The deubiquitinating enzyme USP11 regulates breast cancer progression by stabilizing PGAM5.

Breast cancer is common worldwide. Phosphoglycerate mutase 5 (PGAM5) belongs to the phosphoglycerate mutase family and plays an important role in many cancers. However, research on its role in breast cancer remains unclear. The present investigation highlights the significant expression of PGAM5 in breast cancer and its essential role in cell proliferation, invasion, apoptosis and the regulation of ferroptosis in breast cancer cells. Overexpression or knockdown of ubiquitin-specific protease 11 (USP11) promotes or inhibits the growth and metastasis of breast cancer cells, respectively, in vitro and in vivo. Mechanistically, USP11 stabilizes PGAM5 via de-ubiquitination, protecting it from proteasome-mediated degradation. In addition, the USP11/PGAM5 complex promotes breast cancer progression by activating iron death-related proteins, indicating that the synergy between USP11 and PGAM5 may serve as a predictor of disease outcome and provide a new treatment strategy for breast cancer.

RPLP1 promotes tumor metastasis and is associated with a poor prognosis in triple-negative breast cancer patients.

BACKGROUND: Cancer metastasis is the major reason for cancer related deaths, and the mechanism of cancer metastasis still unclear. RPLP1, a member of a group of proteins known as ribosomal proteins, is associated with tumorigenesis and primary cell immortalization and is involved in cellular transformation. However, the expression and potential function of RPLP1 in TNBC remain unclear. METHODS: The expression of RPLP1 in TNBC tissues and cell lines were detected with Real-Time PCR and western blotting. 81 cases of TNBC tissue samples and adjacent non-tumor tissue samples were tested by immunochemistry to determine the correlation between the RPLP1 expression and clinicopathological characteristics. In vitro, we determined the role and mechanistic pathways of RPLP1 in tumor metastasis in TNBC cell lines. RESULTS: In this study, we detected high levels of RPLP1 expression in TNBC samples and cell lines. RPLP1 is upregulated in triple-negative breast cancer (TNBC) tissues and cells, and high expression levels correlate with an increased risk of recurrence and metastasis. Furthermore, high RPLP1 expression was associated with a poor prognosis and was an independent prognostic marker for TNBC. In RPLP1-induced cancer metastasis, RPLP1 may increase cancer cell invasion, which is likely the result of its effect on the cancer cell epithelial-mesenchymal transition. CONCLUSIONS: Altogether, our findings indicate RPLP1 is a poor prognostic potential biomarker and anti-metastasis candidate therapeutic target in triple-negative breast cancer.

N2O Emission and Hydroxylamine Oxidase (HAO) Activity in a Nitrogen Removal Process Based on Activated Sludge with Three COD/NH4+ Ratios.

This study dealt with nitrous oxide (N2O) emission and hydroxylamine oxidase (HAO) activity of waste sludge in a nitrification and denitrification process employing three carbon nitrogen (C/N) ratios in a sequencing batch reactor (SBR). The experimental results indicated that N2O emission increased dramatically after the C/N ratio in the sludge increased from 6.5 to 9.3, which was greater than the N2O emission at two other C/N ratios (3.5, 6.5). The HAO activity in the anoxic period was higher with all three C/N ratios than in the aerobic period. The results suggest that N2O was produced primarily in the aerobic period and the main source of the N2O emission resulted from denitrification by nitrifying bacteria and aerobic hydroxylamine oxidation. When a relatively deficient carbon source existed, the N2O emission under anoxic conditions was affected by the HAO activity and vice versa. When the HAO activity was relatively high, it was found that more N2O was released.

The far-upstream element-binding protein 2 is correlated with proliferation and doxorubicin resistance in human breast cancer cell lines.

