Application and subgroup analysis of competing risks model based on different lymph node staging systems in differentiated thyroid cancer.

Differentiated thyroid cancer (DTC) is the most common endocrine malignancy, with a rising incidence worldwide. Accurate prognostic models are essential for effective patient management. This study evaluates the prognostic value of various lymph node staging systems in DTC using a competing risks model. We used SEER database records (1998-2016) of 16,527 DTC patients, analyzing N stage, positive lymph node numbers (PLNNs), metastatic lymph node ratio (MLNR), log odds of positive lymph nodes (LODDS), and log odds of the negative lymph node (NLN)/T stage ratio (LONT). Univariate and multivariate analyses in a competing risks model were performed, along with subgroup analyses based on demographic and clinical characteristics. In this study of 16,527 patients with DTC, different lymph node staging systems showed different prognostic correlations in univariate and multivariate analyses. In particular, PLNNs showed significant prognostic correlations in several subgroups. Additionally, PLNNs were more suitable as a lymph node staging system for DTC than LODDS and MLNR in N1 stage subgroups, with an optimal cut-off of 13. Receiver operating characteristic curves, calibration curves and nomograms improved the clinical utility of the prognostic model based on PLNNs. Using competing risks model and subgroup analyses, we found that PLNNs had the best prognostic discriminatory efficacy for patients with DTC, especially those with N1 stage disease, and had an optimal cut-off value of 13.

Ultrasound radiomics based on axillary lymph nodes images for predicting lymph node metastasis in breast cancer.

Objectives: To determine whether ultrasound radiomics can be used to distinguish axillary lymph nodes (ALN) metastases in breast cancer based on ALN imaging. Methods: A total of 147 breast cancer patients with 41 non-metastatic lymph nodes and 109 metastatic lymph nodes were divided into a training set (105 ALN) and a validation set (45 ALN). Radiomics features were extracted from ultrasound images and a radiomics signature (RS) was built. The Intraclass correlation coefficients (ICCs), Spearman correlation analysis, and least absolute shrinkage and selection operator (LASSO) methods were used to select the ALN status-related features. All images were assessed by two radiologists with at least 10 years of experience in ALN ultrasound examination. The performance levels of the model and radiologists in the training and validation subgroups were then evaluated and compared. Result: Radiomics signature accurately predicted the ALN status, achieved an area under the receiver operator characteristic curve of 0.929 (95%CI, 0.881-0.978) and area under curve(AUC) of 0.919 (95%CI, 95%CI, 0.841-0.997) in training and validation cohorts respectively. The radiomics model performed better than two experts' prediction of ALN status in both cohorts (P<0.05). Besides, prediction in subgroups based on baseline clinicopathological information also achieved good discrimination performance, with an AUC of 0.937, 0.918, 0.885, 0.930, and 0.913 in HR+/HER2-, HER2+, triple-negative, tumor sized ≤ 3cm and tumor sized>3 cm, respectively. Conclusion: The radiomics model demonstrated a good ability to predict ALN status in patients with breast cancer, which might provide essential information for decision-making.

Newly found variations of the right posterior portal vein identified radiologically in 1,003 Chinese patients: a cross-sectional study.

Background: The anatomy of the right posterior portal vein (RPPV) plays an important role in planning hepatic resection, living transplantation and interventional radiological procedures, yet the incidence of variations of RPPV without a common trunk in Chinese persons is still unclear. Therefore, we conducted this study and discussed its clinical implications. Methods: A retrospective analysis of multidetector computed tomography (MDCT) scans was performed in 1,933 patients with various abdominal pathologies between September 28, 2018 through May 23, 2019. After excluding 930 patients, a total of 1,003 patients were included in this study. Variations of the RPPV without a common trunk were classified according to classification standards. Results: A total of 1,003 patients were included. RPPV without a common trunk was found in 216 (21.54%, 216/1,003) patients. Among them, we identified three variations of the origin from the right portal vein (RPV): first separate origin of P6, P7, or simultaneous separate origin of P6 and P7, and the incidences of these three variations were 1.50% (15/1,003), 6.58% (66/1,003) and 13.46% (135/1,003), respectively. Among 1,003 patients included in this study, 787 patients (78.46%, 787/1,003) showed that RPPV normally divided into P6 and P7 branches. Conclusions: Variations of the RPPV without a common trunk were not rare in Chinese population. Knowledge of this anatomic variation of the RPPV is extremely important for hepatic and transplant surgeons and interventional radiologists.

Receptor-ligand pair typing and prognostic risk model for papillary thyroid carcinoma based on single-cell sequencing.

