Network Analysis and Basic Experiments on the Inhibition of Renal Cancer Proliferation and Migration by Alpinetin through PI3K/AKT/ mTOR Pathway.

BACKGROUND: Alpinetin, a natural flavonoid, has been shown to have anticancer effects on many tumors. This study investigated the antitumor effect of alpinetin on renal clear cell carcinoma (ccRCC). METHODS: Network Pharmacology analysis was carried out on the targets and molecular mechanisms of alpinetin treating ccRCC. The Annexin V PE/7-AAD kit was used to detect apoptosis. Flow cytometry and Cell Counting Kit-8 (CCK-8) were used to detect cell proliferation and cycle. A 24-well transwell chamber and the ibidi scratch insertion performed cell migration analysis. The protein expression of the target molecule was detected by Western blotting. Nude mouse tumorigenesis assays were used to determine the in vivo antitumor effects of alpinetin. RESULTS: The network pharmacology revealed that GAPDH, HRAS, SRC, EGFR, and AKT1 are the main targets of alpinetin in treating ccRCC, with the PI3K/AKT signaling pathway being the main pathway of action. We found that alpinetin could significantly inhibit the proliferation and migration of ccRCC cells by inducing apoptosis. In addition, alpinetin also inhibited the cycle progression of ccRCC cells by blocking them in the G1 phase. Furthermore, in vivo and in vitro, alpinetin could inhibit the activation of an important pathway involved in the proliferation and migration of ccRCC cells, namely the PI3K/Akt pathway. CONCLUSION: Alpinetin can inhibit the growth of ccRCC cells by inhibiting the activation of the PI3K/Akt pathway and can be a potential anti-cancer drug for ccRCC.

Prophylactic central neck dissection for cN0 papillary thyroid carcinoma: is there any difference between western countries and China? A systematic review and meta-analysis.

Background: Recommendations for the performance of prophylactic central neck dissection (pCND) in patients with clinically node-uninvolved (cN0) papillary thyroid carcinoma (PTC) are not the same. This meta-analysis set out to compare the effectiveness of pCND with total thyroidectomy (TT) in different countries and regions, mainly between western countries and China. Methods: The electronic databases PubMed, EMBASE, and Cochrane Library were searched for studies published until August 2022. The incidence rate of cervical lymph node metastases (LNMs), locoregional recurrences (LRRs), and postoperative complications were pooled by a random-effects model. Subgroup analyses based on different countries and regions were performed. Results: Eighteen studies involving 5,346 patients were analyzed. In the subgroup of western countries, patients undergoing pCND with TT had a significantly lower LRR rate [69/1,804, 3.82% vs. 139/2,541, 5.47%; odds ratio (OR) = 0.56; 95% CI 0.37-0.85] and a higher rate of temporary hypoparathyroidism (HPT) (316/1,279, 24.71% vs. 194/1,467, 13.22%; OR = 2.23; 95% CI 1.61-3.08) than that of the TT alone group, while no statistically significant difference was found in the rate of permanent HPT and temporary and permanent recurrent laryngeal nerve (RLN) injury. In the Chinese subgroup, the pCND with TT group had a significantly higher incidence rate of both temporary HPT (87/374, 23.26% vs. 36/324, 11.11%; OR = 2.24; 95% CI 1.32-3.81) and permanent HPT (21/374, 5.61% vs. 4/324, 1.23%; OR = 3.58; 95% CI = 1.24-10.37) than that of the TT alone group, while no significant difference was detected in the rate of LRR and temporary and permanent RLN injury. Conclusion: Compared with the TT alone for cN0 PTC patients, pCND with TT had a significantly lower LRR rate while having a higher temporary HPT rate in Europe, America, and Australia; however, it showed no significant difference in decreasing LRR rate while having a significantly raised rate of temporary and permanent HPT in China. More population-based results are required to advocate precision medicine in PTC. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO/, identifier CRD42022358546.

A multi-dimensional nomogram to predict non-sentinel lymph node metastases in T1-2HR+ breast cancer.

