Bioreaction-Compatible Bivariate Lanthanide MOF Sensor Enables Stimulus-Multiresponsive Platform for ctDNA On-Site Detection.

Detection of circulating tumor DNA (ctDNA) in liquid biopsy is of great importance for tumor diagnosis but difficult due to its low amount in bodily fluids. Herein, a novel ctDNA detection platform is established by quantifying DNA amplification by-product pyrophosphate (PPi) using a newly designed bivariable lanthanide metal-organic framework (Ln-MOF), namely, Ce/Eu-DPA MOF (CE-24, DPA = pyridine-2,6-dicarboxylic acid). CE-24 MOF exhibits ultrafast dual-response (fluorescence enhancement and enzyme-activity inhibition) to PPi stimuli by virtue of host-guest interaction. The platform is applied to detecting colon carcinoma-related ctDNA (KARS G12D mutation) combined with the isothermal nucleic acid exponential amplification reaction (EXPAR). ctDNA triggers the generation of a large amount of PPi, and the ctDNA quantification is achieved through the ratio fluorescence/colorimetric dual-mode assay of PPi. The combination of the EXPAR and the dual-mode PPi sensing allows the ctDNA assay method to be low-cost, convenient, bioreaction-compatible (freedom from the interference of bioreaction systems), sensitive (limit of detection down to 101 fM), and suitable for on-site detection. To the best of our knowledge, this work is the first application of Ln-MOF for ctDNA detection, and it provides a novel universal strategy for the rapid detection of nucleic acid biomarkers in point-of-care scenarios.

Risk factors for parapharyngeal and retropharyngeal metastases in papillary thyroid cancer: a matched case-control study.

PURPOSE: To analyze risk factors for parapharyngeal (PP) and retropharyngeal (RP) metastases in papillary thyroid cancer (PTC). METHODS: A matched case-control study was conducted, comprising 130 age- and sex-matched cases of PTC. Among these cases, 50 had PP/RP metastases, 50 had central and/or lateral neck lymph node metastases, and 30 showed no lymph node metastases. Preoperative thyroid function test, computed tomography images, and postoperative pathological findings were collected. Associations between cases were assessed using univariate conditional logistic regression analysis, followed by multivariate conditional logistic regression analysis, and backward stepwise selection to predict risk factors for PP/RP metastases. RESULTS: The study found that thyroglobulin was significantly associated with the development of PP/RP metastases [136.10(16.55-312.60) vs. 27.60(10.28-55.62) vs. 8.74(6.35-21.10) P < 0.001]. CONCLUSIONS: The study concludes that thyroglobulin is a significant risk factor for PP/RP metastases in PTC. This finding emphasizes the importance of monitoring thyroglobulin levels in PTC patients to identify those at risk of developing PP/RP metastases.

TERT genetic polymorphism rs2736100 is associated with an aggressive manifestation of papillary thyroid carcinoma.

Objectives: TERT rs2736100 genetic polymorphism is commonly found in human malignancies, indicating its key role in cancer cell transformation. The aim of this study is to investigate the effects of the functional TERT rs2736100 genetic polymorphism on the outcomes of papillary thyroid carcinoma (PTC) patients. Materials and methods: We performed a retrospective study on the relationship between rs2736100 and clinicopathological outcomes of PTC in 500 patients (378 females and 122 males) aged 43.8 ± 11.4 years (range 15-74 years) with a median follow-up of 60 months (range, 1-455 months). Results: TERT rs2736100 genetic polymorphism (TG/GG vs. TT) was significantly associated with several high-risk clinicopathological features such as tumor spread, extrathyroidal extension, central/lateral lymph node metastases, and Stage T III or IV disease. However, in Kaplan-Meier survival analyses, the rs2736100 mutation was unrelated to overall disease-free survival with a log-rank value of p > 0.05. In Cox-regression analyses, the overall survival rate of recurrence/neo-metastasis was related to a larger tumor size, younger age, and tumor spread but unrelated to the rs2736100 mutation. Conclusions and significance: TERT rs2736100 genetic polymorphism mutation is more likely to manifest with aggressive clinicopathological characteristics but cannot worsen prognosis in PTC.

