Cellular senescence primes liver fibrosis regression through Notch-EZH2.

Cellular senescence plays a pivotal role in wound healing. At the initiation of liver fibrosis regression, accumulated senescent cells were detected and genes of senescence were upregulated. Flow cytometry combined with single-cell RNA sequencing analyses revealed that most of senescent cells were liver nonparenchymal cells. Removing senescent cells by dasatinib and quercetin (DQ), alleviated hepatic cellular senescence, impeded fibrosis regression, and disrupted liver sinusoids. Clearance of senescent cells not only decreased senescent macrophages but also shrank the proportion of anti-inflammatory M2 macrophages through apoptotic pathway. Subsequently, macrophages were depleted by clodronate, which diminished hepatic senescent cells and impaired fibrosis regression. Mechanistically, the change of the epigenetic regulator enhancer of zeste homolog2 (EZH2) accompanied with the emergence of hepatic senescent cells while liver fibrosis regressed. Blocking EZH2 signaling by EPZ6438 reduced hepatic senescent cells and macrophages, decelerating liver fibrosis regression. Moreover, the promoter region of EZH2 was transcriptionally suppressed by Notch-Hes1 (hairy and enhancer of split 1) signaling. Disruption of Notch in macrophages using Lyz2 (lysozyme 2) Cre-RBP-J (recombination signal binding protein Jκ) f/f transgenic mice, enhanced hepatic cellular senescence, and facilitated fibrosis regression by upregulating EZH2 and blocking EZH2 abrogated the above effects caused by Notch deficiency. Ultimately, adopting Notch inhibitor Ly3039478 or exosome-mediated RBP-J decoy oligodeoxynucleotides accelerated liver fibrosis regression by augmenting hepatic cellular senescence.

A nomogram for intraoperatively predicting non-sentinel lymph node metastases in early breast cancer patients with positive sentinel lymph nodes.

Background: Individualized decisions are required in early-stage breast cancer patients. We aimed to establish a novel model for predicting non-sentinel lymph node (SLN) metastases in patients with positive SLNs, using preoperative and intraoperative characteristics and inflammatory indicators. Methods: The data of 489 patients with invasive breast cancer were retrospectively collected from Xuanwu Hospital between 2014 and 2021. Among them, 96 patients with at least one positive SLN were used to build the predictive model. Univariate and multivariate analyses were performed to identify the risk factors of non-SLN metastases. A nomogram was developed using these risk factors and was validated by calibration curves. The area under the receiver operating characteristics curve (AUC) and decision curve analyses (DCA) were used to compare our novel nomogram with the Memorial Sloan Kettering Cancer Center (MSKCC) nomogram. Cross-validation was performed for further internal validation of the predictive model. External validation was conducted using another treatment group (n=46 patients) in Xuanwu Hospital. Results: Non-SLN metastases occurred in 42 of the 83 patients with positive SLNs (50.6%). Multivariate stepwise logistic regression indicated that the risk factors were age (P=0.032), number of positive SLNs (P=0.020), number of negative SLNs (P=0.011), resected tumor size (P=0.038), and monocyte count (P=0.012). A predictive model was developed and virtualized by nomogram using these five risk factors. The AUC of our nomogram was 0.867, which was significantly higher than that of the MSKCC model. DCA also showed a superior clinical value for our novel nomogram. After 10-fold cross-validation with 400 times repetitions, the AUC of our model was still 0.830. External validation of our model showed an AUC of 0.727. The model was well-calibrated in the internal and external validation series. Conclusions: A five-factor nomogram was developed for predicting non-SLN metastases in early-stage breast cancer patients. This novel tool exhibited good accuracy and could assist clinicians with intraoperative decisions in breast cancer patients with positive SLNs.

Gelatinous lanthanide coordination polymer with aggregation-enhanced antenna effect for ratiometric detection of endogenous alkaline phosphatase.