Far-upstream element (FUSE)-binding protein 2 (FBP2) was a member of single-stranded DNA-binding protein family; it played an important role in regulating transcription and post-transcription and is involved in the regulation of C-MYC gene expression in liver tumors. However, the role of FBP2 in breast cancer and its mechanism has not been studied yet. Here, we discovered that FBP2 was up-regulated in breast cancer tissues and breast cancer cell lines. Moreover, immunohistochemistry analysis demonstrated that up-regulated FBP2 was highly associated with tumor grade, Ki-67, and poor prognosis, which was an independent prognostic factor for survival of breast cancer patients. At the cellular level, we found that FBP2 was correlated with cell cycle progression by accelerating G1/S transition, and knockdown of FBP2 could weaken cell proliferation, anchorage-independent cell growth, while enhancing the sensitivity of breast cancer cells to doxorubicin. More importantly, we found that activation of PI3K/AKT pathway could phosphorylate FBP2, and then make FBP2 shuttle from cytoplasm into the nucleus, which was the main mechanism of breast cancer cell proliferation and drug resistance. Taken together, our findings supported the notion that FBP2 might via PI3K/AKT pathway influence breast cancer progression and drug resistance, which might provide a new target for the design of anti-cancer drugs for breast cancer patients.

Identification of biomimetic nanoplatform-mediated delivery of si-ISG15 for treatment of triple-negative breast cancer.

Triple-negative breast cancer (TNBC) is recognized as the most malicious form of breast cancer and exhibits an alarming tendency for recurrence, a heightened propensity for metastasis, and an overwhelmingly grim prognosis. Therefore, effective therapy approaches for TNBC are urgently required. In this study, the interferon-stimulated gene 15 (ISG15) expression level was analyzed by bioinformatics and verified by Western blot analysis. The effects of ISG15 on the proliferation and metastasis of TNBC cells were assessed using MTT, Colony formation, EdU, Transwell, and Flow cytometry assays. We also developed a cancer cell-biomimetic nanoparticle delivery system and evaluated its therapeutic efficacy in vivo. In this study, we reported that ISG15 was upregulated in TNBC, and its high expression level correlated with an increased risk of tumorigenesis. Through in vitro and in vivo studies, we discovered that ISG15 knockdown drastically suppressed cell proliferation, invasion, and migration and induced apoptosis in TNBC cells. Our findings revealed that ISG15 was a candidate therapeutic target in TNBC because of its key role in malignant growth and invasion. Moreover, co-immunoprecipitation showed that ISG15 exerted oncogenic functions through its interaction with ATP binding cassette subfamily E member 1 and activated the Janus kinase/signal transducers and activators of the transcription signaling pathway. Furthermore, we created a nanoparticle-based siRNA camouflaged using a cancer cell membrane vesicle delivery system (the CM@NP complex) and confirmed its therapeutic effects in vivo. Our findings confirmed that ISG15 may play a pivotal oncogenic role in the development of TNBC and that CM@siRNA-NP complexes are an effective delivery system and a novel biological strategy for treating TNBC.

Enhanced breast cancer treatment using phototherapy and RNS therapy with macrophage membrane-coated liposomes.

Breast cancer, the predominant malignancy afflicting women, continues to pose formidable challenges despite advancements in therapeutic interventions. This study elucidates the potential of phototherapy, comprising both photothermal and photodynamic therapy (PTT/PDT), as a novel and promising modality. To achieve this goal, we devised liposomes coated with macrophage cell membranes including macrophage-associated membrane proteins, which have demonstrated promise in biomimetic delivery systems for targeting tumors while preserving their inherent tumor-homing capabilities. This integrated biomimetic delivery system comprised IR780, NONOate, and perfluorocarbon. This strategic encapsulation aims to achieve a synergistic combination of photodynamic therapy (PDT) and reactive nitrogen species (RNS) therapy. Under near-infrared laser irradiation at 808 nm, IR780 demonstrates its ability to prolifically generate reactive oxygen species (ROS), including superoxide anion (O2•-), singlet oxygen, and hydroxyl radical (·OH). Simultaneously, NONOate releases nitric oxide (NO) gas upon the same laser irradiation, thereby engaging with IR780-induced ROS to facilitate the formation of peroxynitrite anion (ONOO-), ultimately inducing programmed cell death in cancer cells. Additionally, the perfluorocarbon component of our delivery system exhibits a notable affinity for oxygen and demonstrates efficient oxygen-carrying capabilities. Our results demonstrate that IR780-NO-PFH-Lip@M significantly enhances breast cancer cell toxicity, reducing proliferation and in vivo tumor growth through simultaneous heat, ROS, and RNS production. This study contributes valuable insights to the ongoing discourse on innovative strategies for advancing cancer therapeutics.