The papillary thyroid carcinoma (PTC) microenvironment consists of various cancer and surrounding cells, and the communication between them is mainly performed through ligand-receptor (LR) interactions. Single-cell RNA sequencing (scRNA-seq) has been performed to investigate the role of intercellular communication networks in tumor progression. In addition, scRNA-seq can accurately identify the characteristics of immune cell subsets, which is of great significance for predicting the efficacy of immunotherapy. In this study, the cell-cell communication network was analyzed through LR pairs, and a new PTC molecular phenotype was developed based on LR pairs. Furthermore, a risk model was established to predict patient response to PD-1 blockade immunotherapy. The scRNA-seq dataset was obtained from GSE184362, and the bulk tumor RNA-seq dataset was obtained from The Cancer Genome Atlas. CellPhoneDB was used for cellular communication analysis. LR pair correlations were calculated and used to identify molecular subtypes, and the least absolute shrinkage and selection operator (Lasso) Cox regression was used to develop a risk model based on LR pairs. The IMvigor210 and GSE78220 cohorts were used as external validations for the LR.score to predict responses to PD-L1 blockade therapy. A total of 149 LR pairs with significant expression and prognostic correlation were included, and three PTC molecular subtypes were obtained from those with significant prognostic differences. Then, five LR pairs were selected to construct the risk scoring model, a reliable and independent prognostic factor in the training set, test set, and whole dataset. Furthermore, two external validation sets confirmed the predictive efficacy of the LR.score for response to PD-1 blockade therapy.

Prognostic value of LODDS in medullary thyroid carcinoma based on competing risk model and propensity score matching analysis.

Log odds of positive lymph nodes (LODDS) is an independent prognostic factor for patients with medullary thyroid carcinoma (MTC). However, the optimal cutoff value for LODDS needs to be further confirmed, and previous studies have ignored the prevalent competing events of non-cancer deaths among patients with MTC, thus possibly overestimating the risk of death from cancer. The information of patients with MTC who underwent total thyroidectomy was collected from SEER database. Restricted cubic splines (RCS) were used to determine the optimal cutoff for LODDS. Moreover, patients' overall survival (OS) and disease-specific survival (DSS) were determined using Kaplan-Meier and Cox proportional-hazards model. The competing risk models (CRM) were used to reduce the effect of competing events, and propensity score matching was performed to balance the confounding factors between groups. The cutoff value of LODDS determined by RCS was - 1.004, and a total of 2314 patients with MTC were recruited. In the CRM after PSM, factors such as age over 55 years at diagnosis, being male, treatment with chemotherapy or radiotherapy, unknown tumor size, and LODDS > - 1.004 were significantly associated with poor prognosis of patients both in univariate and multivariate analyses, while the presence of multifocal tumor indicated better prognosis. Patients with MTC who were over 55 years old at diagnosis, were male, received chemotherapy or radiation, had an unclear initial tumor size, and had LODDS > - 1.004 had a worse prognosis than patients with multifocal tumor.

An introduction of an easy-operating and economical technique for tissue microarray preparation.

AIM: Tissue microarray (TMA) is a powerful and effective tool for in situ tissue analysis. However, manual TMA construction methods showed varied qualities. This study aimed to raise a standardised TMA preparation technique that can be easily operated and is economical. METHODS: A sampling needle was used to punch the tissue rods from the donor block and holes in the recipient block. To indicate the dots' positions and ensure vertical punching, a novel auxiliary device made using commercial three-dimensional printing technology was attached. The TMA block was made up of tissue rods and a recipient block. RESULTS: A 77-rod (7×11) TMA block was constructed. The rows and columns were fixed in straight lines. There was no specimen loss during the process of embedding. CONCLUSIONS: An alternative method for the construction of TMA blocks that met the basic requirement of many laboratories and can be effortlessly performed was presented.

High expression of SMARCE1 predicts poor prognosis and promotes cell growth and metastasis in gastric cancer.

Background: Gastric cancer (GC) is one of the most lethal cancers worldwide with a high risk for recurrence and metastasis. Therefore, further understanding of the metastatic mechanism and the development of treatment strategies are required. Although increasing evidence suggests that SWI/SNF Related, Matrix Associated, Actin Dependent Regulator of Chromatin, Subfamily E, Member 1 (SMARCE1) promotes cancer metastasis, its role in GC remains unclear. Materials and methods: GC samples (n=122) were used to investigate the association between SMARCE1 expression, patient clinicopathological features, and prognosis. The expression of SMARCE1 in GC tissues was measured using real-time polymerase chain reaction, western blotting, and immunohistochemistry. MGC-803 and AGS cells were transfected with lentivirus to upregulate or downregulate SMARCE1 expression. The roles of SMARCE1 in GC cell proliferation, migration, and invasion were determined using Cell Counting Kit-8 assay, colony formation assay, wound healing, transwell migration, and invasion assay. Nude mice models were established to observe tumorigenesis. The specific mitogen-activated protein kinase (MAPK) inhibitor U0126 was utilized to verify the involved pathway. Results: SMARCE1 was highly expressed in GC tissues and cell lines. High expression of SMARCE1 was correlated with the malignant clinicopathological characteristics of GC patients, including tumor size, depth of invasion, degree of differentiation, lymph node involvement, and TNM stage (all P<0.05). Kaplan-Meier survival analysis revealed that high SMARCE1 expression predicted poor prognosis in GC patients (P<0.01). Moreover, SMARCE1 was an independent risk factor of poor prognosis (P<0.01). Functional study revealed that overexpression of SMARCE1 markedly promoted the proliferation, migration, and invasion of GC cells in vitro and tumorigenesis in vivo. Furthermore, SMARCE1 activated the MAPK/ERK signaling pathway. U0126 significantly inhibited the SMARCE1-induced proliferation and mobility of GC cells. Conclusion: SMARCE1 promoted growth and metastasis of GC, indicating its potential usefulness as a prognostic biomarker and target for therapeutic intervention against this disease.

