Defective renal excretion and increased production of uric acid engender hyperuricemia that predisposes to gout. However, molecular mechanisms underlying defective uric acid excretion remain largely unknown. Here, we report a rare genetic variant of gout-unprecedented NUMB gene within a hereditary human gout family, which was identified by an unbiased genome-wide sequencing approach. This dysfunctional missense variant within the conserved region of the NUMB gene (NUMBR630H) underwent intracellular redistribution and degradation through an autophagy-dependent mechanism. Mechanistically, we identified the uric acid transporter, ATP Binding Cassette Subfamily G Member 2 (ABCG2), as a novel NUMB-binding protein through its intracellular YxNxxF motif. In polarized renal tubular epithelial cells (RTECs), NUMB promoted ABCG2 trafficking towards the apical plasma membrane. Genetic loss-of-function of NUMB resulted in redistribution of ABCG2 in the basolateral domain and ultimately defective excretion of uric acid. To recapitulate the clinical situation in human gout patients, we generated a NUMBR630H knock-in mouse strain, which showed marked increases of serum urate and decreased uric acid excretion. The NUMBR630H knock-in mice exhibited clinically relevant hyperuricemia. In summary, we have uncovered a novel NUMB-mediated mechanism of uric acid excretion and a functional missense variant of NUMB in humans, which causes hyperuricemia and gout.

Type 1 diabetes is a T cell-mediated autoimmune disease characterised by pancreatic beta cell destruction. In this study, we explored the pathogenic immune responses in initiation of type 1 diabetes and new immunological targets for type 1 diabetes prevention and treatment. We obtained peripheral blood samples from four individuals with newly diagnosed latent autoimmune diabetes in adults (LADA) and from four healthy control participants. Single-cell RNA-sequencing (scRNA-seq) was performed on peripheral blood mononuclear cells to uncover transcriptomic profiles of early LADA. Validation was performed through flow cytometry in a cohort comprising 54 LADA, 17 adult-onset type 2 diabetes, and 26 healthy adults, matched using propensity score matching (PSM) based on age and sex. A similar PSM method matched 15 paediatric type 1 diabetes patients with 15 healthy children. Further flow cytometry analysis was performed in both peripheral blood and pancreatic tissues of non-obese diabetic (NOD) mice. Additionally, cell adoptive transfer and clearance assays were performed in NOD mice to explore the role of this monocyte subset in islet inflammation and onset of type 1 diabetes. The scRNA-seq data showed that upregulated genes in peripheral T cells and monocytes from early-onset LADA patients were primarily enriched in the IFN signalling pathway. A new cluster of classical monocytes (cluster 4) was identified, and the proportion of this cluster was significantly increased in individuals with LADA compared with healthy control individuals (11.93 https://ngdc.cncb.ac.cn/gsa-human/ ) under accession number HRA003649.

Converting genetic information into functional proteins is a complex, multi-step process, with each step being tightly regulated to ensure the accuracy of translation, which is critical to cellular health. In recent years, advances in modern biotechnology, especially the development of cryo-electron microscopy and single-molecule techniques, have enabled a clearer understanding of the mechanisms of protein translation fidelity. Although there are many studies on the regulation of protein translation in prokaryotes, and the basic elements of translation are highly conserved in prokaryotes and eukaryotes, there are still great differences in the specific regulatory mechanisms. This review describes how eukaryotic ribosomes and translation factors regulate protein translation and ensure translation accuracy. However, a certain frequency of translation errors does occur in translation, so we describe diseases that arise when the rate of translation errors reaches or exceeds a threshold of cellular tolerance.

Mitochondrial DNA (mtDNA) is a critical genome contained within the mitochondria of eukaryotic cells, with many copies present in each mitochondrion. Mutations in mtDNA often are inherited and can lead to severe health problems, including various inherited diseases and premature aging. The lack of efficient repair mechanisms and the susceptibility of mtDNA to damage exacerbate the threat to human health. Heteroplasmy, the presence of different mtDNA genotypes within a single cell, increases the complexity of these diseases and requires an effective editing method for correction. Recently, gene-editing techniques, including programmable nucleases such as restriction endonuclease, zinc finger nuclease, transcription activator-like effector nuclease, clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeats–associated 9 and base editors, have provided new tools for editing mtDNA in mammalian cells. Base editors are particularly promising because of their high efficiency and precision in correcting mtDNA mutations. In this review, we discuss the application of these techniques in mitochondrial gene editing and their limitations. We also explore the potential of base editors for mtDNA modification and discuss the opportunities and challenges associated with their application in mitochondrial gene editing. In conclusion, this review highlights the advancements, limitations and opportunities in current mitochondrial gene-editing technologies and approaches. Our insights aim to stimulate the development of new editing strategies that can ultimately alleviate the adverse effects of mitochondrial hereditary diseases.