Visible light photocatalytic degradation of oxytetracycline hydrochloride using chitosan-loaded Z-scheme heterostructured material BiOCOOH/O-gC3N4.

In this work, a Z-scheme heterostructured BiOCOOH/O-gC3N4 material was synthesized and immobilized on chitosan (CTS) to obtain the BiOCOOH/O-gC3N4/CTS photocatalytic material for photocatalytic degradation of oxytetracycline hydrochloride (CTC).Our findings indicate that the composite material BiOCOOH/O-gC3N4, as well as the BiOCOOH/O-gC3N4/CTS composite membrane, displayed a significantly higher efficiency in photocatalytic degradation of CTC compared to BiOCOOH alone, owing to the synergistic effect of adsorption and photocatalysis. Following four cycles of use, the composite material retained around 96 % of its initial photocatalytic degradation activity. The addition of CTS in the photocatalytic material resolved issues such as aggregation and difficult recovery commonly encountered with powder materials, thereby facilitating effective collision between the photocatalytic active sites and CTC. Experimental and theoretical calculations provided confirmation that the combination of BiOCOOH and O-gC3N4 effectively enhanced the light absorption capacity and photocatalytic performance. Furthermore, we investigated the influence of environmental factors such as pH value and anions on the photocatalytic degradation experiment, which offers valuable insights for the application of composite catalysts in wastewater treatment.

Prime editing-mediated correction of the leptin receptor in muscle cells of db/db mice.

Genetic diseases can be caused by monogenic diseases, which result from a single gene mutation in the DNA sequence. Many innovative approaches have been developed to cure monogenic genetic diseases, namely by genome editing. A specific type of genomic editing, prime editing, has the potential advantage to edit the human genome without requiring double-strand breaks or donor DNA templates for editing. Additionally, prime editing does not require a precisely positioned protospacer adjacent motif (PAM) sequence, which offers flexible target and more precise genomic editing. Here we detail a novel construction of a prime editing extended guide RNA (pegRNA) to target mutated leptin receptors in B6.BKS(D)-Leprdb/J mice (db/db mice). The pegRNA was then injected into the flexor digitorum brevis (FDB) muscle of db/db mice to demonstrate in vivo efficacy, which resulted in pegRNA mediated base transversion at endogenous base transversion. Genomic DNA sequencing confirmed that prime editing could correct the mutation of leptin receptor gene in db/db mice. Furthermore, prime editing treated skeletal muscle exhibited enhanced leptin receptor signals. Thus, the current study showed in vivo efficacy of prime editing to correct mutant protein and rescue the physiology associated with functional protein.

SUZ12 depletion suppresses the proliferation of gastric cancer cells.

BACKGROUND/AIMS: SUZ12 and EZH2 are two main components of polycomb repressive complex 2 (PRC2) that is known to be of great importance in tumorigenesis. EZH2 has been reported to play a vital role in pathogenesis of human cancer. However, whether SUZ12 has equivalent roles in tumorigenesis has not been demonstrated. Here, we investigated a possible role of SUZ12 for the proliferation of gastric cancer cells. METHODS: Western-blot analysis was used to detected the levels of SUZ12, H3K27me3, EZH2 and p27 in ten gastric cell lines. SUZ12 was depleted by RNA interference. Cell cycle was detected by flow cytometry. Luciferase assays was to analyze whether miR-200b directly regulate SUZ12. RESULTS: We found that SUZ12 depletion mediated by RNA interference (RNAi) led to a reduction of gastric cell numbers and arrested the cell cycle at G1/S point. As an important G1/S phase inhibitory gene, p27 is re-induced to some extent by SUZ12 knockdown. Furthermore, we demonstrated that SUZ12 was directly downregulated by miR-200b. CONCLUSION: We provide evidence suggesting that SUZ12 may be a potential therapeutic target for gastric cancer.