Aggregation-induced emission (AIE) and antenna effect (AE) are two significant behaviors that have attracted increasing attention. However, it is challenging to achieve the synergistic effect of AIE and AE in luminescent materials for more extensive applications. Here, four gelatinous Ln3+ coordination polymers (Ln-CPs) are synthesized by self-assembly of ciprofloxacin (CIP), adenosine monophosphate (AMP), and Ln3+ ions in aqueous medium. Encouragingly, a remarkable increase in the characteristic fluorescence of Ln3+ and a significant decrease in CIP are observed along with increasing concentration of Ln-CPs, which is attributed to the large aggregates formed by self-assembly that strictly constrain the intramolecular motions of antenna ligands, thereby achieving the aggregation-enhanced AE. More meaningfully, Eu-CP not only shows a rice-like morphology at high aggregation state, but also provides an opportunity for the selective detection of alkaline phosphatase (ALP). A new flower-like polymer is formed upon incubating Eu-CP with ALP, accompanied by the fluorescence quenching of Eu3+ and recovery of CIP, a ratiometric determination of ALP in the range of 0.1-6.0 U·L-1 is thus achieved. Additionally, ALP assay in human serum and bioimaging in living cells have been successfully performed. This research opens a new horizon for the fabrication of Ln3+-based luminescent materials with promising applications.

Myeloid-specific blockade of Notch signaling ameliorates nonalcoholic fatty liver disease in mice.

Rationale: Macrophages play a central role in the development and progression of nonalcoholic fatty liver disease (NAFLD). Studies have shown that Notch signaling mediated by transcription factor recombination signal binding protein for immunoglobulin kappa J region (RBP-J), is implicated in macrophage activation and plasticity. Naturally, we asked whether Notch signaling in macrophages plays a role in NAFLD, whether regulating Notch signaling in macrophages could serve as a therapeutic strategy to treat NAFLD. Methods: Immunofluorescence staining was used to detect the changes of macrophage Notch signaling in the livers of human patients with NAFLD and choline deficient amino acid-defined (CDAA) diet-fed mice. Lyz2-Cre RBP-Jflox or wild-type C57BL/6 male mice were fed with CDAA or high fat diet (HFD) to induce experimental steatohepatitis or steatosis, respectively. Liver histology examinations were performed using hematoxylin-eosin (H&E), Oil Red O staining, Sirius red staining and immunohistochemistry staining for F4/80, Col1α1 and αSMA. The expression of inflammatory factors, fibrosis or lipid metabolism associated genes were evaluated by quantitative reverse transcription (qRT)-PCR, Western blot or enzyme-linked immunosorbent assay (ELISA). The mRNA expression of liver samples was profiled by using RNA-seq. A hairpin-type decoy oligodeoxynucleotides (ODNs) for transcription factor RBP-J was loaded into bEnd.3-derived exosomes by electroporating. Mice with experimental NAFLD were treated with exosomes loading RBP-J decoy ODNs via tail vein injection. In vivo distribution of exosomes was analyzed by fluorescence labeling and imaging. Results: The results showed that Notch signaling was activated in hepatic macrophages in human with NAFLD or in CDAA-fed mice. Myeloid-specific RBP-J deficiency decreased the expression of inflammatory factors interleukin-1 beta (IL1ß) and tumor necrosis factor alpha (TNFα), attenuated experimental steatohepatitis in mice. Furthermore, we found that Notch blockade attenuated lipid accumulation in hepatocytes by inhibiting the expression of IL1ß and TNFα in macrophages in vitro. Meanwhile, we observed that tail vein-injected exosomes were mainly taken up by hepatic macrophages in mice with steatohepatitis. RBP-J decoy ODNs delivered by exosomes could efficiently inhibit Notch signaling in hepatic macrophages in vivo and ameliorate steatohepatitis or steatosis in CDAA or HFD mice, respectively. Conclusions: Combined, macrophage RBP-J promotes the progression of NAFLD at least partially through regulating the expression of pro-inflammatory cytokines IL1ß and TNFα. Infusion of exosomes loaded with RBP-J decoy ODNs might be a promising therapy to treat NAFLD.

Age-related liver endothelial zonation triggers steatohepatitis by inactivating pericentral endothelium-derived C-kit.