Background: Axillary lymph node dissection (ALND) could be omitted for T1-2 breast cancer patients with 1-2 positive sentinel lymph node (SLN) after breast-conserving surgery when radiation is planned. However, whether ALND could be replaced by radiation in patients with 1-3 positive SLNs when no more non-SLN metastasis were observed after mastectomy are still controversial. The aim of our study was to develop and validate a nomogram for predicting the possibility of non-SLN metastasis in T1-2 and hormone receptor (HR) positive breast cancer patients with 1-3 positive SLNs after mastectomy. Methods: We retrospectively reviewed and analyzed the data including the basic information, preoperative sonographic characteristics, and pathological features in breast cancer patients with 1-3 positive SLNs in our medical center between Jan 2016 and Dec 2021. The Chi-square, Fisher's exact test, and t test were used for comparison of categorical and qualitative variables among patients with or without non-SLN metastasis. Univariate and multivariate logistic regression were used to determine the risk factors for non-SLN metastasis. These predictors were used to build the nomogram. The C-index and area under the receiver operating characteristic curve (AUC) was calculated to assess the accuracy of the model. Results: A total of 49 in 107 (45.8%) patients were identified with non-SLN metastasis. In multivariate analysis, four variables including younger age, lower estrogen receptor (ER) expression, higher histological score, and cortex thickening of the lymph nodes were determined to be significantly associated with non-SLN metastasis. An individualized nomogram was consequently established with a favorable C-index of 0.822 and verified via two internal validation cohorts. Conclusions: The current study developed a nomogram predicting non-SLN metastasis for T1-2 and HR+ breast cancer with 1-3 positive SLNs after mastectomy and found that patients in the high-risk group exhibited worse relapse-free survival. The novel nomogram may further help surgeons to determine whether ALND could be omitted when 1-3 positive SLNs were observed in T1-2 and HR+ breast cancer patients.

Analysis of Characteristics, Pathogens and Drug Resistance of Urinary Tract Infection Associated with Long-Term Indwelling Double-J Stent.

Objective: To investigate the characteristics, pathogens and drug resistance of urinary tract infection (UTI) associated with long-term indwelling double-J stent. Methods: The clinical data of 102 patients with urinary tract infection associated with long-term indwelling double-J stent in University-Town Hospital of Chongqing Medical University and Chongqing Traditional Chinese Medicine Hospital from September 2010 to July 2022 were collected retrospectively, and the difference between etiological characteristics were analyzed. Urine and double-J stent samples of patients were collected for pathogen identification and drug sensitivity test. Results: A total of 102 patients, 39 (38.23%) males and 63 (61.77%) females, aged 24-72 years, with a median age of 48 years, were included in this study. Urinary calculi (40.20%) and ureteral stricture (24.50%) were the main causes of urinary tract infection associated with long-term indwelling double-J stent. Among the patients with urinary tract infection caused by double-J stent, female patients were higher than male patients (61.77% vs 38.23%). In terms of positive rate of pathogenic bacteria culture, the rate of double-J stent was higher than that of urine (67.65% vs 35.29%). The main pathogenic bacteria in urine were Escherichia coli (30.55%) of Gram negative bacteria, while the main pathogenic bacteria in double-J stent were enterococcus faecalis (27.53%) of Gram positive bacteria. The resistance rate of Gram positive bacteria in double-J stent to vancomycin, ciprofloxacin, meropenem and piperacillin/tazobactam was significantly higher than that in urine (P<0.05). The resistance rate of Gram negative bacteria in double-J stent to imipenem, cefepime, piperacillin/tazobactam, meropenem and cefoperazone/sulbactam was significantly higher than that in urine (P<0.05). Conclusion: Double-J stent associated urinary tract infection is more common in women than in men. Escherichia coli and Enterococcus faecalis are the main pathogens, and the pathogens show strong drug resistance.

Safety and immunogenicity of the third (booster) dose of inactivated and recombinant protein SARS-CoV-2 vaccine for patients with endocrine-related cancer.

Background: Our study aimed to evaluate the safety and immunogenicity of the third (booster) dose of the COVID-19 vaccine for patients with endocrine-related cancers. Methods: This observational study involved 94 breast cancer patients, 92 thyroid cancer patients, and 123 healthy individuals who had received the third (booster) dose of the COVID-19 vaccine. Data on the adverse effects, serum anti-receptor binding domain (RBD)-immunoglobulin (Ig) G, and neutralizing antibodies (NAbs) were collected prospectively. Results: The serum anti-RBD-IgG and NAb titers were significantly lower for the patients with endocrine-related malignancies than for the healthy controls (3.01 [IQR: 1.11-6.70] vs. 4.19 [1.95-9.11], p = 0.001; 0.23 [0.11-0.52] vs. 0.41 [0.22-0.78], p = 0.001), and the seroconversion rates of anti-RBD-IgG and NAbs showed similar results. The serum antibody titers and seroconversion rates were significantly lower for patients aged ≥65 years with endocrine-related cancers, but there were no significant differences related to gender, vaccine type, or cancer type. Subgroup analysis showed that the antibody titers and seroconversion rates were significantly lower for patients with intermediate to advanced breast cancer, HR-/Her2+ breast cancer, and breast cancer undergoing treatment than for healthy controls. In contrast, breast cancer patients who completed their treatment and those who received endocrine therapy after completing their treatment were not significantly different from healthy controls. The NAbs titers and seroconversion rates were significantly lower for patients with primary thyroid cancer (0.19 [IQR: 0.10-0.46] vs. 0.41 [0.22-0.78], p = 0.003; 55.9 vs. 84.9%, p < 0.001); the seroconversion rates were significantly higher for the patients with combined Hashimoto's thyroiditis than for those without it. Multiple linear regression showed that patients aged ≥65 years who were receiving treatment were at risk of having lower antibody levels. Conclusion: The third (booster) dose of the COVID-19 vaccine is safe and well-tolerated. Our data support a third (booster) dose of the SARS-CoV-2 vaccine for breast and thyroid cancer patients. Breast cancer patients aged ≥65 years who are receiving treatment should be more protected, while thyroid cancer and breast cancer patients who have completed their treatment can be vaccinated like the general population.