ERα promotes SUMO1 transcription by binding with the ERE and enhances SUMO1-mediated protein SUMOylation in breast cancer.

Background: Estrogen plays a crucial role in the tumorigenesis of breast cancer (BC), and epigenetic modification by SUMOylation is essential for cancer development. However, the mechanism underlying estrogen's actions on protein SUMOylation and its effect on BC development are still incompletely understood. Methods: SUMO1 in BC cell lines was verified via real-time quantitative PCR (RT-qPCR) and western blot. Cell proliferation and colony formation assays was also performed to evaluate SUMOylation as mediated by SUMO1. Luciferase activity to examine whether E2 promoted the transcription of SUMO1, and chromatin immunoprecipitation (ChIP) assay to determine the binding of estrogen receptor alpha (ERα) to SUMO1 were conduction, and an animal model was used to evaluate the effects of E2-ERα-enhanced SUMO1 transcription. Results: E2 promoted SUMO1 mRNA and protein expression levels in a dose- and time-dependent manner in ER-positive BC cells; it exerted no influence on SUMO2/3 expression; in E2-induced SUMO1 transcription, ERα, but not ERß, was essential to the process. In addition, E2-ERα upregulated the transcription of SUMO1 by binding with an estrogen-response element half-site (1/2ERE, in the -134 to -123 bp region) of the SUMO1 promoter, and E2-ERα induced SUMO1 transcription-enhanced cellular viability in ER-positive BC cells. To further determine SUMOylation as mediated by SUMO1 in ER-positive BC, we evaluated novel SUMO1 target proteins such as Ras and demonstrated that E2 increased Ras SUMOylation and cellular proliferation by affecting downstream signaling-pathway transduction. Finally, our data revealed that E2-ERα enhanced SUMO1 transcription to promote tumor growth in a BC orthotopic tumor model. Conclusions: Collectively, our results showed that E2 promoted the transcription and protein expression of SUMO1 via ERα binding to a 1/2ERE in the SUMO1 promoter, and that E2-ERα also augmented SUMO1-mediated Ras SUMOylation and mediated cellular responses in ER-positive BC. We therefore achieved significant insights into the mechanism involved in ER-positive BC development and provided a novel target for its treatment.

Heterojunction material BiYO3/g-C3N4 modified with cellulose nanofibers for photocatalytic degradation of tetracycline.

Cellulose nanofibers (CNFs) with large specific surface area and superb adsorption capacity are excellent photocatalyst carriers. In this study, heterojunction powder material BiYO3/g-C3N4 was successfully synthesized for the photocatalytic degradation of tetracycline (TC). The photocatalytic material BiYO3/g-C3N4/CNFs was obtained by loading BiYO3/g-C3N4 on CNFs using electrostatic self-assembly method. BiYO3/g-C3N4/CNFs exhibit a fluffy porous structure and large specific surface area, strong absorption in the visible light range, and the rapid transfer of photogenerated electron-hole pairs. Polymer-modified photocatalytic materials overcome the disadvantages of powder materials that are easy to reunite and difficult to recover. With synergistic effects of adsorption and photocatalysis, the catalyst demonstrated excellent TC removal efficiency, and the composite maintained nearly 90 % of its initial photocatalytic degradation activity after five cycles of use. The superior photocatalytic activity of the catalysts is also attributable to the formation of heterojunctions, and the heterojunction electron transfer pathway was confirmed by experimental studies and theoretical calculations. This work demonstrates that there is great research potential in using polymer modified photocatalysts to improve photocatalyst performance.

The Study of the Transport Mechanism of Isorhynchophylline in Liver.