Doxorubicin (DOX) is an efficacious and widely used drug for human malignancy treatment, but its clinical application is limited due to side effects, especially cardiotoxicity. Our present study revealed that DOX could induce apoptosis in cardiomyocytes. Herein, we screened the dysregulated long noncoding RNAs (lncRNAs) in DOX-treated cardiomyocytes. Notably, overexpression of lncRNA NONMMUT015745 (lnc5745) could alleviate DOX-induced cardiomyocyte apoptosis both in vitro and in vivo. Conversely, silencing lnc5745 promotes cardiomyocyte apoptosis. Moreover, Rab2A, a direct target of lnc5745, possesses a protective effect in DOX-induced cardiotoxicity once knocked down. Importantly, we verified that the p53-related apoptotic signalling pathway was responsible for the lnc5745-mediated protective role against DOX-induced cardiomyocyte apoptosis. Mechanistically, Rab2A interacts with p53 and phosphorylated p53 on Ser 33 (p53 (Phospho-Ser 33)), promotes p53 phosphorylation, thereby activating the apoptotic pathway. Taken together, our results suggested that lnc5745 protects against DOX-induced cardiomyocyte apoptosis through suppressing Rab2A expression, modifying p53 phosphorylation, thereby regulating p53-related apoptotic signalling pathway. Our findings establish the functional mode of the lnc5745-Rab2A-p53 axis in DOX-induced cardiotoxicity. The development of new strategies targeting the lnc5745-Rab2A-p53 axis could attenuate DOX-induced cardiotoxicity, which is beneficial to its clinical anti-tumour application.

Mitochondria are the sites of oxidative metabolism in eukaryotes where the metabolites of sugars, fats, and amino acids are oxidized to harvest energy. Notably, mitochondria store Ca2+ and work in synergy with organelles such as the endoplasmic reticulum and extracellular matrix to control the dynamic balance of Ca2+ concentration in cells. Mitochondria are the vital organelles in heart tissue. Mitochondrial Ca2+ homeostasis is particularly important for maintaining the physiological and pathological mechanisms of the heart. Mitochondrial Ca2+ homeostasis plays a key role in the regulation of cardiac energy metabolism, mechanisms of death, oxygen free radical production, and autophagy. The imbalance of mitochondrial Ca2+ balance is closely associated with cardiac remodeling. The mitochondrial Ca2+ uniporter (mtCU) protein complex is responsible for the uptake and release of mitochondrial Ca2+ and regulation of Ca2+ homeostasis in mitochondria and consequently, in cells. This review summarizes the mechanisms of mitochondrial Ca2+ homeostasis in physiological and pathological cardiac remodeling and the regulatory effects of the mitochondrial calcium regulatory complex on cardiac energy metabolism, cell death, and autophagy, and also provides the theoretical basis for mitochondrial Ca2+ as a novel target for the treatment of cardiovascular diseases.

BRAF T1799A mutation is the most common genetic variation in thyroid cancer, resulting in the production of BRAF V600E mutant protein reported to make cells resistant to apoptosis. However, the mechanism by which BRAF V600E regulates cell death remains unknown. We constructed BRAF V600E overexpression and knockdown 8505C and BCPAP papillary and anaplastic thyroid cancer cell to investigate regulatory mechanism of BRAF V600E in cell death induced by staurosporine (STS). Induced BRAF V600E expression attenuated STS‐induced papillary and anaplastic thyroid cancer death, while BRAF V600E knockdown aggravated it. TMRM and calcein‐AM staining showed that opening of the mitochondrial permeability transition pore (mPTP) during STS‐induced cell death could be significantly inhibited by BRAF V600E. Moreover, our study demonstrated that BRAF V600E constitutively activates mitochondrial ERK (mERK) to inhibit GSK‐3‐dependent CypD phosphorylation, thereby making BRAF V600E mutant tumour cells more resistant to mPTP opening. In the mitochondria of BRAF V600E mutant cells, there was an interaction between ERK1/2 and GSKa/ß, while upon BRAF V600E knockdown, interaction of GSKa/ß to ERK was decreased significantly. These results show that in thyroid cancer, BRAF V600E regulates the mitochondrial permeability transition through the pERK‐pGSK‐CypD pathway to resist death, providing new intervention targets for BRAF V600E mutant tumours.