Parathyroid Hormone Reduction Predicts Transient Hypocalcemia after Total Thyroidectomy: A Single-Center Prospective Study.

OBJECTIVE: We performed this study to investigate the risk factors for postoperative hypocalcemia after total thyroidectomy with central lymph node dissection (CLND). Study Design. This was a single-center prospective study based on 176 consecutive patients who underwent total thyroidectomy for papillary thyroid carcinoma. Setting. Patients were recruited between January 2016 and June 2018. Subjects and Methods. Patients who underwent bilateral (n = 155, bilateral group) and ipsilateral CLND (n = 21) after total thyroidectomy were included. The preoperative and postoperative parathyroid hormone (PTH) and calcium levels were detected. The risk factors for transient hypocalcemia were identified using logistic regression analysis and receiver operating characteristic (ROC) curve analysis. RESULTS: Fifty-one (28.98%) patients developed transient hypocalcemia, and 2 patients (1.14%) developed permanent hypoparathyroidism. There was no difference in the gender ratio or the morbidity of hypocalcemia between the patients who underwent bilateral and ipsilateral CLND. On postoperative day 1, PTH decrease was a risk factor for transient hypocalcemia in the whole cohort (ß = 0.043, OR = 1.044, 95% CI 1.023-1.065, p < 0.001), bilateral group (ß = 0.042, OR = 1.043, 95% CI 1.022-1.064, p < 0.001), and female patients (ß = 0.049, OR = 1.050, 95% CI 1.026-1.075, p < 0.001). Tumor diameter was a risk factor for transient hypocalcemia in female patients (ß = 0.499, OR = 1.647, 95% CI 1.003-2.704, p=0.049). The ROC curve analysis illustrated that 65.58%, 71.00%, and 71.00% PTH level reduction had high accuracy in predicting transient hypocalcemia in the whole cohort, bilateral group, and female patients, respectively (AUC = 0.986, 0.987, and 0.987). CONCLUSION: Asymptomatic female patients with bilateral CLND and a 71.00% PTH level reduction were at a high risk of transient hypocalcemia.

Expression of PIM-1 in salivary gland adenoid cystic carcinoma: Association with tumor progression and patients' prognosis.

Pim-1 proto-oncogene, serine/threonine kinase (PIM-1) phosphorylates a series of substrates to exert its oncogenic function in numerous malignancies. The present study investigated the clinical significance of the PIM-1 protein, apoptosis status and apoptosis-associated proteins, including forkhead box O3a (FOXO3a), B cell lymphoma-2 (BCL-2) and BCL-2-associted agonist of cell death (BAD), were investigated in salivary gland adenoid cystic carcinoma (ACC) tissues. PIM-1 expression levels in 4 pairs of ACC tissues and corresponding normal salivary gland tissues were determined by western blot analysis. PIM-1, FOXO3a, BAD and BCL-2 expression levels in 60 ACC tissues were evaluated by immunohistochemistry (IHC). A terminal deoxynucleotidyl-transferase-mediated dUTP nick end labeling assay was performed to detect the apoptosis status of ACC tissues. PIM-1 was revealed to be highly expressed in ACC tissues compared with adjacent normal tissues. IHC staining results demonstrated high expression ratios of PIM-1, FOXO3a, BCL-2 and BAD [33.33% (20/60), 51.67% (31/60), 51.67% (31/60) and 55% (33/60)], respectively, and significant correlations between the expression of PIM-1 and FOXO3a and BCL-2 (P<0.05). Apoptotic rates were significantly associated with PIM-1, FOXO3a, BCL-2 and BAD expression levels (P<0.05). PIM-1 expression levels were significantly associated with tumor size, lymph node involvement, nerve invasion, distant metastasis and weakly associated with tumor node metastasis stage. Kaplan-Meier survival curves revealed that PIM-1 expression level was significantly associated with disease-free survival of patients with ACC (P=0.009). Cox regression multivariate analysis results revealed that histotype, distant metastasis and apoptotic rate were independent prognosis factors for ACC. Assessment of PIM-1 may be useful in investigating the malignant behaviors of ACC and predicting the outcome of patients with ACC.