To investigate the transport mechanism of isorhynchophylline (IRN) by using the specific inhibitors of organic cation transporters (OCTs) and organic anion transporting polypeptides (OATPs) and attempt illustrate the metabolic mechanism of IRN in the liver. All animals were randomly divided into three groups: control group (only inject IRN), RIF group (inject IRN and rifampicin), and ADR group (inject IRN and adrenalone). The control group was injected with IRN via the caudal vein. The RIF group was injected with rifampicin (RIF) by gavage, and after 1 h, IRN was injected into the caudal vein. Similarly, the ADR group received adrenalone by the caudal vein, and after 0.5 h, IRN was injected into the caudal vein. Thereafter, blood samples were obtained by the heart punctures at 90 min, 180 min, and 300 min following drug administration. Rats were sacrificed at 300 min after drug administration; then, the liver tissue was harvested. The level of IRN was measured by using high-performance liquid chromatography (HPLC), and the Kp values were calculated. After RIF administration (OATPs inhibitors), the Kp value of IRN was slightly decreased when compared with that of the control group. Meanwhile, the Kp value of IRN was dramatically reduced compared to that of the control group following ADR administration (OCTs inhibitors). The results suggested that OCTs have mainly participated in the hepatic uptake process of IRN.

Establishing a predictive model of hypoparathyroidism after total thyroidectomy and central lymph node dissection for postoperative calcium supplementation selectively.

Background: The core goal of this article is to find some meaningful risk factors that can affect the postoperative hypoparathyroidism of thyroid cancer, create an effective prediction model on this basis, and use it to selectively implement routine prophylactic calcium supplementation for patients after thyroid carcinoma surgery. Methods: The clinicopathological characteristics of patients with papillary thyroid carcinoma (PTC) who underwent conventional bilateral total thyroidectomy (TT) + bilateral central lymph node dissection from January 2020 to August 2021 in the Affiliated Hospital of Nantong University were retrospectively analyzed. Firstly, this study analyzed the relationship between postoperative hypocalcemia and hypoparathyroidism. Then, we included many potential risk factors such as gender, age, body mass index (BMI), lateral lymph node dissection (LLND) and so on and also performed univariate and multivariate analysis of the independent risk factors for postoperative hypoparathyroidism in patients, and established a predictive scoring model. Results: Among the 401 patients with PTC, 50.1% developed postoperative hypoparathyroidism. There was significant difference in serum calcium concentration between normal parathyroid group and hypoparathyroidism group after thyroid carcinoma surgery. BMI <24 kg/m2, lateral lymph node dissection, multifocality, and extrathyroidal extension (ETE) were all identified as independent risk factors for postoperative hypoparathyroidism. Based on these independent risk factors, a nine-point risk scoring model was created to firstly assess the postoperative parathyroid function status of patients and then to determine whether routine prophylactic calcium supplementation is needed. Importantly, the area under the curve (AUC) of the risk scoring model is equal to 0.979. Conclusions: At present, prophylactic calcium supplementation after thyroid carcinoma surgery is a controversial postoperative treatment. It should be selectively implemented for high-risk patients with hypoparathyroidism after surgery. Routine prophylactic calcium supplementation is recommended for PTC patients with a score greater than or equal to 5, although there are no clinical symptoms of postoperative hypocalcemia caused by hypoparathyroidism. However, prophylactic calcium supplementation is not recommended for patients with PTC with a score of less than 5; if the patient develops hypocalcemia at the later stage, therapeutic calcium supplementation can then be implemented.

Analyzing the mechanism of nitrous oxide production in aerobic phase of anoxic/aerobic sequential batch reactor from the perspective of key enzymes.