This study aimed to investigate the therapeutic effect of human umbilical cord mesenchymal stem cells (hUCMSCs) on experimental autoimmune thyroiditis (EAT) and the underlying mechanisms by utilizing a porcine thyroglobulin-induced EAT rat model. The rats received four tail vein injections of vehicle or hUCMSCs at an interval of 7 days and were sacrificed on day 28 after the first injection. Hematoxylin and eosin staining and enzyme-linked immunosorbent assays (ELISAs) were used to assess the therapeutic effects of hUCMSCs on EAT. Splenic lymphocytes were isolated from rats, and the proportions of CD4+ T cell subsets were analyzed by flow cytometry. Splenic CD4+ T cells from EAT rats were cocultured with hUCMSCs. A loss-of-function assay for protein tyrosine phosphatase non-receptor type 2 (PTPN2) was performed to explore the involvement of PTPN2/signal transducer and activator of transcription 3 (STAT3) signaling on the therapeutic benefit of hUCMSCs in EAT. hUCMSC treatment significantly alleviated inflammation, reduced serum thyroid antibody levels, and decreased the ratios of IL-17α+/CD25+FOXP3+ cells and serum IFN-γ/IL-4 in EAT rats. Furthermore, hUCMSC treatment upregulated PTPN2 protein expression in splenic lymphocytes of EAT rats as well as enhanced the PTPN2 protein level and attenuated phosphorylation of STAT3 in CD4+ T cells in vitro. Importantly, knockdown of Ptpn2 significantly reversed hUCMSC-mediated suppression of cell proliferation and hUCMSC-induced alterations in the expression of inflammatory cytokines in CD4+ T cells. Thus, hUCMSC treatment alleviates thyroid inflammation and the CD4+ T cell imbalance in EAT via PTPN2/STAT3 signaling, serving as a promising therapeutic approach for autoimmune thyroiditis.

Context How lymph node metastasis (LNM)-associated mortality risk is affected by BRAF V600E in papillary thyroid cancer (PTC) remains undefined. Objective To study whether BRAF V600E affected LNM-associated mortality in PTC. Design, Setting, and Participants We retrospectively analyzed the effect of LNM on PTC-specific mortality with respect to BRAF status in 2638 patients (2015 females and 623 males) from 11 centers in 6 countries, with median age of 46 [interquartile range (IQR) 35-58] years and median follow-up time of 58 (IQR 26-107) months. Results Overall, LNM showed a modest mortality risk in wild-type BRAF patients but a strong one in BRAF V600E patients. In conventional PTC (CPTC), LNM showed no increased mortality risk in wild-type BRAF patients but a robustly increased one in BRAF V600E patients; mortality rates were 2/659 (0.3%) vs 4/321 (1.2%) in non-LNM vs LNM patients (P = 0.094) with wild-type BRAF, corresponding to a hazard ratio (HR) (95% CI) of 4.37 (0.80-23.89), which remained insignificant at 3.32 (0.52-21.14) after multivariate adjustment. In BRAF V600E CPTC, morality rates were 7/515 (1.4%) vs 28/363 (7.7%) in non-LNM vs LNM patients (P < 0.001), corresponding to an HR of 4.90 (2.12-11.29) or, after multivariate adjustment, 5.76 (2.19-15.11). Adjusted mortality HR of coexisting LNM and BRAF V600E vs absence of both was 27.39 (5.15-145.80), with Kaplan-Meier analyses showing a similar synergism. Conclusions LNM-associated mortality risk is sharply differentiated by the BRAF status in PTC; in CPTC, LNM showed no increased mortality risk with wild-type BRAF but a robust one with BRAF mutation. These results have strong clinical relevance.