Aging leads to systemic metabolic disorders, including steatosis. Here we show that liver sinusoidal endothelial cell (LSEC) senescence accelerates liver sinusoid capillarization and promotes steatosis by reprogramming liver endothelial zonation and inactivating pericentral endothelium-derived C-kit, which is a type III receptor tyrosine kinase. Specifically, inhibition of endothelial C-kit triggers cellular senescence, perturbing LSEC homeostasis in male mice. During diet-induced nonalcoholic steatohepatitis (NASH) development, Kit deletion worsens hepatic steatosis and exacerbates NASH-associated fibrosis and inflammation. Mechanistically, C-kit transcriptionally inhibits chemokine (C-X-C motif) receptor (CXCR)4 via CCAAT enhancer-binding protein α (CEBPA). Blocking CXCR4 signaling abolishes LSEC-macrophage-neutrophil cross-talk and leads to the recovery of C-kit-deficient mice with NASH. Of therapeutic relevance, infusing C-kit-expressing LSECs into aged mice or mice with diet-induced NASH counteracts age-associated senescence and steatosis and improves the symptoms of diet-induced NASH by restoring metabolic homeostasis of the pericentral liver endothelium. Our work provides an alternative approach that could be useful for treating aging- and diet-induced NASH.

Dual-response fluorescent probe based on nitrogen-doped carbon dots and europium ions hybrid for ratiometric and on-site visual determination of oxytetracycline and tetracycline.

Tetracyclines residues, particularly oxytetracycline (OTC) and tetracycline (TC), have raised extensive concern because of their serious adverse effects on human health. Herein, a dual-response fluorescent probe based on nitrogen-doped carbon dots (N-CDs) and Eu3+ hybrid (N-CDs-Eu3+) was developed to selectively determine OTC and TC. The N-CDs act as ancillary ligands of Eu3+ and recognition units of OTC/TC, while the Eu3+ ions chelated with N-CDs can also specifically recognize OTC/TC. Upon inclusion of OTC/TC, an enhancement in Eu3+ emission occurs due to the energy transfer from OTC/TC to Eu3+ and the efficient elimination of quenching effect caused by H2O molecule, which is attributed to the incorporation of N-CDs; while the blue fluorescence emitted by the N-CDs decreases under the inner filter effect and static quenching effect caused by OTC/TC. Based on the double and reverse response signals, the ratiometric detection of OTC and TC in the range of 0.1-45 µΜ and 0.1-30 µΜ is achieved with a detection limit of 0.017 and 0.041 µM, respectively. In addition, the noticeable variation in fluorescence color of the probe is integrated with a smartphone-assisted analysis device for the rapid on-site quantitative assay of OTC, where the detection limit is 0.15 µΜ. The results show that this probe performs with excellent specificity and anti-interference for both OTC and TC, and satisfactory detection results are obtained in lake water, milk, and honey samples, thereby confirming that the probe exhibits promising application in food safety and environmental monitoring.

A smartphone-integrated light-up lanthanide fluorescent probe for the visual and ratiometric detection of total phosphorus in human urine and environmental water samples.

Phosphate (Pi) plays an essential role in aquatic ecosystems as well as in physiological processes. Here, a dual-emission probe for the sensitive, specific and visual analysis of Pi is fabricated by coordinating Eu3+ with luminol and 2,6-pyridinedicarboxylic acid (DPA). Pi can significantly enhance the characteristic fluorescence of Eu3+ at 615 nm by promoting energy transfer from DPA to Eu3+ and reducing the quenching effect of water molecule, luminol with inherent emission at 423 nm further enhances the Eu3+ fluorescence. Accordingly, ratiometric detection of Pi can be achieved with the fluorescence ratio F615/F423 as a function of Pi concentration. Linearity between F615/F423 and Pi concentration in the range of 0.1-25 µM is shown, and the limit of detection (LOD, 3σ/K) for Pi is 0.027 µM. In addition, a continuous change in the fluorescence color of the probe from blue to red is observed with increasing Pi concentration under a UV lamp, and a smartphone-based visual method is used for the convenient and effective semi-quantitative determination of Pi. The dual-emission probe has been successfully applied to ratiometric and visual analysis of Pi in human urine and environmental water samples, and adequate results are obtained.

A novel immune checkpoint-related gene signature for predicting overall survival and immune status in triple-negative breast cancer.

