METTL3 facilitates prostate cancer progression via inducing HOXC6 m6A modification and stabilizing its expression through IGF2BP2-dependent mechanisms.

HOXC6 (Homeobox C6) and methyltransferase-like 3 (METTL3) have been shown to be involved in the progression of prostate cancer (PCa). However, whether HOXC6 performs oncogenic effects in PCa via METTL3-mediated N6-methyladenosine (m6A) modification is not yet reported. The Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, transwell, scratch, sphere formation assays were applied for cell growth, invasion, migration and stemness analyses. Glycolysis was evaluated by measuring glucose consumption, lactate generation and ATP/ADP ratio. The N6-methyladenine (m6A) modification profile was determined by RNA immunoprecipitation (Me-RIP) assay. The proteins that interact with PGK1 (phosphoglycerate kinase 1) were confirmed by Co-immunoprecipitation assay. Tumor formation experiments in mice were conducted for in vivo assay. PCa tissues and cells showed highly expressed HOXC6 and METTL3. Functionally, the silencing of HOXC6 or METTL3 suppresses PCa cell proliferation, invasion, migration, stemness, and glycolysis. Moreover, METTL3-induced HOXC6 m6A modification to stabilize its expression. In addition, the m6A reader IGF2BP2 directly recognized and bound to HOXC6 mRNA, and maintained its stability, and was involved in the regulation of HOXC6 expression by METTL3. Furthermore, IGF2BP2 knockdown impaired PCa cell proliferation, invasion, migration, stemness, and glycolysis by regulating HOXC6. Besides that HOXC6 interacted with the glycoytic enzyme PGK1 in PCa cells. In vivo assays further showed that METTL3 silencing reduced the expression of HOXC6 and PGK1, and impeded PCa growth. METTL3 promoted PCa progression by maintaining HOXC6 expression in an m6A-IGF2BP2-dependent mechanism.

Innovative insights: ITLN1 modulates renal injury in response to radiation.

Radiation-induced kidney injury is a common side effect of radiotherapy, as the pelvic region is in close proximity to the kidneys, posing a risk of inducing radiation-induced kidney injury when treating any pelvic malignancies with radiotherapy. This type of injury typically manifests as chronic kidney disease a few months after radiotherapy, with the potential to progress to end-stage renal disease. Radiation-induced damage involves various components of the kidney, including glomeruli, tubules, interstitium, and extracellular matrix. Therefore, investigating its molecular mechanisms is crucial. In this study, we extensively searched literature databases, selecting recent transcriptomic studies related to acute kidney injury (AKI) published in the past decade. We downloaded the raw RNA sequencing datasets GSE30718 and GSE66494 related to AKI from the GEO database and identified that intestinal-type lectin ITLN1 plays a significant role in regulating radiation-induced kidney injury in rats. Differential gene analysis was performed using chip data from the GEO database, and further bioinformatics analysis identified 13 genes that may be involved in regulating kidney injury, with ITLN1 being the most relevant to kidney damage, thus selected as the target gene for this study. Subsequently, a rat model of radiation-induced kidney injury was established for experimental validation, assessing kidney tissue morphology and injury extent through staining observation and immunohistochemical staining. The protective effect of ITLN1 on kidney function was evaluated by measuring changes in rat body weight and blood pressure, serum kidney injury markers, and kidney structure. The experimental results indicate that overexpression of ITLN1 can improve kidney function in rats with radiation-induced kidney injury by activating the Akt/GSK-3ß/Nrf2 signaling pathway, suppressing oxidative stress, cell apoptosis, inflammation, cellular senescence, and fibrosis. This study highlights the significant role of ITLN1 in regulating kidney injury, providing a novel target for future treatments of radiation-induced kidney injury.

[Identification and expression analysis of TCP family members in tobacco (Nicotiana tabacum L.)].

TCP family as plant specific transcription factor, plays an important role in different aspects of plant development. In order to screen TCP family members in tobacco, the homologous sequences of tobacco and Arabidopsis TCP family were identified by genome-wide homologous alignment. The physicochemical properties, phylogenetic relationships and cis-acting elements were analyzed by bioinformatics. The homologous genes of AtTCP3/AtTCP4 were screened, and RT-qPCR was used to detect the changes of gene expression upon 20% PEG6000 treatment. The results show that tobacco contains 63 TCP family members. Their amino acid sequence length ranged from 89 aa to 596 aa, and their protein hydropathicity grand average of hydropathicity (GRAVY) ranged from -1.147 to 0.125. The isoelectric point (pI) ranges from 4.42 to 9.94, the number of introns is 0 to 3, and the subcellular location is all located in the nucleus. The results of conserved domain and phylogenetic relationship analysis showed that the tobacco TCP family can be divided into PCF, CIN and CYC/TB1 subfamilies, and each subfamily has a stable sequence. The results of cis-acting elements in gene promoter region showed that TCP family genes contain low docile acting elements (LTR) and a variety of stress and metabolic regulation related elements (MYB, MYC). Analysis of gene expression patterns showed that AtTCP3/AtTCP4 homologous genes (NtTCP6, NtTCP28, NtTCP30, NtTCP33, NtTCP42, NtTCP57, NtTCP63) accounted for 20% PEG6000 treatment significantly up-regulated/down-regulated expression, and NtTCP30 and NtTCP57 genes were selected as candidate genes in response to drought. The results of this study analyzed the TCP family in the tobacco genome and provided candidate genes for the study of drought-resistance gene function and variety breeding in tobacco.

