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Objective:To Constructing a nomogram based on clinical, ultrasound and BRAF V600E gene for predicting cervical lymph node metastasis in patients with papillary thyroid carcinoma (PTC).Methods:The clinical data of 287 patients with PTC (374 malignant nodules) from December 2019 to December 2021 in the First Affiliated Hospital of Hunan Normal University were analyzed retrospectively. Among them, there were 205 nodes with cervical lymph node metastasis and 169 nodes without cervical lymph node metastasis. The echo type, capsule, boundary, shape, number, diameter, location, cystic and solid properties, aspect ratio, blood flow signal, echo distribution, ultrasonic classification, microcalcification and enlarged lymph nodes were observed by ultrasound. The mutation of BRAF V600E gene was detected by fluorescence polymerase chain reaction. The nomograph model for predicting neck lymph node metastasis in patients with PTC was constructed and validated by R3.6.3 software.Results:Univariate analysis result showed that gender, age, microcalcifications, aspect ratio, morphology, blood flow signal, diameter, echo distribution, enlarged lymph nodes, ultrasound classification and BRAF V600E gene were the risk factors for cervical lymph node metastasis in patients with PTC ( P<0.05 or <0.01). Multivariate Logistic regression analysis result showed that age (<40 years old), ultrasonic classification (≥4a) and diameter (>1 cm) were independent risk factors for cervical lymph node metastasis in patients with PTC ( OR = 2.847, 1.436 and 2.475; 95% CI 1.827 to 4.436, 1.075 to 1.918 and 1.505 to 4.069; P<0.01 or <0.05). The age, ultrasonic classification and diameter were included as predictors for constructing the nomogram model. The receiver operating characteristic curve analysis result shows that the area under the curve predicted by the nomogram model for neck lymph node metastasis in patients with PTC was 0.692 (95% CI 0.631 to 0.753). Conclusions:Nomogram based on age, ultrasonic classification and diameter is of high value in predicting neck lymph node metastasis in patients with PTC.
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Objective:To investigate the clinical features of patients with Langerhans cell histiocytosis (LCH), and analyze the association between BRAF V600E mutation status and clinical features. Methods:A retrospective analysis was carried out for the clinical data of 60 patients with LCH at the Department of Pediatric Oncology, Sun Yat-sen Memorial Hospital between April 2013 and December 2019.Among them, 39 patients undertook BRAF V600E mutation testing, which in paraffin-embedded tissue samples were detected by quantitative real-time PCR (qRT-PCR), and in peripheral blood and/or bone marrow were tested by high-throughput sequencing, for analyzing the correlation between BRAF V600E mutation and clinical characteristics of LCH. Results:(1)Clinical characteristics: the age of 60 LCH patients was (4.08±0.45) years, with 43 male cases and 17 female cases.Patients at young age (≤2 years) and with risk organ (RO+ ) and central nervous system (CNS) risk lesions involvement were concentrated in the multisystem involvement (MS) group ( P<0.05). (2)Therapeutic response after induction therapy: the response to induction therapy was achieved in 28 of 60 treated patients (41.7%) and 32 (53.3%) did not.After excluding stratification factors of treatment regimen, MS ( OR=6.855, 95% CI: 2.077-22.622, P=0.002) and the age≤2 years ( OR=4.944; 95% CI: 1.601-15.275; P=0.005) were risk factors in poor chemotherapy response.RO+ ( OR=8.250, 95% CI: 1.617-42.090, P=0.005) was a significant risk factor for a poor chemotherapy response in JLSG-02 treatment group.Differently, RO+ had no dramatic effect on chemotherapy response in CCHG-LCH-2019 treatment group.(3) BRAF V600E mutation: 39 patients were determined BRAF V600E status, with the positive rate of BRAF V600E mutation in paraffin-embedded tissue samples reaching 70.3%(26 cases). BRAF V600E mutation was not associated with early treatment response, age, sex, MS and RO+ ( P>0.05). However, the positive rate of BRAF V600E in children with MS and CNS risk lesions was higher than the controls, with 76.0% (19 cases) vs.57.1% (8 cases) and 74.1% (20 cases) vs.58.3% (7 cases), respectively.Totally, 3 of 8 cases were positive in bone marrow, with 2 cases of MS, and 1 case of multiple bone invasions, and 1 of 5 cases was positive in peripheral blood, with liver and spleen being involved. Conclusions:LCH patients with age≤2 years, MS and RO+ exhibited a poor response to initial treatment, required for more aggressive treatment strategy.Lesion with activating BRAF V600E mutations suggests that LCH is a clonal disorder.There may be great variability between BRAF V600E mutations and MS as well as CNS risk lesions.In the mutation dataset, part of patients had positive BRAF V600E mutations in bone marrow/peripheral blood.This might suggest a different pathogenesis in such patients, has a certain clinical sense in some aspect.