How the vast majority of nitrous oxide (N2O) in the aerobic zone of nitrogen bio-removal process is produced is still a controversial issue. To solve this issue, this study measured the activities of two key denitrifying enzymes (nitric oxide reductase (Nor) and nitrous oxide reductase (N2OR)) in an A/O SBR with different chemical nitrogen demand (COD)/total nitrogen (TN) ratios. By analyzing the Spearman's correlations between the N2O production, the enzyme activities, and the factors, the main N2O production process was identified. By comparing the activities of these enzymes, this study analyzed the reasons for the N2O production. Results show that Nor activities had a linear relationship with total N2O concentrations (y = 0.34749 + 31.31365x, R2 = 0.83362) and were not affected by COD (r = 0.299, N = 15, P = 0.279 > 0.05), which showed that most of the N2O released and produced came from the autotrophic denitrification. N2OR activities had a positive correlation with COD (r = 0.692, N = 15, P = 0.004 < 0.01), which showed that heterotrophic denitrification played a role as an N2O consumer. Nor activities were much higher than N2OR activities and the gap between them increased when the total N2O concentration increased, showing that the heterotrophic denitrification was difficult to consume all the N2O produced by the autotrophic denitrification. Reducing autotrophic denitrification is the best way to reduce N2O production in aerobic phase.

Analgesic and Anxiolytic Effects of Gastrodin and Its Influences on Ferroptosis and Jejunal Microbiota in Complete Freund's Adjuvant-Injected Mice.

This study investigated the effects of gastrodin (GAS) on analgesic, anxiolytic, ferroptosis, and jejunal microbiota in chronic inflammatory pain mice. The chronic inflammatory pain model of C57BL/6J mice was established by hindpaw injection of complete Freund's adjuvant (CFA). After GAS treatment, thermal hyperalgesia test, mechanical allodynia test, elevated plus-maze (EPMT), and open-field test (OFT) were performed to assess the behavioral changes of pain and anxiety. mRNAs of FTHI, GPX4, HO-1, and PTGS2 and jejunal microbiota were measured by qPCR. In CFA-injected C57BL/6 mice, we found that the mechanical and thermal pain threshold were increased with treatment of GAS. In EPMT, the number of entries in open arms and retention times of open arms were increased by GAS. In the OFT, the time spent in the central area was also increased. Furthermore, GAS enhanced mRNA expressions of FTHI, GPX4, and HO-1 but decreased the expression of PTGS2 in a dose-dependent manner. GAS is effective in the treatment of mice chronic inflammatory pain and anxiety-like behaviors. It may be exhibits potential neuroprotective effects through inhibition of ferroptosis independently of the intestinal microbiota.

Surgical outcomes of different approaches to dissection of lymph nodes posterior to right recurrent laryngeal nerve: a retrospective comparative cohort study of endoscopic thyroidectomy via the areolar approach and via the axillo-breast approach.

Background: The American Thyroid Association (ATA) points out that lymph nodes posterior to right recurrent laryngeal nerve (LN-prRLN) should be routinely dissected. Due to the high risk of nerve injury, the lymph nodes in this area are difficult to dissect thoroughly. Although there are many approaches to endoscopic thyroidectomy, no study has been conducted on which one is more suitable. The purpose of this study was to evaluate the safety, thoroughness, related trauma, and feasibility of two widely used endoscopic thyroidectomy approaches, so as to provide a basis for the surgeon to select a better surgical approach. Methods: This retrospective study included patients who underwent ETA (n=26) and ETAB (n=36). All patients had a pathological diagnosis of papillary thyroid carcinoma (PTC) and underwent endoscopic right thyroidectomy from May 2015 to February 2022 in the Affiliated Hospital of Nantong University. The basic clinical data and surgical outcomes of the two groups were compared. Results: There was no statistical difference between the two groups in basic clinical data and oncological characteristics, which meant that the baseline data of the two groups of patients were comparable. Significant statistical significance was observed in the operation duration (149.38±44.15 vs. 119.22±45.48 min, P=0.011), drainage volume 24 h after operation (95.54±16.79 vs. 54.46±15.11 mL, P<0.001), visual analog score (VAS) 24 h after operation (3.69±1.44 vs. 2.25±1.32, P<0.001), hospitalization duration after the operation (3.19±0.75 vs. 2.25±0.44 days, P<0.001), number of lymph node dissections after right recurrent laryngeal nerve resection (0.96±1.08 vs. 2.06±1.77, P=0.007), and number of lymph node metastases after right recurrent laryngeal nerve resection (0.12±0.33 vs. 0.58±1.00, P=0.025). Besides, there was no significant difference in the numbers of central lymph node dissections and central lymph node metastases. Conclusions: Our study indicated that compared with ETA, ETAB may perform a more efficient dissection of the LN-prRLN based on less surgical trauma, which could provide a basis for the surgeon to select a better surgical approach.