Objective: To study the clinical efficacy of inhibition of sodium-glucose cotransporter 2 (SGLT2) in the treatment of type 2 diabetics. Methods: A total of 92 type 2 diabetics were selected. They were equally divided into two groups according to a random number table method: control group (conventional hypoglycemic treatment) and observation group (dapagliflozin). Glycated hemoglobin (HbA1c), Urine glucose/Urine creatinine level, waist circumference, weight, HDL-C, LDL-C, TC, TG, drug-related adverse events, FPG and 2 h postprandial blood glucose were analyzed and compared. Results: After treatment, the HbA1c of both groups was decreased, and the HbA1c level of the observation group was lower; the Urine glucose/Urine creatinine levels of both groups was increased, and the Urine glucose/Urine creatinine levels of the observation group was higher (P < 0.05). The waist circumference and body weight of both groups were lower, and the waist circumference and body weight of the observation group was lowest (P < 0.05). The LDL-C, TC, and TG in both groups were decreased, and the observation group was lower; HDL-C in both groups was higher, and the observation group was highest (P < 0.05). The incidence of drug-related adverse events in the control group (23.91%) was significantly higher than observation group (6.52%) (P < 0.05). Compared with the control group, the FPG of the observation group was lower (P < 0.001), and the FPG level of both groups has a tendency to decrease with time (P < 0.001). Compared with the control group, the 2 h postprandial blood glucose of the observation group was lower (P < 0.001), and the 2 h postprandial blood glucose of the two groups 2 h after a meal had a tendency to decrease with time (P < 0.001). Conclusions: SGLT2 inhibitors have a significant clinical effect on type 2 diabetes patients.

Ribosomal S6 kinase 4 (RSK4) is a putative tumor suppressor gene which is inactivated by epigenetic events in a number human malignancies; however, its role in papillary thyroid carcinoma (PTC) remains largely unknown. The aim of this study was to explore the methylation status of the RSK4 promoter in PTC, and to determine its potential role in thyroid carcinogenesis. Reverse transcription‑quantitative PCR (RT‑qPCR) and western blot analyses were performed to examine the RSK4 mRNA and protein levels, respectively. Methylation‑specific PCR (MSP) and bisulfite genomic sequencing (BGS) were used to analyze methylation status of the RSK4 gene. Sanger sequencing was further carried out to detect the BRAF V600E mutation. Cell proliferation assay was finally performed to evaluate the role of hypermethylation in the growth of PTC cells. The association between RSK4 methylation and the clinicopathological characteristics of patients with PTC was assessed. The methylation frequency of the RSK4 promoter in PTC tissues was higher than that in paired paracancerous tissues. Coincidentally, the RSK4 mRNA levels were also downregulated in PTC tissues when compared with the paracancerous counterparts. The hypermethylation of RSK4 was associated with tumor size and lymph node metastasis. Furthermore, the BRAF V600E mutation may influence RSK4 expression and methylation. Moreover, RSK4 hypermethylation was observed in thyroid cancer cell lines, which was consistent with a lack of RSK4 expression. Upon the 5‑Aza‑deoxycytidine treatment of thyroid cancer cells, RSK4 expression was significantly upregulated, while cell proliferation was inhibited. On the whole, the findings of the present study demonstrate that the hypermethylation of the RSK4 promoter may be one of the mechanisms responsible for the poor RSK4 expression in PTC. Thus, these data suggest that RSK4 may serve as a molecular target for the early diagnosis and treatment of PTC.

目的 通过分析四个中国家系家族性甲状腺乳头状癌(FPTC)透明质酸结合蛋白2(HABP 2)基因G534E位点突变情况,探讨HABP2基因G534E位点突变与FPTC发病的相关性.方法 对4个先证者家系中的11例PTC患者及其家系成员的临床特点及实验室资料进行分析,采用PCR技术及Sanger基因测序对先证者及其家系成员进行目的基因的检测.结果 在四个家系中,PTC患者、多发性结节性甲状腺肿患者和正常亲属中均未检测到HABP2基因G534E位点的阳性突变.结论 HABP2基因G534E位点突变可能不是我国FPTC人群的易感基因,需要进一步验证其他基因位点的突变情况.