The Value of Ki-67 Labeling Index in Central Lymph Node Metastasis and Survival of Papillary Thyroid Carcinoma: Evidence From the Clinical and Molecular Analyses.

BACKGROUND: Recent evidence suggests that the Ki-67 labeling index is associated with lymph node metastasis and the prognosis of papillary thyroid carcinoma (PTC). METHODS: We retrospectively evaluated the clinicopathological features of consecutive PTC patients between Jan 2019 and Oct 2020 in our medical center. The molecular analysis was also conducted by using the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) program. The Chi-square test was performed for the comparison of variables between patients with central lymph node metastasis (CLNM) and not. Besides, univariate and stepwise multivariate logistic regression analyses were further used to determine the risk factors for CLNM in PTC. RESULTS: Our results showed that male gender (odd ratio (OR) = 3.02; 95% CI: 1.81-5.04), tumor size >1 cm (OR = 2.81; 95% CI: 1.84-4.29), multifocality (OR = 2.08; 95% CI: 1.31-3.30, and Ki-67 labeling index (>3% and ≤5%: OR = 1.20; 95% CI: .73-1.97; >5%: OR = 3.85; 95% CI: 1.62-9.14) were independent risk factors for CLNM. After excluding the patients with harvested central lymph nodes <3, increased Ki-67 labeling index was still associated with the number of CLNM and the lymph node ratio. Additionally, the expression level of Ki-67 was significantly correlated with a higher N stage and worse disease-free survival in TCGA and validated GSE60542 datasets. CONCLUSIONS: Higher Ki-67 labeling index (>5%) is significantly associated with the CLNM in PTC patients, like other indicators of the male gender, larger tumor size, and multifocality. Besides, the Ki-67 was also determined to be associated with CLNM and DFS in PTC patients, which may act as an important molecular marker in PTC.

Novel scoring system combined with a virtual reality technique for the preoperative evaluation of the stone-free status after flexible ureteroscopy: the H.L.P.E.S. score.

OBJECTIVE: The original S.O.L.V.E. scoring system was modified using virtual reality technology, and a new H.L.P.E.S scoring system was constructed to improve the accuracy of predicting the stone-free rate after flexible ureteroscopy. METHODS: We retrospectively analyzed clinical and virtual reality data of 150 patients with renal calculi who underwent flexible ureteroscopy at the First Affiliated Hospital of Chongqing Medical University, Chongqing, China, from September 2019 to January 2022. Factors affecting the stone-free rate were evaluated in univariate and multiple logical regression analyses. Factors were divided by cut-off value under the receiver-operating characteristic curve and scored accordingly to a well-known international scoring system. Area under the curve predicted the stone-free rate. The accuracy and superiority of the stone-free rate after flexible ureterorenoscopy was compared between this scoring system and the S.O.L.V.E, R.I.R.S, T.O.HO, and RUSS scores. RESULTS: Multiple logistic regression showed that the stone surface area, renal pelvis volume, and length of the calyces funnel were correlated with stone-free rate (P < 0.01, P = 0.021, P = 0.019, respectively). The H.L.P.E.S. score included stone surface area (1-2 points), renal pelvis volume (1-2 points), length of calyces funnel (1-2 points), pelvic calyceal height (1-2 points), and essence of stone (1-2 points). The area under the receiver-operating characteristic curve of H.L.P.E.S. score was 0.927, which was higher than the S.O.L.V.E., R.I.R.S., T.O.HO, and RUSS scores. CONCLUSION: H.L.P.E.S. scoring can effectively predict the stone-free rate after flexible ureteroscopy for renal calculi and is superior to other scoring systems.

Clinical significance of multi-genic assay in identifying aggressive papillary thyroid carcinoma.