Prophylactic central lymph node dissection performed selectively with cN0 papillary thyroid carcinoma according to a risk-scoring model.

Background: This study aimed to explore the risk factors of central lymph node metastasis (CLNM) in patients with clinical central lymph node-negative papillary thyroid carcinoma (PTC), and emphasize the guidance of the risk scoring model for prophylactic central lymph node dissection (pCLND) in patients with clinical lymph node-negative (cN0) PTC. Methods: A total of 582 patients with cN0 PTC who underwent unilateral/bilateral thyroidectomy and prophylactic central lymph node dissection (pCLND) in the Affiliated Hospital of Nantong University from January 2020 to February 2021 were retrospectively analyzed. Univariate and multivariate analyses were performed to determine the risk factors of cN0 PTC. According to the independent risk factors of patients with cN0 PTC, a risk-scoring model was established. Then, the rationality of this risk scoring model was verified by additional clinical data of 112 patients with cN0 PTC in the Affiliated Hospital of Nantong University from March 2021 to April 2021. Results: Among 582 cases of cN0 PTC, 53.6% of the patients with cN0 had CLNM. The independent risk factors for CLNM in patients with cN0 PTC included male gender, <45 years of age, tumor with a maximum diameter of ≥1.0 cm, tumor location: middle/lower poles of the thyroid gland, multifocality, and extrathyroidal extension (ETE), and some ultrasound features, such as intra-nodular vascularity, microcalcification, irregular shape, and infiltrative margin. According to independent risk factors, a 24-point risk scoring model was established to predict CLNM in patients with cN0 PTC. Conclusions: Currently, prophylactic central neck lymph node dissection is a controversial operation, which should be selectively performed only for high-risk patients with cN0 PTC. For cN0 PTC patients with scores ≥14 and high-risk patients, even if no CLNM is found before surgery, routine prophylactic CLND is recommended. In addition, for cN0 PTC patients with a score of fewer than 14 points, it is recommended to perform fine-needle aspiration (FNA) before surgery, carefully assess the condition of the central lymph nodes, and then select the best surgical plan based on the results of the assessment.

Comprehensive analysis of the relationship between the ferroptosis and tumor-infiltrating immune cells, mutation, and immunotherapy in breast cancer.

Background: Ferroptosis is a kind of programmed cell death that is characterized by iron dependence. It differs from apoptosis, necrosis, autophagy, pyroptosis, and other types of cell death. Some studies have found that most of the genes involved in the regulation of ferroptosis or act as markers of ferroptosis are related to the poor prognosis of cancer patients. Methods: This study evaluated the expression, mutation, and copy number variation (CNV) of 60 previously reported ferroptosis genes in breast cancer samples from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Unsupervised clustering of breast cancer samples with ferroptosis genes was performed, followed by enrichment analysis with Gene Set Variation Analysis (GSVA), mutation display, and correlation analysis of clinical characteristics. Based on the analysis of differences among groups, the ferroptosis-related genes were identified, and the consistent clustering of breast cancer samples was performed. The characteristic genes were screened by stochastic forest algorithm and COX analysis, and a ferroptosis score (ferr.score) model was constructed to evaluate the prognosis of breast cancer patients. Results: Copy number amplification and deletion of ferroptosis genes are common in breast cancer. Breast cancer patients grouped by ferroptosis gene clusters showed significant differences in survival, immune cell infiltration, and enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways. The ferroptosis-related differential genes were identified by comparison among clustering groups of ferroptosis gene. Characteristic genes were screened from these ferroptosis-related differential genes to construct the ferr.score model. The scoring model could accurately distinguish and predict the survival prognosis and immunotherapy efficacy in breast cancer patients. Conclusions: Ferroptosis plays an important role in the occurrence and development of tumors. According to the ferr.score model, the breast cancer samples can be divided into two groups with significantly different prognoses. These results provide novel insights and ideas for immunotherapy in breast cancer patients.