The aim of this cross-sectional study was to investigate whether abnormal glycemic status (diabetes and prediabetes) is associated with increased prevalence of thyroid disorders in North China. A total of 10,860 adult participants were enrolled based on the REACTION study [Ning G. J Diabetes. 2012;4(2):172–173]. They were divided into three groups by glucose levels according to the American Diabetes Association 2010 criteria. The basic characteristics of the study population were analyzed. The prevalence and effects of thyroid disorders, cardio-cerebrovascular disease, and risk factors of thyroid disorders, among the study groups, were investigated. The study population comprised 2994 diabetic patients (27.57%) and 6386 prediabetic patients (58.80%), with the mean age of 55.37 ± 8.82 years. Compared to those subjects with normal glucose status (8.11%), thyroid disorders were more common in the diabetic (15.97%) and prediabetic patients (13.48%) (P < 0.001). Impacts of female gender related to the presence of thyroid disorders were even more profound in the diabetic patients than in those with other glucose status. The results indicate that screening for thyroid disorders among patients with diabetes or prediabetes should be regularly performed, especially in females.

The aim of this cross-sectional study was to investigate whether abnormal glycemic status (diabetes and prediabetes) is associated with increased prevalence of thyroid disorders in North China. A total of 10,860 adult participants were enrolled based on the REACTION study [Ning G. J Diabetes. 2012;4(2):172-173]. They were divided into three groups by glucose levels according to the American Diabetes Association 2010 criteria. The basic characteristics of the study population were analyzed. The prevalence and effects of thyroid disorders, cardio-cerebrovascular disease, and risk factors of thyroid disorders, among the study groups, were investigated. The study population comprised 2994 diabetic patients (27.57%) and 6386 prediabetic patients (58.80%), with the mean age of 55.37 +/- 8.82 years. Compared to those subjects with normal glucose status (8.11%), thyroid disorders were more common in the diabetic (15.97%) and prediabetic patients (13.48%) (P < 0.001). Impacts of female gender related to the presence of thyroid disorders were even more profound in the diabetic patients than in those with other glucose status. The results indicate that screening for thyroid disorders among patients with diabetes or prediabetes should be regularly performed, especially in females.

目的 探讨核糖体蛋白激酶A6(RSK4)在甲状腺乳头状癌(PTC)中的表达及其与临床病理因素的关系.方法 收集200例PTC(A组)和对应的癌旁组织(B组),40例甲状腺良性结节(C组)及其结节旁组织(D组)新鲜标本,提取相应RNA,并逆转录成cDNA,采用实时荧光定量PCR(RT-qPCR)检测RSK4的表达量,并分析其与PTC患者临床病理因素的关系.结果 A组中RSK4表达量显著低于B组(Z=-9.658,P＜0.01)和C组(Z=-5.648,P＜0.01),但C组RSK4表达量与D组比较差异无显著性(P＞0.05).肿瘤直径＞1 cm的PTC组织中RSK4表达量显著低于≤1 cm组(Z=-3.528,P＜0.01),有包膜侵犯的PTC组织中RSK4表达量显著低于无包膜侵犯组(Z=-2.822,P＜0.01).结论 RSK4在PTC组织中低表达,且与肿瘤大小、包膜侵犯相关,可能成为PTC诊断和预后评估的潜在生物标志物.

Objective To detect the methylation status of ribosomal S6 kinase 4 ( RSK4 ) in papillary thyroid carcinoma ( PTC) and to study its correlation with mRNA expression and clinical features. Methods 134 cases PTC tissues and corresponding paracancerous thyroid tissues were collected. DNA methylation status of RSK4 gene in PTC tissues and corresponding paracancerous tissues were analyzed by methylation specific PCR and bisulfite genomic sequencing, and mRNA expression was detected by quantitative realtime PCR. The relationship of DNA methylation status with mRNA expression and clinical features was analyzed. Results The methylation rate of RSK4 in PTC tissues was significantly higher than that in paracancerous tissues by methylation specific PCR ( P<0.05) and bisulfite genomic sequencing ( P<0. 01 ) . The RSK4 mRNA level in PTC tissues was significantly lower than that in paracancerous tissues ( P<0.01) . In PTC tissues, RSK4 mRNA expression in methylation group was lower than that in unmethylation group ( P<0.01) . The RSK4 mRNA level in PTC of methylation was lower than that in paracancerous tissues of methylation ( P<0. 01);the RSK4 mRNA level in PTC of unmethylation was also lower than that in paracancerous tissues of unmethylated ones ( P<0. 01 ) . There was relationship of RSK4 hypermethylation with lymphatic metastasis and TNM grade ( P<0. 05 ) . Serum concentration, of thyroid stimulating hormone, thyroid peroxidase antibody, and thyroglobulin antibody in methylation group were higher than those in unmethylation group ( P<0.05) . Conclusion The hypermethylation of RSK4 promoter's CpG islands might be one of the mechanisms for poor expression of RSK4 in PTC, which may serve as a molecular target of early diagnosis and treatment.