Background: Triple-negative breast cancer (TNBC) is a highly aggressive subtype and only some of patients could benefit from the immunotherapy. The present study aims to investigate the expression pattern and prognostic value of immune checkpoint genes (ICGs) in TNBC and develop a novel ICGs-signature to predict the prognosis and immune status in TNBC. Methods: ICGs expression profiles and clinical characteristics of TNBC samples were obtained from The Cancer Genome Atlas (TCGA) and the Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) database. The least absolute shrinkage and selection operator (LASSO) Cox regression analysis was employed to construct a multi-gene signature for predicting the prognostic outcome. The risk scores were calculated based on the coefficients of each ICG in LASSO-Cox regression model. The median score was considered as the cut-off value to divide the TNBC patients into a high-risk group and a low-risk group. The Kaplan-Meier survival curves were generated to further explore the association between the risk scores and prognostic outcomes. Finally, single sample gene set enrichment analysis (ssGSEA) was conducted to evaluate the immune status and immunophenoscore (IPS) score was used for the quantitative evaluation of tumor immunogenicity. Results: PDCD1, PDCD1LG2 and KIR3DL2 were included in the ICGs-signature model and the risk scores were calculated for each sample according to the coefficients in LASSO-Cox regression. Patients in high-risk group were associated with unfavorable prognosis. The receiver operating characteristic (ROC) curves showed the area under the curve (AUC) values for predicting 1-, 2- and 3-year overall survival (OS) by ICGs-signature were 0.925,0.822 and 0.835, respectively. The adaptive immunity cells and innate immunity cells were significantly abundant in the low-risk group, and low-risk patients tended to have higher IPS scores of PD-1, CTLA4, PD-L1 and PD-L2. Conclusions: A novel ICGs-signature was developed and validated, which may be not only served as a robust prognostic marker, but also a potential indicator reflecting immunotherapy response.

A Nomogram Based on Clinicopathological and Ultrasound Imaging Characteristics for Predicting Cervical Lymph Node Metastasis in cN0 Unilateral Papillary Thyroid Microcarcinoma.

Objective: The aim of this study was to establish a practical nomogram for preoperatively predicting the possibility of cervical lymph node metastasis (CLNM) based on clinicopathological and ultrasound (US) imaging characteristics in patients with clinically node-negative (cN0) unilateral papillary thyroid microcarcinoma (PTMC) in order to determine a personal surgical volume and therapeutic strategy. Methods: A total of 269 consecutive patients diagnosed with cN0 unilateral PTMC by postoperative pathological examination from January 2018 to December 2020 were retrospectively analyzed. All the patients underwent lobectomy or thyroidectomy with routine prophylactic central lymph node dissection (CLND) and were divided into a CLNM group and a non-CLNM group. Using logistic regression, the least absolute shrinkage and selection operator (LASSO) regression analysis was applied to determine the risk factors for CLNM in patients with unilateral cN0 PTMC. A nomogram including risk-factor screening using LASSO regression for predicting the CLNM in patients with cN0 unilateral PTMC was further developed and validated. Results: Risk factors identified by LASSO regression, including age, sex, tumor size, presence of extrathyroidal extension (ETE), tumor diameter/lobe thickness (D/T), tumor location, and coexistent benign lesions, were potential predictors for CLNM in patients with cN0 unilateral PTMC. Meanwhile, age (odds ratio [OR] = 0.261, 95% CI.104-0.605; P = 0.003), sex (men: OR = 3.866; 95% CI 1.758-8.880; P < 0.001), ETE (OR = 3.821; 95% CI 1.168-13.861; P = 0.032), D/T (OR = 72.411; 95% CI 5.483-1212.497; P < 0.001), and coexistent benign lesions (OR = 3.112 95% CI 1.407-7.303; P = 0.007) were shown to be significantly related to CLNM by multivariant logistic regression. A nomogram for predicting CLNM in patients with cN0 unilateral PTMC was established based on the risk factors identified by the LASSO regression analysis. The receiver operating characteristic (ROC) curve for predicting CLNM by nomogram showed that the area under the curve (AUC) was 0.777 and exhibited an excellent consistency. Conclusions: A nomogram based on clinical and US imaging characteristics for predicting the probability of CLNM in patients with cN0 unilateral PTMC was developed, which showed a favorable predictive value and consistency. Further prospective research to observe the oncological outcomes is necessary to determine whether the nomogram could potentially guide a personalized surgical volume and surgical approach.

Development and validation of a novel 14-gene signature for predicting lymph node metastasis in papillary thyroid carcinoma.