BACKGROUND: A minority of papillary thyroid carcinoma (PTC) is highly aggressive, with rapid progression and a poor prognosis. This study investigated the ability of multi-genic assay to identify patients with aggressive PTC. PATIENTS AND METHODS: A total of 117 PTC patients treated at The First Affiliated Hospital of Chongqing Medical University with clinicopathological data and multi-genic assay results and 389 patients with complete data from The Cancer Genome Atlas (TCGA) database were included. The chi-square test was used to analyze the relationship between the multi-genic assay results and clinicopathological characteristics. Univariate and multivariate regression analyses were used to analyze the impact of various factors on prognosis. RESULTS: The median follow-up times of the local and TCGA cohorts were 30 months and 34 months, respectively. The results showed that central lymph node metastasis (P = 0.036), lateral lymph node metastasis (P = 0.003) and mutations in genes other than BRAFV600E (P = 0.002) were significantly associated with disease-free survival (DFS) in the local cohort, while the analysis of TCGA data showed that mutations in genes other than BRAFV600E were significantly related to poor prognosis (P = 0.029). According to univariate and multivariate analyses, mutations in genes other than BRAFV600E (P = 0.021) and lateral lymph node metastasis (P = 0.022) were independent factors for postoperative recurrence, as well as, mutations in genes other than BRAFV600E were an independent factor of survival (P = 0.047). CONCLUSIONS: The multi-genic assay was able to identify aggressive PTC, providing an effective biological basis for surgical management and postoperative treatment.

Is Age ≥ 55 years an optimal cutoff point among patients with differentiated thyroid Microcarcinoma? a preliminary study.

PURPOSE: Age ≥ 55 years is regarded as a pivotal component of TNM stage classification in differentiated thyroid carcinoma (DTC). However, whether this cutoff point is still adaptable for differentiated thyroid microcarcinoma (DTMC) is rarely investigated. METHODS: We reviewed and analyzed the data of DTC patients aged ≥ 55 years from the Surveillance, Epidemiology, and End Results (SEER) database. Univariate and multivariate Cox regression analyses were used to determine the potential risk factors of cancer-specific survival (CSS) in DTMC patients aged ≥ 55 years. The Kaplan-Meier survival curves were used to estimate CSS probability. Receiver operating characteristic (ROC) curves were used to analyze the best age cutoff point for DTMC. RESULTS: Among the DTMC patients, there was no significant difference in the 1-, 3-, 5-, and 7-year CSS probability between the 55-59 years and 60-64-years subgroup (p = 0.72). The ROC curves indicated that 65 years, 65 years, and 64 years were the cutoff age point of 3-, 5-, and 7-year CSS probability in DTMC patients, respectively. Besides, N1b (Hazard ratio (HR) = 3.90, 95% Confidence interval (CI): 2.01-7.57; p < 0.001), extrathyroidal extension (HR = 2.53, 95 %CI: 1.39-4.62; p = 0.002), and M1 (HR = 11.42, 95 %CI: 5.04-25.90; p < 0.001) were the independent risk factors in CSS of DTMC patients. CONCLUSIONS: Our results suggested age at diagnosis ≥ 55 years is not the best cutoff point in stratifying the stage of the DTMC patients. On the contrary, those patients aged above 65 years have a significantly lower probability of CSS, which perhaps should be taken into consideration for treatment decision-making.

Overexpression of TP53INP2 Promotes Apoptosis in Clear Cell Renal Cell Cancer via Caspase-8/TRAF6 Signaling Pathway.

Clear cell renal cell cancer (ccRCC) is a tumor of high malignancy, which can escape apoptosis. The tumor protein p53-inducible nuclear protein 2 (TP53INP2), known as an autophagy protein, is the essential part for autophagosome formation and sensitizes cells to apoptosis. Our study is aimed at exploring the role of TP53INP2 in ccRCC. We have identified the autophagy-related genes (ARGs) of differential expression in ccRCC patients with the help of the TCGA database by bioinformatics analysis. Our assays of quantitative real-time polymerase chain reaction (qRT-PCR) and western blot were for the determination on the both levels of mRNA and protein. Overexpression of TP53INP2 on cellular proliferation, migration, and apoptosis of ccRCC was verified in the ways of performing CCK-8, wound scrape, transwell and flow cytometry assays in vitro, and a mice tumor model in vivo. Transmission electron microscopy was used to measure autophagy formation. The underlying mechanisms of TP53INP2 on ccRCC were determined via coimmunoprecipitation. TP53INP2 was found highly associated with an outcome of worse overall survival (OS) in Kaplan-Meier curves, and this parameter in ccRCC tissues was also lower than the normal tissues. Overexpression of TP53INP2 inhibited ccRCC cellular proliferation, migration, and invasion, as well as the tumor growth of mice. Those cells treated with autophagy inhibitor chloroquine (CQ) or TP53INP2 increased the apoptosis rate. TP53INP2 promoted autophagy formation and elevated the ratio of LC3 II/LC3 I. However, TP53INP2 did not significantly decrease the p-mTOR level. In addition, TP53INP2 activates the expressions of caspase-3, caspase-8, and PARP. Caspase-8 and TNF receptor associated factor 6 (TRAF6) were found to bind to each other in the presence of TP53INP2. TP53INP2 induces apoptosis in ccRCC cells through caspase-8/TRAF6 pathway, rather than the autophagy-dependent pathway.

