aspirin increases the risk of HP infection in the elderly, but does not affect the eradication rate and one-year recurrence rate of bismuth-containing quadruple therapy.Therefore, periodic screening and eradication of HP should be performed.

Objective To solve the problem of large data volume,multiple dimensions and low value for assisting traditional Chinese medicine clinical diagnosis in home health perception layer devices.Methods Based on the principles of traditional Chinese medicine diagnosis,this paper divides home health data into three types:complementary,redundant,and collaborative,and proposes a solution for data fusion at the levels of device data,home events,and traditional Chinese medicine symptoms.Results The proposed data fusion solution in this paper enables the data collected by various devices in the home environment to work together,extracts home data that is more valuable for traditional Chinese medicine diagnosis,and reduces the real-time pressure on the home network bandwidth caused by the sensors on the home side.Conclusion The construction of an open IoT ecosystem for home health based on multiple devices is a huge project,which includes the construction of perception layer hardware,data cleaning,fusion,normalization,labeling,modeling,and other aspects.This paper focuses on the idea of home health data fusion,which can provide directions for cleaning up heterogeneous data from multiple sources at home and also provide ideas for subsequent data labeling and modeling with traditional Chinese medicine characteristics,thus providing more valuable decision-making assistance for traditional Chinese medicine clinical practice.