BACKGROUND: There is still no reasonably accurate method of preoperatively predicting central lymph node metastasis (LNM), and it is essential to develop an effective evaluation model for predicting LNM in papillary thyroid carcinoma (PTC) patients. METHODS: PTC samples were collected from The Cancer Genome Atlas database. Candidate genes were identified as continuously upregulated or downregulated genes in the process of N0 to N1a and N1a to N1b. The least absolute shrinkage and selection operator (LASSO) regression analysis was used to construct the predictive model for LNM. Multivariate logistic regression analysis was performed to screen the potential factors related to LNM, and a nomogram was established. The risk score of the gene signature model for predicting disease-free survival (DFS) was evaluated by Kaplan-Meier analysis. RESULTS: A 14-gene signature was developed by LASSO regression for predicting LNM based on 69 differential expression genes (DEGs) that were continuously upregulated or downregulated in the progress of PTC. The receiver operating characteristic (ROC) curves of the 14-gene signature predicting LNM, central LNM and lateral LNM were generated. The area under the ROC (AUC) values were 0.806 [95% confidence interval (CI): 0.7608-0.8815], 0.755 (95% CI: 0.6839-0.8263) and 0.821 (95% CI: 0.7608-0.8815). The nomogram's C-index value, including the 14-gene signature and other potential risk factors, was 0.786 (95% CI: 0.7296-0.8425), and the calibration exhibited fairly good consistency with the perfect prediction. Based on the 14-gene risk score, high-risk PTC patients had a worse DFS. CONCLUSIONS: A novel 14-gene signature was developed for predicting LNM in PTC patients. The risk score also correlated with DFS in PTC patients.

Analysis of sentinel lymph node biopsy and non-sentinel lymph node metastasis in invasive ductal and invasive lobular breast cancer: a nationwide cross-sectional study (CSBrS-001).

BACKGROUND: Information regarding the implementation of sentinel lymph node biopsy (SLNB) in invasive lobular carcinoma (ILC) is scarce, and whether ILC patients with 1-2 positive sentinel lymph nodes (SLNs) can be omitted from axillary lymph node dissection (ALND) remains controversial. This study aimed to compare involvement of SLNs and non-SLNs between patients with invasive ductal carcinoma (IDC) and ILC. METHODS: We retrospectively collected the clinical and pathological data of invasive breast cancer patients from 37 medical centers in China from January 2018 to December 2018. The number of resected SLNs, positive rate of SLNs, and non-SLNs metastasis were compared between patients with IDC and ILC. RESULTS: A total of 6,922 patients were included, comprising 6,650 with IDC (96.1%) and 272 with ILC (3.9%). No difference was observed in the number of resected SLNs between patients with IDC and ILC (IDC: 4.0±1.9 vs. ILC: 3.9±1.6, P=0.352). The positive rate of SLNs was significantly higher in patients with IDC than that in patients with ILC (19.3% in IDC vs. 12.9% in ILC, P=0.008). The difference in positive rate of SLNs between IDC and ILC was mainly attributed to macro-metastasis. For patients with positive SLNs who received ALND, and those with 1-2 positive SLNs, the metastatic rate of non-SLNs in the ILC group was higher than that in the IDC group (for patients with positive SLNs: 50.0% in ILC vs. 39.9% in IDC, P=0.317; for patients with 1-2 positive SLNs: 45.4% in ILC vs. 34.8% in IDC, P=0.366), but the differences were not statistically significant. CONCLUSIONS: Patients with ILC had similar number of resected SLNs and lower positive rate of SLNs compared to those with IDC. In participants with 1-2 positive SLNs, the ILC group had an increased tendency for non-SLNs metastasis compared with the IDC group. Surgeons may need to be more cautious about omitting ALND for ILC patients with 1-2 positive SLNs.

Risk Factors of Central Lymph Node Metastasis in Papillary Thyroid Microcarcinoma and the Value of Sentinel Lymph Node Biopsy.