Safety and Immunogenicity of Inactivated and Recombinant Protein SARS-CoV-2 Vaccines in Patients With Thyroid Cancer.

Background: This study aimed at assessing the safety and immunogenicity of SARS-CoV-2 vaccines in patients with thyroid cancer. Methods: This observational study included thyroid cancer patients between April 1, 2021, and November 31, 2021, in the Second Affiliated Hospital of Chongqing Medical University. All participants received at least one dose of the SARS-CoV-2 vaccine. SARS-CoV-2 IgG was tested, and the interval time between the last dose and humoral response test ranged from <1 to 8 months. The complications after SARS-CoV-2 vaccines were recorded. Results: A total of 115 participants at least received one dose of SARS-CoV-2 vaccines with a 67.0% IgG-positive rate. Among them, 98 cases had completed vaccination, and the positivity of SARS-CoV-2 IgG antibodies was 96% (24/25) with three doses of ZF2001. SARS-CoV-2 IgG antibodies' positivity was 63.0% (46/73) of two doses of CoronaVac or BBIBP-CorV vaccine. Additionally, after 4 months of the last-dose vaccination, the IgG-positive rate (31.6%, 6/19) significantly decreased in thyroid cancer patients. The IgG-positive rate (81.0%, 64/79) was satisfactory within 3 months of the last-dose vaccination. Ten (10.2%) patients had side effects after SARS-CoV-2 vaccination. Among them, two (2.0%) patients had a fever, five (5.1%) patients had injection site pain, one (1.0%) patient felt dizzy, and one patient felt dizzy and had injection site pain at the same time. Conclusion: SARS-CoV-2 vaccines (CoronaVac, BBIBP-CorV, and ZF2001) are safe in thyroid cancer patients. The regression time of SARS-CoV-2 IgG is significantly shorter in thyroid cancer patients than in healthy adults. Therefore, a booster vaccination dose may be earlier than the systematic strategy for thyroid cancer patients.

Identifying and Validating of an Autophagy-Related Gene Signature for the Prediction of Early Relapse in Breast Cancer.

BACKGROUND: Compelling evidence has demonstrated the pivotal role of autophagy in the prognosis of breast cancer. Breast cancer (BC) patients with early relapse consistently exhibited worse survival. METHODS: The autophagy-related genes were derived from the Human Autophagy Database (HADb) and high-sequencing data were obtained from The Cancer Genome Atlas (TCGA). Discrepantly expressed autophagy genes (DEAGs) between early relapse and long-term survival groups were performed using the Linear Models for Microarray data (LIMMA) method. Lasso Cox regression analysis was conducted for the selection of the 4-gene autophagy-related gene signature. GSE42568 and GSE21653 databases were enrolled in this study for the external validation of the signature. Then patients were divided into high and low-risk groups based on the specific score formula. GSEA was used to discover the related signaling pathway. The Kaplan-Meier curves and the receiver operating characteristic (ROC) curves were used to evaluate the discrimination and accuracy of the 4-gene signature. RESULTS: A signature composed of four autophagy-related mRNA including APOL1, HSPA8, SIRT1, and TP73, was identified as significantly associated with the early relapse in BC patients. Time-dependent receiver-operating characteristic at 1 year suggested remarkable accuracy of the signature [area under the curve (AUC = 0.748)]. The risk score model based on the autophagy-related signature showed favorable predicting value in 1-, 2-, and 3-year relapse-free survival (RFS) in training and two validating cohorts. The GSEA displayed gene sets were remarkably enriched in carcinogenic activation pathways and autophagy-related pathways. The nomogram involving three variables (progesterone receptor status, T stage, and 4-gene signature) exhibited relatively good discrimination with a C-index of 0.766. CONCLUSIONS: Our study establishes an autophagy-related 4-gene signature that can effectively stratify the high-risk and low-risk BC patients for early relapse. Combined with the clinicopathological variables, the signature could significantly help oncologists tailor more efficient treatment strategies for BC patients.

Preoperative and pathological predictive factors of central lymph node metastasis in papillary thyroid microcarcinoma.