TGFß1-Induced LncRNA UCA1 Upregulation Promotes Gastric Cancer Invasion and Migration.

According to recent studies, long noncoding RNA urothelial carcinoma associated 1 (UCA1) is involved in the development and progression of many malignant tumors, including gastric cancer (GC). We validated the detailed role of UCA1 in human GC cell lines and GC tissues so as to determine its exact function and the underlying mechanism of GC invasion and migration. In our research, lncRNA-UCA1 was specifically upregulated in GC tissues and cell lines, and augmented GC cell proliferation, and invasive and migratory capabilities. High UCA1 expression in GC was related with poorer prognosis (poorer invasion depth, lymph node metastasis, advanced TNM [T is for the original (primary) tumor, N for nearby (regional) lymph nodes that are involved, and M for distant metastasis] stage, and shorter overall survival). Epithelial mesenchymal transition (EMT), associated with malignancy of cancers, was reported to be responsible for invasion and migration of cancer cells. Transforming growth factor ß1 (TGFß1)-induced EMT was well evaluated. UCA1 silence reduced the protein levels of EMT-related factors, vimentin and snail, while promoted E-cadherin and zonula occludens-1 protein levels in GC cells; the effect of UCA1 could be partly restored by TGFß1 treatment. Taken together, UCA1 might regulate the tumor proliferation, invasion, and metastasis under TGFß1 induction. Taken together, UCA1 might present a potential oncogenic factor by promoting GC cell proliferation, invasion, and migration. UCA1 could serve as a novel biomarker for prognosis and a novel therapeutic target of GC treatment.

Inhibition effect of Peg-IFNα-2b and Imatinib alone or combination on imatinib-resistant gastrointestinal stromal tumors cell lines.

To investigate the inhibition effect of polyethylene glycol interferon α-2b and imatinib alone or combination on imatinib-resistant GIST cell lines, and to explore the possible mechanism. Imatinib resistant GIST cell lines (GIST-R) were exposured to either Peg-IFNα-2b or imatinib alone or combination. The proliferative inhibition rates and the combination index (CI) values of GIST-R cells were detected by MTT assay. The apoptotic rates of GIST-R cells were detected by flow cytometry. The expression levels of phospho-mammalian target of rapamycin (p-mTOR), and Bcl-2 of GIST-R cells were analyzed by Western blot. GIST-R cells presents remarkable resistance to imatinib, and the resistance index (RI) were (P<0.05). And The proliferative inhibition rate and the apoptotic rate of GIST-R cells in combination of Peg-IFNα2b and Imatinib group were higher than those in Peg-IFNα-2b or imatinib alone group (P<0.05). The CI value of Peg-IFNα-2b and imatinib was less than each alone, which had a synergistic effect (CI=0.63). As compared with the control (GIST-R cells without any treatment), the expression levels of p-mTOR and Bcl-2 proteins of GIST-R cells in combination of Peg-IFNα-2b and imatinib group were decreased (P<0.01). The combination of Peg-IFNα-2b and imatinib generats a synergistic effect in GIST-R cells, and reversal of drug resistance. This effect may be related with apoptosis and down-regulation of the expression of p-mTOR.

Isolated caudate lobe resection for huge hepatocellular carcinoma (10 cm or greater in diameter).