Purpose To test whether the prognostic risk of male sex in papillary thyroid cancer (PTC) is determined by BRAF V600E and can thus be stratified by BRAF status. Patients and Methods We retrospectively investigated the relationship between male sex and clinicopathologic outcomes in PTC, particularly mortality, with respect to BRAF status in 2,638 patients (male, n = 623; female, n = 2,015) from 11 centers in six countries, with median age of 46 years (interquartile range, 35-58 years) at diagnosis and median follow-up time of 58 months (interquartile range, 26-107 months). Results Distant metastasis rates in men and women were not different in wild-type BRAF PTC but were different in BRAF V600E PTC: 8.9% (24 of 270) and 3.7% (30 of 817; P = .001), respectively. In wild-type BRAF PTC, mortality rates were 1.4% (five of 349) versus 0.9% (11 of 1175) in men versus women ( P = .384), with a hazard ratio (HR) of 1.59 (95% CI, 0.55 to 4.57), which remained insignificant at 0.70 (95% CI, 0.23 to 2.09) after clinicopathologic multivariable adjustment. In BRAF V600E PTC, mortality rates were 6.6% (18 of 272) versus 2.9% (24 of 822) in men versus women ( P = .006), with an HR of 2.43 (95% CI, 1.30 to 4.53), which remained significant at 2.74 (95% CI, 1.38 to 5.43) after multivariable adjustment. In conventional-variant PTC, male sex similarly had no effect in wild-type BRAF patients; mortality rates in BRAF V600E patients were 7.2% (16 of 221) versus 2.9% (19 of 662) in men versus women ( P = .004), with an HR of 2.86 (95% CI, 1.45 to 5.67), which remained significant at 3.51 (95% CI, 1.62 to 7.63) after multivariable adjustment. Conclusion Male sex is a robust independent risk factor for PTC-specific mortality in BRAF V600E patients but not in wild-type BRAF patients. The prognostic risk of male sex in PTC can thus be stratified by BRAF status in clinical application.

BackgroundPrecise risk stratification-based treatment of solitary intrathyroidal papillary thyroid cancer (SI-PTC) that is larger than 1.0 cm and 4.0 cm or less is undefined.MethodsA genetic-clinical risk study was performed on BRAF V600E in 955 patients (768 women and 187 men) with SI-PTC, with median age of 46 years and median clinical follow-up time of 64 months at 11 medical centers in six countries. The chi-square test or, for analyses with small numbers, Fisher's exact test was performed to compare recurrence rates. Recurrence-free probability was estimated by Kaplan-Meier (KM) analysis, and the independent effect of BRAF mutation on the recurrence was analyzed by Cox regression and Cox proportional hazard analyses. All statistical tests were two-sided.ResultsRecurrence of SI-PTC larger than 1.0 cm and 4.0 cm or less was 9.5% (21/221) vs 3.4% (11/319) in BRAF mutation vs wild-type BRAF patients, with a hazard ratio (HR) of 3.03 (95% confidence interval [CI] = 1.46 to 6.30) and a patient age- and sex-adjusted hazard ratio of 3.10 (95% CI = 1.49 to 6.45, P = .002). Recurrence rates of SI-PTC larger than 2.0 cm and 4.0 cm or less were 16.5% (13/79) vs 3.6% (5/139) in mutation vs wild-type patients (HR = 5.44, 95% CI = 1.93 to 15.34; and adjusted HR = 5.58, 95% CI = 1.96 to 15.85, P = .001). Recurrence rates of SI-PTC larger than 3.0 cm and 4 cm or less were 30.0% (6/20) vs 1.9% (1/54) in mutation vs wild-type patients (HR = 18.40, 95% CI = 2.21 to 152.98; and adjusted HR = 14.73, 95% CI = 1.74 to 124.80, P = .01). Recurrences of mutation-positive SI-PTC were comparable with those of counterpart invasive solitary PTC, around 20% to 30%, in tumors larger than 2.0 cm to 3.0 cm. BRAF mutation was associated with a statistically significant decrease in recurrence-free patient survival on KM analysis, particularly in SI-PTC larger than 2.0 cm and 4.0 cm or less. Similar results were obtained in conventional SI-PTC. The negative predictive values of BRAF mutation for recurrence were 97.8% (95% CI = 96.3% to 98.8%) for general SI-PTC and 98.2% (95% CI = 96.3% to 99.3%) for conventional SI-PTC.ConclusionsBRAF V600E identifies a subgroup of SI-PTC larger than 1.0 cm and 4.0 cm or less, particularly tumors larger than 2.0 cm and 4.0 cm or less, that has high risk for recurrence comparable with that of invasive solitary PTC, making more aggressive treatment reasonable.