OBJECTIVE: The aim of this study was to identify clinical and pathological markers of CLNM in persons with clinical lymph node-negative papillary thyroid microcarcinoma(PTMC). MATERIALS AND METHODS: Retrospective data were analyzed from 804 PTMC clinically negative patients who were receiving thyroid surgery in the First Affiliated Hospital at Chongqing Medical University from January 2017 to December 2018. The CLNM-positive and CLNM-negative groups were categorised according to histological evidence of the central lymph node involvement, statistically, risk variables for CLNM were found. RESULTS: 324 (40.3%) individuals were diagnosed with CLNM. Sex (P=0.001), age at diagnosis (P<0.001), tumour size(P=0.029), microcaccificities presence (P=0.003), capsules discontinuity(P=0.002), multi-focality(P=0.001) and (ETE)extrathyroidal extension (P < 0.001) differed substantially from one positive CLNM group to the next. For multivariate analyses, women (odds ratio [OR] = 0.489), age [OR = 0.540] are the independent protective factors for CLNM; micro-cacification presence (OR = 1.511), discontinuity of capsules (OR= 2.056), multifocality(OR=1.486) and ETE(OR=10.613) are the independent risk factors for CLNM. Feature curves of the receiver were built and the AUC is 0.763. 32.1% percent (80 patients) of the 249 patients who did not have any of the four risk variables got CLNM. This contrasted with the incidence of CLNM in this research, which was as high as 49.1%. CONCLUSIONS: CLNM has been connected with female sex, age - within 45 years, microcacification occurrences, capsule discontinuity, multifocality and extrathyroid expansion. The patients may benefit from the surgical decision of pCLND whether there are risk factors combined.

Risk factors and prediction model of level II lymph node metastasis in papillary thyroid carcinoma.

Introduction: Surgical management of lateral lymph nodes in papillary thyroid carcinoma, especially at level II, remains controversial. This study aimed to investigate the risk factors for level II lymph node metastasis in patients with papillary thyroid carcinoma and establish a prediction model to estimate the metastatic risk. Materials and methods: A total of 768 patients with papillary thyroid carcinoma underwent thyroidectomy and central plus lateral lymph node dissection, including levels VI, II, III, and IV, at the First Affiliated Hospital of Chongqing Medical University from January 2016 to December 2018. Data on the clinicopathological characteristics were collected and analyzed. Univariate and multivariate analyses were performed to identify risk factors for level II lymph node metastasis. Subsequently, a predictive model was established based on the results of the multivariate analyses. Results: The level II lymph node metastatic rate was 34.11% with the following features: largest tumor diameter >20 mm (Odds ratio=1.629, P=0.026), located in the upper pole (Odds ratio=4.970, P<0.001), clinical lymph node-positive (clinical central lymph node-positive: Odds ratio=1.797; clinical lateral lymph node-positive: Odds ratio=1.805, P=0.008), vascular invasion (Odds ratio=6.759, P=0.012), and rate of central lymph node metastasis (Odds ratio=2.498, P<0.001). Level III lymph node metastasis (Odds ratio=2.749, P<0.001) and level IV lymph node metastasis (Odds ratio=1.732, P=0.007) were independent of level II lymph node metastasis predictors. The prediction model's areas under the receiver operating characteristic curve were 0.815 and 0.804, based on bootstrapping validation. Level II lymph node metastasis was associated with the tumor-free survival rate of patients with papillary thyroid carcinoma (P<0.001). Conclusions: Largest tumor diameter >20 mm, located in the upper pole, clinical lymph node-positive, vascular invasion, rate of central lymph node metastasis, and levels III and IV lymph node metastases were independent level II lymph node metastasis predictors. We developed a prediction model for level II lymph node metastasis. Overall, level II lymph node metastasis dissection should be individualized according to clinicopathological data both preoperatively and intraoperatively.

Risk Factors, Prognostic Factors, and Nomogram for Distant Metastasis in Breast Cancer Patients Without Lymph Node Metastasis.

Background: Lymph node negative (N0) breast cancer can be found coexisting with distant metastasis (DM), which might consequently make clinicians underestimate the risk of relapse and insufficient treatment for this subpopulation. Methods: The clinicopathological characteristics of N0 breast cancer patients from the Surveillance, Epidemiology, and End Results (SEER) database between January 2010 and December 2015 were retrospectively reviewed. Multivariate logistic and Cox analyses were used to identify independent risk factors in promoting DM and the 1-, 3-, and 5- year cancer-specific survival (CSS) in this subpopulation. Result: Seven factors including age (<40 years), tumor size (>10 mm), race (Black), location (central), grade (poor differentiation), histology (invasive lobular carcinoma), and subtype (luminal B and Her-2 enriched) were associated with DM, and the area under curve (AUC) was 0.776 (95% CI: 0.763-0.790). Moreover, T1-3N0M1 patients with age >60 years at diagnosis, Black race, triple-negative breast cancer subtype, no surgery performed, and multiple DMs presented a worse 1-, 3-, and 5-year CSS. The areas under the ROC for 1-, 3-, and 5- year CSS in the training cohort were 0.772, 0.741, and 0.762, respectively, and 0.725, 0.695, and 0.699 in the validation cohort. Conclusion: The clinicopathological characteristics associated with the risk of DM and the prognosis of female breast cancer patients without lymph node metastasis but with DM are determined. A novel nomogram for predicting 1-, 3-, 5- year CSS in T1-3N0M1 patients is also well established and validated, which could help clinicians better stratify patients who are at a high-risk level for receiving relatively aggressive management.