CAPN8 involves with exhausted, inflamed, and desert immune microenvironment to influence the metastasis of thyroid cancer.

Background: Thyroid cancer (THCA) is the most prevalent malignant disease of the endocrine system, in which 5-year survival can attain about 95%, but patients with metastasis have a poor prognosis. Very little is known about the role of CAPN8 in the metastasis of THCA. In particular, the effect of CAPN8 on the tumor immune microenvironment (TIME) and immunotherapy response is unclear. Material and methods: Multiome datasets and multiple cohorts were acquired for analysis. Firstly, the expression and the prognostic value of CAPN8 were explored in public datasets and in vitro tumor tissues. Then, hierarchical clustering analysis was performed to identify the immune subtypes of THCA according to the expression of CAPN8 and the activities of related pathways. Subsequent analyses explored the different patterns of TIME, genetic alteration, DNA replication stress, drug sensitivity, and immunotherapy response among the three immune phenotypes. Finally, five individual cohorts of thyroid cancer were utilized to test the robustness and extrapolation of the three immune clusters. Results: CAPN8 was found to be a significant risk factor for THCA with a markedly elevated level of mRNA and protein in tumor tissues. This potential oncogene could induce the activation of epithelial-mesenchymal transition and E2F-targeted pathways. Three subtypes were identified for THCA, including immune exhausted, inflamed, and immune desert phenotypes. The exhausted type was characterized by a markedly increased expression of inhibitory receptors and infiltration of immune cells but was much more likely to respond to immunotherapy. The immune desert type was resistant to common chemotherapeutics with extensive genomic mutation and copy number variance. Conclusion: The present study firstly explored the role of CAPN8 in the metastasis of THCA from the aspects of TIME. Three immune subtypes were identified with quite different patterns of prognosis, immunotherapy response, and drug sensitivity, providing novel insights for the treatment of THCA and helping understand the cross-talk between CAPN8 and tumor immune microenvironment.

SEC61G promotes breast cancer development and metastasis via modulating glycolysis and is transcriptionally regulated by E2F1.

Breast cancer is the most common cancer in women and its incidence rates are rapidly increasing in China. Understanding the molecular mechanisms of breast cancer tumorigenesis enables the development of novel therapeutic strategies. SEC61G is a subunit of the endoplasmic reticulum translocon that plays critical roles in various tumors. We aimed to investigate the expression and function of SEC61G in breast cancer. By analyzing The Cancer Genome Atlas breast cancer cohort, we found that SEC61G was highly expressed in breast cancer and predicted poor prognosis of breast cancer patients. Overexpression of SEC61G and its prognostic role was also confirmed in the Nanjing Medical University (NMU) breast cancer cohort. Functionally, we demonstrated that knockdown of SEC61G suppressed breast cancer cell proliferation, migration, invasion, and promoted breast cancer cell apoptosis in vitro. Xenograft breast tumor model revealed that knockdown of SEC61G inhibited breast tumor development in vivo. Furthermore, we demonstrated that SEC61G positively regulated glycolysis in breast cancer cells. Mechanistically, we showed that transcription factor E2F1 directly bound to the promoter of SEC61G and regulated its expression in breast cancer cells. SEC61G overexpression antagonized the effect of E2F1 knockdown in regulating breast cancer cell proliferation, invasion, and apoptosis. Finally, we demonstrated that the E2F1/SEC61G axis regulated glycolysis and chemo-sensitivity of Herceptin in breast cancer cells. Taken together, these results of in vitro and in vivo studies demonstrate that SEC61G promotes breast cancer development and metastasis via modulating glycolysis and is transcriptionally regulated by E2F1, which might be utilized as a promising therapeutic target of breast cancer treatment.