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Objective:To investigate the value of BRAF V600E mutation combined with 2015 American Thyroid Association (ATA) Guidelines ultrasound (US) pattern in fine-needle aspiration (FNA) cytology of thyroid nodules with atypia of undetermined significance or follicular lesion of undetermined significance (AUS/FLUS).Methods:This study retrospectively enrolled 96 consecutive patients with 101 AUS/FLUS thyroid nodules who underwent preoperative US, FNA, and BRAF V600E mutation analysis. All AUS/FLUS nodules were classified based on US pattern-based risk stratification of 2015 ATA Guidelines. With postoperative pathology as the gold standard, the diagnostic value of BRAF V600E mutation, US pattern and the combination of two methods were compared.Results:Postoperative pathology confirmed 33 benign nodules and 68 malignant nodules. The mutation rates of BRAF V600E in AUS/FLUS nodules was 51.5%. The sensitivity, specificity, and accuracy of BRAF V600E in the diagnosis AUS/FLUS nodules were 72.1%, 90.9% and 78.2%, respectively. The ROC curve demonstrated that the best cut-off of US pattern was high suspicion. The sensitivity, specificity, and accuracy of US pattern in the diagnosis of AUS/FLUS nodules were 63.2%, 81.8% and 69.3%, respectively. The accuracy of US pattern in determining AUS/FLUS nodules without BRAF V600E mutation was 70.6%. The sensitivity, specificity, and accuracy of the combination of two methods in the differential diagnosis of AUS/FLUS nodules were 89.7%, 75.8%, and 85.1%, respectively. The combination had the highest sensitivity ( P<0.05). Conclusions:BRAF V600E mutation has a good diagnostic value for differentiating benign and malignant AUS/FLUS nodules. Combined with US pattern, the differential diagnostic value for AUS/FLUS nodules without BRAF V600E mutation can be improved, and the sensitivity can be raised.
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Objective To evaluate the diagnostic efficacies of contrast-enhanced ultrasound(CEUS) combined with BRAF V600E mutation detection in ultrasound-guided fine-needle aspiration cytology of thyroid nodules with atypia of undetermined significance . Methods A total of 129 thyroid nodules underwent the examinations of CEUS and BRAF V600E mutation were analyzed retrospectively . With surgical pathology as the gold standard ,ROC curve was used to investigate the diagnostic values of CEUS , BRAF V600E and the combination of the two methods . Results The sensitivity ,specificity and accuracy of CEUS and BRAF V600E gene detection for thyroid nodules with atypia of undetermined significance diagnosed by ultrasound-guided fine-needle aspiration biopsy were 86 .7％ ,83 .3％ ,85 .3％ and 72 .0％ , 100％ ,83 .7％ ,respectively . The sensitivity and accuracy of CEUS were higher than those of BRAF V 600E gene detection ( all P < 0 .001 ) ,however its specificity was lower than BRAF V 600E with statistically significance( P < 0 .001) ,those of the combined test of CEUS and BRAF V600E mutation analysis were 94 .7％ ,83 .3％ ,89 .9％ ,respectively . The combination of two methods had the highest diagnostic efficacy , with statistically difference ( P <0 .001) ,and the area under the ROC curve ( AUC) was higher than that for each test(0 .951 vs 0 .860 vs 0 .901) . Conclusions The combined test of CEUS and BRAF V600E mutation has a higher diagnostic efficacy for cytologically indeterminate thyroid nodules compared with CEUS or BRAF V600E mutation alone .