Prediction Model of Pathologic Central Lymph Node Negativity in cN0 Papillary Thyroid Carcinoma.

BACKGROUND: Most patients with papillary thyroid carcinoma (PTC) have an excellent prognosis. Although central lymph node invasion is frequent, management via central lymph node dissection (CLND) remains controversial. The present study retrospectively investigated independent predictors of pathologic central lymph node negativity (pCLN-) and established a prediction model for pCLN- in clinical lymph node negativity (cN0) PTC. METHODS: A total of 2,687 patients underwent thyroid surgery for cN0 PTC from 2013 to 2018 at the First Affiliated Hospital of Chongqing Medical University, and lobectomy plus ipsilateral CLND was the basic surgical extent. Clinicopathological characteristics were reviewed and analyzed. Univariate and multivariate analyses were performed to identify factors related to pCLN-. A prediction model was established based on the results of multivariate analyses. RESULTS: The pCLN- rate was 51.5% (1,383/2,687). Multivariate analysis revealed that sex, age, thyroid stimulating hormone (TSH), size, location, laterality, unifocality and extrathyroidal extension negativity (ETE-) were independent predictors of pCLN-. The nomogram showed good discriminative ability (C-index: 0.784 and 0.787 in derivation and validation groups, respectively) and was well calibrated. We quantified the clinical usefulness of the nomogram by decision curve analysis. The median length of follow-up was 30 (range 12- 83) months, and 190 cases were lost, with a follow-up rate of 92.9% (2,497/2,687). Of the 2,687 patients included, 21 (0.8%) experienced recurrence. CONCLUSION: This nomogram, which integrates available preoperative clinicopathological features and intraoperative frozen biopsy outcomes, is a reliable tool with high accuracy to predict pCLN- in cN0 PTC.

FOXO1 inhibits prostate cancer cell proliferation via suppressing E2F1 activated NPRL2 expression.

Previous studies in our lab suggest that nitrogen permease regulator 2-like (NPRL2) upregulation in prostate cancer is associated with malignant behavior and poor prognosis. However, the underlying mechanisms of NPRL2 dysregulation remain poorly understood. This study aimed to explore the transcription factors (TFs) contributing to NPRL2 dysregulation in prostate cancer. Potential TFs were identified using prostate tissue/cell-specific chromatin immunoprecipitation (ChIP)-seq data collected in the Cistrome Data Browser and Signaling Pathways Project. Dual-luciferase assay and ChIP-qPCR assay were conducted to assess the binding and activating effect of TFs on the gene promoter. Cell Counting Kit-8 and colony formation assays were performed to assess cell proliferation. Results showed that E2F1 is a TF that bound to the NPRL2 promoter and activated its transcription. NPRL2 inhibition significantly alleviated E2F1 enhanced cell proliferation. Kaplan-Meier survival analysis indicated that E2F1 upregulation was associated with unfavorable progression-free survival and disease-specific survival. FOXO1 interacted and E2F1 in both PC3 and LNCaP cells and weakened the binding of E2F1 to the NPRL2 promoter. Functionally, FOXO1 overexpression significantly slowed the proliferation of PC3 and LNCaP cells and also decreased E2F1 enhanced cell proliferation. In summary, this study revealed a novel FOXO1/E2F1-NPRL2 regulatory axis in prostate cancer. E2F1 binds to the NPRL2 promoter and activates its transcription, while FOXO1 interacts with E2F1 and weakens its transcriptional activating effects. These findings help expand our understanding of the prostate cancer etiology and suggest that the FOXO1/E2F1-NPRL2 signaling axis might be a potential target.

NPRL2 reduces the niraparib sensitivity of castration-resistant prostate cancer via interacting with UBE2M and enhancing neddylation.