Isolated caudate lobectomy for huge hepatocellular carcinoma (HCC) (10 cm or greater in diameter) is a technically demanding surgical procedure that entails the surgeon's experience and precise anatomical knowledge of the liver. We describe our clinical experiences and evaluate the results of partial or total isolated caudate lobectomy for HCC larger than 10 cm in the caudate lobe. En bloc excisions combined with adjacent hepatic parenchyma (as part of extended hepatectomies) were excluded. Twenty-seven patients were included in the study (24 male, three3 female). Median age was 43 years (range, 18 to 81 years). All primary diagnoses were HCC. Twenty-one patients had surgical margins lesser than 1 cm. Tumor embolus within the main trunk of the portal vein was found in five patients by intraoperative ultrasound. Median operative time was 288 minutes (range, 160 to 310 minutes), and estimated intraoperative blood loss was 2260 mL (range, 200 to 7000 mL). Median blood transfusion was 1460 mL (range, 0 to 7200 mL). Postoperative morbidity rate was 44.4 per cent. There were no postoperative deaths. Overall survival rates at 1, 3, and 5 years were 80.2, 52.1, and 27.1 per cent, respectively. Nineteen patients (70.4%) had tumor recurrence as of the last follow-up. The recurrence lesion was treated in most of these patients. Isolated caudate lobectomy for huge HCC is a technically demanding but safe procedure, although the procedure is sometimes extremely difficult.

Null-margin bisegmentectomy VII-VIII for hepatocellular carcinoma in cirrhotic patients.

BACKGROUND/AIMS: Preservation of functional liver parenchyma should be a priority in hepatic surgery to avoid postoperative liver failure and enhance the opportunity to perform repeat resection in case of tumor recurrence. METHODOLOGY: A tumor localized in segments VII, VIII and adhering to or compressing the middle hepatic vein sometimes indicates a need to perform bisegmentectomy VII-VIII without surgical margin. From June 2006 to June 2011, fourteen patients with such a tumor underwent null-margin bisegmentectomy VII-VIII in our hospital. We retrospectively review our experience with this uncommon and technique-challenging hepatic resection. RESULTS: Mean intraoperative blood loss was estimated to be 300 mL and only four patients required blood transfusions less than 4U each person. Mean postoperative hospitalization was 11.2 days. Postoperative complications were encountered in 28.5% of patients and there was no postoperative mortality. Median overall and disease-free survivals were 35 and 23 months, respectively. CONCLUSIONS: The lack of ability to obtain an adequate surgical margin should not be considered as a contraindication for hepatectomy of HCC. In patients with impaired liver functional reserve and with right superiorly located tumors, the preservation of the middle hepatic vein should take priority and null-margin bisegmentectomy VII-VIII for HCC should be recommended.

[Three-dimensional cell culture and histology of vasculogenic mimicry in hepatocellular carcinoma].

BACKGROUND & OBJECTIVE: Vasculogenic mimicry (VM) is a blood supply pattern in malignant tumors. Mimic vessel walls are formed by tumor cells, not by endothelial cells. Previous studies of VM are limited on malignant tumors with high aggression and bi-directional differentiation potentials, but seldom on epithelium-originated tumors. This study was to observe VM in hepatocellular carcinoma (HCC) by 3-dimensional culture of HCC cell line HepG2 and immunohistochemistry. METHODS: Three-dimensional culture system of HepG2 cells was constructed to observe VM. The presence of VM in 15 specimens of HCC was observed by immunohistochemical and histological double staining of CD31 and PAS, or Ferritin and PAS. RESULTS: During 3-dimensional culture, HepG2 cells stretched out thin and long apophyses at the second day, and linked each other to form wreath and net-work structure at the seventh day. All of the 15 HCC simples showed CD31-positive for endothelial cells, and Ferritin-positive for tumor cells. The immunohistochemical and histological double staining also proved that VM, formed by CD31-negative and Ferritin-positive tumor cells, existed in 7 of the 15 simples of HCC. Tumor cells were separated from the tubes by PAS-positive matter like basement membrane. Red blood cells could be seen in the tubes. The number of VM was less in well differentiated simples than in poorly differentiated samples; CD31-positive tumor cells were observed in poorly differentiated samples. CONCLUSIONS: HepG2 cells have the capacity of self-metamorphosing and vasculorizing. Tumor cells can obtain oxygen and nutrition through VM.

Cloning and Analysis of Intron 3 of Human Telomerase Reverse Transcriptase (hTERT) Gene.

Total RNA was extracted from telomerase-positive cell line SPC-A, and reverse transcribed to cDNA. The long template PCR was performed using this cDNA as template and the hTERT cDNA specific oligonucleotides as primers. A long fragment of about 2.2 kb was produced as well as a short fragment of about 150 bp. The long fragment was purified from the gel, cloned into T-easy vector and sequenced from two directions. The sequencing result and homologous comparison indicated that the fragment contained the intron 3 of hTERT gene. Further assay showed that the precursor of this fragment is premature mRNA (pre-mRNA) of hTERT gene and the copy number varied among different cell lines, as verified in this study. These results suggest the feasibility of cloning the intron of eucaryotic gene by the combination of reverse transcription and long template PCR system. And the different splicing efficiency of the intron 3 from the hTERT pre-mRNA was also implied from this assay.