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OBJECTIVE: To investigate the correlation between clinicopathological features and BRAF~(V600E) gene mutation in papillary thyroid carcinoma(PTC). METHODS: The data of 290 cases of PTC admitted from August 2016 to May 2018 in Beijing Tongren Hospital,Capital Medical University were analyzed retrospectively. All the cases of PTC were examined by BRAF~(V600E) immunohistochemistry after operation. Mutation positive group included 192 cases. Mutation negative group included 59 cases and partial mutation group included 39 cases. The cases of partial mutantion were excluded. The clinicopathological differences between the two groups were compared. RESULTS: By statistical analysis,there was a statistical difference between two groups in the invasion of the capsule(P<0.05). In the two groups(microcarcinoma and non-microcarcinoma),there was the same result.There was no statistical difference in other indexes. CONCLUSION: In patients with PTC,BRAF~(V600E) gene mutation is not associated with the clinicopathological features of the PTC. Patients with non-BRAF~(V600E) gene mutations are more likely to invade the membranes of microcarcinomas and non-microcarcinomas.
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Objective To study the screening of trace amount mutation of BRAF V 600E gene for avoiding the appearance of ineffective treatment in cancer patients .Methods The internal competitive amplification fragments were used to improve the inhibition of wild-type blocking (WTB) probe on wile-type BRAF V600E gene to increase the detection efficiency of BRAF V600E genotype of trace amount mutation occurrence .Re-sults When the template DNA concentration was 50 -200 ng/μL ,the constructed trace amount gene muta-tion real time fluorescence quantitative detection method could completely block the amplification of the wild-type BRAF V600E gene .The sensitivity of this assay reached as high as 0 .1％ ,which was in line with the sen-sitivity requirement for the gene trace amount mutation detection technique .In the colorectal biopsy tissues from 50 cases of suspected colorectal cancer ,8 cases (16 .0％ ) of BRAF V600E gene trace amount mutation were detected by using this constructed method ,which had higher detection rate .Conclusion The constructed gene trace amount mutation detection method can make the rapid ,simple and low cost quantitative analysis for BRAF V600E gene trace amount mutation in clinical samples .
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Background and purpose:The BRAF V600E mutation is a highly attractive drug target. Therefore, determining the BRAF gene mutation status of patients is essential in order to assess patients' eligibility for targeted BRAF geneinhibitor therapy. The aim of this study was to validate the utility of immunohistochemistry to rapidly obtain the BRAF gene mutation status. This study aimed to analyze the correlation of the BRAF V600E gene mutation and VE1 protein ex-pression with the clinical pathological characteristics in papillary thyroid carcinoma (PTC).Methods:The mutation status of BRAF V600E was detected by DNA sequencing. Immunohistochemistry was used to detect the expression of BRAF V600E protein in 108 cases of PTC, 54 cases of thyroid adenoma and 54 cases of normal thyroid tissue.Results:The gene mutation rate of BRAF V600E is 67.6%, and VE1 protein expression rate is 64.8% in 108 cases of PTC. The differences were statistically significant compared with thyroid adenoma and goiter (P<0.05), but have no correlation with the clinical pathological characteristics.Conclusion:BRAF V600E gene mutation and VE1 protein expression are useful biomarkers for the pathological diagnosis of PTC. High consistency was observed between the immunohistochemical staining results and the DNA sequencingresults of BRAF V600E gene mutations. Immunohistochemical technique detecting the BRAF V600E protein expression can effectively reflect indirectly BRAF V600E gene mutation status in PTC.BRAF V600E gene mutation has no contribution to the development of papillary thyroid carcinoma.