Objective: The present study aims to investigate the risk factors of central lymph node metastasis (CNM) in papillary thyroid microcarcinoma (PTMC) and evaluate the predictive value of sentinel lymph node biopsy (SLNB) during surgery. Methods: The clinicopathological data of 179 patients with PTMC staging in cN0 and with SLNB performed were analyzed retrospectively. Positive sentinel lymph node ratio (PSLNR) and additional positive lymph node (APLN) were analyzed in cases with positive SLNB. The efficiency of SLNB was investigated. ROC curves were plotted to evaluate the predictive value of PSLNR for APLN. Results: Cumulative maximum diameter of tumors (CMD) (P = 0.041) and capsule involvement (CI) (P = 0.014) were independent risk factors for central lymph node metastasis. The SLNB success rate was 97.28%, and the incidence of CNM was 31.28%. The sensitivity, specificity, false positive rate (FPR), false negative rate (FNR), positive predictive value (PPV), and negative predictive value (NPV) of SLNB to evaluate CNM and APLN were 82.14 vs. 61.54%, 100 vs. 80.39%, 0 vs. 19.61%, 17.86 vs. 38.46%, 100 vs. 34.78%, and 92.48 vs. 92.48%, respectively. For cases with positive SLNB, subgroup analysis was performed according to APLN. The PSLNRs of true and false positive groups were 0.4620 ± 0.1744 and 0.2425 ± 0.1355, respectively (P < 0.001). Analyzing the predictive value of PSLNR by the ROC curve, the optimal diagnostic cutoff point was 0.2917 [AUC = 0.861 (95% CI: 0.757, 0.966), P < 0.001], and the sensitivity, specificity, FPR, FNR, PPV, and NPV of PSLNR were 87.50, 73.33, 26.67, 12.50, 63.64, and 91.67%, respectively. Conclusion: CMD and CI are independent risk factors for central lymph node metastasis in PTMC. SLNB has good predictive value for CNM. For cases with positive SLNB, PSLNR could be used to predict the presence of APLN, which may provide a theoretical basis for intraoperative lymph node dissection.

Carbon nanoparticle-guided intraoperative lymph node biopsy predicts the status of lymph nodes posterior to right recurrent laryngeal nerve in cN0 papillary thyroid carcinoma.

BACKGROUND: The lymph nodes in the right central compartment can be divided into 2 parts by the right recurrent laryngeal nerve (RLN), and there is a lack of an accurate and convenient method for assessing metastases in the lymph node posterior to the right recurrent laryngeal nerve (LN-prRLN) in cN0 thyroid papillary carcinoma patients. METHODS: Patients diagnosed with cN0 thyroid papillary carcinoma and underwent intraoperative carbon nanoparticle-guided lymph node biopsy from January 2017 to November 2020 at the Center for Thyroid and Breast Surgery of Xuanwu Hospital were retrospectively analyzed. The intraoperative frozen section examination and postoperative LN-prRLN status should have been comprehensively recorded. The participants were divided into the LN-prRLN positive group and LN-prRLN negative group according to their recorded LN-prRLN status. RESULTS: In total, 189 cases (LN-prRLN positive group, n=30; LN-prRLN negative group, n=159) were included in the analysis. The univariate and multivariate regression analyses revealed that the number of metastatic lymph nodes during intraoperative lymph node biopsy was the potential predictor for LN-prRLN metastasis [odds ratio (OR): 1.320, 95% confidence interval (CI): 1.057 to 1.649, P=0.014]. The receiver operating characteristic (ROC) curve showed that the area under the curve (AUC) reached 0.7 upon a combined analysis of multiple lymph node statuses located at pre-laryngeal (Delphian), pre-tracheal, and para-tracheal lymph nodes ipsilateral to the tumor in predicting the metastasis of LN-prRLN, and the cut-off value was 0.5. CONCLUSIONS: Number of metastatic lymph nodes in intraoperative biopsy was an indicator of LN-prRLN metastasis in cN0 thyroid carcinoma patients. Patients staging in cN0 with negative intraoperative lymph node status might be considered not to require LN-prRLN dissection during central lymph nodes dissection.

A 10-Gene Signature for Predicting the Response to Neoadjuvant Trastuzumab Therapy in HER2-Positive Breast Cancer.