Decreased Nrf2 protein level and low sperm quality in intractable spermatocystitis.

ABSTRACT: To investigate the molecular etiology of low sperm quality in patients with intractable spermatocystitis, spermatozoa samples from patients with persistent hematospermia undergoing transurethral seminal vesiculoscopy and healthy volunteers were utilized. Spermatozoa samples were collected from the seminal vesicles through transurethral seminal vesiculoscopy or by masturbation ejaculation. Sperm quality was analyzed by a WLJY-9000 color semen analysis system. Measurement of tumor necrosis factor alpha (TNFα) and interleukin-6 (IL-6) in the seminal plasma was performed using enzyme-linked immunosorbent assay (ELISA). Measurement of H2O2 in the seminal plasma was performed with a hydrogen peroxide kit. The protein levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and phosphorylated-Nrf2 (p-Nrf2) were measured by western blot analysis and immunofluorescence assays. Low sperm quality parameters and increased levels of inflammatory cytokines (TNFα, IL-6, and H2O2) in the seminal plasma were detected among the semen samples from the patients with persistent hematospermia. Nrf2 and p-Nrf2 were strongly expressed in the nucleus and periphery of human sperm cells, according to the results of the immunofluorescence assays. The protein levels of Nrf2 and p-Nrf2 were significantly lower in the spermatozoa samples from patients with persistent hematospermia than in those from healthy volunteers with normal sperm motility. The results suggested that Nrf2 signaling might play a role in the low sperm quality of patients with intractable spermatocystitis.

Role of skip N2 lymph node metastasis for patients with the stage III-N2 lung adenocarcinoma: a propensity score matching analysis.

PURPOSE: Recent studies have indicated some differences in the prognosis of patients with stage III-N2 lung adenocarcinoma, and the prognosis of patients with skip N2 lymph node metastasis (SKN2) is good. This study grouped patients with stage III-N2 lung adenocarcinoma by propensity score matching (PSM) to evaluate the impact of SKN2 on the prognosis of these patients. METHODS: The clinical data for patients who underwent radical lobectomy and had a postoperative pathological diagnosis of stage III-N2 lung adenocarcinoma at our centre from 2016 to 2018 were collected, and PSM was performed at a ratio of 1:1. RESULTS: A total of 456 patients were enrolled in this study. After PSM, 112 patients were included in the SKN2 group, and 112 patients were included in the non-SKN2 group. When comparing the SKN2 group with the non-SKN2 group, the 3-year OS rate was (71.4% vs. 12.5%, p < 0.001), and the 3-year DFS rate was (35.7% vs. 5.4%, p < 0.001). It is further divided into four groups:single-station SKN2 (N2a1),Multi-station SKN2 (N2a2),single-station non-SKN2 (N2b1) and Multi-station non-SKN2 (N2b2).The 3-year OS and DFS rates of skip lymph node metastasis were better than those of non-skip lymph node metastasis(OS:N2a1 vs. N2b1 68.4% vs. 23.5%,p < 0.001;N2a2 vs. N2b2 73.0% vs. 7.7%,p < 0.001)(DFS:N2a1 vs. N2b1 68.4% vs. 5.9%,p < 0.001;N2a2 vs. N2b2 62.2% vs. 5.1%,p < 0.001), regardless of the number of N2 station(OS:N2a1 vs. N2a2 68.4% vs. 73.0%,p = 0.584;N2b1 vs. N2b2 23.5% vs. 7.7%,p = 0.051). On multivariate analysis, sex (p = 0.008) ,Vascular tumour thrombus(p = 0.047),size(p = 0.002)and SKN2 (p < 0.001) were independent predictors of OS. CONCLUSION: For patients with stage III-N2 lung adenocarcinoma, the prognosis of SKN2 patients is better than non-SKN2 patients', and SKN2 may be used as an important factor in the N2 subgroup classification in future TNM staging.

Adjuvant chemotherapy can benefit the survival of stage I lung adenocarcinoma patients with tumour spread through air spaces after resection: Propensity-score matched analysis.