Skip metastasis in papillary thyroid carcinoma is difficult to predict in clinical practice.

BACKGROUND: Cervical lymph node metastases are very common in papillary thyroid cancer (PTC), and typically spread in a predictable stepwise fashion in clinical practice. However, lateral lymph node metastasis (LLNM) without central lymph node metastasis (CLNM) as skip metastasis is not rare in PTC. The aim of this study was to investigate the incidence, risk factors and pattern of skip metastasis in PTC. METHODS: A total of 271 patients with PTC and suspicious LLN diagnosed by pre-operation examinations who underwent total thyroidectomy and central lymph node dissection plus lateral lymph node dissection between January 2008 and December 2011 were enrolled in this study. Clinicopathological features were collected, and the pattern of cervical lymph node metastasis and skip metastasis were analyzed. RESULTS: The LLNM rate was 74.9% (203/271, diagnosed by postoperative pathology examination) and significantly associated with extrathyroid extension (ETE), primary tumor located at the upper pole, and CLNM (p < 0.05). The skip metastasis rate was 14.8% (30/203) and was more frequently found in microcarcinoma patients, especially when the primary tumor size was ≤0.5 cm (p = 0.001 OR = 12.9). However, skip metastasis was unrelated to the remaining factors examined. CONCLUSION: Small cancers with a pre-operation diagnosis of LLNM are more likely to have skip metastases, especially when the primary tumor size is less than 0.5 cm in diameter; however, this type of metastasis appears to develop in a random fashion. Thus, additional research is needed to identify potential predictive factors, such as a primary tumor located at the upper pole.

The significance of Delphian lymph node in papillary thyroid cancer.

AIM: To investigate the clinical significance of the DLN metastasis in papillary thyroid cancer (PTC). METHODS: A total of 231 PTC patients who underwent first surgical treatment in the Department of Hand and Neck Surgery of Zhejiang Cancer Hospital from January 2013 to June 2014 were enrolled. The relationship between Delphian lymph node (DLN) metastasis and patient age, gender, tumor size, tumor number, unilateral or bilateral, capsular invasion, pretracheal and paratracheal node metastasis, and lateral node metastasis was analyzed. RESULTS: Within 231 cases, 69 showed DLN, but only 19 (8.23%) were found with metastasis. In the univariate analysis, DLN metastasis was significantly associated with tumor size (P = 0.023), capsular invasion (P = 0.001), pretracheal or paratracheal node metastasis (P = 0.003) and lateral node metastasis (P = 0.001), while there were no significant correlation between DLN metastasis and gender (P = 0.976), age (P = 0.976), tumor number (P = 0.234) and unilateral or bilateral (P = 0.724). In the multivariate analysis, capsular invasion was an independent risk factor of DLN metastasis (P < 0.05, odds ratio = 10.15). CONCLUSION: Capsular invasion is an independent risk factor of DLN metastasis and DLN metastasis could be used as a predictor of lateral node metastasis. The dissection of DLN in PTC patients is recommended and lateral lymph node should be evaluated for patients with DLN positive.

Risk factors analyses for lateral lymph node metastases in papillary thyroid carcinomas: a retrospective study of 356 patients.

Objective: The aim of this study was to investigate the incidence and risk factors for lateral lymph node metastasis (LLNM) in patients with papillary thyroid carcinoma (PTC). Subjects and methods: 356 patients diagnosed with PTC who underwent total thyroidectomy and central lymph node dissection and lateral lymph node dissection between January 2005 and December 2011 were enrolled. The relation between LLNM and clinicopathological features such as gender, age, tumor size, tumor spread, psammoma bodies, tumor multifocality, extrathyroidal extension (ETE), unilateral or bilateral disease, tumor primary location and central lymph node metastases (CLNM) was analyzed. Results: The rate of LLNM was 75.0%. In the univariate analysis, it was significantly associated with age, tumor size, tumor spread, extrathyroidal extension, primary tumor location and central lymph node metastasis (p < 0.05). In contrast, in the multivariate analysis, it was significantly associated with primary tumor location, central lymph node metastasis (p < 0.05) and tumor size > 1.5 cm with p = 0.05 but was unrelated to the other factors. Conclusion: Patients with PTC, with the primary tumor located in the upper part of the lobe and positive central compartment lymph node metastasis with a tumor size > 1.5 cm diameter are more likely to have LLNM. Therefore, more meticulous evaluations including the lateral lymph nodes should be performed before surgery.