BACKGROUND AND PURPOSE: Dual-target therapy may increase the incidence of adverse events and cause economic burden to patients with human epidermal growth factor receptor 2 (HER2)-positive breast cancer. It is necessary to identify the patients who could benefit greatly from a single-target neoadjuvant therapy in order to avoid overtreatment of patients. PATIENTS AND METHODS: The baseline transcriptome data and clinical characteristics of patients with HER2-positive breast cancer who received neoadjuvant trastuzumab therapy were obtained from the Gene Expression Omnibus database. Least absolute shrinkage and selection operator (LASSO) regression analyses were used to construct the predictive model for pathologic complete response (pCR). RESULTS: A 10-gene signature model for predicting pCR rate after neoadjuvant trastuzumab therapy was constructed by LASSO regression. The areas under the receiver operating characteristics (ROC) curves in the training set and validation set were 0.896 (95% confidence interval [CI], 0.8165-0.9758) and 0.775 (95% CI, 0.5402-1), respectively. The result of logistic regression analysis showed that the risk score calculated by the 10-gene signature model was a potential predictor for pCR. Among the 10-gene signature, TFAP2B, SUSD2, AQP3, MUCL1, and ANKRD30A were found to be predictors for worse relapse-free survival (RFS) in patients with HER2-positive breast cancer, whereas MGP, YIF1B, ANKRD36BP2, and FBXO6 were found to be predictors for favorable RFS. CONCLUSION: A novel 10-gene signature that could predict the response of neoadjuvant anti-HER2 therapy in patients with HER2-positive breast cancer was developed, and the risk score of the 10-gene signature could be calculated to guide the selection of anti-HER2 therapy regimens.

Profilin 2 (PFN2) promotes the proliferation, migration, invasion and epithelial-to-mesenchymal transition of triple negative breast cancer cells.

BACKGROUND: Triple negative breast cancer (TNBC) is the most aggressive subtype with the worst prognosis. The role of profilin 2 (PFN2) in TNBC is very controversial. The current study is to explore the role of PFN2 in TNBC. METHODS: PFN2 expression in TNBC and normal breast tissues were evaluated by immunohistochemical analysis. The association between PFN2 expression and prognosis in TNBC patients was analyzed from the TCGA database. A cell counting kit-8 (CCK8) assay was employed to investigate the effects of PFN2 in TNBC cell proliferations. The migration and invasion capability of TNBC cells was evaluated by transwell assays. Western blot was performed to assess the related protein expression of TGF-ß/Smad signaling and epithelial to mesenchymal transition. Finally, TNBC xenografts were established to determine the tumorigenicity in vivo using female Nod/Scid mice. RESULTS: PFN2 is upregulated in TNBC and the higher expression was associated with worse survival. CCK8 assays and Transwell assays demonstrated that PFN2 promoted the proliferation, migration and invasion of TNBC cells. Smad2 and Smad3 were upregulated in PFN2 overexpressing TNBC cells, which further induced the process of epithelial­to­mesenchymal transition. Similarly, the overexpressing PFN2 TNBC cells exhibited stronger tumorigenicity in vivo. CONCLUSIONS: Higher PFN2 expression is associated with a worse 10-year overall survival and relapse-free survival in breast cancer patients, as well as worse 10-year relapse-free survival in TNBC patients. PFN2 promotes the proliferation, migration and invasion of TNBC cells by regulating epithelial-to-mesenchymal transition.

Role of intraoperative neuromonitoring of recurrent laryngeal nerve in thyroid and parathyroid surgery.

OBJECTIVE: To investigate whether intraoperative neuromonitoring (IONM) has a significant advantage in reducing the incidence of recurrent laryngeal nerve (RLN) injury. METHODS: Patients who underwent thyroid and parathyroid surgery from October 2012 to December 2017 at the Center for Thyroid and Breast Surgery of Xuanwu Hospital were retrospectively analyzed. They were divided into the IONM group and visualization alone group (VA group) according to whether IONM was used. RESULTS: In total, 1696 nerves at risk of injury (IONM group, n = 1104; VA group, n = 592) were included in the analysis. Among the high-risk nerves, permanent damage occurred in no cases in the IONM group but in one case in the VA group. Because the higher proportion of central lymph node metastasis caused difficulties in central cervical lymph node dissection and identification of the RLN, the patients undergoing lateral cervical lymph node dissection in the VA group had a significantly higher risk of postoperative RLN injury (11.76% vs. 0.00%). CONCLUSION: IONM technology has advantages in protection of the RLN, especially in high-risk nerves and patients with a high proportion of central lymph node metastasis who require central and lateral cervical lymph node dissection.