Background: It is still unclear whether stage I lung adenocarcinoma patients with tumour spread through air spaces (STAS) can benefit from postoperative adjuvant chemotherapy (ACT) after lobectomy. This study investigated the effect of ACT on the postoperative survival of patients with stage I (STAS+) lung adenocarcinoma. Methods: We retrospectively analysed the clinical data of stage I (STAS+) invasive lung adenocarcinoma patients who underwent lobectomy in the Department of Thoracic Surgery of our hospital from January 1, 2013 to January 1, 2016. Propensity score matching (PSM) was performed to group patients to investigate whether ACT could lead to better prognosis of patients. Results: A total of 593 patients with stage I (STAS+) lung adenocarcinoma were enrolled. The study after PSM included 406 patients. Kaplan-Meier survival analysis showed the experimental group had a better 3-year recurrence-free survival (RFS) rate (p = 0.037) and the 5-year RFS rate (p = 0.022) than the control group. It also had higher 5-year overall survival (p = 0.017). The multivariate analysis by Cox proportional hazard regression model showed that stage I STAS+ lung adenocarcinoma patients with lymphatic vessel invasion (HR: 1.711, 95% CI: 1.052-2.784; p = 0.045), vascular invasion (HR: 5.014, 95% CI: 3.154-7.969; p < 0.001), and visceral pleural invasion (HR: 2.086, 95% CI: 1.162-3.743; p = 0.014), and without ACT (HR: 1.675, 95% CI: 1.043-2.689; p = 0.033) had a significant survival disadvantage. Conclusion: ACT can boost the postoperative survival of patients with stage I (STAS+) lung adenocarcinoma.

Clinical characteristics and prognostic value of EGFR mutation in stage I lung adenocarcinoma with spread through air spaces after surgical resection.

The clinical data of stage I invasive lung adenocarcinoma patients with spread through air spaces (STAS) who underwent lobectomy from January 1, 2013 to January 1, 2016 at the Department of Thoracic Surgery of Hebei Medical University were analyzed retrospectively, and statistical analysis was carried out to explore their clinical features and prognostic value of EGFR mutation. A total of 280 patients were included in the study cohort, and EGFR mutations were detected in 154 patients. EGFR mutations were more common in non-smokers (p=0.045), females (p<0.001), without vascular tumor thrombus (p=0.037), and histological subtype LPA/APA/PPA (p=0.001). Multivariate analysis of the Cox risk regression model showed that EGFR gene mutation (p=0.807) was not an independent influencing factor of recurrence-free survival (RFS), but EGFR mutation was an independent influencing factor of overall survival (OS) (p=0.012), and OS of patients with EGFR mutation was better. The EGFR mutation also significantly increased the progression-free survival (PFS) of relapsed patients (p<0.001), but the PFS of relapsed EGFR mutation patients who received adjuvant chemotherapy after the operation was worse than that of patients who did not receive adjuvant chemotherapy (p=0.029). EGFR gene mutation is not a risk factor for postoperative recurrence in patients with stage I lung adenocarcinoma with STAS but the 5-year survival rate of patients with EGFR gene mutation is better than that of wild-type. Postoperative adjuvant chemotherapy for patients with EGFR mutation should be carefully considered.

The role of PERK and IRE1 signaling pathways in excessive fluoride mediated impairment of lymphocytes in rats' spleen in vivo and in vitro.

Fluoride is capable of inducing immunotoxicity, but its molecular mechanisms remain elusive. This study aimed to explore the roles of Protein kinase receptor-like ER kinase (PERK) and inositol requiring enzyme 1 (IRE1) signaling pathways in excessive fluoride-induced immunotoxicity, focusing on the regulatory roles of these two pathways in cell division and apoptosis. Firstly, we assessed the changes in cell division and apoptosis in rats exposed to 0, 50, or 100 mg/L fluoride, and detected the expression of PERK and IRE1 signaling-related proteins in spleen. Additionally, to validate the role of these two pathways, we evaluated the changes in cell division and apoptosis of primary lymphocytes from rat's spleen to 4 mM fluoride after knockdown of PERK and IRE1 in vitro. In vivo results confirmed that fluoride inhibited cell division, promoted the apoptosis and resulted in histological and ultrastructural abnormalities of rat spleen. In addition, fluoride induced activation of the PERK and IRE1 signalings and the associated apoptosis. Moreover, the in vitro results further verified the findings in vivo that fluoride activated these two signalings in B lymphocytes. Importantly, after knockdown of PERK and IRE1 in lymphocytes, the cell division ability was restored, and apoptosis decreased in fluoride-treated lymphocytes; the results correlated well with the expression of PERK and IRE1 signaling-related proteins, thus confirming the pivotal role of these pathways in immunosuppression by excessive fluoride. This study indicates that the mechanisms underlying the deleterious effects of fluoride on immune system are related to activation of the PERK and IRE1 signaling pathways.

Analyzing axial stress and deformation of tubular for steam injection process in deviated wells based on the varied (T, P) fields.

The axial stress and deformation of high temperature high pressure deviated gas wells are studied. A new model is multiple nonlinear equation systems by comprehensive consideration of axial load of tubular string, internal and external fluid pressure, normal pressure between the tubular and well wall, and friction and viscous friction of fluid flowing. The varied temperature and pressure fields were researched by the coupled differential equations concerning mass, momentum, and energy equations instead of traditional methods. The axial load, the normal pressure, the friction, and four deformation lengths of tubular string are got ten by means of the dimensionless iterative interpolation algorithm. The basic data of the X Well, 1300 meters deep, are used for case history calculations. The results and some useful conclusions can provide technical reliability in the process of designing well testing in oil or gas wells.