Risk factors analyses for lateral lymph node metastases in papillary thyroid carcinomas: a retrospective study of 356 patients

ABSTRACT Objective The aim of this study was to investigate the incidence and risk factors for lateral lymph node metastasis (LLNM) in patients with papillary thyroid carcinoma (PTC). Subjects and methods 356 patients diagnosed with PTC who underwent total thyroidectomy and central lymph node dissection and lateral lymph node dissection between January 2005 and December 2011 were enrolled. The relation between LLNM and clinicopathological features such as gender, age, tumor size, tumor spread, psammoma bodies, tumor multifocality, extrathyroidal extension (ETE), unilateral or bilateral disease, tumor primary location and central lymph node metastases (CLNM) was analyzed. Results The rate of LLNM was 75.0%. In the univariate analysis, it was significantly associated with age, tumor size, tumor spread, extrathyroidal extension, primary tumor location and central lymph node metastasis (p < 0.05). In contrast, in the multivariate analysis, it was significantly associated with primary tumor location, central lymph node metastasis (p < 0.05) and tumor size > 1.5 cm with p = 0.05 but was unrelated to the other factors. Conclusion Patients with PTC, with the primary tumor located in the upper part of the lobe and positive central compartment lymph node metastasis with a tumor size > 1.5 cm diameter are more likely to have LLNM. Therefore, more meticulous evaluations including the lateral lymph nodes should be performed before surgery.

Functional evaluation of TERT-CLPTM1L genetic variants associated with susceptibility of papillary thyroid carcinoma.

TERT is the catalytic subunit of telomerase which plays an essential part in cellular immortality by maintaining telomere integrity. TERT is commonly over-expressed in human malignancies, indicating its key role in cell transformation. The chromosome 5p15.33 TERT-CLPTM1L region has been associated with susceptibility of multiple cancers via a genome-wide association approach. However, the involvement of this locus in papillary thyroid carcinoma (PTC) etiology is still largely unknown. We analyzed 15 haplotype-tagging single nucleotide polymorphisms (htSNPs) of the TERT-CLPTM1L region in a two stage case-control design. After genotyping 2300 PTC patients and frequency-matched 2300 unaffected controls, we found that TERT rs2736100 genetic variant is significantly associated with elevated PTC risk. Ex vivo reporter gene assays indicated that the PTC susceptibility rs2736100 polymorphism locating in a potential TERT intronic enhancer has a genotype-specific effect on TERT expression. Correlations between rs2736100 genotypes and tissue-specific TERT expression supported the regulatory function of this genetic variant in vivo. Our data demonstrated that the functional TERT rs2736100 SNP as a novel genetic component of PTC etiology. This study, together with recent studies in other cancers, unequivocally establishes an essential role of TERT in cancers.

Whole exome sequencing identifies lncRNA GAS8-AS1 and LPAR4 as novel papillary thyroid carcinoma driver alternations.

Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer. However, we know little of mutational spectrum in the Chinese population. Thus, here we report the identification of somatic mutations for Chinese PTC using 402 tumor-normal pairs (Discovery: 91 pairs via exome sequencing; validation: 311 pairs via Sanger sequencing). We observed three distinct mutational signatures, evidently different from the two mutational signatures among Caucasian PTCs. Ten significantly mutated genes were identified, most previously uncharacterized. Notably, we found that long non-coding RNA (lncRNA) GAS8-AS1 is the secondary most frequently altered gene and acts as a novel tumor suppressor in PTC. As a mutation hotspot, the c.713A>G/714T>C dinucleotide substitution was found among 89.1% patients with GAS8-AS1 mutations and associated with advanced PTC disease (P = 0.009). Interestingly, the wild-type lncRNA GAS8-AS1 (A713T714) showed consistently higher capability to inhibit cancer cell growth compared to the mutated lncRNA (G713C714). Further studies also elucidated the oncogene nature of the G protein-coupled receptor LPAR4 and its c.872T>G (p.Ile291Ser) mutation in PTC malignant transformation. The BRAF c.1799T>A (p.Val600Glu) substitution was present in 59.0% Chinese PTCs, more frequently observed in patients with lymph node metastasis (P = 1.6 × 10(-4)). Together our study defines a exome mutational spectrum of PTC in the Chinese population and highlights lncRNA GAS8-AS1 and LPAR4 as potential diagnostics and therapeutic targets.

Potential role for carbon nanoparticles identification and preservation in situ of parathyroid glands during total thyroidectomy and central compartment node dissection.

OBJECTIVE: To determine the potential role of intraoperative carbon nanoparticles (CN) injections for identification and preservation of parathyroid glands, thereby reducing the postoperative hypocalcaemia. METHODS: 100 patients with thyroid cancer who underwent total thyroidectomy and central compartment node dissection (CCND) were randomly assigned to receive intraoperative injection of (CN) or not for identifying and preserving normal parathyroid glands. RESULTS: There was no significantly difference for preoperative and postoperative parathyroid hormone (PTH) levels between the CN and control group (P>0.05). The levels of albumin-adjusted serum calcium (AASC) before surgery and at day 1 and 1 month after surgery did not reach the significant difference between the two groups (P>0.05). However, the patients in CN group had the higher level of AASC at day 3 after surgery than those in control group (P=0.044). Transient postoperative hypoparathyroidism occurred in 24 (48%) patients in CN group and 28 (56%) in control groups, respectively (P=0.423). The incidence of transient postoperative hypocalcemia was 20% (10/50) in CN group and 24% (12/50) in control groups, respectively (P=0.629). CONCLUSIONS: Carbon nanoparticles can make the thyroid gland and the central lymph node black-stained, but no-stained for parathyroid glands. After rapidly identifying parathyroid and distinguishing it from thyroid and lymph nodes by carbon nanoparticles, complete lymph node dissection and preservation of parathyroid glands become feasible during total thyroidectomy with neck lymph node dissection. After identification, strict adherence to capsular dissection remains essential for safe preservation in situ of the parathyroid glands and their blood supply.

Adenovirus-mediated interferon-γ gene therapy induced human pancreatic carcinoma Capan-2 cell apoptosis in vitro and in vivo.

Pancreatic cancer is one of the most lethal human malignancies with a very low 5-year survival rate, which highlights urgent needs for more effective therapeutic strategies. In this study, we examined the potential therapeutic effects of an adenovirus encoding human interferon gamma (Ad-IFNγ) on pancreatic carcinoma cells Capan-2 in vitro and in vivo. The results indicated that Ad-IFNγ could significantly inhibit tumor cell growth via inducing cell apoptosis. After infection, IFNγ expressed durably and stably in xenografts, predominantly in tumor tissue, while much less in blood and liver. Thus, adenovirus-mediated intratumoral injection of human IFNγ gene could be an effective gene therapeutic system for the treatment of pancreatic carcinoma.

Finite element model of human ear reconstruction through micro-computer tomography.

CONCLUSIONS: The validated finite element model of human middle ear can be used as a tool in hearing loss research. OBJECTIVE: To establish an accurate finite element model of the external ear canal and middle ear through micro-computer tomography and test its feasibility in the human. METHODS: A fresh specimen of human temporal bone was scanned by micro-computer tomography. The finite element model was established based on the images, and then the simulation results were compared with published experimental data. RESULTS: After a series of simulations were conducted, the simulated results were compatible with the published experimental data and this confirms the validation of the finite element model.