Objective In recent years,the incidence of papillary thyroid carcinoma(PTC)increased gradually and the recurrence and metastasis of PTC patients are also increasing.The aim of this study was to investigate the effect of BRAFV600E mutation on the expression of HMGB1 in PTC,to explore the mechanism of BRAF gene affecting the development and prognosis of PTC,and then guiding clinical practice of precise therapy.Methods The preoperative serum and postoperative fresh tissue were collected from 44 cases of PTC patients in our hospital from September,2015 to December,2015.According to BRAFV600E mutations divided into positive group and negative group.Western blot,immunohistochemistry and fluorescence quantitative PCR were used to detect the level of HMGB1 in cancer tissues.The level of HMGB1 in serum was tested by ELISA method.Western blot was performed using Image J software.The relative quantitative method was used to calculate the fluorescence quantitative PCR data.The concentration of HMGB1 protein in serum was calculated by ELISA Calc regression/fit calculation program.All the data were analyzed by SPSS 20.0.Western blot,fluorescence quantitative PCR and ELISA data were analyzed by Chi-square test,Mann-Whitney-Wilcoxon test and independent sample t test.The relationship between lymph node metastasis and extrathyroid infiltration and the occurrence of BRAFV600E mutation was analyzed by chi-square test.Results In PTC cancer tissues,HMGB1 was mainly located in the cytoplasm,the levels of HMGB1mRNA and protein in positive group were lower than that of the negative group(Z =2.117,P ＜0.01,x2 =19.989,P ＜ 0.05),and the change did not show in peripheral blood(t =1.135,P ＞ 0.05).The expression of HMGB1 mRNA and protein in PTC with lymph node metastasis was lower than those without lymph node metastasis(Z =-2.216,P ＜0.05;t =-2.217,P ＜ 0.05),and so as in the extrathyroid infiltration(Z =-2.267,P ＜ 0.05;t =-3.885,P ＜ 0.01).Conclusion BRAFV600E mutations may accelerate the malignant development of PTC by down regulating the expression of HMGB1.

BACKGROUND:Papillary thyroid cancer (PTC) is the most common differentiated thyroid cancer and is responsible for 80-90% of thyroid cancer cases. Despite typically excellent prognoses, these subclinical low-risk cancers are often treated aggressively by surgical thyroidectomy. Consequently, the objective of this study was to generate a prognostic matrix to be used prior to PTC intervention.METHODS:In this study, 80 PTC patients were assessed. Following adjustment for sex, logistic regression analysis showed that BRAFV600E mutation, transforming growth factor beta (TGF-β) expression, age, and tumor size are risk factors that can affect tumor clinical stage (p < 0.05). Based on the results of this analysis, we generated a matrix that incorporated 4 variables: patient age, tumor size, BRAFV600E mutation, and TGF-β expression.RESULTS:We observed that the corresponding area under curve was as high as 0.91. The sensitivity and specificity of the model were 94.74 and 83.61%, respectively. These values are significantly higher than those generated from single indexes.CONCLUSION:As a result of this analysis, it is hoped that the resultant matrix can be utilized during clinical diagnosis and treatment prior to thyroid nodule surgery.