In this study, we explored the regulatory effects of nitrogen permease regulator 2-like (NPRL2) on niraparib sensitivity, a PARP inhibitor (PARPi) in castrate-resistant prostate cancer (CRPC). Data from The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) program were retrospectively examined. Gene-set enrichment analysis (GSEA) was conducted between high and low NRPL2 expression prostate adenocarcinoma (PRAD) cases in TCGA. CCK-8 assay, Western blot analysis of apoptotic proteins, and flow cytometric analysis of apoptosis were applied to test niraparib sensitivity. Immunofluorescent (IF) staining and co-immunoprecipitation (co-IP) were conducted to explore the proteins interacting with NPRL2. Results showed that the upregulation of a canonical protein-coding transcript of NPRL2 (ENST00000232501.7) is associated with an unfavorable prognosis. Bioinformatic analysis predicts a physical interaction between NPRL2 and UBE2M, which is validated by a following Co-IP assay. This interaction increases NPRL2 stability by reducing polyubiquitination and proteasomal degradation. Depletion of NPRL2 or UBE2M significantly increases the niraparib sensitivity of CRPC cells and enhances niraparib-induced tumor growth inhibition in vivo. NPRL2 cooperatively enhances UBE2M-mediated neddylation and facilitates the degradation of multiple substrates of Cullin-RING E3 ubiquitin ligases (CRLs). In conclusion, this study identified a novel NPRL2-UBE2M complex in modulating neddylation and niraparib sensitivity of CRPC cells. Therefore, targeting NPRL2 might be considered as an adjuvant strategy for PARPi therapy.

Clinical implications of Delphian lymph node metastasis in papillary thyroid carcinoma.

BACKGROUND: As is known, identifying risk factors precisely for lymph node metastasis (LNM) plays a vital role in initial treatment for papillary thyroid carcinoma (PTC). Nonetheless, whether Delphian lymph node (DLN) metastasis has value in predicting LNM remains an open question. This study covered a sample of 1,575 patients, which is the largest sample group so far, aiming to assess the predictive validity of DLN metastasis in PTC. METHODS: This retrospective cohort study was conducted with 1,575 eligible PTC patients who underwent thyroid operation between July 2013 and December 2018 and clinicopathologic parameters of patients with DLN metastasis were compared with those without DLN metastasis. RESULTS: The incidence of DLN metastasis, according to our research samples, is 24.4% (384/1,575 patients). And results show that DLN positivity was closely associated with adverse prognostic factors including younger age, larger tumor size, extrathyroid extension, tumor location in the isthmus or upper lobe of the thyroid, number of LNM >5, higher recurrence. After carefully adjusting important confounding factors, we find that in multivariate logistic regression analyses, DLN metastasis is an independent predictor for both central LNM (CLNM, adjusted OR =7.81, P<0.001) and lateral LNM (LLNM, adjusted OR =3.40, P<0.001). Moreover, the stratified analyses also show convincing evidence of a positive correlation between DLN metastasis and LNM in levels II-IV in the vast majority of subgroups. CONCLUSIONS: The present study suggests that DLN metastasis is an independent risk factor for CLNM and LLNM of levels II-IV. The cervical lymph nodes should be meticulously evaluated to guide tailored treatment during operation in patients with DLN involvement.

Multi-gene assay and clinical characteristics research in papillary thyroid carcinoma.

BACKGROUND: To investigate the significance of multi-gene assay in papillary thyroid carcinoma (PTC) patients in clinical practice. METHODS: From April to December 2019, medical records of 68 patients with PTC after the initial surgery were retrospectively collected and analyzed in terms of the relations between gene mutations and clinicopathological characteristics. RESULTS: RET/PTC rearrangement was not detected in BRAF V600E mutation patients (P<0.001). Besides, compared with wild-type patients, BRAF V600E mutation was associated with significantly older age (P=0.001) and a higher rate of extrathyroid invasion (P=0.023). Significantly higher BRAF V600E mutation rates were found in clinical lymph node-negative (P=0.041) and non-metastatic lateral lymph nodes (P=0.027) patients as RET/PTC rearrangement was associated with younger age (P=0.001) and the increasing metastatic number of lymph nodes (P=0.020). Compared to other gene mutations, the multivariate analysis showed that larger tumor size [odds ratio (OR), 8.831; 95% CI: 1.971-35.578; P=0.004], the BRAF V600E mutation alone(OR, 10.567; 95% CI: 1.748-63.873; P=0.010) or in combination with one additional gene mutation (OR, 8.654; 95% CI: 1.453-68.603; P=0.041), and Hashimoto's thyroiditis (OR, 0.112; 95% CI: 0.025-0.499; P=0.004) were all independent predictors for the prevalence of ETE. CONCLUSIONS: BRAF V600E mutation was associated with older age and the aggressiveness of PTC but was independent of lymph node metastasis (LNM). RET/PTC rearrangement suggested more LNM in young patients with PTC. BRAF V600E mutation combined with other gene mutations, namely, multi-gene mutations, could indicate a higher aggressiveness in PTC.